Algae and Aquatic Biomass For A Sustainable Production of 2nd Generation Biofuels

Proposalfulltitle:
Algaeandaquaticbiomassforasustainableproductionof2ndgenerationbiofuels
Proposalacronym:
AquaFUELs
Typeoffundingscheme:
Cooperation
Theme5Energy
Taxonomy,BiologyandBiotechnology
Nameofthecoordinatingperson:
Dr.RaffaelloGarofalo
Coordinatoremail:
ebb@ebbeu.org
Coordinatorphone: +3227632477
Coordinatorfax:
+3227630457
REV
Date
Organisation
Beneficiaries
involved
FINAL
20/05/2011
NatasciaBiondi,
MarioTredici
UNIFI
UNIFI
Dissemination
level
PU
AquaFUELsTaxonomy,BiologyandBiotechnology
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Tableofcontents
INTRODUCTION ................................................................................................................................................. 5
1
1.1
IMPORTANCE OF ALGAE AND AQUATIC BIOMASS FOR BIOFUELS ......................................................................... 6
1.1.1 Suitability of algae as biomass producers ..................................................................................................... 6
1.1.2 Sustainability, the strategic advantage of algal biofuels ............................................................................... 6
1.2
RATIONALE OF THE DOCUMENT ......................................................................................................................... 8
1.3
TARGET GROUPS ................................................................................................................................................ 8
1.4
PROBLEMS INCURRED ........................................................................................................................................ 9
1.5
COMMON ERRONEOUS "MYTH" .......................................................................................................................... 9
2
CRITERIA FOR STRAIN SELECTION ............................................................................................................ 10
2.1
PRODUCTIVITY ................................................................................................................................................. 10
2.2
ROBUSTNESS .................................................................................................................................................... 10
2.3
HARVESTABILITY ............................................................................................................................................. 10
2.4
BIOMASS COMPOSITION ................................................................................................................................... 10
2.5
PROCESSABILITY / EXTRACTABILITY ................................................................................................................ 11
2.6
ADDED VALUE OF CO-PRODUCTS ..................................................................................................................... 11
2.7
LOCAL ORIGIN OF STRAINS ............................................................................................................................... 11
3
BIOLOGY OF ALGAE....................................................................................................................................... 12
3.1
CYANOBACTERIA ............................................................................................................................................. 12
3.2
CHLOROPHYTA (GREEN ALGAE)...................................................................................................................... 15
3.3
RHODOPHYTA (RED ALGAE)............................................................................................................................ 17
3.4
HETEROKONTOPHYTA ...................................................................................................................................... 18
3.4.1 Phaeophyceae (Brown algae)...................................................................................................................... 19
3.4.2 Eustigmatophyceae ..................................................................................................................................... 21
3.4.3 Other classes ............................................................................................................................................... 21
3.5
LABYRINTHULEA (PHYLUM HETEROKONTA) ................................................................................................... 21
3.6
BACILLARIOPHYTA (DIATOMS)........................................................................................................................ 22
3.7
HAPTOPHYTA ................................................................................................................................................... 24
3.8
DINOPHYTA (DINOFLAGELLATES) ................................................................................................................... 24
3.9
OTHER ALGAL GROUPS..................................................................................................................................... 26
4
BIOTECHNOLOGY OF ALGAE ...................................................................................................................... 28
4.1
INTRODUCTION ................................................................................................................................................ 28
4.2
CULTIVATION SYSTEMS ................................................................................................................................... 29
4.2.1 Open ponds ................................................................................................................................................. 30
4.2.2 Photobioreactors ......................................................................................................................................... 31
Main photobioreactors designs ...............................................................................................................................................32
Polyethylene bags and vertical columns.............................................................................................................................32
Tubular PBR ......................................................................................................................................................................33
Flat photobioreactors (panels) ............................................................................................................................................35
4.2.3 Sustainability of different cultivation systems............................................................................................ 35

4.3
HARVESTING METHODS.................................................................................................................................... 36
4.4
BIOTECHNOLOGY OF THE MAJOR MICROALGAL GROUPS .................................................................................. 37
4.4.1 Cyanobacteria ............................................................................................................................................. 37
4.4.2 Chlorophyta (Green Algae)......................................................................................................................... 38
4.4.3 Rhodophyta (Red Algae) ............................................................................................................................ 39
4.4.4 Heterokontophyta........................................................................................................................................ 39
4.4.5 Labyrinthulea (phylum Heterokonta).......................................................................................................... 40
4.4.6 Bacillariophyta (Diatoms)........................................................................................................................... 40
4.4.7 Haptophyta.................................................................................................................................................. 41
4.4.8 Dinophyta (Dinoflagellates)........................................................................................................................ 41
4.5
BIOTECHNOLOGY AND USES FOR MACROALGAE .............................................................................................. 41
4.6
BIOTECHNOLOGY OF OTHER AQUATIC BIOMASS ............................................................................................. 43
5
SYMBOLOGY.................................................................................................................................................... 44
6
REFERENCES .................................................................................................................................................... 45
7
PROKARYOTIC MICROALGAE ..................................................................................................................... 51
7.1
CYANOBACTERIA ............................................................................................................................................. 51
7.1.1 Arthrospira sp. (common name spirulina) .................................................................................................. 51
7.1.2 Phormidium sp............................................................................................................................................ 54
7.1.3 Anabaena sp................................................................................................................................................ 57
7.1.4 Synechococcus sp........................................................................................................................................ 60
7.1.5 Synechocystis sp.......................................................................................................................................... 62
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8
EUKARYOTIC MICROALGAE........................................................................................................................ 64
8.1
CHLOROPHYTA ................................................................................................................................................ 64
8.1.1 Ostreococcus sp. ......................................................................................................................................... 64
8.1.2 Tetraselmis sp ............................................................................................................................................. 66
8.1.3 Botryococcus braunii.................................................................................................................................. 70
8.1.4 Chlamydomonas reinhardtii ....................................................................................................................... 72
8.1.5 Haematococcus pluvialis ............................................................................................................................ 76
8.1.6 Dunaliella sp............................................................................................................................................... 79
8.1.7 Chlorococcum sp. ....................................................................................................................................... 83
8.1.8 Neochloris oleoabundans............................................................................................................................ 86
8.1.9 Scenedesmus sp........................................................................................................................................... 92
8.1.10 Desmodesmus sp. ........................................................................................................................................ 97
8.1.11 Chlorella sp. ............................................................................................................................................... 99
8.1.12 Parietochloris incisa................................................................................................................................. 109
8.1.13 Prototheca sp. ........................................................................................................................................... 111
8.2
RHODOPHYTA ................................................................................................................................................ 113
8.2.1 Porphyridium cruentum ............................................................................................................................ 113
8.3
BACILLARIOPHYTA ........................................................................................................................................ 115
8.3.1 Benthic diatoms (Amphora; Amphiprora; Cylindrotheca; Navicula; Nitzschia) ...................................... 115
Amphora sp. ..........................................................................................................................................................................115
Amphiprora hyalina ..............................................................................................................................................................116
Cylindrotheca sp. ..................................................................................................................................................................116
Navicula sp. ..........................................................................................................................................................................117
Nitzschia dissipata ................................................................................................................................................................118
8.3.2 Phaeodactylum tricornutum...................................................................................................................... 126

8.3.3 Chaetoceros muelleri................................................................................................................................ 126
8.3.4 Cyclotella cryptica.................................................................................................................................... 136
8.3.5 Odontella aurita........................................................................................................................................ 139
8.3.6 Skeletonema sp.......................................................................................................................................... 141
8.3.7 Thalassiosira pseudonana ........................................................................................................................ 143
8.4
EUSTIGMATOPHYCEAE (PHYLUM HETEROKONTOPHYTA) .............................................................................. 146
8.4.1 Monodus subterraneus.............................................................................................................................. 146
8.4.2 Nannochloropsis sp................................................................................................................................... 148
8.5
HAPTOPHYTA ................................................................................................................................................. 153
8.5.1 Isochrysis sp.............................................................................................................................................. 153
8.5.2 Pavlova sp................................................................................................................................................. 156
8.6
DINOPHYTA ................................................................................................................................................... 159
8.6.1 Crypthecodinium cohnii............................................................................................................................ 159
8.7
LABYRINTHULOMYCETES .............................................................................................................................. 162
8.7.1 Schizochytrium sp. .................................................................................................................................... 162
8.7.2 Thraustochytrium sp. ................................................................................................................................ 164
8.7.3 Ulkenia sp. ................................................................................................................................................ 166
9
MACROALGAE............................................................................................................................................... 167
9.1
CHLOROPHYTA .............................................................................................................................................. 167
9.1.1 Caulerpa sp............................................................................................................................................... 167
Caulerpa racemosa ...............................................................................................................................................................167
Caulerpa taxifolia .................................................................................................................................................................168
9.1.2
Ulva sp. ..................................................................................................................................................... 170
Ulva lactuca..........................................................................................................................................................................170
Ulva rigida............................................................................................................................................................................170
9.1.3
9.1.4
Cladophora sp. ......................................................................................................................................... 178

Codium sp. ................................................................................................................................................ 180
Codium fragile ......................................................................................................................................................................180

Codium parvulum..................................................................................................................................................................180
9.2
RHODOPHYTA ................................................................................................................................................ 183
9.2.1 Chondrus crispus ...................................................................................................................................... 183
9.2.2 Mastocarpus stellatus ............................................................................................................................... 185
9.2.3 Grateloupia turuturu................................................................................................................................. 187
9.2.4 Palmaria palmata ..................................................................................................................................... 189
9.2.5 Solieria chordalis...................................................................................................................................... 191
9.3
PHAEOPHYCEAE (PHYLUM HETEROKONTOPHYTA) ........................................................................................ 193
9.3.1 Alaria esculenta ........................................................................................................................................ 193
9.3.2 Undaria pinnatifida .................................................................................................................................. 195
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Ascophyllum nodosum .............................................................................................................................. 197

Fucus sp. ................................................................................................................................................... 199
Fucus serratus.......................................................................................................................................................................199
Fucus spiralis........................................................................................................................................................................199
Fucus vesiculosus..................................................................................................................................................................200
9.3.5
9.3.6
Himanthalia elongata ............................................................................................................................... 202

Cystoseira sp............................................................................................................................................. 204
Cystoseira baccata................................................................................................................................................................204
Cystoseira tamariscifolia ......................................................................................................................................................205
9.3.7
9.3.8
9.3.9
Halidrys siliquosa ..................................................................................................................................... 207

Sargassum muticum .................................................................................................................................. 208
Laminaria, Saccharina, Saccorhiza.......................................................................................................... 210
Laminaria sp. ........................................................................................................................................................................210

Laminaria digitata ...........................................................................................................................................................210
Laminaria hyperborea......................................................................................................................................................211
Laminaria ochroleuca......................................................................................................................................................212
Saccharina latissima .............................................................................................................................................................212
Saccorhiza polyschides .........................................................................................................................................................213
10 OTHER AQUATIC BIOMASS ........................................................................................................................ 221

10.1
EGERIA DENSA ................................................................................................................................................ 221
10.2
EICHHORNIA CRASSIPES................................................................................................................................... 223
10.3
ELODEA CANADENSIS ...................................................................................................................................... 226
10.4
LAGAROSIPHON MAJOR .................................................................................................................................... 228
10.5
LEMNA MINOR ................................................................................................................................................. 231
11 CONCLUDING REMARKS............................................................................................................................. 236
ANNEX I ................................................................................................................................................................... 237
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1 Introduction
Algae are a group of organisms that have been generally described as photoautotrophic unicellular or
multicellular,mainlywaterdwellingorganismslackingcomplexmorphologicalorganization.Historically,the
prokaryotic bluegreen algae, or cyanobacteria (Class Cyanophyceae), are often included in discussing
microalgae,andindeedsomecyanobacterialspecies(Arthrospiraorspirulina)holdaprominentpositionin
thebiotechnologicalexploitationofmicroalgae.
Thereareseveralmaingroupsofmicroalgaedifferinginbiochemicalconstituents,ultrastructure,and
life cycle. Some of the characteristics traditionally used for algae classification are the nature of their
photosyntheticpigments,storageproducts,cellwall,presenceorabsenceof flagellaand thenumber of
membranessurroundingthechloroplast.
More recently classification has been based on comparisons of specific DNA sequences, leading to
major revisions in classification of many groups of alga. Recent molecular genetic studies confirmed that
photoautotrophic eukaryotes belong to several highly diverse groups of organisms and are the result of
different and independent events of secondary endosymbioses. As a consequence algae belong to
geneticallywidelydivertinggroupsoforganismsoftencloserrelatedtononphotosyntheticorganismsthan
to more distant algal clades (Fig. 1). This fact requires due attention when developing tools such as
transformationorgeneticengineeringetcformicroalgae.
The most recent results on algal taxonomy, are summarized in detail by the Tree of Life project
(http://tolweb.org/tree/)andAlgaeBase(http://www.algaebase.org/),bothprovidinguptodatetaxonomic
informationconcerningclassificationofalgalspecies,thatiscontinuouslybeingupdatedandrevisedinlight
ofnewestresultsobtainedbymoleculargeneticapproachessuchasDNAsequencecomparisons.
Figure 1Phylogenetictreeoftheeukaryoticorganisms(modifiedfromTreeofLifeProject
http://tolweb.org/tree/).
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Microalgae reproduction occurs primarily by vegetative (asexual) cell division, although sexual
reproductioncanoccurinmanyspeciesunderappropriategrowthconditions.
1.1 Importanceofalgaeandaquaticbiomassforbiofuels
1.1.1
Suitabilityofalgaeasbiomassproducers
Microalgaeareconsideredfastgrowingphotosyntheticorganismsandhavebeenreportedtoreachshort
termmaximalsummerproductivitiesof5060gperm2perdayinCO2enrichedracewaypondsinHawaii
and California. This corresponds to transformation of 56% of incoming light energy into biomass. Such
numbers, as well as productivity data from lab scale experiments have promoted the reputation of
microalgae as prime candidates for providing unlimited amounts of cheap biomass as food, fodder or
energy.Furthermoremanyalgalstrainscanproducelargeamountsofoilorlipidlikestorageproductsthat
can easily be converted into biodiesel (Sheehan et al., 1998). This has been used by some to combine
maximalbiomassproductivitywithmaximaloilcontent,yieldingphantastic oilproductivitynumbersthat
havebeenexploitedforfundingintensiveresearchonalgalbiofuelsproduction.Potentialoilproductivities
of over 100 tons/ha per year were initially predicted. However, none of the large scale long term
experiments ever reached the high productivity projections. Current productivity obtained in large scale
operations range from 40 60 tons of algal biomass production per ha and year, with conservative
projectionsanticipatingupto100tonsofbiomass,or30tonsofbiodieselperhaandyearinsubtropicalor
tropical,sunnyclimates(Scottetal.,2010).
Five groups of microalgae were classified as high priority for biofuel production by the US microalgal
species program ASP (Sheehan et al., 1998): diatoms (Class Bacillariophyceae), green algae (Class
Chlorophyceae), goldenbrown algae (Class Chrysophyceae), prymnesiophytes or haptophytes (Class
Prymnesiophyceae), and eustigmatophytes (Class Eustigmatophyceae). However, different classes of
macroalgae,aswellasfurtheryetlessstudied microalgalgroupsmayturnouttobe equallyrelevantfor
successfulbiomassproductionfromalgae.
Otheraquaticbiomasssuchaswaterlentils(Lemna),waterhyacinth,Elodeaandothershavealsobeen
considered for potential biofuel production due to their significant productivity and their usefulness in
treatingpollutednutrientrichwaterbodies.
1.1.2
Sustainability,thestrategicadvantageofalgalbiofuels
LandUseCurrentbiofuelssuchasoilfromsoybean,palm,andrapeseed,orethanolfromcornorwheat,
sufferedfromserioussetbacksrevealedbyrecentanalysisshowingtheiradverseecologicalimpactandlow
greenhousegasreductionpotential.ArecentstatementbyUNEPDirector,A.Steiner,reads:"biofuelsfrom
palm oil grown by Indonesia might never be deemed to be sustainable", due to ongoing destruction of
tropical forests for expanding palm oil production. In addition, with yields of less than 5005,000 L of
biodiesel per hectare (Johnston et al., 2009), those crops require enormous areas of scarce arable land,
waterandfertilizerandaregenerallyhighlyworkintensive.Lifecycleassessments(LCA)indicatethatno,or
verylow,reductionsingreenhousegasemissionscanbeachievedusingsuchbiofuels(Zahetal.,2007),and
iftheyarebeingproducedfollowingconversionofnaturalecosystemstheirGHGemissionssurpassthose
offossilfuelsforyearstocome(Fargioneetal.,2008).Themajorimpact,landuse,isoftennotadequately
considered if the strategic implications of expanding biofuels production are taken into account
(Searchingeretal.,2008):
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Supplyingtheworld's2030liquidfueldemandfromdedicatedbiofuelcropswouldconsumeallormost
oftheavailablelandintheappropriateclimaticzonesincludingmostremainingnaturalecosystemsevenif
themostproductivebiofuelscropssuchaspalmoil,Miscanthuscellulosicethanolorsugarcaneethanolor
anoptimizedmixofthosecropswereplanted.Evenifadoublingofyieldsisachievedinthenext20years,
around half of the worlds remaining intact ecosystems will have to be sacrificed to cover the projected
liquidfueldemand.
Onlyalgaecanprovidesufficientliquidfuelsonafewpercentofavailabledrylandareas.Algalbiomass
if managed properly, may be produced on unproductive desert land, under utilization of ocean or waste
water by exploiting and recycling of waste nutrients from municipal and agricultural sources, since no
health concerns need to be considered for biofuel production. Algal biofuel production is thus
complementary to ongoing efforts to grow lignocellulosic biomass in areas with good soil and water
resources,becausethemicroalgaeareprojectedtobegrowninthoseareaswherethelignocellulosicoroil
seedscropswillnotperformwell(Brown,M.Lewis,BiodieselfromMicroalgae:ComplementarityinaFuel
Development Strategy, NREL, http://www.nrel.gov/docs/legosti/old/5715.pdf ). Algae may produce
sufficientbiofuelsonlessthan10%ofavailabledrylandsevenundercurrentproductivityestimates,often
usingthemostunproductiveareaslikesaltflatsordegradeddrylandsoils.Theymayalsodeliveradditional
environmentalservicessuchaswastewatertreatmentorexhaustgasdetoxification.
Table 1Comparisonoflanduseimpactofvariousbiofuelcrops.*Forallcropsitisassumedthat50%ofbiomass
energyisusedforprocessenergyandallnutrientswillberecycledtothemaximumpossible,accordingtothestate
oftheartofsugarcaneethanolproduction.InMiscanthusthis50%areremovedfromtheclaimedethanolyield,
since no leftover biomass is available for process energy. The crops marked in italics, as well as algae, are
experimentalasnoactualproductioninthelargescalehasbeendemonstrated.(LandareasarederivedfromIto
andOikawa,2004;biofuelproductivitiesarederivedfromJohnstonetal.,2009).
Landtype
Area
(miokm2)
Tropicalandsubtropical
evergreenforest
Tropicalandsubtropical
dryforest
TropicalSavanna,
Woodland
Midlattitudeforests,
abandonedcroplands
Warm
Shrubland/grasslandor
desert
NaturalProductivity
(tonsofcarbonfixedper
Cropand
biofuelyield
hectareandyear)
(tonsperha/GJ
perha)
Palmoil
(5/189)
Jatrophaoil
(1.5/56.7)
%areaof
corresponding
ecosystemrequired
tocover2030
demand
10.5
10.7
110%!!!
4.7
7.67
6.7
6.65
Caneethanol
(4.34/116)
270%
14
5.30
Miscanthus
cellulosic
ethanol*
(4.4/120)
95%
33
13.50
Algaeoil
(20/756)
5.48.2%
765%!!
Other Environmental Services Interestingly, even under desert conditions the water footprint of algal
biomassproductionislowerthanthatofirrigatedmaizeethanolproduction,calculatedonthewaterinput
per unit of energy created. Our estimates suggest that it requires less than 0.5 million liters of water to
produce one 1 ton of dry algal biomass, and this may be waste or seawater. It can take about 3 million
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liters of water to produce 1 ton of rice and about 2 million liters to produce 1 ton of soybeans
(www.clw.csiro.au/issues/water/water_for_food.html
and
www.gdrc.org/uem/footprints/water
footprint.html). However, significant investments into (waste)water and CO2 infrastructure would be
requiredtoachievethenecessaryglobalalgaebiomassproductionpotential.
Properly planned algal biomass production facilities may recover and reuse most of the nutrients
applied, minimizing eutrophication impact, and this in contrast to intensive agriculture where nutrient
runoffandescapingnitricoxideposeseriousproblems.
In fact, algae have been shown to be able to treat successfully any kind, even the most problematic,
formsofwastewater.Pesticideuseinalgalcultivationisexpectedtobeminimal.Evenasmallproportionof
thealgalbiomassrequiredforenergypurposeswouldprovidesufficientproteintoreplaceallthesoybean
cultivation capacity installed for feed production, resulting in reduced deforestation or 'negative indirect
landusechanges'(Searchingeretal.,2008).
While algal biomass production may never achieve the low production costs of other agricultural
commodities, full accounting of above and additional environmental services may result in a balance
favouringthealgalfuels.Thispointrequiresintensiveinvestigationprogressingfarbeyondcurrentlyused
LCA models (Stephenson et al., 2010), starting with defining those production parameters and system
boundariesthatwillactuallydelivertheabovementionedenvironmentaladvantages.
1.2 Rationaleofthedocument
Thisdocumentsummarizesinshortcurrentviewsandprospectsonthepotentialcontributionofalgaeto
biofuelproduction.
Recentpredictionsandcalculations,bothattheEUlevelandintheUS(AUSDARegionalRoadmapto
MeetingtheBiofuelsGoalsoftheRenewableFuelsStandardby2020,USDAStrategicBiofuelsProduction
report, June 2010), do not incorporate a significant algae potential into their projections for the next 10
years. This is dictated by the fact that algae experts and external observers disagree about the true
potentialofalgalbiofuelsproductionrelatingtoeconomicandenvironmentalsustainability,andanygiven
timeframeforachievingcompetitivealgalbiofuelproductionisspeculativeatthebest.
Given the high complexity of algal taxonomy and evolutionary relationships, this document was
conceivedasaninstrumenttoplacethealgaethathavearisenaninterestforbiofuelproductionwithinthe
correctframe.Thelistofalgaeproposedisbasedontheliteratureconcerningbiofuelproduction,onthe
commercially produced algae and on the feedback from the questionnaire in deliverable 1.1. Detailed
descriptionofbiotechnologyisprovidedonlyforpivotaltaxaorgroupoftaxa,thatareactuallyproducedat
least at pilot scale. These taxa represent the reference model for those taxa that are not currently
exploited.
Itwasbeyondthescopeofthisdocument toproposeanykindofneworrevisedtaxonomyofalgae.
The classification reported is based on AlgaeBase (http://www.algaebase.org/) and Tree of Life project
(http://tolweb.org/tree/).
The other aquatic biomasses species reported are all invasive weeds that have been proposed as
biofuel crops. The classification reported is based on ITIS Catalogue of Life 2010
(http://www.catalogueoflife.org/annualchecklist/2010/search/all) and US Department of Agriculture
PLANTSDatabase(http://plants.usda.gov/).
1.3 Targetgroups
Since the major gaps in knowledge concerning algal biofuels are the lack of operating pilot scale and
commercial algal biofuels production facilities, the target audience for this report are all interested
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stakeholders in Europes energy, agricultural and environmental policy, such as policy makers, NGOs,
research infrastructures, interested industries and financial institutions. The conclusion of this report
shouldbeanappealforrapidandsignificantinvestmentsintoalgaerelatedbiomassproductionintheform
of public or privately managed pilot or production facilities with the aim of testing a multitude of
operational parameters for increasing yields and sustainability while reducing production costs of algal
biomassasmuchaspossible.TakingintoaccountthelongtermmultitrillionEuroperyearenergyandby
product market, this report aims at convincing all involved potential stakeholders and investors to
designinglongtermstrategiesforfundinganalgaebasedfuelandbiomassdevelopmentprogramthatmay
providethenecessaryinsightontheirundoubtedlyhugepotential.
1.4 Problemsincurred
It is a disturbing fact that today no results on true sustained biomass production yields for biofuels
production are available, and no functional biofuel production plant is accessible anywhere around the
world.Thusallrecentpublicationsinthefield,beityields,economicsorLCA,remainpurespeculationand
demandgreatestcareintheirinterpretation.
During the last 10 years algal biofuels companies, driven by large investments from venture capital,
haveaimedtodemonstrateapotentialforrapidlyachievingeconomicprofitability.Thistrendleadtohigh
degreesofsecrecysurroundingmysteriousprocessesdevelopedwhosetechnicalandscientificsoundness
cannotbeconfirmednordiscardedduetolackingaccesstorawdataandfacilities.Thesummarypresented
belowthereforereliesonpublicationsandpatentsreleasedbymostlyacademicinstitutionsandafewopen
minded companies feeling that little in terms of technology and biology in the process of algal biomass
production deserves this degree of secrecy. Nevertheless it cannot be excluded that certain secret
breakthroughsmayhavebeenachievedrecentlythatwouldputthestateoftheartsignificantlyaheadof
whatisbeingpresentedhere.
1.5 Commonerroneous"myth"
Incontrasttooftenvoicedopinions,algaearenotsignificantlymoreefficientinbiomassproductionthan
otherplantsgrownunderoptimalconditions.Themostcommonerroriscomparingbiomassdoublingtime
or specific growth rates, which indicate the rate of biomass accumulation under exponential growth
conditions, where indeed algae and cyanobacteria may multiply several times per day. However, those
conditions are possible under very low biomass densities only that are not applicable to large scale algal
cultivation since actual biomass produced per day is in fact lower than in cultures with higher biomass
densities, where all the incoming light is captured by algae and used for photosynthesic biomass
production.
If such doubling times of exponentially growing cultures are being applied to denser cultures (which
couldbedonewithheterotrophicorganismsbyincreasingthefoodinput)infactveryeasilyfantasticdaily
growth rates can be assumed. However other than in heterotrophic culture conditions the one and only
energysourceforgrowthofalgaeisincominglightenergythatistransformedwithanefficiencyofaround
3%intobiomass.Underabsolutelyoptimizedconditionsintermsoftemperature,lightintensity,mixingand
CO2 supply, higher photosynthetic efficiencies of up to 7% may be achieved, however under exponential
increase in bioreactor and maintenance costs that are generally claimed not to be covered by increased
yield,andalsorequirefarhigherenergyinputsleadingtoanegativeratiobetweenenergyinputandgainin
formofalgalbiomassforexampleintubularphotobioreactors(Jorqueraetal.2010).
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2 Criteriaforstrainselection
The objective was to agree on criteria for both micro and macro algae on which species can be
selected for their suitability for biofuel production. These criteria should ideally be quantitatively
measurable.Forallcriteriaoneshouldkeepanoutdoor,largescalesysteminmind,becauseforlab
(scale)experimentationsdifferentcriteriamayapply.
2.1 Productivity
Thisincludesproductivityofbiomassandofspecificbiomasscomponents(e.g.lipids).Inordertobe
able to compare algae with traditional crops and with each other in terms of productivity, the most
objective criterion is to use photosynthetic efficiency as a measure for productivity. Basically this
meansthe%ofavailablelight(energy)thatisconvertedintobiomassorspecificbiomasscomponents.
Inthiswayallspeciesandallcultivationsystemscanbecompared.
2.2 Robustness
Thisisarathervaguetermwhichincludesresistancetomanyextremeconditions.Thiscriterioncanbe
bestassessedusingtableinwhichtheresistancetotheseseveralconditionsisscored.
Table2Conditionsofrobustness.
Condition
pH
Oxygenconcentration
Temperature
Salinity
Organiccontaminants
Relevantfor
Reduceriskofinfection
CO2transfer
Closedphotobioreactors
Outdoorcultivation
Openwatercultivation
Range
i.e.<4and>10
>20%
Large range to accommodate
day/night and seasonal fluctuation
(e.g.1040C)
Cultivation in fresh / sea / e.g010%salinity
brackishwater
Reduceriskofinfection
Ability
to
grow
on Concentration
of
organic
wastewater/fluegas
contaminants that still allows good
growth
2.3 Harvestability
For microalgae this will mainly be the sedimentation rate and the possibilities for induced or auto
flocculation.
Formacroalgaethisincludesthepossibilitiesformechanicalharvestingorharvestingbyhand.
2.4 Biomasscomposition
Thisshouldincludeabreakdownofthetotalbiomasscompositionin:
totalcaloricvalueofthebiomass(forburningit),
%lipidsandlipidcomposition(forbiodiesel),
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% starch and carbohydrate composition (for bio ethanol and to identify higher value
byproducts(i.e.agar),
%proteinandproteincomposition(soluble/insolubleforfood/feedpurposes),
presenceofheavymetalsortoxins(specification).
2.5 Processability/extractability
This should include relevant aspects for biorefinery, such as the cell volume, thickness/toughness of
thecellwallandthepresenceoftoughfibers(macroalgae)andthemoisturecontent.Ameasurefor
thiscouldbetheenergyinputpergramofdryweightnecessaryforfullbiorefinery.
2.6 Addedvalueofcoproducts
Does the organism produce any by or coproduct that have an intrinsic added value, such as
carotenoids.Thisisimportanttoreducethecostsofthefinalbiofuelproduct.Hereaspecificationof
thecompoundsandtheirexpectedaddedvaluepergramofdrybiomassshouldbeindicated.
2.7 Localoriginofstrains
Theuseoflocallyselectedstrainsmaybeofsignificancebothforeaseofmanagementandforreasons
of sustainability Based on criteria of the 'Roundtable on Sustainable Biofuels' (http://rsb.epfl.ch/).
Nonnative potentially invasive biofuels crops should not be used in open cultivation systems, and
adherence to this rule will require the identification and use of locally isolated algal strains.
Furthermore such strains may have unique adaptations to the local climate, water and possible
parasitesthatimportedorevenlaboratorygrownstrainsmaynothave.
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3 Biologyofalgae
Algae are an assemblage of organisms that have been generally described as photoautotrophic
unicellular or multicellular organisms lacking complex morphological organization, and as such have
been reclassified several times in recent biology according to technical advances on the basis of
differences in subcellular organization, or later of molecular genetic characteristics that will allow,
whensufficientdatawillbeavailable,precisedeterminationofevolutionaryrelationships.Historically,
algaeincludedalsocyanobcateria,prokaryoticoxygenicphototrophs.
Many microalgae grow quite rapidly and their reproduction occurs primarily by vegetative
(asexual) cell division, although sexual reproduction can occur in many species under appropriate
growth conditions. There are several main groups of microalgae, which differ primarily in pigment
composition, biochemical constituents, ultrastructure, and life cycle. Five groups were of primary
importance:diatoms(Bacillariophyta),greenalgae(Chlorophyta),PrymnesiophytaorHaptophytaand
Eustigmatophytatogetherwiththeprokaryoticbluegreenalgae,orcyanobacteria.
Recent molecular genetic studies confirmed that photoautotrophic eukaryotes belong to several
highly diverse groups of organisms and are the result of different and independent events of
secondary endosymbioses. As a consequence algae are a genetically widely diverting group of
organisms, a fact that will require due attention when developing tools such as transformation or
geneticengineeringetc.
3.1 Cyanobacteria
Cyanobacteria are prokaryotic photoautotrophic microorganisms that can be found in almost every
environment, from oceans to freshwater to bare rock to soil. Though the prokaryotic Cyanobacteria
(commonlyreferredtoasbluegreenalgae)weretraditionallyincludedas"algae"inoldertextbooks,
manymodernsourcesregardthisasoutdatedastheyarenowconsideredtobebacteria.
Classification The cyanobacteria were traditionally classified by morphology according to the
International Code of Botanical Nomenclature into five orders: Chroococcales, Pleurocapsales,
Oscillatoriales, Nostocales and Stigonematales. Starting from the 1970s cyanobacteria have been
classifiedalsoaccordingtotheInternationalCodeofNomenclatureofBacteriaandtheyweretreated
in the Bergeys Manual of Systematic Bacteriology Volume 3 of the 1989 edition, then updated in
volume I of the edition of 2004, where cyanobacterai are subdivided in five subsections, IV,
correspondingto theordersoftheBotanicalCode, exceptthattheProchloraleshavebeenincluded
within the cyanobacteria and precisely in the I subsection (Komrek and Anagnostidis, 1986;
AnagnostidisandKomrek,1988;CastenholzandWaterbury,1989;KomrekandAnagnostidis,1989;
Anagnostidis and Komrek, 1990; Castenholz, 2001;Oren, 2004; Wilmotte and Herdman, 2001;
HerreroandFlores(eds),2008).
CellstructureThefirsttwosubsectionsincludeunicellularcyanobacteria(CastenholzandWaterbury,
1989;Castenholz,2001).ThemembersofChroococcalesareunicellularcyanobacteriathatreproduce
bybinaryfissionorbudding.Cellsarecoccoidorrodshapedandcanvaryinlengthfrom0.5to30m.
Division can occur in one to three successive planes, so that cells can be single or in colonies. The
classic taxonomic criterion has been the cell morphology and the plane of cell division. In
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Pleurocapsales, cyanobacteria reproduce by multiple fission which generates small spherical cells
namedbaeocytes,thatcanbemotileornotaccordingtothegenus(HerdmanandRippka,1988b).In
unicellular forms there is only multiple fission, whereas in colonial forms after binary fission on
differentplanessomeofthecellsundergomultiplefission.
The remaining sections include filamentous cyanobacteria. In Oscillatoriales (Castenholz and
Waterbury, 1989; Castenholz, 2001), the cells are uniseriately arranged and do not form specialized
cells(akinetesandheterocysts).Theyreproducebybinaryfissioninasingleplane.Filamentdiameter
varies from 0.4 to 100 m. Outside the cell wall a sheath may be present. In this case terminal
hormogonia of short length can glide out of the sheath and eventually form new sheaths. New
filaments(ortrichomes)areoriginatedfromfragmentationincorrespondencetoadeadcellorcertain
cells(necridialcells)arepurposelydestinedtodie.InNostocalesandStigonematales(Castenholzand
Waterbury,1989;Castenholz,2001)thecellshavetheabilitytodifferentiatecellslikeheterocystsand
akinetes.Nostocalesarefilamentouscyanobacteriadividingonlybybinaryfissioninoneplane,though
somegeneraproducefalsebranching.Filamentsmaybecomposedofcellsofuniformdiameterorby
cellswithdecreasingdiametertowardstheendofthefilament(taperingtrichomes).Heterocystsmay
beterminalandintercalaryoronlyterminalindifferentgenera.Motiletrichomes(hormogonia)canbe
formedfordispersioninsomegenera(HerdmanandRippka,1988b).Resistancecell(akinetes)maybe
formed under unfavorable conditions (Herdman and Rippka, 1988a). Stigonematales, unlike
Nostocales, includes species with truly branched trichomes. Within this class there is the maximum
degreeofcomplexityanddifferentiationofallthecyanobacterialgroups.Longitudinalorobliquecell
division occurs in addition to tranverse division, so that periodic true branching and, in some cases,
multiseriatetrichomesareformed.Hormogoniamaybeformed,evenifreproductionoccursmainlyby
random breakage of the trichome. Akinets can be produced in some genera. Heterocysts are both
intercalaryandterminal. Celldiametervarieswithinatrichomeassecondarybranchesareusually
narrower.
Incyanobacteria(CastenholzandWaterbury,1989),generationtimesareusuallyhigherthan24h,
thoughsomeunicellularandoscillatorianstrainscanduplicatein4h.Somegeneracanhavecomplex
morphogenetic cycles including filamentation, aseriate phases, dispersal through hormogonia and
akinetesproduction.
Cyanobacteria share the basic cell characteristics with the other Bacteria (Stanier and Cohen
Bazire,1977;CastenholzandWaterbury,1989).ThecellwallisoftheGramnegativetype,thoughthe
peptidoglycan layer is considerably thicker than in the other Gramnegative bacteria. Many
cyanobacteria have a sheath or glycocalix or capsule or gel, mucilage or slime outside the outer
membrane of the cell wall, mainly composed of polysaccharides. Some sheaths can have a
microfibrillar structure and can become laminated with aging of the trichome. Some cyanobacteria
(andmanyhormogonia)cancontaingasvesiclesthatallowbuoyancyinthewatercolumn.
Cyanobacteria have an elaborate and highly organized system of internal membranes which
function in photosynthesis (thylakoids) (Stanier and CohenBazire, 1977; Castenholz and Waterbury,
1989).Thelipophilicpigmentschlorophylla(bothreactioncentersandantenna)andphotosynthetic
carotenoids are located within the thylakoids, while the hydrophilic antenna pigments
(allophycocyaninAPC, phycocyanin PC and, where they are present, phycoerythrin PE or
phycoerythrocyaninPEC)arelocatedinthephycobilisomeswhichareattachedtotheoutsideofthe
thylakoidmembranes.Thephycobilisomeishaemidiscoidalandiscomposedofstacksofbiliproteinsin
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the order (from inside to outside) APC, PC, PE orPEC. Genera belonging to the former group of the
Prochlorales lack phycobilisomes and have chlorophyll b as antenna pigment. The cyanobacterium
Acaryochloris marina has been reported to contain chlorophyll d instead of chlorophyll a as light
harvesting pigment, so that its photosynthetic process depends on farred light (710718 nm)
(Miyashitaetal.,2003).
Thereservecarbohydrateisglycogen(StanierandCohenBazire,1977;CastenholzandWaterbury,
1989). Cyanobacteria contain also cyanophycin, a nitrogen reserve polymer made of arginine and
asparticacid,polyphosphategranulesandcarboxisomes,thatareacellreserveofthephotosynthesis
keyenzymerubisco(ribulose1,5biphosphatecarboxylase).Somecyanobacteriaalsocontainpoly
hydroxybutyrategranules.
PhysiologyPhotosynthesisincyanobacteria(Wolk,1973;StanierandCohenBazire,1977;Castenholz
and Waterbury, 1989)uses water as an electron donor and produces oxygen as a byproduct. This
wateroxidizing process is accomplished by coupling the activity of photosystem (PS) II and I (Z
scheme).Underanaerobicconditionssomegenera(belongingtotheIandIIIsubsections)areableto
carry out anoxygenic photosynthesis using only PS I to carry out cyclic photophosphorylation and
obtainATP,andusingelectrondonorsotherthanwater(hydrogensulfideorthiosulphate)(Cohenet
al.,1975;Garlicketal.,1977).CarbondioxideisreducedtoformcarbohydratesviatheCalvincycle.
During respiration reduced NADP is obtained through the pentose phosphate cycle. The plasma
membrane contains only components of the respiratory chain, while the thylakoid membrane hosts
bothrespiratoryandphotosyntheticelectrontransport.
Mostcyanobacteriaareobligatephotoautotrophs,butsomespeciescangrowasheterotrophsin
thedarkattheexpenseofglucose,fructoseorsucrose.Underanaerobicconditions,somespeciescan
performlactatefermentation(OrenandShilo,1979).
Nitrogen fixation occurs both in heterocystous cyanobacteria and in some nonhetrocystous
cyanobacteria. To avoid contact of nitrogenase with oxygen (and then its permanent inactivation)
theselattercyanobacteriaadoptatemporalseparationbetweenthephotosyntheticandthenitrogen
fixation processes (Bergman et al., 1997). Increased respiration rates allow to control the oxygen
concentrationinsidethecell,duetodiffusion,necessarytocarryoutcellmetabolism.Inheterocystous
forms,thenitrogenfixationprocessisspatiallyseparatedfromtheoxygencphotosynthesis.Nitrogen
fixationiscarriedoutinspecializedcells,theheterocysts(AdamsandDuggan,1999).Thesehavemany
characteristics that allow to reduce diffusion of oxygen, such as a thick cell wall surrounded by a
complexexternalenvelopeandareorganizationofthephotosyntheticapparatus:lackofPSIItoavoid
internal oxygen production, presence of PS I to obtain ATP through cyclic photophosphorilation.
Reducingpowerisobtainedfromvegetativecellsintheformofsugars.Molecularnitrogenisfixedinto
ammoniaandimmediatelyconvertedtoorganicform,usuallyasglutamine.Asnitrogenfixationisa
veryenergyconsumingprocess,nitrogeneseisproducedandheterocystsaredifferentiatedonlyinthe
absenceofcombinednitrogenintheenvironementsurroundingthecell.
EcologyCyanobacteriaaretheonlygroupoforganismsthatareabletoreducenitrogenandcarbon
in aerobic conditions, a fact that may be responsible for their evolutionary and ecological success
(WhittonandPotts(eds),2000).Theycontributesignificantlytoglobalecologyandtheoxygencycle.
The large amounts of oxygen in the atmosphere originally derive from the activities of ancient
cyanobacteria.ThetinymarinecyanobacteriumProchlorococcuswasdiscoveredin1986(Chisholmet
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al.,1988)and,togetherwiththepicoplanktoniccyanobacteria,accountsforuptohalfoftheprimary
productionofwaters,fromoligotrophicopenoceantoestuarineecosystems.
Duetotheirabilitytofixnitrogeninaerobicconditionstheyareoftenfoundassymbionts(Raiet
al. (eds), 2002) with a number of other groups of organisms such as fungi (lichens), corals,
pteridophytes (Azolla), angiosperms (Gunnera), protists (including some diatoms), and sponges. The
ricepaddiesofAsiarelyonnitrogenfixingcyanobacteriaasfertilizers,bothfreebiomassandsymbiont
tothefernAzolla.
Theycanoccurasplanktoniccells(thankstothebuoyancyability)orformphototrophicbiofilmsin
freshwaterandmarineenvironments,theyoccurindampsoil,oreventemporarilymoistenedrocksin
deserts(WhittonandPotts(eds),2000).Someliveinthefurofsloths,providingaformofcamouflage.
Aquatic cyanobacteria are probably best known for the extensive and highly visible blooms that can
forminbothfreshwaterandmarineenvironments.Theassociationoftoxicitywithsuch bloomshas
frequentlyledtotheclosureofrecreationalwaterswhenbloomsareobserved.Certaincyanobacteria
producecyanotoxins(ChorusandBartram,1999)includingneurotoxins,hepatotoxins,cytotoxins,and
endotoxins. Examples of cyanotoxins are anatoxina, anatoxinas, aplysiatoxin, saxitoxin,
cylindrospermopsin,microcystins,nodularin.Thesetoxinscanbedangeroustohumansandanimals.
Several cases of human poisoning have been documented. Recent studies suggest that significant
exposuretohighlevelsofBMAAanonproteicaminoacidproducedbymanycyanobacteriacouldbe
amongthecausesofneurodegenerativediseasessuchasAmyotrophicLateralSclerosis.
Genome sequencing The unicellular cyanobacterium Synechocystis sp. PCC6803 was the third
prokaryote and first photosynthetic organism whose genome was completely sequenced (Kaneko et
al. 1996) . It continues to be an important model organism. Today over 40 complete cyanobacterial
genomes are known (www.ncbi.nlm.nih.gov). The smallest genomes have been found in
Prochlorococcusspp. (1.7 Mb)andthe largestinNostocpunctiforme(9Mb). ThoseofCalothrixspp.
areestimatedat1215Mb,aslargeasyeast.
RelationshiptochloroplastsChloroplastsfoundineukaryotes(algaeandplants)likelyevolvedfrom
an endosymbiotic relation with cyanobacteria. This endosymbiotic theory is supported by various
structural and genetic similarities. Primary chloroplasts are found among the "true plants" or green
plantsaswellasamongtheredalgaeandglaucophytes,marinespecieswhichcontainphycobilins.It
now appears that these chloroplasts probably had a single origin, in an ancestor of the clade called
Primoplantae.Otheralgaelikelytooktheirchloroplastsfromtheseformsbysecondaryendosymbiosis
oringestion.
3.2 Chlorophyta(GreenAlgae)
The green algae are a large group of algae from which the embryophytes (higher plants) emerged
(Jeffrey et al., 2004). The group including both green algae and embryophytes is monophyletic (and
often just known as kingdom Plantae). The green algae include unicellular and colonial flagellates,
usuallybutnotalwayswithtwoflagellapercell,aswellasvariouscolonial,coccoid,andfilamentous
forms. In the Charales, the closest relatives of higher plants, full differentiation of tissues occurs
(Thomas,2002).Thereareabout6,000speciesofgreenalgae.Manyspecieslivemostoftheirlivesas
singlecells,whileotherspeciesformcoloniesorlongfilaments.
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Some species of green algae, particularly of genera Trebouxia and Pseudotrebouxia

(Trebouxiophyceae),canbefoundinsymbioticassociationswithfungitoformlichens.Ingeneralthe
fungalspeciesthatpartnerinlichenscannotliveontheirown,whilethealgalspeciesisoftenfound
livinginnaturewithoutthefungus.
Prasinophyceae are a class of primitive eukaryotic marine green algae (Sym and Pienaar, 1993).
Their best known genus is Ostreococcus, which is considered to be the smallest (ca. 0.95 m) free
living eukaryote and which has been detected in marine samples around the world (Courties et al.,
1994). Prasinophyceae are thought to have low cellular complexity, that is, they possess single,
multipleornoflagellaandcontainonlyasinglechloroplastandasinglemitochondrion.Theyalsohave
verysmallgenomesforaeukaryote(about12Mbp),andthegenomesoftwoOstreococcusspecies,
tauriiandlucimarinus,havebeencompletelysequenced.Ithasbeensuggestedthataprasinophyceae
like flagellate was the ancestor to Chlorophyta and Streptophyta (Kapraun, 2007). A study of
photosyntheticgenesequencediversity(rbcL)intheGulfofMexicoindicatedthatPrasinophyceaeare
particularly prevalent at the Subsurface Chlorophyll Maximum (SCM) (Warwick et al., 2003) and
several different ecotypes of Ostreococcus have been detected in the environment (Guillou et al.,
2004).Theseecotypesaredistinguishedbytheiradaptationtolightintensities.
The Chlorophyceae are one of the classes of green algae, distinguished mainly on the basis of
ultrastructural morphology. For example the chlorophycean CW clade, and chlorophycean DO clade,
are defined by the arrangement of their flagella. Members of the CW clade have flagella that are
displaced in a "clockwise" (CW, 17 o'clock) direction eg. Chlamydomonadales. Members of the DO
cladehaveflagellathatare"directlyopposed"(DO,126o'clock)e.g.Sphaeropleales.
They share many similarities with the higher plants, including the presence of asymmetrical
flagellated cells, the breakdown of the nuclear envelope at mitosis, and the presence of
phytochromes,flavonoids,andthechemicalprecursorstothecuticle(Ravenetal.,2005).
CellstructureAlmostallformshavechloroplasts.Thesecontainchlorophyllsaandb,givingthema
bright green colour (as well as the accessory pigments beta carotene and xanthophylls), and have
stackedthylakoids(vandenHoeketal.,1995).Allgreenalgaehavemitochondriawithflatcristae.The
storage product for members of this group is true starch, amylose, and amylopectin (1,4linked
polyglucans),andisfoundinsidethechloroplasts.Thestarch(seenaswhitishgranuleswiththeTEM)
can often be observed surrounding the pyrenoid, a distinct spherical structure embedded in the
chloroplast. There may be more than one pyrenoid or the prenoid is not always present (e.g.,
AnkistrodesmusandTetraedron)orthepyrenoidislacking.Inmostrepresentativetaxa,thecellsare
surrounded by a cellulose cell wall (Wehr and Sheath, 2003). Some taxa may also have chitin or
sporopollenin deposited on the wall. This gives added strength and is thought to help prevent
desiccation. Some taxa have wall ornamentation, such as scales, a rough texture, thick walls with
distinctlayers,warts,ridges,andspines.TheVolvocalesusuallyhavecellwalls,loricae,orgelatinous
matrices and the main component of the cell walls is glycoprotein, rather than cellulose. The
flagellatedgreenmicroalgaecanhavefromonetoeightisokontflagella.
The Chlorophyta macroalgae share the following common characteristics: flagella of swimming
cells in pairs or multiples of two; stellate structure linking nine pairs of microtubules at basal body
transition zone; thylakoids single or stacked; plastid with two membranes without periplastid
endoplasmic reticulum; starch inside plastid; glycolate dehydrogenase present; cell wall, when
present,ofcellulose;celldivisionwithoutphragmoplast.
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OriginThechloroplastsofgreenalgaeareboundbyadoublemembrane,sopresumablytheywere
acquired by direct endosymbiosis of cyanobacteria. A number of cyanobacteria show similar
pigmentation,butthisappearstohavearisenmorethanonce,andthechloroplastsofgreenalgaeare
nolongerconsideredcloselyrelatedtosuchforms.Instead,thegreenalgaeprobablyshareacommon
originwiththeredalgae.
PhylogenyTheordersoutsidetheChlorophytaareoftengroupedasthedivisionCharophyta,which
isparaphyletictohigherplants,togethercomprisingtheStreptophyta.SometimestheCharophytais
restricted to the Charales, and a division Gamophyta is introduced for the Zygnematales and
Desmidiales.InoldersystemstheChlorophytamaybetakentoincludeallthegreenalgae,buttaken
asabovetheyappeartoformamonophyleticgroup.
OneofthemostbasalgreenalgaeistheflagellateMesostigma,althoughitisnotyetclearwhetherit
issistertoallothergreenalgae,orwhetheritisoneofthemorebasalmembersoftheStreptophyta.
ReproductionMostgreenalgaecanproliferatevegetativelybycelldivision,oftenthemothercellcan
divide into up to 16 offspring before releasing them. They often can profliferate sexually whereby
haploid algae cells of opposing mating type (containing only one copy of their DNA) can fuse with
otherhaploidcellstoformdiploidzygotes.Theycanalsofollowareproductioncyclecalledalternation
of generations. Reproduction varies from fusion of identical cells (isogamy) to fertilization of a large
nonmotile cell by a smaller motile one (oogamy). However, these traits show some variation, most
notablyamongthebasalgreenalgae,calledprasinophytes.
Some taxa produce motile cells (planospores). Planospores may be asexual zoospores or sexual
gametes.Aplanospores(nonmotilecells)maybealsoproduced.
When filamentous algae conjugate, they form bridges between cells, and leave empty cell walls
behindthatcanbeeasilydistinguishedunderthelightmicroscope.
The species of Ulva are reproductively isomorphic, the diploid vegetative phase is the site of
meiosis and releases haploid zoospores, which germinate and grow producing a haploid phase
alternatingwiththevegetativephase.
3.3 Rhodophyta(RedAlgae)
The Rhodophyta are a distinct eukaryotic lineage characterized by the accessory photosynthetic
pigments phycoerythrin, phycocyanin and allophycocyanin arranged in phycobilisomes, and the
absenceofflagellaandcentrioles(Woelkerling,1990).Thisisalargeassemblageofbetween2500and
6000 species in about 670 largely marine genera (Woelkerling, 1990) that predominate along the
coastalandcontinentalshelfareasoftropical,temperateandcoldwaterregions(Lning,1990).Red
algaeareecologicallysignificantasprimaryproducers,providersofstructuralhabitatforothermarine
organisms,andtheirimportantroleintheprimaryestablishmentandmaintenanceofcoralreefs.Red
algaearecommonandwidespread,andecologicallyimportant.
Cell structure Red algae have a number of general characteristics that in combination distinguish
themfromothereukaryoticgroups:
absenceofflagellaandcentrioles,
florideanstarchasastorageproductandthestorageofstarchinthecytoplasm,
phycoerythrin,phycocyanin,andallophycocyaninasaccessorypigments,
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unstackedthylakoidsinplastids,
nochloroplastendoplasmicreticulum.
Therhodophytaexhibitthefollowingcommoncharacteristics:theyareunicellulartomulticellular(up
to 1 m), mostly freeliving but in some cases parasitic or symbiotic, with chloroplasts containing
phycobilins. Cell walls are made of cellulose with mucopolysaccharides (mainly agars and
carrageenans)penetratedinmanyredalgaebyporesmostlyblockedbyproteins(complexreferredto
aspitconnections).Theirmitochondriahaveflatcristaesometimesassociatedwithformingfacesof
dictyosomes. Thylakoids are single, with phycobilisomes, plastids with peripheral thylakoid. During
mitosis, the nuclear envelope mostly remains intact but some microtubules of spindle extend from
noncentriolarpolarbodiesthroughpolargapsinthenuclearenvelope.
Phylogeny Traditionally the red algae were divided into two Classes the Bangiophyceae and
Florideophyceae.AlternativelyasingleClass,theRhodophyceaeandtwoSubclasses,Bangiophycidae
andFlorideophycidaeareused.BasedonultrastructureandmolecularevidencetheBangiophyceaeis
now accepted as a paraphyletic group, while the Florideophyceae is considered to be monophyletic
basedontwosynapomorphiccharacterspresenceofafilamentousgonimoblastandtetrasporangia
(GarbaryandGabrielson,1990[andreferenceswithin],Raganetal.,1994).
ReproductionTheyusuallyhaveseparatedphasesofvegetativegrowthandsexualreproduction.
3.4 Heterokontophyta
The Heterokontophyta are a major line of eukaryotes. Most are algae, ranging from the giant
multicellularkelptotheunicellularforms.Thenameheterokontsreferstothemotilelifecyclestage,
inwhichtheflagellatecellspossesstwodifferentshapedflagella(Leipeetal.,1994;Patterson,1989).
Cell structure Heterokont algae are surrounded by four membranes, which are counted from the
outermosttotheinnermostmembrane.Thefirstmembraneiscontinuouswiththehost'schloroplast
endoplasmic reticulum, or cER. The second membrane presents a barrier between the lumen of the
endoplasmicreticulumandtheprimaryendosymbiontorchloroplast,whichrepresentsthenexttwo
membranes, within which the thylakoid membranes are found. This arrangement of membranes
suggest that heterokont chloroplasts were obtained from the reduction of a symbiotic red algal
eukaryote, which had arisen by evolutionary divergence from the monophyletic primary
endosymbiotic ancestor that is thought to have given rise to all eukaryotic photoautotrophs. The
chloroplasts usually contain chlorophyll a and chlorophyll c, and usually the accessory pigment
fucoxanthin,givingthemagoldenbrownorbrownishgreencolor.
Many heterokonts are unicellular flagellates, and most others produce flagellate cells at some
point in their lifecycle, for instance as gametes or zoospores. The name heterokont refers to the
characteristic form of these cells, which typically have two unequal flagella. The anterior or tinsel
flagellum is covered with lateral bristles or mastigonemes, while the other flagellum is whiplash,
smooth and usually shorter, or sometimes reduced to a basal body. The flagella are inserted
subapicallyorlaterally,andareusuallysupportedbyfourmicrotubulerootsinadistinctivepattern.
Mastigonemes are manufactured from glycoproteins in the cell's endoplasmic reticulum before
beingtransportedtoitssurface.Whenthetinselflagellummoves,thesecreateabackwardscurrent,
pulling the cell through the water or bringing in food. The mastigonemes have a peculiar tripartite
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structure, which may be taken as the defining characteristic of the group, thereby including a few
protiststhatdonotproducecellswiththetypicalheterokontform.Theyhavebeenlostinafewlines.
OriginsMostbasalheterokontsarecolorless.Thissuggeststhattheydivergedbeforeacquisitionof
chloroplasts within the group. Fucoxanthincontaining chloroplasts are also found among the
haptophyta. These two groups may have a common ancestry, and possibly also a common
phylogenetic history with cryptophyta. This may be interpreted as suggesting that the ancestral
heterokont was an alga, and all colorless groups arose through loss of the secondary endosymbiont
anditschloroplast.
Phylogeny As noted above, classification varies considerably. Originally the heterokont algae were
treatedastwodivisions,firstwithinthekingdomPlantaeandlatertheProtista.
In this scheme, however, the Chrysophyceae are paraphyletic to both other groups. As a result,
various members have been given their own classes and often divisions. Recent systems often treat
these as classes within a single division, called the Heterokontophyta, Chromophyta or Ochrophyta.
Thisisnotuniversal,howeverforinstanceRoundetal.(1990)treatthediatomsasadivision.
Thediscoverythatoomycetesandhyphochytridsarerelatedtothesealgae,ratherthanfungias
previouslythought,hasledmanyauthorstoincludethemamongtheheterokonts.Shoulditturnout
that they evolved from colored ancestors, the group would be paraphyletic in their absence. Once
again, however, usage varies. Patterson (1999) named this extended group the stramenopiles,
characterizedbythepresenceoftripartitemastigonemes,mitochondriawithtubularcristae,andopen
mitosis. He used the stramenopiles as a prototype for a classification without Linnaean ranks. Their
composition has been essentially stable, but their use within ranked systems varies. CavalierSmith
(1981)treatstheheterokontsasidenticalincompositionwiththestramenopiles;thisisthedefinition
followed here. He has proposed placing them in a separate kingdom Chromalveolata, together with
thehaptophytes,cryptomonadsandalveolates.Thisisoneofthemostcommonrevisionstothefive
kingdom system, but has not been generally adopted, partly because some biologists doubt their
monophyly. A few treat the Chromalveolata as identical in composition with the heterokonts, or list
themasakingdomStramenopila.
3.4.1
Phaeophyceae(Brownalgae)
The class of the Phaeophyceae (Guiry and Guiry, 2007), or brown algae, is a large group of mostly
marine multicellular algae, including many seaweeds of colder Northern Hemisphere waters. Brown
algaeareuniqueamongheterokontsindevelopingintomulticellularformswithdifferentiatedtissues,
but they reproduce by means of flagellate spores and gametes, which closely resemble other
heterokontcells.Geneticstudiesshowtheirclosestrelativestobetheyellowgreenalgae.
Theyplayanimportantroleinmarineenvironmentsbothasfood,andforthehabitatstheyform.
ForinstanceMacrocystis,amemberoftheLaminarialesorkelps,mayreach60minlength,andforms
prominent underwater forests. Another example is Sargassum, which creates unique habitats in the
tropical waters of the Sargasso Sea. Many brown algae such as members of the order Fucales are
commonlyfoundalongrockyseashores.Somemembersoftheclassareusedasfoodforhumans.
Worldwide there are about 15002000 species of brown algae. Some species are of sufficient
commercialimportance,suchasAscophyllumnodosum,thattheyhavebecomesubjectsofextensive
researchintheirownright(Senn,1987;vandenHoek,1995).
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Algal structure Filamentous, syntagmatic or parenchymatous; cell wall present, containing alginate
compounds and cellulose; plasmodesmata or pores between cells in parenchymatous forms;
chloroplasts with girdle lamella; outer chloroplast endoplasmic reticulum membrane with direct
membrane connection to the outer nuclear envelope membrane; plastid DNA with ringtype
genophore; eyespots present or absent; plastid pigments include chlorophylls a and c1 and c2,
fucoxanthin, and violaxanthin; swimming cells with two flagella usually inserted laterally, one
anteriorly directed, one posteriorlydirected; usually four microtubular kinetosome roots but no
striated kinetosome root (rhizoplast); flagellar transitional helix typically with 6 gyres located above
themajortransitionalplate;noparaflagellarrod;littletosubstantialtissuedifferentiationoccurringin
parenchymatous forms.; macroscopic or microscopic, some polysiphonous; some form crusts,
cushionsorarehollowandothersgrowtoformlargeleatheryfronds(Jones,1962).
Evolutionary history Phaeophyta evolved from the phaeothamniophyceae between 150 and 200
million years ago. Claims that earlier (Ediacaran) fossils are brown algae have since been dismissed
(Loeblich,1974;Medlinetal.,1997;Lee,2008).Thelineagesofbrownalgaedivergedinthefollowing
order,fromoldesttoyoungest:Dictyotales;Sphacelariales;Cutleriales;Desmarestiales;Ectocarpales;
Laminarales; Fucales. Theiroccurrence asfossilsisraredue totheirgenerally softbodiedhabit,and
scientists continue to debate the identification of some finds. Only a few species of brown algae
depositsignificantquantitiesofmineralsinoraroundtheircellwalls.Otheralgalgroups,suchasthe
red algae and green algae have a number of calcareous members, which are more likely to leave
evidenceinthefossilrecordthanthesoftbodiesofmostbrownalgae.Miocenefossilsofasoftbodied
brownmacroalgae,Julescrania,havebeenfoundwellpreservedinMontereyFormationdiatomites,
butfewotherdubiouslyassignedfossils,particularlyofolderspecimensareknowninthefossilrecord
(Coyeretal.,2001).
LifecycleThelifecycleshowsgreatvariabilityfromonegrouptoanother.Howeverthelifecycleof
Laminaria consists of the diploid generation, that is the large kelp well known to most people. It
producessporangiafromspecialisedmicroscopicstructures,thesedividemeiotically(meiosis)before
theyarereleased.Astheyarehaploidthereareequalnumbersofmaleandfemalespores(Thomas,
2002). With the exception of the Fucales all brown algae have a life cycle which consists of an
alternationbetweenhaploidanddiploidforms.
EcologyBrownalgaehaveadaptedtoawidevarietyofmarineecologicalnichesincludingthetidal
splashzone,rockpools,thewholeintertidalzoneandrelativelydeepnearshorewaters.Theyarean
important constituent of some brackish water ecosystems, and four species are restricted to life in
freshwater(Lee,2008).AlargenumberofPhaeophyceaeareintertidalorupperlittoral,andtheyare
predominantly cool and cold water organisms that benefit from nutrients in up welling cold water
currentsandinflowsfromland;Sargassumbeingaprominentexceptiontothisgeneralisation.Brown
algaegrowinginbrackishwatersarealmostsolelyasexual(Lee,2008).
ChemistryBrownalgaehavea13Cvaluebetween20.810.5,incontrastwithredalgaeand
greens.Thisreflectstheirdifferentmetabolicpathways(Fletcheretal.,2004).
They have cellulose walls with alginic acid; fucoidin also important in amorphous section of cell
walls.Afewspecies(ofPadina)calcifywitharagoniteneedles(Lee,2008).
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3.4.2
Eustigmatophyceae
Eustigmatophyceae(Hibberd,1981;Andersenetal.,1998)areasmallgroup(7genera;12species)of
eukaryoticalgaethatincludesmarine,freshwaterandsoillivingspecies(vandenHoeketal.,1995).In
termsofecology,eustigmatophyceaeoccurasphotosyntheticautotrophsacrossarangeofsystems.
Mostgenera liveinfreshwater(Fawley,2007)orinsoil,althoughNannochloropsis(GuiryandGuiry,
2007)containsmarinespeciesofpicophytoplankton(24m).
CellstructureAlleustigmatophyceaeareunicellular,withcoccoidcellsandpolysaccharidecellwalls.
They contain one or more yellowgreen chloroplasts, which contain chlorophyll a and the accessory
pigments violaxanthin and carotene. Eustigmatophyte zoids (gametes) possess a single or pair of
flagella, originating from the apex of the cell. Unlike other heterokontophyta, eustigmatophyceae
zoids do not have typical photoreceptive organelles (or eyespot); instead, an orangered eyespot
outsideofthechloroplastislocatedattheanteriorendofthezoid.
3.4.3
Otherclasses
Yellowgreen algae or Xanthophyceae are an important group of heterokont algae. Most live in
freshwater,butsomearefoundinmarineandsoilhabitats.Theyvaryfromsinglecelledflagellatesto
simple colonial and filamentous forms. Xanthophyceae chloroplasts contain the photosynthetic
pigmentschlorophylla,chlorophyllc,Carotene,andthexanthophyllsvaucheriaxanthin,duatoxantin
and diadinoxanthin. Unlike other heterokonts, their chloroplasts do not contain fucoxanthin, which
accounts for their lighter colour. Its storage polysaccharide is chrysolaminarin. Xanthophyceae cell
wallsareproducedofcelluloseandhemicellulose.Theyappeartobetheclosestrelativesofthebrown
algae (Stace, 1991). Recent ultrastructural and molecular phylogenetic DNA (nuclear and plastid)
research shows that the Mischococcales might be paraphyletic, and the Tribonematales and
Botrydiales polyphyletic, and suggests two orders at most be used until the relationships within the
divisionaresorted(Adletal.,2005).
The golden algae or Chrysophyceae are a large group of algae found mostly in freshwater.
Originallytheyweretakentoincludeallsuchformsexceptthediatomsandmulticellularbrownalgae,
but since then they have been divided into several different groups based on pigmentation and cell
structure. They are now usually restricted to a core group of closely related forms, distinguished
primarily by the structure of the flagella in motile cells, also treated as an order Chromulinales. It is
possiblemembershipwillberevisedfurtherasmorespeciesarestudiedindetail.Mostmembersare
unicellular flagellates, with either two visible flagella, as in Ochromonas, or sometimes one, as in
Chromulina. Most genera have no cell covering, some have loricae or shells. Some members are
generallyamoeboid,withlongbranchingcellextensions,thoughtheypassthroughflagellatestagesas
well. Other members are nonmotile. Cells may be naked and embedded in mucilage, such as
Chrysosaccus,orcoccoidandsurroundedbyacellwall,asinChrysosphaera.Afewarefilamentousor
evenparenchymatousinorganization,suchasPhaeoplaca.
3.5 Labyrinthulea(phylumHeterokonta)
Thraustochytrids are exclusively marine heterotrophic protists that feed nonphagotrophically as
saprobes,epibiontsonalgae(microandmacroalgae)ormorerarelyasparasitesofmicroalgae(suchas
Skeletonema)andanimals. Theseunicellulareukaryoticprotistsareacommoncomponentofmarine
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microbialconsortia.Whethertoincludeornotthisgroupamongalgaeisstillobjectofdebateinthe
scientificcommunity,butduetotheirphylogeneticvicinityandtothefactthatproductsfromthese
organismareonthemarketasalgalproductstheyarediscussedinthisdocument.
CellstructureThraustochytriidaepresentanectoplasmicnet(EN)thatformsabranchednetworkof
plasma membrane extensions, associated with an organelle termed the bothrosome or
sagenogenetosome (sagenogen) at the periphery of the cell. The EN contributes to the increased
surfaceareaofthecellandcontainshydrolyticenzymesthataresurfaceboundoraresecretedinto
thesurroundingmedium,helpinginthedigestionoforganicmaterial.TheENalsoattachesthecellsto
surfacesand,inthecaseofthraustochytrids,penetratesorganicparticles(RaghuKumar,2002).Their
cellwallsarecomposedofnoncellulosicscalesandcontainsulphatedpolysaccharides,predominantly
ofgalactoseorfucose,andproteins(LeanderandPorter,2001;RaghuKumar,2002).Vegetativestages
ofthraustochytridsconsistofsinglecellswhichareglobosetosubglobose,measuring4to20min
diameter,mostlygrowingepibionticallyonvarioussubstrata.
Phylogeny The Labyrinthulomycota are a group of microorganisms of uncertain taxonomic
collocation.Theywereplacedamongfungiduetotheirfunctionalecologyandmorphology.Theywere
thenplacedinthegroupofOomycetesduetothepresenceofbiflagellatezoospores.(RaghuKumar,
1996).CavalierSmithetal.(1994)includestheLabyrinthulomycetesinthephylumHeterokonta,based
on the analysis of the 18S rRNA. The Class Labyrinthulea is then subdivided in the two subclasses
ThraustochytridaeandLabyrinthulidae.
Reproduction The Labyrinthulomycetes possess simple, asexual life cycles. Most thraustochytrids
reproduce by means of zoospores which possess a long anterior, tinsel flagellum and a short,
posterior,whiplashflagellum.Themodeofproductionofzoosporesvariesbetweengeneraandforms
themajortaxonomiccriterion(RaghuKumar,2002).Thecytoplasmiccontentsofthematurecell,the
sporangium,dividedirectlyintozoosporesinthegenusThraustochytrium.Thecytoplasmescapesas
one amoeboid mass, prior to zoospore division in the genus Ulkenia. The genus Schizochytrium is
characerisedbysuccessivebipartitionofavegetativecell,resultinginaclusterofcells,eachofwhich
develops into a zoosporangium or zoospore. Species within the genera are primarily defined by the
numberofproliferatingbodiesandthesizeandshapeofthesporangia.
3.6 Bacillariophyta(Diatoms)
Despitetheabundanceanddiversityofdiatomsinnature,fewspeciesareculturedinaquacultureor
for biotechnology relevant products (Lebeau and Robert, 2003). Further, only a handful of diatoms
have been studied and often only one or a few strains without any information on intraspecific
variation. There is a need to identify new diatom strains with as much positive characteristics as
possible or to breed or select for improved strains. Therefore, a basic knowledge of physiology,
ecologyandtaxonomyisimportant.
Diatomsarethemostspeciesrichandproductivegroupofeukaryoticalgae.Overacomparatively
short evolutionary time (< 250 Ma) (Sims et al., 2006; Sorhannus, 2007), they have diversified into
hundredsofgeneraandperhaps200.000extantspecies(Mann,1999).Theyareextremelyabundant
in all aquatic ecosystems, occurring in the plankton and benthos of marine and fresh waters (as
freeliving organisms or as endosymbionts in e.g. dinoflagellates and foraminifers), and in terrestrial
environments,suchasdampsoilsandmoistsurfacesofrocksandplants,fromthetropicstothepolar
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regions (Round et al., 1990). Diatoms have global ecological significance in the carbon and silicon
cyclesandprobablyprovide2025%ofglobalphotosyntheticfixationofcarbonandcontributeupto
40%ofthetotaloceanicprimaryproduction(Fieldetal.,1998;Falkowskietal.,1998).
Cell structure Diatoms are unicellular (10200 m), although they can also exist as colonies. The
hallmarkofthediatomcellisitsintricatelyshapedandornamentedsilicacellwall,calledthefrustule,
which consist of two overlapping thecae (Greek diatomos, meaning cut in half ), each in turn
consistingofavalveandanumberofhooplikeorsegmentalgirdlebands.Thefrustulesshowawide
diversity in shape, form and ornamentation, which has been the basis of the traditional diatom
taxonomy.Thiscellwallisencasedinanorganicmatrixthatisrichinproteinsandsugars.
Major pigments of diatoms are chlorophylls a and c, carotene, fucoxanthin, diatoxanthin and
diadinoxanthin. Diatoms are primarily photosynthetic. A few, however, are obligate heterotrophs,
whileotherscanliveheterotrophicallyintheabsenceoflight,providedanappropriateorganiccarbon
source is available. The main storage compounds of diatoms are lipids (TAGs) and a 1,3linked
carbohydrateknownaschrysolaminarin.
LifecycleDiatomshaveadiplonticlifecyclewithaprolongedvegetativephaseduringwhichthecells
divide mitotically. Their unique cell wall structure (two overlapping halves) and division pattern in
which new cell wall components are formed within the parental cell, cause diatoms to gradually
reducetheircellsizeinthecourseofthemitoticpartofthelifecycle(MacDonaldPfitzerrule)(Round
et al., 1990). Cell size is restored through the development of a special expanding cell called the
auxospore, which normally results from sexual reproduction (Chepurnov et al., 2004). Sexual events
arevitaltoreestablishtheinitialcellsizeandtoavoidbecomingtoosmallforsurvival(Chepurnovet
al., 2004). As genetic recombination is also achieved through sexual reproduction, the obligatory
natureofsexintheirlifecyclemaybelinkedwiththeevolutionaryandecologicalsuccessofdiatoms.
ClassificationDiatomsarerecognizedasasinglewelldefinedgroupattheclassordivisional(called
Bacillariophyta)level(MannandEvans,2007;Kooistraetal.,2007).
Diatoms have historically been divided into two classes or orders, the centric and the pennate
diatoms. Most centric diatoms have valves with a radial symmetry (circular or shortly elliptical,
triangular, or polygonal) while pennate diatoms are mostly elongate and have a bilateral symmetry.
Sexualreproductionincentricsisusuallyoogamous,withmotileuniflagellatesperm.Pennatediatoms
are usually isogamous with amoeboid gametes which may be differentiated biochemically and
behaviourally (Chepurnov et al., 2004). The pennate diatoms include araphid and raphid forms. The
raphe,aslitopeninginthecellwall,allowforactiveglidingalongsurfaces(Roundetal.,1990).
Arecentreevaluationofdiatomphylogeny(MedlinandKaczmarska,2004)hasdivideddiatomsinto
three classes: the Coscinodiscophyceae includes radial centric diatoms, the Mediophyceae includes
multipolar centrics plus some radial centric diatoms, and the Bacillariophyceae includes pennate
diatoms.Membersoftheseclassesdifferwithregardstocellshape,presence/absenceofaraphe(in
pennate diatoms), structure and arrangement of the Golgi apparatus and chloroplast pyrenoid and
modeofsexualreproduction(MedlinandKaczmarska,2004).
Traditionally, classification at the order level and below was almost exclusively based on
morphologicalfeaturesofthesiliceouscellwall(Roundetal.,1990).Speciesboundarieshavelargely
been based on discontinuities in morphological and ultrastructural characters of the frustule. The
advent of molecular genetic studies have shown that many morphologicallydefined species are, in
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fact,assemblagesofcrypticspecies;thatis,assemblagesofgeneticallyandbiologicallydistinct,albeit
often closely related species. Cryptic diversity has been discovered throughout the diatom diversity
(reviewedine.g.MannandEvans,2007;Kooistraetal.,2007).
The prevalence of cryptic species has important consequences for understanding patterns in
diatomdiversityandbiogeography,theirevolutionaryhistoryandtheirrelationshiptoenvironmental
conditions. Good knowledge of species limits is also a prerequisite for further studies at the species
leveland below,includingwithinspeciesgenetic, physiologicalandbiochemicalvariation,whichcan
haveimportantconsequencesincommercialusesofdiatoms.
GenomesequencingTheentiregenomesofthecentricdiatom,Thalassiosirapseudonana(34Mb),
(Armbrustetal.,2004)andthepennatediatom,Phaeodactylumtricornutum(27Mb),(Bowleretal.,
2008) have been sequenced. A draft sequence is now available for the polar species Fragilariopsis
cylindrus (80 Mb) and the toxic coastal species Pseudonitzschia multiseries (300 Mb) (Armbrust,
2009).
Oneoftheoutcomesofthesequencingprojectsthusfarisrecognitionoftheuniquecombination
of genes and metabolic pathways that distinguish diatoms from the evolutionarily distinct plant and
animal lineages. Enormous amounts of diversity are encapsulated within diatoms. For example, T.
pseudonanaandP.tricornutumprobablydivergedfromoneanotheronlyabout90millionyears(Ma)
ago,yettheirgenomesareaboutasdifferentasthoseofmammalsandfish,whichdivergedabout550
Maago(Bowleretal.,2008).
3.7 Haptophyta
TheHaptophyta,alsonamedPrymnesiophyta,aresometimesclassifiedasHaptophyceae(Satohetal.,
2009).However,althoughthephylogeneticsofthisgrouphasbecomemuchmorewellunderstoodin
recentyears,thereremainssomedisputeoverwhichtaxonlevelismostappropriate.
CellstructureThechloroplastsarepigmentedsimilarlytothoseoftheheterokonts(Anderson,2004),
butthestructureoftherestofthecellisdifferent,sotheymaybeseparatelines,whosechloroplasts
arederivedfromsimilarendosymbionts.
Thecellstypicallyhavetwoslightlyunequalflagella,bothofwhicharesmooth,andauniqueorganelle
called haptonema, which is superficially similar to a flagellum but differs in the arrangement of
microtubules and in its use. The name comes from the Greek hapsis, touch, and nema, thread. The
mitochondriahavetubularcristae.
Examples and classification The bestknown haptophytes are coccolithophores, which have an
exoskeleton ofcalcareousplatescalledcoccoliths.Coccolithophoresaresomeofthemostabundant
marinephytoplankton,especiallyintheopenoceanandareextremelyabundantasmicrofossils.Other
planktonic haptophytes include Chrysochromulina and Prymnesium, which periodically form toxic
marinealgalblooms,andPhaeocystis,thebloomsofwhichcanproduceunpleasantfoamwhichoften
accumulatesonbeaches.Bothmolecularandmorphologicalevidencesupportstheirdivisionintofive
orders.
3.8 Dinophyta(Dinoflagellates)
Dinophytaordinoflagellatesarecommonorganismsinalltypesofaquaticecosystems.Roughlyhalfof
thespeciesinthegrouparephotosynthetic(GainesandElbrchter,1987),theotherhalfisexclusively
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heterotrophic and feeds via osmotrophy and phagotrophy. As a consequence, they are prominent
membersofboththephytoplanktonandthezooplanktonofmarineandfreshwaterecosystems.They
are also common in benthic environments and in sea ice. Approximately 4500 species assigned to
morethan550generahavebeendescribed,nearlythreequartersofthegeneraandmorethanhalfof
thespeciesbeingfossil.Oftheca.2000livingspecies,morethan1700aremarineandabout220are
from freshwater (Taylor et al., 2008). Between the years 2000 and 2007 three new dinoflagellate
families, 22 new genera, and 87 new species were described (Centre of Excellence for Dinophyte
TaxonomyCEDiT).Recentmolecularanalyseshaveshownthattherearelargenumbersofundescribed
dinoflagellatespeciesinenvironmentslikemarinepicoplankton(e.g.MoreiraandLpezGarca,2002;
Worden,2006)orassymbionts(zooxanthellae)inmanytypesofprotistsandinvertebrateslikecorals
(CoffrothandSantos,2005).
Thephotosyntheticandmixotrophic(utilisingbothinorganiccarbonandorganiccarbonsources)
species are very important players in oceanic carbon cycles, and some cause harmful (toxic) algal
blooms when cell densities reach exceedingly high levels (Taylor, 1987). Many photosynthetic
dinoflagellatesarealsoconsumersofbacteriaandothermicroeukaryotes(Stoecker,1999).
Cell structure Dinoflagellates have two flagella with independent beating pattern that confers a
peculiarrotattoryswimmingwhirlingmotion.Flagellaareinsertedapically(desmokonttype)orthey
emerge from a region close to the midpoint ventral side of the cell (dinokont type) (Barsanti and
Gaultieri,2006).Aroundthecelltheypresentalayerofflatvescicelssurroundingthecells:thesecan
beemptyorfilledwithcellulose(armoreddinoflagellates).Theypossesschlorophyllsa,b,c1andc2
andfucoxanthin,peridinin,dinoxanthin.Dinoflagellateshavenovelcytoskeletalandnuclearfeatures
(e.g. permanently condensed chromosomes) that make them very distinctive among eukaryotes
(Fensomeetal.,1999).Dinoflagellatesmorphologicalfeaturesinclude:
A system of abutting membranous sacs, called alveoli, positioned beneath the plasma
membrane(synapomorphy);thealveoliarefilledwithcellulosicmaterial.
Distinctmicroporesthroughthecellsurfacethatfunctioninpinocytosis(synapomorphy).
Extrusiveorganelles.
Closedmitosis.
Tubularmitochondrialcristae.
Complexorganellesfoundinthegroupincludestructuresreminiscentofafullfledgedvertebrate
eye(butinaunicellularorganismthatlacksabrain),nematocystscomparabletothoseofcnidarians,
andabewilderingarrayofplastidtypesinthephotosyntheticforms.Dinoflagellatesexistasplasmodia
(i.e.multinucleateorganisms),biflagellatedcells,coccoidstagesandeven,inonesmallgroup,ascell
arraysthatapproachmulticellularity.
Genetically,dinoflagellatesarealsounique.Thenucleusofalargemajorityofdinoflagellates(the
socalled dinokaryotes) is so different from other eukaryotic nuclei that it has been given its own
name, the dinokaryon. Dinokarya lack nucleosomes, and DNA content is orders of magnitude larger
thanthatofothereukaryoticcells,forexamplethoseofhumans.Thesedinokaryadivideviaaunique
form of mitosis. Recent research is starting to show just how unique dinoflagellate genetic systems
are. For example, gene products of all dinoflagellate nuclei (not only dinokarya) are processed in a
unique way: a spliced leader is transspliced to all mRNA molecules. The genomes of plastids and
mitochondriaofthegrouparealsounique:theyareatomized(i.e.thegenomeissplitintoverysmall
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fragments), and gene content is much, much lower than that of comparable organelles in other
organisms.
Ecological significance Dinoflagellates are perhaps best known as causers of harmful algal blooms
(webpages about this topic: ISSHA, WHOI, IOC). About 7580% of toxic phytoplankton species are
dinoflagellates(Cembella,2003),andtheycauseredtidesthatoftenkillfishand/orshellfisheither
directly,becauseoftoxinproduction,orbecauseofeffectscausedbylargenumbersofcellsthatclog
animal gills, deplete oxygen, etc. (Smayda, 1997). Dinoflagellate toxins are among the most potent
biotoxinsknown.Theyoftenaccumulateinshellfishorfish,andwhentheseareeatenbyhumansthey
cause diseases like paralytic shellfish poisoning (PSP), neurotoxic shellfish poisoning (NSP), diarrheic
shellfishpoisoning(DSP)andciguatera(LehaneandLewis,2000).Theyalsohavebeenlinkedtomajor
human health concerns, especially in estuarine environments (Pfiesteria). Some syndinians, notably
Hematodinium,areparasitesofeconomicallysignificantcrustaceanspecies.
Themainecologicalsignificanceofdinoflagellateslieselsewhere,though.Theyaresecondonlyto
diatomsasmarineprimaryproducers.Asphagotrophicorganismstheyarealsoimportantcomponents
ofthemicrobialloopintheoceansandhelptochannelsignificantamountsofenergyintoplanktonic
foodwebs.Aszooxanthellae,dinoflagellateshaveapivotalroleinthebiologyofreefbuildingcorals.
3.9 Otheralgalgroups
Euglenophyta (or euglenoids) are one of the bestknown groups of flagellates, commonly found in
freshwater especially when it is rich in organic materials, with a few marine and endosymbiotic
members.Manyeuglenidshavechloroplastsandproduceenergythroughphotosynthesis,butothers
feedbyphagocytosisorstrictlybydiffusion.Euglenoidsaredistinguishedmainlybythepresenceofa
pellicle, which is composed of proteinaceous strips underneath the cell membrane, supported by
dorsal and ventral microtubules. This varies from rigid to flexible, and gives the cell its shape, often
giving it distinctive striations. In many euglenoids the strips can slide past one another, causing an
inchingmotioncalledmetaboly.Otherwisetheymoveusingtheflagella.
Cryptophyta are aquatic unicellular photosynthetic eukaryotes that inhabit both marine and
freshwater environments. Plastids are very diverse in pigmentation. In addition to being important
from an ecological perspective, the cryptophytes are of pivotal significance in terms of our
understanding of endosymbiosis and the evolution of plastids. This is because cryptophyta acquired
photosynthesis by the process of secondary (i.e., eukaryoteeukaryote) endosymbiosis, having
engulfed and assimilated a red algal endosymbiont at some time during their evolutionary past
(ArchibaldandKeeling,2002;Bhattacharyaetal.,2003;Gouldetal.,2008). Asaresult,cryptophyta
are extremely complex from a genetic and cell perspective, having a fourmembranebound plastid
andfourdistinctDNAcontainingcellularcompartments,theplastid,mitochondrion,hostnucleusand
endosymbiont nucleus, the latter being referred to as the nucleomorph. The cryptophyte
nucleomorph genome is highly reduced in structure and coding capacity, and the focus of ongoing
research aims at understanding the pattern and process of secondary endosymbiosis (Gilson and
McFadden,2002;Archibald,2007).
Glaucophyta, also known as glaucocystophytes or glaucocystids, are a small group of freshwater
microscopic algae (Keeling, 2004). The glaucophytes are of interest to biologists studying the
development of chloroplasts, because some studies suggest that they may be similar to the original
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alga type which led to green plants and red algae (Keeling, 2004, Kim and Graham, 2008). The
chloroplasts of glaucophytes are known as cyanelles. Unlike other eukaryotic plastids they have a
peptidoglycan layer which is believed to be a relic of the endosymbiotic origin of plastids from
cyanobacteria (Keeling, 2004). Glaucophytes contain the photosynthetic pigment chlorophyll a
(Keeling, 2004). Along with red algae and cyanobacteria they harvest light via phycobilisomes,
structures consisting largely of phycobiliproteins. Glaucophytes have mitochondria with flat cristae,
andundergoopenmitosiswithoutcentrioles.Motileformshavetwounequalflagella,whichmayhave
fine hairs and are anchored by a multilayered system of microtubules, both of which are similar to
formsfoundinsomegreenalgae.Thereareonly13speciesofglaucophytesknown,noneofwhichare
particularlycommoninnature.
Chlorarachniophyta are green amoebas living in sea water sample. The green color is caused by
chloroplasts, which are permanently housed in the amoeba cell. Although the chloroplasts contain
chlorophylls a and b, chlorarachniophyta are phylogenetically distinct from other chlorophyll b
containingeukaryotes(ChlorophytaandEuglenophyta),andconstituteanindependentphylum,which
wasestablishedonlytwodecadesago(HibberdandNorris,1984).Thegroupconsistsoffourgenera
andfivespecies,andtherearealsoseveralspecieswaitingtobenamed(Ishidaetal.,1999;2000).
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4 Biotechnologyofalgae
4.1 Introduction
Ediblebluegreenmicroalgae,includingthecyanobacteriaNostocandArthrospira(formerlySpirulina),have
beenpartofthehumandietsincemanycenturies.However,themoderneraofmicroalgalbiotechnology
beginsintheearly1950s,whenthefirstalgaemassculturesinphotobioreactors(PBR)wereattempted.
Today, between 8,000 and 10,000 tons of algae biomass are produced annually (Pulz and Gross, 2004;
Becker, 2007), mainly for use as human food supplements and animal feed. Table 3 shows the present
statusofcommercialproductionandthemainapplicationareasofmicroalgae(Tredicietal.,2009).Table4
showsthecomparisonofseveralalgaeproductionsystems.
Microalgae biomass is either harvested from natural habitats or obtained through more or less
controlledcultivationprocessescarriedoutmainlyinlargeopenpondsorlagoons.Commercialproduction
inphotobioreactors(PBR)islimitedtoafewhundredsoftons(seebelow).
Table3Commercialproductionofmicroalgae(fromTredicial.,2009).
The commercial production of microalgae is mainly limited to species belonging to the genera
Arthrospira, ChlorellaandDunaliella that,duetotheirhighgrowthrateorofaselectivegrowthmedium
that limits contamination, can be grown in large open ponds. These systems are easier to operate, less
expensive, and more durable than large closed reactors (Tredici, 2004). However, the majority of
microalgaedoesnotrequireaspecificgrowthenvironmentoraselectivemediumandcannotbecultivated
for prolonged periods in outdoor open systems because of contamination. Photobioreactors provide a
closedenvironmentfortheculture(limitingdirectfalloutandinvasionbyunwantedspecies)andabetter
control of culture parameters (pH, temperature, pO2, etc.) which ensure the dominance of the desired
species(Tredici,2004).
To exploit the large biodiversity of microalgae and cyanobacteria, lowcost and scalable PBR are a
necessity, either to be used alone or in combination with open ponds, in this latter case to produce the
inocularequiredtomaintaintheselectedspeciesintheponds.
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Table4Comparisonofseveralalgaeproductionsystems(fromTredicietal.,2009).
4.2 Cultivationsystems
As all microorganisms, microalgae can be cultivated in three modes, in batch, semicontinuous or
continuouscultivation.
Batch cultivation relates to inoculation of the bioreactor, provided with fresh cultivation medium, with
sufficient biomass to induce immediate, rapid and vigorous growth until some component required for
growthbecomeslimiting.Forphotoautotrophicallygrowingalgaethisisoftentheamountofincominglight
energy.Whileoftencalledlightlimitation,thiseffectseemsrathertobebasedoncellularrespiorationof
verydenseculturesduringthenightequalingthephotosyntheticcarbonfixationrateandthuslossofnet
biomassaccumulation.
More sophisticated batch cultivation may rely on addition of limiting amounts of nutrients such as
nitrogen, so that the cells enter a stress phase and will accumulate high concentrations of oil or other
metabolitesbeforethegrowthprocesscomestoacompletearrest.
Semicontinuouscultivationrelatestoharvestingofacertainproportionofbiomasseveryfewdayswhile
resupplying the reactor with water and essential nutrients for inducing a new growth cycle. This avoids
complete restarting of the reactor with fresh algae and medium, which can be a work intensive and
expensive process. Boussiba et al. (1987, 1988) have applied this process for studying growth and oil
production by Nannochloropsis and Isochrysis. Limiting nitrogen supply allowed creating biomass with
average30%oilcontentoveratwomonthsperiod.
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SimilarlythebioreactorfacilityatQetura(Israel)reliesonsemicontinuouscultivationforproductionof
green Haematococcus biomass, while the second growth stage, the stress phase inducing astaxanthin
biosynthesis,ishandledasbatchculture.
Continuous cultivation: continuous cultivation is widely used for cultivation of heterotrophic organisms
whereby all reactor parameters as well as cell density are tightly supervised and controlled and growing
biomass is continuously harvested to maintain optimal growth conditions for extended periods of time.
While this mode can yield the best possible biomass yields, production of high concentrations of oil or
carotenoidsrequiringstressconditionsisnotpossible.
4.2.1
Openponds
Openshallowponds(Fig.2)mixedandaeratedbyanygivenmethodhavebeenthemethodofchoicesofar
for producing biotechnologically relevant amounts of algal biomass. CO2 is inserted as required. Cultures
are mixed in circular ponds using circulation ponds, or in the oval so called raceway ponds using paddle
wheelstomovethemediumaround.MajorproducersofArthrospira(Spirulina)andDunaliellainCalifornia,
Hawaii,AustraliaandIsraelhavebeenrelyingonthismethodforover30years,takingadvantageofthefact
that the growth media of those two algal species (basic or highly saline respectively) are unsuitable for
competing algal species. Currently, commercial cultivation of microalgae is limited to a few microalgal
species that are cultivatedin open ponds. A raceway pond is an economic open system for microalgae
cultivation. It consists of acircuit of parallel tunnels placed at ground level, in which the microalgae
suspensionflowsgentlyandismovedbyapaddlewheel.Thesystemisbuiltinconcreteorditchduginto
the ground and may be lined with white plastic sheets. Evaporation is significant as well as temperature
fluctuations,andrainfallscandilutetheavailablenutrients.
High level of contamination by predators and other fast growing heterotrophs have restricted
thecommercialproductionofalgaeinopenculturesystemstoonlythoseorganismsthatcangrowunder
extremeconditions.ExamplesareDunaliellasalina(highsalinity)andArthrospiraplatensis(Spirulina)(high
alkalinity).However,BGUismaintaininganopenNannochloropsiscultureoutdoorswithoutanysignificant
contaminationproblems,andSeambiotichassimilarlybeenabletosuccessfullymaintainNannochloropsis
cultureseveninclosevicinityoftheocean.
The Algal Biofuels Program of the DOE (Sheehan et al., 1998) has also concluded that open raceway
ponds would be the method of choice for algal biomass production, whereby spontaneously developing
localalgalpopulationsmightbethebiomassofchoiceforsimplifyingtheproductionprocess.
Figure2Examplesofmicroalgaeopencultivationsystems:a)Boardmixedponds,UNIFI,Italy(1980);b)Pilotracewaypond,
Necton,Portugal(1996);c)Racewayponds,HainanDIC,China(1996);d)Cyanotech,Hawaii,USA(2008);e)Fluegasfixation
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projectofSeambioticLtd,Israel;f)Racewaypond,sEarthrisefarms,California,USA(1982);g)Racewaysof2000m2,designedby
BGUandinoperationatEilat;Israel;h)PondsinoperationatParryAgroIndustries,India(totalpondsarea53ha)(2006).
4.2.2
Photobioreactors
Photobioreactors (PBRs) (some examples are reported in Fig. 3) can be defined as culture systems for
phototrophsinwhichagreatproportionofthelightdoesnonimpingedirectlyontheculturesurface,but
hastopassthroughthetransparentreactorswallstoreachthecultivatedcells.Consequently,PBRsdonot
allow,orstronglimit,directexchangeofgasesandcontaminantswiththeenvironment(Tredici,2004).
Reactorsusedforthecultivationofmicroalgaeareeithernaturallyorartificiallyilluminated,andmay
beopenorclosedsystems.Opensystemswithnaturallyilluminatedlargesurfaceareasincludeessentially
raceways ponds. PBRs are closed cultivation systems to grow microalgae under photoautotrophic
conditions.Severaltypesofphotobioreactors(PBRs)havebeendesignedandexperimentedwithsincethe
late 1940s, when algal cultures were first considered the ideal solar technology to produce in a cost
effectivemannerbiomassandproteinonalargescaleandsavethehungrypartofhumanity(Tredicietal.,
2010).Theycanbeclassifiedonthebasisofbothdesignandmodeofoperation.Indesignterms,themain
categoriesofreactorsare(1)flatortubular;(2)horizontal,inclined,verticalorspiral;and(3)manifoldor
serpentine. The most used designs are flatplates (vertical or inclined), tubular reactors (serpentine or
manifoldwithhorizontal,verticalorinclinedarrangement),airbubbledplasticbagsandannularcolumns.
Someofthem,especiallyinsmallscale,areartificiallyilluminatedwithfluorescentorothertypesoflamps.
ThetwomajorcategoriesofPBRsrecentlyproposedforbiofuelsatrelativelylargescaleareflatandtubular
reactors(Tredicietal.,2010).
As PBR are more expensive to install and operate, intensive study is being performed in an effort to
reducetheircostandthusfacilitatetheiruseespeciallyforlowvalueproductslikealgaloils.Allparameters
(e.g. nutrients, light regime, gas exchange) are maintained to realize optimal culture conditions. Costs of
PBRs are much higher, but contamination is much lower compared to open systems (Tredici, 2004). PBR
design and engineering is still a very active field of research today, since closed culture systems are
necessary to grow photosynthetic microbes and exploit them as a source of aquaculture feeds, food
additives, specialty chemicals, cosmetics and are preferred by industry as research tools for biofuel
production.
Inairbubbledplasticbagsandannularcolumnsthecostofoperationmaybeveryhighduetotheneed
of mixing to provide the necessary mass transfer. This operation requires more than 100 W.m3
(approximately 2000 MJ ha1 day1) or up to 50% of the energy stored in the biomass [Bassi et al.,
UnpublishedData].
Withinallthephotobioreactortypesdescribed,tubularPBRsarethemostpopulardesign(Chisti,2007;
Tredicietal.,2010).Theiroperationcostsarenormallysignificantlyhigherthanthoseofponds.Mixingin
tubular reactors to achieve sufficient fluid velocities and Reynolds numbers may require even higher
energyinputs.Besides,thereistheneedforcooling,whichisgenerallyprovidedbywater(evenseawater)
sprayingorbyinsertionofacoolingserpentineintheculture.
FlatPanelReactorsconsistofarectangulartransparentboxwithadepthofonly1to5cm.Accordingly,
thisreactortypeallowsgrowthofhighcelldensities.Verticalreactorsinterceptsunraysatlargeanglesand
dilute light compared with horizontal ponds. Besides, their rear surface receives mostly diffuse and
reflectedradiationoflowintensity.Forthisreason,verticalPBRsaremoreefficientthanhorizontalponds
intermsofsolarenergyutilization(Tredicietal.,2010).
OneofthemainproblemsassociatedwithPBRscultureisthattheformationofbiofilmsofbacteria,or
the cultivated algae themselves, at the transparent walls of the tubes considerably reduce light
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penetration.Biofilmsareminimizedbymaintaininghighlyturbulentflow,usingsmallparticlesandregularly
cleaningwithhydrogenperoxide.
PBRshavemanyadvantagesoveropenponds,someofthemwerereferredinthemostrecentarticleof
Tredici(Tredicietal.,2010),andaredescribedbelow:
theyareclosed,makingiteasiertomaintainanunialgalcultureandreducingevaporation;
due to a higher surfacetovolume ratio, PBRs may attain higher volumetric productivities
thanponds and adopt higher cell concentrations, which reduce the costs for medium
preparationandhandlingandthatforharvesting;
PBRsprovideamoreaccuratecontrolofcultureparameters;
theultimateandmostimportantadvantageofPBRs,however,isthattheypermittocultivatealgal
speciesthatcannotbegrowninopenponds.
Photobioreactorspayheavilyfortheiradvantages.Ingeneral,PBRsaremuchmoreexpensivetobuild
thanponds,butsimplelowcostsystemscanalsobedesigned.Mostconstrainsrequirefurther
investigation, specially lowcost materials for construction and lower energy requirements, but also
overheating,biofouling,oxygenaccumulation,difficultyinscalingupandcelldamagebyshearstress.
Figure3Examplesofmicroalgaeclosecultivationsystems:ab)GreenWallPanelsandtubularphotobioreactors
fromUNIFIUniversitdegliStudidiFirenzeandF&MFotosintetica&Microbiologicas.r.l.,Italy;c)FlatPanel
photobioreactorfromNecton,S.A.,Portugal;de)TubularphotobioreactorwithfluegasfluxuationfromAlgaFuel,
S.A.,Portugal;fg)TubularphotobioreactorsfromAlgaTechnologies,Israel;h)FlatPanelatBenGurionUniversity
BGU,Israel.
Mainphotobioreactorsdesigns
Polyethylenebagsandverticalcolumns
Polyethylenebags(sleeves)suspendedfromaframeworkorsupportedwithinameshframeandmixedby
air bubbling are the most common cultivation devices used in hatcheries for the production of algal
biomass (Fig. 4). From 50 to 500 L in volume, such reactors are mostly used indoors with artificial
illumination.Theyarecurrentlyusedbydifferentcompanies(e.g.KeturaKibbutz,Israel;GreenSea,France;
NOVAgreen GmbH, Germany) for the cultivation of selected species for the cosmetic, food or
pharmaceuticalmarkets.Althoughsleevereactorsareinexpensive,theirlowsurfacetovolumeratio(S/V),
biofoulingandtheneedofaverylargenumberofunitsforlargescaleproductionlimittheirapplications.
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The company Proviron developed inflatable bags with multiple vertical panes (ProviAPT)
(http://www.proviron.com/algae/GB/algae_solution_working.php)(Fig.4).Thebig,translucent,plasticbag
isfilledwithwatercontainingthemultipleverticalpanes.ThealgaegrowinthesepaneswhileCO2richair
isbubbledthrough.Duetothewaterthereactorisselfsupportingandthewaterbuffersthetemperature
efficientlybetweendayandnight.Itisacontinoussytemwithcontinousharvesting.
Vertical columns are made of rigid transparent cylinders (typically 22.5 m in height and 3050 cm in
diameter),withmixingachievedbyairbubblingorbyanairlift.Theyareextensivelyusedinhatcheriesand
suffer from the same limitations of sleeves. After the first design by Helm, vertical cylinders internally
illuminatedhavebeendevelopedattheUniversityofFlorence.Thereactor,namedannularcolumn(Fig.4),
ismadeoftwoconcentrictransparentcylindersofdifferentdiametersealedatthebasetoformaculture
chamberwithmuchhigherS/Vcomparedtocompletelyfilledcolumns.Lampsorfluorescenttubescanbe
placedinsidetheinner cylinder.Annularcolumnsof 30230Linvolumeare currently commercialized by
Fotosintetica&MicrobiologicaS.r.l.(Italy),aspinoffcompanyoftheUniversityofFlorence.Thesmallsize
andrelativelyhighcostofthisreactorlimititsuseforlargescaleproduction.
Figure4PolyethylenebagsandAnnularColumns.Topleft:PolyethylenebagsatNecton,SAAlgarve,Portugal;
Topright:AnnularColumnsatDepartmentofAgriculturalBiotechnologyoftheUniversityofFlorence,Italy;
Bottom:Inflatablebagswithmultipleverticalpanes(Proviron,Belgium).
TubularPBR
MostofthePBRadoptedincommercialplantsareofthetubulartype.Thiscategorycanbesubdividedinto
threemainsubgroups:i)serpentine,ii)manifoldandiii)helicalPBR(Fig.5,6).
SerpentinePBR,firstlydesignedbyTamiyainthe1950s,consistofstraighttubesconnectedbyUbends
toformaflatloop(thephotostage)thatmaybearrangedeitherverticallyorhorizontally.
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Figure5Horizontaltubularphotobioreactors.MeraPharmaceuticals,IncUSA(left);Serpentinephotobioreactor.
AlgaeLinkNVTheNetherlands(right).
ManifoldPBRaremadeofparalleltubesconnectedattheendsbytwomanifolds.Themainadvantages
of these systems over serpentine reactors are the reduction of head losses and lower oxygen
concentrations,twofactorsthatfacilitatescaleuptoindustrialsize.Ahorizontalmanifoldreactorisused
byAlgatechnologiesLtd(Israel)forthecommercialproductionofH.pluvialis.TheNearHorizontalTubular
Reactor(NHTR),amanifoldreactorwithtubesinclinedfrom5to20tothehorizontalsothatmixingcould
be achieved by air bubbling was developed by Tredici and coworkers (Tredici et al., 1999). A 20m long
NHTRwasexperimentedwithattheUniversityofHawaii(USA).TwosmallNHTRunitsarecurrentlyusedby
ENISpA(Italy)todevelopCO2abatementstrategieswithmicroalgae(Pedronietal.,2004).Asimilardesign,
the triangular airlift reactor, consisting of a series of riser tubes, gas separators and downcomer tubes
arrangedinatriangularconfigurationwasusedbyGreenFuelTechnologiesCo(USA)fortheabatementof
greenhouse gases. Rather common, both at pilot scale and in commercial plants, are manifold
photobioreactorsarrangedfencelike.
Figure6Firstontheleft:TriangularPhotobioreactor.GreenFuelTechnologiesCoUSA.Middleandrightfigures:
BioFencesystems.VariconAquaSolutionsLtdUK
TheBioFence,developedbyAppliedPhotosyntheticsLtd(UK)inthelate1990s,consistsofanarrayof
transparenttubesrackedtogetherinafencelikestructureinwhichtheculturesuspensioniscirculatedby
a centrifugal pump or by an airlift. BioFence systems are currently commercialized by Varicon Aqua
SolutionsLtd(UK).Biofencephotobioreactorsfrom10to35000LarealsodistributedbyBBraunBiotech
International GmbH (Germany). Industrial scale plants based on this design are operated by Bioprodukte
ProfSteinbergGmbH(Germany)fortheproductionofChlorellaandbyAlgatechnologiesLtd(Israel)forthe
growthofH.pluvialis.
Helical photobioreactors (biocoils), consisting of smalldiameter flexible tubes wound around an
uprightstructurehavealsobeenmuchexperimentedandcommercializedinthepast.
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Flatphotobioreactors(panels)
Flat photobioreactors (panels) have not been used for mass cultivation of algae until recently. The first
versionofthealveolarpaneldevelopedbyTredicietal.in1988,inwhichmixingwasachievedbyapump,
was brought to industrial level by Pulz and coworkers (Pulz and Scheibenbogen, 1998). This reactor is
commercializedbyBBraunBiotechInternationalGmbH(Germany)insizesvaryingfrom10to2000L.Inthe
mid1990s,Richmondandcoworkersdevelopedglasspanels(withoutalveoli)ofvariouswidths(Huetal.,
1996). Glass panels are highly transparent, easy to clean and resistant to weathering. However, weight,
fragility and cost discourage their use for large scale plants. In the early 2000s the concept of the
disposablepanelwasdevelopedandpatentedbytwogroupsworkingindependentlyinItalyandIsrael.A
disposable panel is a flat reactor consisting of a plastic culture chamber enclosed in a rectangular metal
frameorcage.
Themainadvantagesofthesesystemsarethelowconstructioncost,thecapacitytobescaledup(Fig.
7) and a disposable culture chamber for the cultivation of those microalgae which suffer from
contamination. These systems are today successfully used in some hatcheries for microalgae production
both in Israel and Italy. The green wall panel is commercialized by Fotosintetica & Microbiologica Srl
(Italy).
Figure7ComparisonbetweenopenpondsandPBRproductivityandenergyoutput(SourceProfTredici).
4.2.3
Sustainabilityofdifferentcultivationsystems
The present attempts to develop largescale microalgae production systems for biofuel generation stem
fromtheperceivedinadequaciesoftraditionalfeedstockusedforbiofuelproduction.
Microalgaehaveadvantagesovertraditionalenergycrops,suchasthepotentialforhighyieldbiomass
production on marginal land or in seawater. Thus with microalgae, biofuel production without landuse
conflictsbecomesfeasible.
AnotheradvantageofmicroalgaeistheirabilitytocaptureCO2frompowerorindustrialplants.Overall
microalgalbasedbiofuelisclaimedtoreduceenvironmentalimpact(Chisti,2007).Thusworldwidethereis
great hope that the cultivation of microalgae for energy generation can contribute to sustainable energy
supplyofthefuture.
However,sofarnooverallsustainabilityassessmenthasbeen conductedon thebasisof datafroma
microalgaedemonstrationplantswiththesizeof10hectaresandhighproductivitiesof100tonsha1year1.
Preliminary assessments are indicating that with present technologies the energetic inputs of microalgae
production could exceed the energetic output. In closed simplebuild reactors more than 3 W m2 of
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electricalenergyareusedforoperatingthecultivation.Thiscomesclosetotheamountactuallyharvested
at the end. However, it has to be taken into account, that in nowadays commercial algae cultivation, a
positiveenergybalanceisnottargetedduetothehighpricesachievedfortheproducts.
LCA and sensitive analysis can help to identify at which points of the production chain technological
progressisneededandhowthedemonstrationplantcouldbedesignedinamoresustainableway.
Alongdisputeconcerningtheoptimalmodesofcultivationforalgalbiofuelsproductionmayhavebeenlaid
torestduringthelasttwoyearsthankstoanumberofanalysesdeterminingtheenvironmentallifecycle
impactofdifferentcultivationmodes.
Several LCA analyses agree in one major conclusion, that even under optimistic yield assumptions
tubular photobioreactors require far more energy to run than the algal biomass being produced. More
simplepanelreactorsfaresomewhatbetter,butstillareunabletoreturntheamountofenergyrequiredto
constructandrunthem.Ontheotherhandopenracewaypondsassumingyieldsof20gm2ormorecan
returnsignificantlymoreenergyintheformofbiomassthanhastobeinvestedforbuilding,runningand
maintainingtheponds(Joqueraetal.,2010;Stephensonetal.,2010).
Thisdifferencebecomesevenmorepronouncedwhentakingintoaccountthataboveanalyseshavenot
incorporatedenergyexpensesforcoolingorheatingwhichcanbetremendousinphotobioreactors,butare
notrequiredinopenponds.
4.3 Harvestingmethods
Algaeharvestinganddewateringprocessesmayaccountfor2030%ofthetotalproductioncostforalgal
biomass (Uduman et al., 2010; Pienkos et al., 2009). In order to remove large quantities of water and
processlargealgalbiomassvolumes,aharvestingmethodinvolvingseveralstepsisrequired(Mataetal.,
2010).Theconcentrationofalgalbiomassistypically0.51.0gL1inopenpondsandcanreach510gL1in
closedsystems.At1gL1algalbiomass,1000kgofwatermustbeprocessedtoobtain1kgofdrybiomass.
Most common harvesting methods include sedimentation, centrifugation, filtration, ultrafiltration,
sometimeswithanadditionalflocculationsteporwithacombinationofflocculationflotation.Thechoice
ofwhichharvestingtechniquetousedependsonthespeciesofmicroalgaeandthefinalproductdesired.
Microalgal properties that simplify harvesting are large cell size, high specific gravity compared to the
medium and reliable autoflocculation. Moreover the optimum harvesting method should have minimum
energy requirements and be as economical as possible. The main harvesting techniques were recently
describedbyUdumanetal.(2010),andaresummarizedinTable5.
Centrifugation is seen as the most efficient recovery technique, yet the energy and capital costs
associatedareunappealing(Udumanetal.,2010).Filterpressesoperatingunderpressureorvacuumcan
beusedtorecoverlargequantitiesofbiomass,butforsomeapplicationsfiltrationcanberelativelyslow
whichmaybeunsatisfactory.Alternatively,membranemicrofiltrationandultrafiltrationareotherpossible
alternativestoconventionalfiltrationforrecoveringalgalbiomass,whicharemoresuitableforfragilecells
andsmallscaleproductionprocesses.Flocculationisalsoaveryefficienttechniquethatiscurrentlyapplied
inindustry.Microstrainersareanattractiveharvestingmethodbecauseoftheirmechanicalsimplicityand
availability in large unit sizes (Mata et al., 2010). Nevertheless, a significant engineering research effort
aimedatdevelopingmorecosteffectivealgalharvestingtechniqueswillberequired.
BGU and Algatech (Israel) have designed successful harvesting and processing procedures for
Haematococcus biomass that are based on sedimentation in large funnels and works without energy
requirementsatover90%efficiency,creatingaconcentratedcellslurrythatcanbefurtherdewateredand
dried. Flocculation of Nannochloropsis and Isochrysis has also been successfully applied (Boussiba et al.,
1987,1988).Evodos(TheNetherlands),leaderofWP7,hasestablisheda100%subsidiary,calledEvodos
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Algae Technologies B.V. to market algae harvesting machines. Evodos technology allows over 90% algae
separation due to a patented innovation which results in improved fluid dynamics and produces a paste
(30% dry weight) with a positive energy balance. The high speed rotating separation device (rotor) is
equippedwithflexiblecurvedvanes,allowingthesmallestparticlestosettlequickly.
Table5.Harvestingtechniques.
4.4 Biotechnologyofthemajormicroalgalgroups
4.4.1
Cyanobacteria
Some cyanobacteria are easily transformed genetically and have been under investigation for enhancing
photosyntheticproductivity,hydrogenproductionetc.
Algenolhastakenadvantageofthetransformabilityofcyanobacteriatocreateacyanobacterilastrain
transformingphotosyntheticallyproducedsugarintoethanol,whichisexcretedintothegrowthmedium.
This technology has attracted significant funding for establishing of a large pilot plant using especially
designedbioreactors.
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Figure8http://www.algenolbiofuels.com/overview.htm.
Aninterestingapplicationofthetransformabilitytechniqueswascloningand expressionofBTitoxins
into Anaebaena for creating a biological mosquito control agent, though release of this organism into
naturehasnotbeenlicensedsofar(Xiaoqiangetal.,1997).
Cyanobacteria can be thermophilic and occur in all watery environments including toxic hotsprings.
This ability has been exploited to establish a revolutionary exhaust gas scrubbing methodology with
thermophiliccyanobacteriaisolatedfromYellowstonehotspringsgrowingdirectlyonpanelslitbysunlight
concentrated and delivered via light guides, in chambers fed by exhaust gas
(http://www.greencarcongress.com/2005/12/greenshift_lice.html).
Recent Researches have also hinted at cyanobacteria possible application to the generation of clean
andgreenenergyviaconvertingsunlightdirectlyintoelectricity.
A number of important advances have occurred in cyanobacterial biotechnology in the recent years.
Worldwideattentionisdrawntowardscyanobacteriafortheirpossibleuseinmariculture,asfood,feed,
fuel, fertilizers, colourants, and for the production of various secondary metabolites including vitamins,
toxins,enzymes,pharmaceuticalsandpharmacologicalprobes,andforpollutionabatement(Tredicietal.,
2009). Only a few cyanobacterial strains have been wellcharacterized or exploited commercially. Some
cyanobacteria are sold as food, notably Aphanizomenon flosaquae and Arthrospira platensis (Spirulina).
Spirulina is being grown commercially (and marketed as dietary supplements) mostly in open raceway
ponds in Asia and USA, while short precommercial cultivation was also performed by Ben Gurion
UniversityforKoorindustriesinIsrael(Vonshak,personalcommunication).
4.4.2
Chlorophyta(GreenAlgae)
A significant number of green algal species are being exploited for production of pigments (Dunaliella,
Haematococcus) or neutraceuticals (Chlorella), and biological remediation (Chlorella, Scenedesmus),
whereby the vast metabolic capacity of green algae holds a great promise for identification of further
commerciallyviableproductionprocesses.Furthermore,Chlamydomonasreinhardtiiisthemostadvanced
toolforstudyingmolecular,geneticandbiochemicalaspectsofmicroalgae.
Chlorella, Dunaliella and Haematococcus provide the vast majority of algal biomass produced
worldwide, exclusively as nutraceutical or health products. Chlorella is being cultivated in open ponds in
Asia(e.g.http://www.taiwanchlorella.com/)orinEurope'slargestclosedphotobioreactorfacilityinKlotze,
Germany (http://www.algomed.de/index.php). The protein, vitamin and mineral rich dry biomass is
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marketedasfoodadditive.ThecapacityofDunaliellatosurviveinDeadSeawaterandtoaccumulatehigh
concentrations of carotene and glycerol has early on caught the attention of scientists, and a range of
productionfacilitiesinopenlagoons,e.g.inAustralia(http://www.nutrakol.com/pdf/Dunaliella.pdf),Asia
or in raceway ponds in Eilat, Israel (http://www.wondercare.co.in/nature.htm) have been operating
successfully.Haematococcuspluvialisisproducinglargeamountsofthehighlyvaluedpigmentastaxanthin
that is a potent antioxidant with significant health benefits reported. Astaxanthin accumulation requires
cultivationundernutrientstress,andmajorproducerstodayarefoundinAsia,AustraliaandHawaii(Fig.2
d), where the algae are cultivated in raceway ponds (http://www.cyanotech.com/), or Israel, where
astaxanthinisproducedinalargephotobioreactorfacilityinQetura(Fig.3f/g;http://www.algatech.com/).
Other production processes for various pigments from species such as Scenedesmus (lutein)
Chlorococcum(astaxanthin)orChlorellazofingensis(both)(DelCampoetal.,2004)havebeeninvestigated
butnotachievedcommercialdeployment.
Inallinstancesdescribed,alternativeproductseitherfromlandplantsorfromchemicalsynthesisare
significantlycheapertoproducethanthealgalproduct,sothattodatetheonlymarketnicheforsuchalgal
productsisthehealthmarket.
Asignificantnumberofspeciesorgeneraarebeingconsideredasbiofuelfeedstock,duetothefactthat
numerousgreenalgaespeciesareveryfastgrowingandareabletoaccumulatehighconcentrationsofoil
usefulforbiodieselproduction.AbouthalfthestrainstestedbyHuetal.(2008)aregreenalgae,asare11
out of 30 tested by Rodolfi et al. (2009) Their high lipid productivity, accessability to genetic engineering
and capability to grow rapidly in the dark fermenting organic substrates has made Chlorella the primary
platform for custom made biofuels production via heterotrophic fermentation by biofuels company
Solazyme.
4.4.3
Rhodophyta(RedAlgae)
Several Red Algae species have significant biotechnology potential. Some of them are economically
importantassourceofagarsandcarrageenans.Forthisreason,extensivefarmingandnaturalharvestof
redalgaeoccursinnumerousareasoftheworld(seemacroalgalbiotechnology).
The unicellular red algae Porphyridium is producing significant amounts of LCPUFA and medicinally
activepolymers.Upto44%ofitstotalfattyacidsconsistofthehighvaluelongchainPUFAEPA(Cohenand
Cohen, 1991). Besides red microalgae in general, Porphyridium cruentum produces highly sulfonated
extracellular polysaccharides with strong and specific activity against animal viruses such as Herpes
(Huheiheletal.,2002).
4.4.4
Heterokontophyta
Thisgroupincludeavarietyofalgaldivisionsofhighestsignificancetobiotechnology,withsignificanceto
aquaculture,foodandbiofuelsindustriesaswellasthenutraceuticalmarket.
Among the microalgae the most important group from the biotechnological point of view is
represented by the eustigamotphytes. Nannochloropsis is considered a promising alga for industrial
applications because of its ability to accumulate high levels of polyunsaturated fatty acids
(eicosapentaenoicacid).Itismainlyusedasfeedforfishlarvaeandrotifers(seedetailedformattheendof
the document). Nannochloropsis is also used as an ingredient in cosmetic products (Tredici et al., 2009).
Nannochloropsis has also been proposed as feedstocks for biodiesel production because of its ability to
accumulate up to 60% lipid under nitrogen starvation (Rodolfi et al., 2009). Monodus is a freshwater
eustigamtophytewhichisalsorichineicosapentaenoicacid(EPA).IthasbeencultivatedtostudyitsEPA
productionunderlaboratoryconditionsandoutdoorsindifferenttypesofphotobiorectorsunderdifferent
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conditions.InSpainproductivitiesof0.16and0.54gL1d1wereobtainedina57Lbubblecolumnandina
75L helical photobioreactor, respectively, with a maximum EPA productivity of 9 mg L1 d1 (Lu et al.,
2002). In Israel, Hu et al. (1997) in a flat inclined modular photobioreactor made of four 14L units,
obtainedabiomassproductivityforMonodusofabout1.4gL1d1andanEPAproductivityof59mgL1d1
attheoptimalcelldensityof4gL1.ForitsEPAcontentMonodussubterraneusisusedforthepreparation
of nutraceutical products together with the DHArich primnesiophyte Isochrysis galbana by Nikken
SohonshaCo.(Japan).Recently,afreshwaterxantophytealga,Trachydiscusminutus,hasbeenproposedas
asourceofEPAduetoitslargecontentofthisfattyacid(upto>35%oftotalfattyacids)(ezankaetal.,
2010).
4.4.5
Labyrinthulea(phylumHeterokonta)
Thraustochytrids are a commercial source of polyunsaturated fatty acids, in particular doicosahexaenoic

acid (DHA). DHArich oils are produced through fermentation from the marine thraustochytrids
Schizochytrium and Ulkenia by Martek Co. (USA) and Nutrinova (Germany), respectively, for nutritional
supplementsandanimalfeeds(Lewisetal.,1999;Tredicietal.,2009).Duringcommercialproductionthere
are two phases: first, biomass production and then lipid induction. Glucose is generally the preferred
substratetogrowtheseorganimsms,thoughthereisalotofvariabilityamongstrainsconcerningthebest
carbonandnitrogensource.DriedSchizochytriumhasGRASstatusforuseasfeedtobroilerchickensand
layinghenfeedinordertoenhanceDHAinthemeatandeggs (Raghukumar,2008).Schizochytrium cells
areproducedbyAquafauna,Inc(USA)tobeincludedinfeedforaquaculture,inparticulartoenrichinDHA
rotifersandArtemia.MostthraustochytridsproduceoneormoreofotherPUFAs,namelyarachidonicacid,
docosapentaenoic acid, eicosapentaenoic acid and docosatetraenoic acid and, although they can be
considered as a source of these molecules, there is not an actual exploitment. The DHA biosynthetic
pathway in thraustochytrids has presently attracted attention because of efforts by many researchers to
producerecombinantplantsoryeastscapableofproducingPUFAs.Thraustochytridsarealsoconsidereda
potential source of carotenoids such as carotene and the xanthophylls astaxanthin and canthaxanthin
(Raghukumar,2008).
4.4.6
Bacillariophyta(Diatoms)
Diatoms are used for various biotechnological applications, mainly at the laboratory scale, and in
aquaculture at a higher scale for commercial outlets. Examples of applications are: (1) silicon production
originating from frustules, for technological applications in nanotechnology, pollution remediation or as
foodinaquaculturethankstothelipidandaminoacidrichalgalcontent,(2)intracellularmetabolitesthat
accumulatedincells,e.g.lipids,particularlyeicosapentaenoicacid(EPA)forpharmaceuticalapplications,or
amino acids for cosmetic applications, and (3) extracellular metabolites released into the medium, e.g.
variouspigments(forchickenandfishfeeds)andantibiotics(LebeauandRobert,2003).
Indiatoms,themajorfattyacidsareC16:0andC16:1(Huetal.,2008).Themostnutritionallyrelevant
biomolecules produced by diatoms are verylongchain polyunsaturated fatty acids (vlcPUFAs) like
arachidonic acid (AA) (C20:46), eicosapentaenoic acid (EPA) (C20:53), docosahexaenoic acid (DHA)
(C22:63)andotheromega3fattyacids(Bozarthetal.,2009).
Theaveragetotallipidcontentofanoleaginousdiatomis22.7%DCWwhenmaintainedundernormal
growthconditions,whereasatotallipidcontentof44.6%DCWisachievableunderstressconditions(Huet
al., 2008). A general trend towards accumulation of lipids, particularly TAG, in response to nitrogen
deficiency has been observed in various algal taxa including diatoms. However, in diatoms, silicon is an
equally important nutrient that affects cellular lipid metabolism (Roessler, 1988). Phosphorus limitation
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also results in increased lipid content, mainly TAG (Reitan et al., 1994). Typically, high light intensity
decreasestotalpolarlipidcontentwithaconcomitantincreaseintheamountofneutralstoragelipids.High
light also alters fatty acid synthesis to produce more of the saturated and monounsaturated fatty acids
thatmainlymakeupneutrallipids.Ingeneral,thetotallipidcontentofcellsincreasewithage.Anincrease
inTAGsisoftenobservedduringstationaryphase(Huetal.,2008).
4.4.7
Haptophyta
EconomicallyimportanthaptophytesincludePavlovalutheriandIsochrysissp.,whicharewidelyusedinthe
aquaculture industries due to their high docosahexaenoic acid (DHA) content. They are commercially
producedbyseveralcompaniestobesoldasfeedforbivalveshellfishlarvaeandpostlarvae,shrimplarvae
androtiferenrichemntasconcentrates(e.g.64to90gL1),aloneormixedwithotheralgalspecies(Tredici
etal.,2009).DuetothehighDHAcontent,Isochrysisgalbanaisused,mixedwithMonodussubterraneus,
for the production of nutraceuticals by Nikken Sohonsha Co. (Japan). Recently, exctracts of Isochrysis sp.
havebeenpatentedforapplicationincosmetics,asingredientsforskinandhairhealthproducts(Herrmann
etal.,2009).
ThesuitabilityofIsochrysisforproducingoilforbiodieselproductionhasbeenconfirmedinpilotsized
racewaypondsinIsrael(Boussibaetal.,1988).
4.4.8
Dinophyta(Dinoflagellates)
A few photoautotroph Dinoflagellate species have been evaluated for lipid productivity. The most
prominentapplicationsforDinoflagellatesinvolvesheterotrophicspeciesthataregrowninfermentorsfor
production of PUFArich oil or biofuels. Commercial cultivation of the heterotrophic dinoflagellate
Cryptechodinium cohnii to obtain a docosahexaenoic acid (DHA)rich oil to be used as an ingredient in
nutraceuticalproductsiscarriedoutinfermentorsbyMartekCo.intheUS.
Dinoflagellatesareamongthemajortoxinproducersknown.Besidesthehugenegativeimpactofthese
toxinsontheecosystemswhentheyareproducedinlargeamountsfromalgaeblooms,anapplicationcan
be foreseen. Dinoflagellate, as well as cyanobacteria, toxins are used in medical studies in attempts to
understandtheirmodesofactionandassesstheirtherapeuticpotentialorthatoftheiranalogs.Saxitoxins,
areconsideredsafe,effective,longactingtopicalanesthetics(GarciaCamachoetal.,2007).Thesaxitoxin
related tetrodotoxin, a nonpeptide neurotoxin, is being developed by WEX Pharmaceuticals Inc.
(www.wexpharma.com) for treatment of cancerrelated pain as an alternative to opioids. Okadaic acid is
consideredamodelpotentneurotoxinforanalyzingthetherapeuticeffectsofatypicalantipsychoticdrugs
in treatment of cognitive impairment and neuropathological changes of schizophrenia and other
neurodegenerativediseases(GarciaCamachoetal.,2007).
Dinoflagelltesproducesmanybioactivemoleculeswithinterestinganticancerproperties,thatpresently
havebeenevidencedonlyatlabscreeningphase.
4.5 Biotechnologyandusesformacroalgae
Current uses and biotechnology of Macroalgae far exceeds those of Microalgae, with Macroalgae being
harvested for food, feed or biocompounds (agar) all across the world, with major concentrations in East
Asia. Three distinct algal classes, Chlorophyta, Rhodophyta, and Phaeophyta, have macroalgae
representatives,andallclassesarebeingexploitedeconomically.
Macroalgaebasedfuelsholdgreatpromise,directlyrelatedtothepotentialtoproducemorebiomass
perunitareainayearthananyotherformofbiomass.ThemainChlorophytaspeciesrelevanttobiofuel
usesarebelongingtothegenusofUlvaandCaulerpa.Theredalgaespeciesrelevanttobiofuelusesmainly
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belongtotheorderofGigartinales,HalymenialesandPalmariales.Thebrownalgaerelevanttobiofueluses
aremainlybelongingtotheorderofFucales(includingrepresentantofthefollowingfamilies:Alariaceae,
Fucaceae,Himanthaliaceae,Sargasseae),LaminarialesandTilopteridales.
Sewagecanbetreatedwithalgae,reducingtheneedforgreateramountsoftoxicchemicalsthanare
alreadyused.Algaecanbeusedtocapturefertilizersinrunofffromfarms.Whensubsequentlyharvested,
theenrichedalgaeitselfcanbeusedasfertilizer.AgriculturalResearchServicescientistsfoundthat6090%
ofnitrogenrunoffand70100%ofphosphorusrunoffcanbecapturedfrommanureeffluentsusinganalgal
turf scrubber (ATS). Scientists developed the ATS, which are shallow, 100foot raceways of nylon netting
wherealgaecoloniescanform,andstudieditsefficacyforthreeyears.Theyfoundthatalgaecanreadilybe
used to reduce the nutrient runoff from agricultural fields and increase the quality of water flowing into
rivers,streams,andoceans.Theenrichedalgaeitselfalsocanbeusedasafertilizer.Researcherscollected
anddriedthenutrientrichalgaefromtheATSandstudieditspotentialasanorganicfertilizer.Theyfound
thatcucumberandcornseedlingsgrewjustaswellusingATSorganicfertilizerastheydidwithcommercial
fertilizers.Airdriedalgaefromanalgalturfscrubber(ATS)capturedmostofthenitrogenandphosphorus
in manure runoff (United States Department of Agriculture, Agricultural Research Service,
www.ars.usda.gov/is/AR/archive/may10/algae0510.htm).
Severalredmacroalgaespeciesareusedasfood,suchasDulse(Palmariapalmata)andPorphyrainthe
British Isles, or nori in Asia. Cultivation is relatively simple and began in Japan more than 300 years ago.
Majorproductsfrommacroalgaetodayare:
Agar,agelatinoussubstancederivedfromredalgae,hasanumberofcommercialuses.
Alginates:Between100,000and170,000wettonsofMacrocystisareharvestedannuallyinCaliforniafor
alginateextractionandabalonefeed.
Fertilizer:Forcenturiesseaweedhasbeenusedasafertilizer.Thiskindoforetheyoftengatherandlayon
great heapes, where it heteth and rotteth, and will have a strong and loathsome smell; when being so
rotten they cast on the land, as they do their muck, and thereof springeth good corn, especially barley.
Afterspringtydesorgreatrigsofthesea,theyfetchitinsacksonhorsebackes,andcariethesamethree,
four,orfivemiles,andcastitontheland,whichdothverymuchbetterthegroundforcornandgrass.
Nutrition: Naturally growing seaweeds are an important source of food, especially in Asia. They provide
manyvitaminsincluding:A,B1,B2,B6,niacinandC,andarerichiniodine,potassium,iron,magnesiumand
calcium.
Algaearenationalfoodsofmanynations:Chinaconsumesmorethan70species,includingfat
choy, a cyanobacterium considered a vegetable; Japan, over 20 species; Ireland, dulse; Chile,
cochayuyo.Laverisusedtomake"laverbread"inWaleswhereitisknownasbaralawr;inKorea,
gim;inJapan,noriandaonori.ItisalsousedalongthewestcoastofNorthAmericafromCalifornia
toBritishColumbia,inHawaiiandbytheMoriofNewZealand.Sealettuceandbadderlocksarea
saladingredientinScotland,Ireland,GreenlandandIceland.Dulseisagoodsourceofmineralsand
vitamins, contains all trace elements and has a high protein content. Dulse is commonly used in Ireland,
Iceland, Atlantic Canada and the Northeast United States. Dulse can be used both fresh or sundried
(ground to flakes or powder). Finely diced, it can also be used as a flavour enhancer in place of
monosodiumglutamate.
Stabilizing substances: Carrageenan, from the red alga Chondrus crispus, is used as a stabiliser in milk
products.
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4.6 BiotechnologyofOtherAquaticBiomass
Aquaticvascularplantscanrepresentattractivebiomassproductionalternativesespeciallywhenintegrated
into treatment of eutrophicated water bodies. A large number of often invasive water plants, such as
WaterHyacinth,Lemna,variousreeds,Elodea,Egeriaetcbecomeapestinnutrientrichwaterscloggingup
waterwaysandsuffocatingnativefloraandfauna.Ontheotherhandsuchplantsprovidehighproductivity
under significant nutrient removal capacity. Lacking success with microalgae has caused former algae
pioneer PetroAlgae to switch technology to cultivation of "microcrops" (Lemna) on nutrient rich lagoons
(http://www.petroalgae.com/index.php),offeringcertainadvantagessuchasamaturetechnology,easier
harvesting(Fig.9)andprocesscontrolattheexpenseofsomewhatreducedproductivity.
Figure9RemovalofalgaebloomsandwaterhyacinthatHertebeesportdam,SouthAfrica
(http://www.dwa.gov.za/harties/).
Remediationeffortsofpollutedwaterbodiesingeneralcanprovidesignificantamountsoffreewaste
biomass from such water plant material (see Hartebeesport dam http://www.dwa.gov.za/harties/) that
maybeutilizedforfuelsproduction.Alternatively,biologicalwatertreatmentusingconstructedwetlands
canalsoprovidesignificantamountsofwastebiomassthatmaybeutilizedforbiofuelsproduction.
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5 Symbology
Symbologymightbeusedtoallowquickunderstandingofthefollowingsections.
SYMBOLOGYofthelevelofuseofthealga
Thissymbolindicatesthatthealgahasapotentialapplicationandiscurrentlyusedat
pilotexperimentallevelforbiofuels
Thissymbolindicatesthatthealgahasapotentialapplicationforbiofuels,thoughthere
isnopilotproduction
Thissymbolindicatesthatthealgaiscommerciallyproducedandavailableinlarge
quantities
D
Potentiallyinterestingforbiodieselproduction
E
Potentiallyinterestingforbioethanolproduction
H
Potentiallyinterestingforbiohydrogenproduction
B
Interestingforbiomassproduction
PIV
Pivotaltaxonforbiotechnologydescription
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7 Prokaryoticmicroalgae
7.1 Cyanobacteria
7.1.1
Arthrospirasp.(commonnamespirulina)
Figure10Arthrospirajenneri
Picturefromhttp://wwwcyanosite.bio.purdue.edu
SYMBOLS:
B,PIV
TAXONOMY
BotanicalCode
Phylum
Class
Order
Family
Genus
Species
Cyanobacteria
Cyanophyceae
Oscillatoriales
Phormidiaceae
Arthrospira
MicrobiologicalCode
Group
Subsection
Family
Genus
Cyanobacteria
3
3.1
Arthrospira
ImportantSpecies
A.fusiformis,A.jenneri,A.maxima,A.platensis
BIOLOGY
Arthrospira(Spirulina)isamicroscopicbluegreenalgaintheshapeofaspiralcoil,livingbothinseaand
fresh water. Spirulina is the common name for human and animal food supplements produced primarily
fromtwospeciesofcyanobacteria:Arthrospira platensis,andArthrospiramaxima. Thoughreferred toas
'algae' because they are aquatic organisms capable of photosynthesis, cyanobacteria are not related the
eukaryoticalgae(Vonshak,1997).
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Arthrospira is cultivated around the world, and is used as a human dietary supplement, as well as a
wholefood,andisavailableintablet,flake,andpowderform.Itisalsousedasafeedsupplementinthe
aquaculture,aquarium,andpoultryindustries(Ciferri,1983).
Arthrospira are freefloating filamentous cyanobacteria characterized by cylindrical, multicellular
trichomesinanopenlefthandhelix.TheyoccurnaturallyintropicalandsubtropicallakeswithhighpHand
high concentrations of carbonate and bicarbonate. Arthrospira platensis occurs in Africa, Asia and South
America,whereasArthrospiramaximaisconfinedtoCentralAmerica.Thesespecieswereonceclassifiedin
thegenusSpirulina.TheyareinfactArthrospira;nevertheless,theoldertermSpirulinaremainsinusefor
historicalreasons.SpirulinawasfoundinabundanceatLakeTexcocobyFrenchresearchersinthe1960s,
butthereisnoreferencetoitsusethereasadailyfoodsourceafterthe16thcentury.Thefirstlargescale
Spirulinaproductionplant,runbySosaTexcoco,wasestablishedthereintheearly1970s.
SpirulinamayhaveanevenlongerhistoryinChad,asfarbackasthe9thcenturyKanemEmpire.Itisstill
in daily use today, dried into cakes called dih, harvested from small lakes and ponds around Lake Chad
(Abdulqaderetal.,2000).
Biochemicalcompositionofalgaeandaquaticbiomass:mainconstituents
Spirulina contains an unusually high amount of protein with, between 55% and 77% of dry weight
(Babadzhanovetal.,2004;Tokusogluetal.,2003).Itisacompleteproteinsourcecontainingallessential
amino acids, though with reduced amounts of methionine, cysteine, and lysine when compared to the
proteinsofmeat,eggs,andmilk.Itis,howeversuperiortotypicalplantprotein,suchasthatfromlegumes.
Spirulinaisrichingammalinolenicacid(GLA),andalsocontainsalphalinolenicacid(ALA),linoleicacid(LA),
stearidonicacid(SDA).SpirulinaalsocontainsvitaminB1(thiamine),B2(riboflavin),B3(nicotinamide),B6
(pyridoxine), B9 (folic acid), vitamin C, vitamin D, vitamin A, and vitamin E. Spirulina is a rich source of
potassium, and also contains calcium, chromium, copper, iron, magnesium, manganese, phosphorus,
selenium,sodium,andzinc.
Spirulina contains many pigments, including chlorophyll a, betacarotene, echinenone, myxoxanthophyll,
zeaxanthin, canthaxanthin, diatoxanthin, 3'hydroxyechinenone, betacryptoxanthinand, oscillaxanthin,
plusthephycobiliproteinscphycocyaninandallophycocyanin(Leemaetal.,2010)
BIOTECHNOLOGY
Culturemedia
Spirulina can live in a wide range of media with elevated salinity and high alkalinity. See
http://www.antenna.ch/en/malnutrition/growyourownspirulina.html)foranexemplarygrowthmedium.
Cultivationmethods
SinceSpirulinacultivationdoesnotincludeastressphase,thealgaecanbecultivatedinsemicontinuous
modewithadaily,constantamountsbeingharvestedmaintainingthecultureinastablestate.Theoptimal
temperatureforgrowthis3538Cwhiletheminimumis1520C.
Productionsystem
Spirulina is being produced in natural open water bodies of suitable temperature, salinity and alkalinity,
andinracewaypondscultivationusingappropriategrowthmedia.MostcultivatedSpirulinaisproducedin
openchannelracewayponds,withpaddlewheelsusedtoagitatethewaterculture.Thelargestcommercial
producers of Spirulina are located in the United States, Thailand, India, Taiwan, China, Pakistan, Burma
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(a.k.a. Myanmar) and Chile (Borowitzka, 1992; 1999; Vonshak (ed.), 1997). Paddle wheels speed in the
orderof20cms1hasbeenrecommended.
Spirulinacultivationmaybecarriedoutinclosedandopensystems.Thefirstoneinvolvelaboratoryand
pilotscale photobioreactors (Vonshak (ed.), 1997; Tredici and Chini Zittelli, 1998), not used in industrial
production.
CO2supplyisnotnormallyrequiredasthehighpHfacilitatesuptakeofCO2 fromtheatmosphere,and
sodiumsodiumbicarbonatemayserveascarbonsource.
HarvestingmethodsandBiomassprocessing
Spirulina is forming large aggregates and can be harvested easily using nets. Filtration is simply
accomplishedbypassingtheculturethroughafineweavecloth,usinggravityasthedrivingforce.Synthetic
fibrecloth(especiallypolyamideorpolyester)withameshsizeofabout30to50micronsisthepreferred
filteringmediumdevice.Supportingthefiltrationclothbyafinenetwillacceleratesomewhatthefiltration
andprotecttheclothagainstrupturing,butasimplebagmadefromtheclothworkswellalso.Thebiomass
isdriedusuallyspraydriedandtransformedtopillsdirectly.
Scalinguplimitation
For a health product contaminating contamination by small animals can pose a serious problem. Since
Spirulinapreferstemperaturesabove30degreescentigrade,reducedtemperaturesduringwinterevenin
subtropicalclimatesrequiretemporarybreaksintheproductionprocess.
HIGHLIGHTSINBIOTECHNOLOGY
Spirulinaisoneofonlytwoalgalspecies(togetherwithChlorella)thathasbeencultivatedfordecadesina
varietyofpondsandbioreactors.
Productionpriceisestimatedat$5perkg,anddriedalgaearesoldasfoodandfeedadditivesonly.
References
Abdulqader, G., Barsanti, L., Tredici, M. (2000) Harvest of Arthrospira platensis from Lake Kossorom (Chad) and its
householdusageamongtheKanembu.JournalofAppliedPhycology12:493498.
Babadzhanov A.S., Abdusamatova N., Yusupova F.M., Faizullaeva N., Mezhlumyan L..G., Malikova M.K.. (2004) Chemical
compositionofSpirulinaplatensiscultivatedinUzbekistan.ChemistryofNaturalCompounds.40:276279.
Borowitzka M.A. (1992) Algal biotechnology products and processes matching science and economics. Journal of Applied
Phycology3:267279.
Borowitzka M.A. (1999) Commercial production of microalgae: ponds, tanks, tubes andfermenters. Journal of Biotechnology 70:
313321.
CiferriO.(1983)Spirulina,theediblemicroorganism.MicrobiologicalReviews47:551578.
GarrityG.M.,BooneD.R.,CastenholzR.W.(eds)(2004)BergeysManualofSystematicBacteriology,SecondEdition,Volume1,The
ArchaeaandthedeeplybranchingandphototrophicBacteria,Springer.
Leema J.T.M., Kirubagaran R., Vinithkumar N.V., Dheenan P.S., Karthikayulu S. (2010) High value pigment production from
Arthrospira(Spirulina)platensisculturedinseawater.BioresourceTechnology101:92219227.
TokusogluO.,UnalM.K.(2003)Biomassnutrientprofilesofthreemicroalgae:Spirulinaplatensis,Chlorellavulgaris,andIsochrysis
galbana.JournalofFoodScience68:11441148.
Vonshak, A. (ed.) (1997) Spirulina platensis (Arthrospira): Physiology, Cellbiology and Biotechnology. London: Taylor & Francis,
1997.
.
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7.1.2
Phormidiumsp.
Figure11Phormidiumautumnale
Picturefromhttp://botany.natur.cuni.cz/skaloud/Cyano/Phoaut.htm
SYMBOLS:D,H
TAXONOMY
BotanicalCode
Phylum
Class
Order
Family
Genus
Species
Cyanobacteria
Cyanophyceae
Oscillatoriales
Phormidiaceae
Phormidium
MicrobiologicalCode
Group
Subsection
Family
Genus
Cyanobacteria
3
3.1
Lyngbya(includingPhormidium)
RelatedSpecies
Thereare635species(andinfraspecific)namesinthedatabaseatpresent,ofwhich161havebeenflagged
ascurrentlyacceptedtaxonomically.
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BIOLOGY
Trichomes are isopolar, more or less straight, coiled or waved, usually 214 m wide, uniserial, never
branched,composedofcylindricaluptoslightlybarrelshapedcells,moreorlessisodiametricalorslightly
shorter or longer than wide, constricted or unconstricted at the cross walls, not attenuated and bent or
screwliketwistedtowardstheends,motile(waving,creeping,oscillations)withinandoutofsheaths.End
cells widely rounded, attenuated or pointed, sometimes with calyptra. Sheaths develop facultatively in
differentfrequenciesonlyinsuboptimalconditions,orindependenceonchangingenvironmentalfactors,
orregularlyinallconditions.Sheathsaretubelike,firm,colourless,joinedtothetrichomes,notlayered,
openedattheends,containingalwaysonlyonetrichome.Cellcontentusuallybluegreen,rarelybrownish,
pinkish or violet, sometimes modifications with stable PE:PC ratio occur; thylakoids situated
perpendicularlytothecellwall(radiallyinacrosssection).Celldivisioncrosswise,perpendicularlytothe
longaxisofatrichome,daughtercellsgrowuptotheoriginalsizebeforethenextdivision.Allcellscapable
ofdivisionwiththeexceptionofapicalones.Reproductionbyhormogonia,whichseparateattheendparts
oftrichomesbyhelpofnecridiccellsorbyfragmentationofwholetrichomeswithnecridiccells.
Rarely solitary filaments, usually in mats on different aeric or water substrates (soil, wet rocks, mud,
waterplants,stonesandwoodsinbothstagnantandstreamingwaters),somespeciesoccurinthemarine
littoral.Severalspeciesareknownfromextremehabitats(thermalsprings,desertsoils,etc.),fewofthem
takepartinthebiolithogenicprocessesandformtravertinecrustsinlimestonewaterbiotopes.
BIOTECHNOLOGY
Phormidiumhasbeenproposedbymanyauthorsfortreatmentofdifferenteffluents(swine,aquaculture,
municipal, industrial) due to its ability to remove nutrients and to degrade pollutants as phenols and
hydrocarbons (CaizaresVillanueva et al., 1994; Dumas et al., 1998; Satheesh Kumar et al., 2009;
Shashirekhaetal.,1997).
Phormidium extracts have shown antimicrobial activity in many screenings (Biondi et al., 2008;
RodrguezMeizosoetal.,2008)andseveralbioactivemoleculeshavebeenindividuated,suchasantiHIV
glycolipidsandhierridin,amoleculewithantiplasmodialactivity(Gustafsonetal.,1989;Papendorfetal.,
1998).AbioactiveAntarcticP.priestleyiwascultivatedundercontinuousilluminationatlowintensityina
10L bubbled reactor obtaining productivies of 95 mg L1 day1 (Biondi et al., 2008). At present, extracts
from Phormidium persicinum have been included in cosmetic products as skin rejuvenator (Morvan and
Valee,2007,2010).
Though the studies have all been limited to laboratory scale, Phormidium have been considered for
hydrogen production (Patel and Madamwar, 1994; Prabaharan and Subramanian, 1996) and also for
biodieselproduction(Franciscoetal.,2010).
References
Biondi N., Tredici M.R., Taton A., Wilmotte A., Hodgson D.A., Losi D., Marinelli F. (2008) Cyanobacteria from benthic mats of
Antarcticlakesasasourceofnewbioactivities.JournalofAppliedMicrobiology105:105115.
CaizaresVillanueva R.O., Ramos A., Corona A.I., Monroy O., De La Torre M., GomezLojero C., Travieso L. (1994) Phormidium
treatmentofanaerobicallytreatedswinewastewater.WaterResearch28:18911895.
DumasA.,LalibertG.,LessardP.,delaNoeJ.(1998)BiotreatmentoffishfarmeffluentsusingthecyanobacteriumPhormidium
bohneri.AquaculturalEngineering17:5768.
Francisco E.C., Neves D.B., Jacoblopes E., Franco T.T. (2010) Microalgae as feedstock for biodiesel production: Carbon dioxide
sequestration,lipidproductionandbiofuelquality.JournalofChemicalTechnologyandBiotechnology85:395403.
Gomont M. (1892'1893'). Monographie des Oscillaries (Nostocaces homocystes). Annales des Sciences Naturelles, Botanique,
Series716:91264,Plates17.[BIOLOGYsection]
ArchaeaandthedeeplybranchingandphototrophicBacteria,Springer.[BIOLOGYsection]
GustafsonK.R.,CardellinaIIJ.H.,FullerR.W.,WeislowO.S.,KiserR.F.,SnaderK.M.,PattersonG.M.L,BoydM.R(1989)AIDSantiviral
sulfolipidsfromcyanobacteria(bluegreenalgae).JournaloftheNationalCancerInstitute81:12541258.
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KomarekJ.(1992)in:http://www.cyanodb.cz/Phormidium[BIOLOGYsection]
MorvanP.Y.,ValleeR.(2007)Effectofmicroalgalextractsonthioredoxinexpressioninhumanskincellsandtheirprotectionfor
skin.IFSCCMagazine,10(2):119126.
MorvanP.Y.,ValleeR.(2010) Newsolutionforbrighterandyoungerskin.PersonalCareMagazine,June2010.
PapendorfO.,KonigG.M.,WrightA.D.(1998)HierridinBand2,4dimethoxy6heptadecylphenol,secondarymetabolitsfromthe
cyanobacteriumPhormidiumectocarpiwithantiplasmodialactivity.Phytochemistry49:23832386.
Patel S., Madamwar D. (1994) Photohydrogen production from a coupled system of Halobacterium Halobium and Phormidium
valderianum.InternationalJournalofHydrogenEnergy19:733738.
PrabaharanD.,SubramanianG.(1996)OxygenfreehydrogenproductionbythemarinecyanobacteriumPhormidiumvalderianum
BDU20041.BioresourceTechnology57:111116.
RodrguezMeizoso I., Jaime L., Santoyo S., Cifuentes A., GarcaBlairsy Reina G., Seorns F.J., Ibez E. (2008) Pressurized fluid
extractionofbioactivecompoundsfromPhormidiumspecies.JournalofAgriculturalandFoodChemistry56:35173523.
SatheeshKumarM.,MuralitharanG.,ThajuddinN.(2009)Screeningofahypersalinecyanobacterium,Phormidiumtenue,forthe
degradationofaromatichydrocarbons:naphthaleneandanthracene.BiotechnologyLetters31:18631866.
ShashirekhaS.,UmaL.,SubramanianG.(1997)PhenoldegradationbythemarinecyanobacteriumPhormidiumvalderianumBDU
30501.JournalofIndustrialMicrobiology&Biotechnology19:130133.
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7.1.3 Anabaenasp.
Figure12Anabaenasiamensis40x
PicturefromUNIFI
SYMBOLS:H
TAXONOMY
BotanicalCode
Phylum
Class
Order
Family
Genus
Species
Cyanobacteria
Cyanophyceae
Nostocales
Nostocaceae
Anabaena
MicrobiologicalCode
Group
Subsection
Family
Genus
Cyanobacteria
4
4.1
Anabaena
Species
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BIOLOGY
Anabaena trichomes are untapered with constrictions at the crosswalls. They can be straight, curved or
helical.Thecellsarecylindrical,sphericalorbarrelshapedand,usually,broaderthanlongerwithawidthof
210m.Theterminalcellmayberound,taperedorconical.Heterocystsareintercalary,terminalorboth
andtheishape,usuallysphericaltobarrelshaped,maybeconicalwhenterminal.Akinetesareformedwith
a speciesdependent position in the trichome. No trichome sheath, but often a mucilaginous covering, is
present.Trichomesareusuallymotile.Reproductionisbytrichomefragmentation.Gasvesiclesarepresent
inanumberofspecies,especiallyplanktonic.
Anabaena is a main component of freshwater and saline lake phytoplankton. Marine species also
occur, but never dominate phytoplankton community. Several Anabaena species are able to produce
toxins:theneurotoxinsanatoxina(A.flosaquae,A.circinalis,A.mendotae,A.planktonica,Anabaenasp.),
anatoxina(S) (A. flosaquae), saxitoxin (A. circinalis), and BMAA (A. variabilis, Anabaena sp.) and the
hepatotoxinsmicrocystinLR(A.flosaquae,A.circinalis,A.lemmermannii,A.subcylindrica,Anabaenasp.)
andcylindrospermopsin(A.bergii,A.lapponica).
Anabaena has been the model organism for studies on cyanobacterial nitrogen fixation, including
heterocyst structure and nitrogen fixation genes. This genus includes both obligate photoautotrophs and
facultativeheterotrophs.
BIOTECHNOLOGY
Besides toxins Anabaena can produce a number of bioactive molecules, such as enzyme inhibitors (e.g.,
circinamide, anabaenopeptilides), cardioactive molecules (e.g., anabaenopeptins, pawainaphycin C), and
antimicrobial agents (e.g., bastadins, laxaphycins, spiroidesin), that may have a potential interest for the
pharmaceuticalindustry.
Freshwater Anabaena strains are cultivated in Allen and Arnon (1955) growth medium or BG110
(Rippka et al., 1979), while marine strains are usually cultivated in ASP2 medium without nitrogen (Van
Baalen,1962).
Outdoor cultures of Anabaena have been performed, to verify the production potential of
biotechnologicallyinterestingpolysaccharides,ina1m2deepopenpondobtainingbiomassproductivities
(with a 10cm deep culture) from 9 to 23 g m2 day1 depending on the season, with a photosynthetic
efficiencyaround2.4%andina55Lairlifttubularphotobioreactor,obtainingvolumetricproductivitiesof
0.4gL1day1correspondintto910gm2day1inwinter(Morenoetal.,2003).Huetal.(1996)studiedthe
effect of light path length, mixing mode, cell density and dilution rate on productivity of Anabaena
siamensis using a flat inclined modular photobioreactor, obtaining with air bubbling at the optimal cell
densityof3.20.7gL1maximumproductivitiesofca1.43gL1day1withalightpathof2.6cm.
ResearchonhydrogenproductionbyAnabaenastartedinthe1970s(BenemannandWeare,1974)and
afternumerouslaboratoryresearches(LopesPintoetal.,2002),hasledtosomesmallscaleexperiments
outdoors. In a 4.35L volume photobioreactor made of 10mm internal diameter PVC tubing with A.
variabilis mutant (PK84) Tsygankov et al. (2002) obtained a very low efficiency of conversion from light
energy into biomass (maximum 0.82%) and into hydrogen (maximum 0.094%). With the same
photobioreactor, Lindlblad et al. (2002) obtained a maximum conversion efficiency of 0.042% using the
hydrogen uptake deficient mutant AMC 414 of Anabaena PCC 7120. Using a gastight box with outside
dimensionsof136.8x108.6x13cmandinternaldimensionsof128.4x100.2x13 cm,containg20L of
culture and 8 L of glass beads and ca 158L of gas phase, Smith and Lambert (1981) obtained with
Anabaenacylindricaasustainedhydrogenproductionforoveronemonth.
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References
AllenM.B.,ArnonD.I.(1955)Studiesonnitrogenfixingbluegreenalgae.1.GrowthandnitrogenfixationbyAnabaenacylindrica
Lemm.PlantPhysiology30:366372.
BenemannJ.R.,WeareN.M.(1974)HydrogenevolutionbynitrogenfixingAnabaenacylindricacultures.Science184:174175.
Bornet,.andFlahault,C.(1886'1888').RevisiondesNostocaceshtrocystescontenuesdanslesprincipauxherbiersdeFrance.
AnnalesdesSciencesNaturelles,Botanique,SeptimeSrie7:177262.[BIOLOGYsection]
Burja A.M., Banaigs B., AbouMansour E., Burgess J.G., Wright P.C. (2001) Marine cyanobacteria a prolific source of natural
products.Tetrahedron57:93479377.
CastenholzR.W.(1989)FamilyI.Nostocaceae.GenusI.AnabaenaBorydeSt.Vincent1822..In:StaleyJ.T.,BryantM.P.,PfennigN.,
Holt J.G. (eds.) Bergeys Manual of Systematic Bacteriology, Volume 3, Williams & Wilkins, Baltimore, pp. 17831785.
[BIOLOGYsection]
HuQ.,GutermanH.,RichmondA.(1996)AFlatInclinedModularPhotobioreactorforoutdoormasscultivationofphotoautotrophs.
BiotechnologyandBioengineering,51:5160.
JaiswalP.,KumarSinghP.,PrasannaR.(2008)Cyanobacteriabioactivemoleculesanoverviewoftheirtoxicproperties.Canadian
JournalofMicrobiology54:701717.
LindbladP.,ChristenssonK.,LindbergP.,FedorovA.,PintoF.,TsygankovA.(2002)PhotoproductionofH2bywildtypeAnabaena
PCC7120andahydrogenuptakedeficientmutant:fromlaboratoryexperimentstooutdoorculture.InternationalJournalof
HydrogenEnergy27:12711281.
LopesPintoF.A.,TroshinaO.,LindbladP.(2002)Abrieflookatthreedecadesofresearchoncyanobacterialhydrogenevolution
InternationalJournalofHydrogenEnergy27:12091215.
Moreno J., Vargas M.A., Rodrguez H., Rivas J., Guerrero M.G. (2003) Outdoor cultivation of a nitrogenfixing marine
cyanobacterium,Anabaenasp.ATCC33047.BiomolecularEngineering20:191197.
RippkaR.,DeruellesJ.,WaterburyJ.B.,HerdmanM.,StanierR.Y.(1979)Genericassignmentsstrainhistoriesandpropertiesofpure
culturesofcyanobacteria.JournalofGeneralMicrobiology111:161.
Sivonen K., Brner T. (2008) Bioactive compounds produced by cyanobacteria. In: Herreo A., Flores E. (eds.) The Cyanobacteria.
MolecularBiology,GeneticsandEvolution,CaisterAcademicPress,Norfolk(UK),pp.159191.
Smith G.D., Lambert G.R. (1981) An outdoor biophotolytic system using the cyanobacterium Anabaena cylindrica B629.
BiotechnologyandBioengineering23:.213220.
Tsygankov A.A., Fedorov A.S., Kosourov S.N., Rao K.K. (2002) Hydrogen production by cyanobacteria in an automated outdoor
photobioreactorunderaerobicconditions.BiotechnologyandBioengineering80:777783.
VanBaalenC.(1962)Studiesonmarinebluegreenalgae.BotanicaMarina4:197201.
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7.1.4 Synechococcussp.
Figure13Synechococcussp.100x
PicturefromUNIFI
SYMBOLS:H,E
TAXONOMY
BotanicalCode
Phylum
Class
Order
Family
Genus
Species
Cyanobacteria
Cyanophyceae
Synechococcales
Synechococcaceae
Synechococcus
MicrobiologicalCode
Group
Subsection
Family
Genus
Cyanobacteria
1
1.1
Synechococcus
Species
Thereare75species(andinfraspecific)namesinthedatabaseatpresent,ofwhich34havebeenflaggedas
currentlyacceptedtaxonomically.
BIOLOGY
Cellsaresolitaryorgroupedinmicroscopicormacroscopic,irregularclusters,butnotformingmucilaginous
colonies; cells sometimes in short series of pseudofilamentous formations with 24 (up to 20) cells.
Mucilageisabsentorveryfine,colourless,homogeneous,diffluent,aroundsinglecells.Cellsarelongoval
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orcylindrical,sometimesseveraltimeslongerthanwide,straight,arcuateorsigmoid,1.5uptomorethan
20mlongand0.46(upto11)mwide.Thylakoidsareparietal.Cellsdividebybinaryfission,transversely
(alwaysinoneplaneinsuccessivegenerations,perpendiculartothelongitudinalaxis)intotwoisomorphic
or different (after asymmetric binary fission) daughter cells, which occasionally remain joined for longer
period in pairs. Different cyanobacteria commonly designated as Anacystis nidulans and Agmenellum
quadruplicatumbelongtothisgenus(=Synechococcusnidulans).TheSynechococcusgroupisaprovisional
assemblagewhichcanbedefinedasunicellularcoccoidtorodshapedcyanobacteriathatdividebybinary
fissioninasingleplane.Itissubdividedintoseveralclusters.MostoftheSynechococusgroupmembersare
obligatephotoautotrophs.
Synechococcus species occur in ecologically extreme conditions (e.g. thermophilic Synechococcus
lividus,S.bigranulatus).Severalspeciesgrowwithinmatsandcoloniesofotheralgae,orformfinecolonies
on wet substrates (mud, wood, stones, etc.) Interesting picoplanctic or planktic species were found in
oceansaswellasinfreshwaterreservoirs(e.g.,S.rhodobactron).
BIOTECHNOLOGY
Synechococcushasbeeninvestigatedasahydrogenproducer(KumazawaandMitsui,1994;LopesPintoet
al., 2002). Its main biotechnological interest is its transformability, that allowed insertion of genes from
other bacteria, in order to enchance production of naturally occurring molecules such as poly
hydroxybutyrate (Miyake et al., 2000; Nishioka et al., 2001), interesting for bioplastic production, or
exogenousmoleculessuchasethanol(DengandColeman,1999)orisobutyraldehyde(Atsumietal.,2009),
aprecursorofbutanolapotentialfuel,thatcouldbeobtaineddirectlyfromCO2andsunlight.
References
Atsumi S., Higashide W., Liao J.C. (2009) Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Nature
DengM.D.,ColemanJ.R.(1999)Ethanolsynthesisbygeneticengineeringincyanobacteria.AppliedandEnvironmentalMicrobiology
65:523528.
HerdmanM.,CastenholzR.W.,ItemanI.,WaterburyJ.B.,RippkaR.(2001)TheArchaeaandthedeeplybranchingandphototrophic
bacteria. In: Boone D.R., Castenholz R.W. (eds.) Bergey's Manual of Systematic Bacteriology, 2nd edn. Springer Verlag:
Heidelberg.[BIOLOGYsection]
KomrekJ.(1992)in:http://www.cyanodb.cz/Synechococcus[BIOLOGYsection]
KomrekJ.,AnagnostidisK.(1998)Cyanoprokaryota1.Teil:Chroococcales.In:EttlH.,GrtnerG.,HeynigH.,MollenhauerD.(eds.)
SsswasserfloravonMitteleuropa19/1,GustavFischer,JenaStuttgartLbeckUlm.[BIOLOGYsection]
Kumazawa S., Mitsui A. (1994) Efficient hydrogen photoproduction by synchronously grown cells of a marine cyanobacterium,
Synechococcussp.MiamiBG043511,underhighcelldensityconditions.BiotechnologyandBioengineering44:854858.
Miyake M., Takase K., Narato M., Khatipov E., Schnackenberg J., Shirai M., Kurane R., Asada Y. (2000) Polyhydroxybutyrate
productionfromcarbondioxidebycyanobacteria.AppliedBiochemistryandBiotechnology8486:9911002.
Ngeli C. (1849) Gattungen einzelliger Algen, physiologisch und systematisch bearbeitet. Neue Denkschriften der Allg.
SchweizerischenGesellschaftfrdieGesammtenNaturwissenschaften10:iviii,1139,plsIVIII.[BIOLOGYsection]
NishiokaM.,NakaiK.,MiyakeM.,AsadaY.,TayaM.(2001)Productionofpolyhydroxybutyratebythermophiliccyanobacterium,
Synechococcussp.MA19,underphosphatelimitedconditions.BiotechnologyLetters23:10951099.
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7.1.5 Synechocystissp.
Figure14Synechocystissp.PCC6308
Picturefromwww.ibvf.cartuja.csic.es/
SYMBOLS:H,E
TAXONOMY
BotanicalCode
Phylum
Class
Order
Family
Genus
Species
Cyanobacteria
Cyanophyceae
Synechococcales
Merismopediaceae
Synechocystis
MicrobiologicalCode
Group
Subsection
Family
Genus
Cyanobacteria
1
1.1
Synechocystis
Species
BIOLOGY
Cells are solitary, spherical or widely oval, after division hemispherical and for a short time the two cells
remainattachedtogether.Cellsarewithoutmucilageorwithnarrow,fine,colourlessandusuallydiffluent
and indistinct mucilaginous envelopes. In several strains the cell walls contain a special Slayer with
characteristical hexagonal substructure. It never forms colonies. Cell division occurs by binary fission
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(mainlybypinching)intotwomorphologicallyequaldaughtercells,whichreachtheoriginalglobularshape
before next division; cells always divide in two perpendicular planes in successive generations. If
mucilaginousenvelopesareformedaroundthecells,theysplittogetherwithdividingcells.Synechocystis
has been widely investigated from the physiology and genetic point of view and some strains (e.g.,
SynechocystisPCC6803)areamongthemostwellknowncyanobacteria.
Severalspeciesareplanktonicinfreshwaterreservoirsorinsalineorseawaters,othergrowinpoolsof
thermalandmineralsprings,ofbrackishswampsorinmoors.
BIOTECHNOLOGY
Its biotechnology is very similar to that of Synechoccoccus. It is a potential producer of
polyhydroxyalkanoates(Sudeshetal.,2001;Wuetal.,2001).Ithasbeenstudiedforhydrogenproduction
(LopesPinto et al., 2002; Angermayr et al., 2009) as well as, being easily transformable, for ethanol
productionafterbeinggeneticengineered(DexterandFu,2009).
References
Angermayr A., Hellingwerf K.J., Lindblad P., Teixeira de Mattos M.J. (2009) Energy biotechnology with cyanobacteria. Current
OpinioninBiotechnology20:257263.
DexterJ.,FuP.(2009)Metabolicengineeringofcyanobacteriaforethanolproduction.EnergyandEnvironmentalScience2:857
864.
KomrekJ.(1992)in:http://www.cyanodb.cz/Synechocystis[BIOLOGYsection]
KomrekJ.,AnagnostidisK.(1998)Cyanoprokaryota1.Teil:Chroococcales.In:EttlH.,GrtnerG.,HeynigH.,MollenhauerD.(eds.)
SsswasserfloravonMitteleuropa19/1,GustavFischer,JenaStuttgartLbeckUlm.[BIOLOGYsection]
SauvageauC.(1892)Surlesalguesd'eaudoucerecoltesenAlgriependantlesessiondelasocietbotaniqueen1982.Bulletinde
laSocitBotaniquedeFrance34:104128.[BIOLOGYsection]
SudeshK.,TaguchiK.,DoiY.(2001)CancyanobacteriabeapotentialPHAproducer?RIKENReview42:7576.
Wu G.F., Wu Q.Y., Shen Z.Y. (2001) Accumulation of polyhydroxybutyrate in cyanobacterium Synechocystis sp. PCC6803.
BioresourceTechnology76:8590.
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8 Eukaryoticmicroalgae
8.1 Chlorophyta
8.1.1 Ostreococcussp.
Figure15Ostreococcustaurii
Figure16Ostreococcuslucimarinus
http://genome.jgi
psf.org/Ostta4/Ostta.jpg&imgrefurl=http://genome.jg
ipsf.org/Ostta4/Ostta4.home
http://bioinformatics.psb.ugent.be/plaza/img/organisms/Ostreococcus_lu
cimarinus
SYMBOLS:
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Prasinophyceae
Mamiellales
Mamiellaceae
Ostreococcus
RelatedSpecies
O.taurii,O.lucimarinus
BIOLOGY
Ostreococcus tauri and related species are the smallest known eukaryotes. They are similar to flattened
spheresinshapenearly1mindiameter.ThemoststrikingfeatureofO.tauriandrelatedspeciesistheir
minimal cellular organization: a naked cell, lacking flagella, with a single chloroplast and mitochondrion,
andasmallgenome(12Mbp).Ostreococcusisagloballyabundantpicoeukaryoteintheeuphoticzone.
Recent work has shown that smallsubunit rDNA sequences of Ostreococcus from cultures and
environmental samples cluster into four different clades that are likely distinct enough to represent
different species. Light and nutrient conditions experienced by surface and deep isolates could be the
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driving factors behind their genetic divergence. Comparative analysis of Ostreococcus sp. will help to
understandnichedifferentiationinunicellulareukaryotesandevolutionofgenomesizeineukaryotes.
BIOTECHNOLOGY
Atpresent,nobiotechnologicaldevelopmenthasbeenreportedforthisalga.However,itoneofthemost
studiedalgaeforgeneticanalysis(Hallmann,2007)anditisimportantforthepossibilitiestohaveaninsight
inmetabolicandgeneticpathwaysofmetaboliteproductionandgrowthprocesses.
References
Palenik B., Grimwood J., Aerts A. et al. (2007) The tiny eukaryote Ostreococcus provides genomic insights into the paradox of
planktonspeciation.ProceedingsoftheNationalAcademyofSciencesoftheUSA104:77057710.[BIOLOGYsection]
Rodrguez F., Derelle E., Guillou L., Le Gall F., Vaulot D., Moreau H. (2005) Ecotype diversity in the marine picoeukaryote
Ostreococcus(Chlorophyta,Prasinophyceae).EnvironmentalMicrobiology7:853859.[BIOLOGYsection]
HallmannA.(2007)Algaltransgenicsandbiotechnology.TransgenicPlantJournal1:8198.
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8.1.2
Tetraselmissp
Figure17Tetraselmischuii
CNRS,StationBiologiquedeRoscoff
http://planktonnet.awi.de
SYMBOLS:D,E,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Prasinophyceae
Chlorodendrales
Chlorodendraceae
Tetraselmis
RelatedSpecies
T. alacris, T. apiculata, T. ascus, T. astigmatica, T. chuii, T. convolutae, T. cordiformis, T. desikacharyi, T.
gracilis, T. hazeni, T. impellucida, T. inconspicua, T. levis, T. maculata, T. marina, T. micropapillata, T.
rubens,T.striata,T.suecica,T.tetrabrachia,T.tetrathele,T.verrucosa,T.wettsteinii.
BIOLOGY
Structuralandmorphologicalfeatures
Tetraselmisisamarinegreenflagellate.Thefourflagellaofthisalga,insertedinananteriordepressionof
thecell,arecoveredbyscalesofdifferenttypes:pentagonal,rodshapedandhairscalesand,onlyinsome
strains,knottedscales(BarsantiandGualtieri,2006).Mostspeciesofthegenususuallyareencounteredas
solitary,freeswimming,thecatecells.Cellwallorthecaisformedbythefusionofscalessimilartothose
found covering the flagella (Nozaki, 2003). Cells are ovoid or ellipsoidal, somewhat compressed
equatorially. In the case of Tetraselmis suecica cells have an average size of 10 x 8 m and a weight of
about200pg.Thechloroplastissingle,cupshapedwithonepyrenoid(inthespecieswereitispresent)and
astigma.Theasexualreproductionisbybipartitionwithinthetheca,whilesexualreproductionisunknown
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(Nozaki, 2003). It is a very common component in inshore marine environments, tide pools in particular,
buttherearealsosevenfreshwaterspecies(Nozaki,2003).Tetraselmisisaveryrobustmicroorganismable
to resist to extreme pH, salinity and temperature and to adapt to rapid changes in environmental
conditions.Thisfeaturemakesitparticularlysuitableforoutdoormasscultivation.Amongthe50species
known,themostwidelyusedarethemarineT.suecica,T.chuiandT.tetrathele.
Grosscompositionunderoptimalandstressedconditions
Tetraselmissuecicahasahighproteincontent(upto4050%).Carbohydrateisabout20%andlipidabout
20%ofthecelldryweight.Undernutrientstress(nitrogenorphosphorusdeprivation)Tetraselmissuecica
accumulatescarbohydrates.
Renaud et al. (1999) report for Tetraselmis sp. a content of 2630% protein, 1314% lipid, 89%
carbohydrateand1417%ashaswellasabout60%ofpolyunsaturatedfattyacidsoverthetotalfattyacid
content.
BIOTECHNOLOGY
Tetraselmis offers a valuable source of protein, bioactive compounds, antioxidants, vitamins, sterols and
polyunsaturated fatty acids for human and animal consumption. One the most important applications of
Tetraselmisisinaquacultureforrearingzooplanktonandlarvalstagesofmarinefish,bivalvemolluscsand
crustaceans(MullerFeugaetal.,2003;Tredicietal.,2009).Thegenushasbeenfoundtohaveantibacterial
activity towards important aquaculture pathogens (Austin et al., 1992) and it was also proposed as
probiotic(Tredicietal.,2009,IriantoandAustin,2002).Duetoitshighcontentofgoodqualityprotein(40
50%d.wt),Tetraselmissuecicabiomasscouldrepresentanalternativeingredientforanimalfeed.Because
ofitshigh contentofvitamin E(0.130.25gkg1),Tetraselmishasalsobeenproposedasasourceofthis
vitaminaspreservativeinfoods,additiveinanimalfeedandsunscreenincosmetics(CarballoCrdenaset
al.,2003).Activeingredientsextractedfromthismicroalgaarecurrentlyusedinthedevelopmentofnovel
cosmeticformulationsinfluencinggrowthofhumanhairand/orpigmentationofhumanskin(Pertileetal.,
2010). An emerging use of Tetraselmis is for carbon biofixation in combination with biofuels production
(biodiesel,bioethanol)(Tredici,2010).
CultureMedia
F medium (Guillard and Ryther, 1962) is widely used for cultivation of Tetraselmis. The microalga can be
grownalsoinnaturalseawaterintegratedwithnutrients.
Cultivationmethods
IngeneralTetraselmisiscultivatedunderautotrophicconditions.Somespecieshavealsoheterotrophicor
mixotrophic capacity. Intensive cultivation of Tetraselmis has been carried out in open raceway ponds
andindifferentkindsofclosedphotobioreactors(PBR).InHawaiiin24m2flumesLawsandBerning(1991)
obtained a productivity of 1520 g C m2 day1 with photosynthetic efficiencies of 910% with Tetraselmis
suecica. In pilotscale open ponds in Southern Italy, Tetraselmis tetrathele has reached productivities of
about 30 g m2 day1 during the summer, with photosynthetic efficiencies in the PAR region around 5%
(Materassi et al., 1983). A parallel cultivation of Tetraselmis suecica in pilotscale open ponds and near
horizontal tubular reactors has shown similar productivities for the two systems, about 26 g m2 day1
(Pedroni et al., 2004). Current methods for its culture rely on batch, semicontinuous or continuous
cultivation. At present, culture methods used in hatcheries for Tetraselmis production use mainly
polyethylenebagsandtransparentglassfibrecylinders(upto500L)usuallykeptindoorswithartificiallight
(Fulks and Main, 1991). Advances have recently been made in the field of photobioreactor technology
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(Tredici et al., 2010) that improved microalgae productivity in comparison to the traditional culture
systems. Among these systems annular columns (Chini Zittelli et al., 2006) and disposable flat panels
(Tredicietal.,2010)havebeenandarecurrentlyusedinlaboratoryandatpilotscaleoutdoors.Duringthe
summer in Central Italy in an experiment reproducing a full scale plant arrangement, in 120L annular
columnsatadailydilutionrateof40%,Tetraselmissuecicaattainedanaveragevolumetricproductivityof
0.46gl1day1,andanoverallarealproductivityof36.3gm2day1withaphotosyntheticefficiencyinthe
PAR region of 9.4% (Chini Zittelli et al., 2006). Tetraselmis has been also grown at industrial scale under
heterotrophicconditionsinfermenterswithyieldsinexcessof100gL1day1(Dayetal.,1991).
Harvestingmethods
Tetraselmissettlesspontaneouslyandcanbeharvestedinfunnels,thoughcentrifugationisafasteranda
moreefficientharvestingmethod.
Upscalinglimitations
AlthoughTetraselmisisaveryrobustmicroorganismabletoresisttoextremepH,salinityandtemperature
andtoadapttotherapidchangesinenvironmentalconditions,toobtainhighproductivitieshighamounts
ofenergyaregenerallyrequiredforculturemixingduetothehighsedimentationrateofthemicroalga.
Majorcharacteristicsofthisgenusarerobustnessandhighproductivityinoutdoormasscultures
References
AustinB.,BauderE.,StobieM.B.C.(1992)InhibitionofbacterialfishpathogensbyTetraselmsissuecica.JournalofFishDiseases15:
5561.
BarsantiL.,GualtieriP.(2006)Algae.Anatomy,Biochemistry,andBiotechnology.Taylor&Francis,BocaRaton.
CarballoCrdenas E.C., Tuan P.M., Janssen M., Wijffels R.H. (2003) Vitamin E (tocopherol) production by marine microalgae
DunaliellatertiolectaandTetraselmissuecicainbatchcultivation.BiomolecularEngineering20:139147.
Chini Zittelli G., Rodolfi L., Biondi N., Tredici M.R. (2006) Productivity and photosynthetic efficiency of outdoor cultures of
Tetraselmissuecicainannularcolumns.Aquaculture261:932943.
Day J.G., Edwards A.P., Rodgers G.A. (1991) Development of an industrialscale process for the heterotrophic production of
microalgalmolluscfeed.BioresourceTechnology38:245249.
Fulks W., Main K.L. (eds.) (1991) Rotifer and Microalgae Culture Systems. Proceedings of a U.S.Asia Workshop. The Oceanic
Institute,Honolulu,Hawaii,USA.
Guillard R.R.L., Ryther J.H. (1962). Studies of marine planktonic diatoms. I. Cyclotella nana (Hustedt) and Detonula confervacea
(Cleve).CanadianJournalofMicrobiology8:229239.
IriantoA.,AustinB.(2002)ProbioticsinaquacultureJournalofFishDiseases25:633642.
Laws E.A., Berning J.L. (1991) A Study of the energetics and economics of microalgal mass culture with the marine chlorophyte
Tetraselmissuecica:Implicationsforuseofpowerplantstackgases.BiotechnologyandBioengineering37:936947.
Materassi R., Tredici M.R., Milicia F., Sili C., Pelosi E., Vincenzini M., Torzillo G., Balloni W., Florenzano G., Wagener K. (1983)
Developmentofaproductionsizesystemforthemasscultureofmatinemicroalgae.In:PalzW.,PirrwitzD.(eds)Energy
frombiomass.RiedelPublishingCompany,Boston,SeriesE,vol.5,pp.150158.
MullerFeugaA.,RobertR.,CahuC.,RobinJ.,DivanachP.(2003)Usesofmicroalgaeinaquaculture.In:Stottrup,J.G.,McEvoy,L.A.
(Eds.),LiveFeedsinMarineAquaculture.Blackwell,Oxford,pp.253299.
Nozaki H. (2003) Flagellated green algae. In: Wehr J.D., Sheath R.G. (eds.) Freshwater algae of North America. Ecology and
classification.Elsevier,Amsterdam,pp.225252.
Pedroni P.M., Lamenti G., Prosperi G., Ritorto L., Scolla G., Capuano F., Valdiserri M. (2004) Enitecnologie R & D project on
microalgae biofixation of CO2: outdoor comparative tests of biomass productivity using flue gas CO2 from a NGCC power
plant.ProceedingsofSeventhInternationalConferenceonGreenhouseGasControlTechnologies(GHGT7),59September
2004,Vancouver,Canada.
PertileP.,ZanellaL.,HermmannM.,HolgerJ.,GaeblerS.(2010)ExtractsofTetraselmissp.USpatent2010/0143267.
Renaud S.M., Thinh L.V., Parry D.L. (1999) The gross chemical composition and fatty acid composition of 18 species of tropical
Australianmicroalgaeforpossibleuseinmariculture.Aquaculture170:147159.
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Throndsen,J.(1996)Theplanktonicmarineflagellates.In:Tomas,C.R.(ed)Identifyingmarinephytoplankton.AcademicPress,San
Diego,pp.591730
Tredici M.R. (2010) Photobiology of microalgae mass cultures: understanding the tools for the next green revolution. Biofuels 1,
143162.
Tredici M.R.,BiondiN.,Chini ZittelliG.,PonisE.,RodolfiL.(2009)Advancesinmicroalgalcultureforaquaculturefeedandother
uses. In: Burnell G., Allan G., (eds.) New Technologies in Aquaculture: Improving production efficiency, quality and
environmentalmanagement.WoodheadPublishingLtd,Cambridge,UK,andCRCPressLLC,BocaRaton,FL,USA,pp.610
676.
Tredici M.R., Chini Zittelli G., Rodolfi L. (2010) Photobioreactors In: Flickinger M.C., Anderson S. (eds) Encyclopedia of Industrial
Biotechnology:Bioprocess,Bioseparation,andCellTechnology.JohnWiley&Sons,Inc.,Hoboken,NJ,USA.Vol6,pp.3821
3838.
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8.1.3
Botryococcusbraunii
Figure18Botryococcusbraunii
http://erenovable.com/wpcontent/uploads/2007/02/BotryococcusbrauniiAlga.jpg
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Chlorococcales
DictyosphaeriaceaeWest1916,Botryococcaceae(Wille1909)
Botryococcus
Botryococcusbraunii
RelatedSpecies
B. australis, B. balkachicus, B. braunii, B. calcareous, B. canadensis, B. comperei, B. coorongianus, B.
fernandoi,B.giganteus,B.neglectus,B.pila,B.protuberans,B.sudeticus,B.terribilis,B.terricola.
BIOLOGY
Botryococcus braunii is a green colonial microalga. Colonies, under microscope observation, exhibit a
typical morphology characterised by a botryoid organisation of individual pyriformshaped cells held
together by a refringent matrix containing lipids. Oil droplets can be excreted from the matrix by the
pressure of a coverglass. Ultrastructural studies reveal that the matrix surrounding the basal part of the
cells consists of outer walls originating from successive cellular divisions. Furthermore, the bulk of B.
braunii hydrocarbons are stored in these outer walls Colonies frequently are compounded by
interconnectingstrandsoftoughmucilagebetweenclustersofcells.Cellshaveacupshapedplastidwitha
naked pyrenoidlike body. Found in phytoplankton and metaphyton of ponds and lakes. Geographically
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widespread,itisrarelyabundant,thoughithastheabilitytoformblooms,sometimesenduringovermany
years,likeintheDarwinReservoirinAustralia.
Botryococcus braunii produces various types of ether lipids and hydrocarbons up to 60% of biomass.
Accordingtohydrocarbonproduction,B.brauniiissubclassifiedintothreechemicalraces.AlgaeinraceA
produceessentiallynalkadieneandtrienehydrocarbons,oddcarbonnumberedfromC23to,algaeinrace
B produce triterpenoid hydrocarbons, C30C37 botryococcenes and C31C34 methylated squalenes and
algae in race L produce a single tetraterpenoid hydrocarbon, lycopadiene. 18S rRNA sequences of four
strains of B. braunii belonging to the three chemical races established that these strains formed a
monophyleticgroup.
BIOTECHNOLOGY
Most studies concerning the influence of various factors on the production of biomass and
hydrocarbonswerecarriedoutinthelaboratory.InthethreechemicalracesofB.braunii,thehydrocarbon
productivity,aswellastheetherlipidsynthesis,wasshowntobeoptimalduringtheexponentialphaseof
growthanddoesnotoccurinnitrogenandphosphorousdeficientmedia(MetzgerandLargeau,2005).
Entrapment of B. braunii colonies in calcium alginate beads exhibits some interesting advantages by
comparison to free suspension cultures: enhancement in chlorophyll photosynthetic activity, protection
against photoinhibition towards high irradiance and an increase in hydrocarbon production despite a
decreaseintherateofbiomassproduction;however,thelackofstabilityofcalciumalginatebeadsovera
longperiodisnotfavourabletotheiruseinculturesonalargescale(MetzgerandLargeau,2005).
There are no reports on successful largescale cultivation. Experimental cultures under natural
illumination in tubular photobioreactors, immersed in water for cooling, have been conducted up to a
volumeof200L(GudinandChaumont,1984).In1000Lracewayponds,averagebiomassproductivityof89
mg L1 day1 were achieved, with a lower hydrocarbon content compared to indoor cultures, with B.
mahabali(Dayanandaetal.,2010).Outdoorculturesareeasilycontaminatedbyotheralgae(Metzgerand
Largeau,2005).SmallscalecommercialproductionofB.brauniiiscarriedoutinPortugalbyA4FAlgafuel
S.A.
Botryococcusisabletogrowondifferenttypesofeffluentsunderlaboratoryconditions(Metzgerand
Largeau, 2005; Shen et al., 2008), although very slowly: e.g., on secondary treated domestic sewage its
productivitywasofabout30mgL1day1(Sawayamaetal.,1994;Sydneyetal.,2011).
References
Dayananda C., Kumudha A., Sarada R., Ravishankar G.A. (2010) Isolation, characterization and outdoor cultivation of green
microalgaeBotryococcussp.ScientificResearchandEssays5:24972505.
GudinC.,ChaumontD.(1984)Solarbiotechnologystudyanddevelopmentoftubularsolarreceptorsforcontrolledproductionof
photosynthetic cellular biomass for methane production and specific exocellular biomass. In: Palz W., Pirrwitz D. (eds.)
Energyfrombiomass,serieE,vol5.Reidel,Dordrecht,pp184193.
MetzgerP.,LargeauC.(2005)Botryococcusbraunii:arichsourceforhydrocarbonsandrelatedetherlipids.AppliedMicrobiology
andBiotechnology66:486496.[AlsoforBIOLOGYsection]
SawayamaS.,InoueS.,YokoyamaS.(1994)ContinuouscultureofhydrocarbonrichmicroalgaBotryococcusbrauniiinsecondarily
treatedsewage.AppliedMicrobiologyandBiotechnology41:729731.
Shen Y., Yuan W., Pei Z., Mao E. (2008) Culture of microalga Botryococcus in livestock wastewater. Transactions of the ASABE
(AmericanSocietyofAgriculturalandBiologicalEngineers)51:13951400.
SydneyE.B.,daSilvaT.E.,TokarskiA.,NovakA.C.,deCarvalhoJ.C.,WoiciecohwskiA.L.,LarrocheC.,SoccolC.R.(2011)Screeningof
microalgae with potential for biodiesel production and nutrient removal from treated domestic sewage. Applied Energy:
doi:10.1016/j.apenergy.2010.11.024.
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8.1.4
Chlamydomonasreinhardtii
Figure19Chlamydomonasreinhardtii
http://sandwalk.blogspot.com/2007/10/genomeofchlamydomonasreinhardtii.html
SYMBOLS:H,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Volvocales
Chlamydomonadaceae
Chlamydomonas
RelatedSpecies
NCBINucleotideSequences
As of 2 June 2009, nucleotide sequence data are available at http://www.ncbi.nlm.nih.gov/Genbank for
265,283samplesidentifiedasChlamydomonas.
Taxonomic identifications of these samples to genus (or species) levels unaccompanied by explicit
indication of voucher specimens may not be verifiable on morphological/anatomical grounds and
consequentlybeoflittleornotaxonomicvalue.
Numbers of names and species: there are 1163 species (and infraspecific) names in the database at
present,ofwhich382havebeenflaggedascurrentlyacceptedtaxonomically.
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BIOLOGY
Chlamydomonasreinhardtiiisaunicellulargreenalgaabout1020mindiameterwithtwoflagella.The
cell wall is made of hydroxyprolinerich glycoproteins. C. reinhardtii has a single, large, cupshaped
chloroplast,alargepyrenoid,andastigma("eyespot")thatsenseslight.
C. reinhardtii is primarily used as a model organism in biology. C. reinhardtii can grow
photoautotrophicallyt,orheterotrophicallyinthedarkwhensuppliedwithacetate.Thishasbeenexploited
forcreationofthousandsofmutantsaffectedinthephotosyntheticprocess(www.chlamy.org).
Due to its sexual and vegetative growth, heterotroph and photoautotroph growth cycles, genetic
mappingofphotosyntheticmarkersbecamepossibleearlyonandmadeChlamydomonasaprimetoolfor
studying the genetics of photosynthesis, among others revealing maternal inheritance of chloroplast
encodedgenes(Harris,1989).
The alga as such is not a prime target for biofuels production. However, access to the numerous
mutantsandgenetictransformationtechnologieshavemadethisalgaaprimetargetforbiofuelsresearch
(mainlyforbiohydrogen).
Vegetative cells of the reinhardtii sspecies are haploid and have 17 small chromosomes. Haploid
gametesdevelopunder nitrogen starvation.Two matingtypesmt(+)andmt()thenfusetoforma diploid
zygote.Thezygoteundergoesmeiosis andreleasesfourflagellatedhaploidcellsthatresumethevegetative
lifecycle.
Underidealgrowthconditions,cellsmaysometimesundergotwoorthreeroundsof mitosisbeforethe
daughtercellsarereleasedfromtheoldcellwallintothemedium.Thus,asinglegrowthstepmayresultin
4to16daughtercellspermothercell.Inpresenceofacetatethecellsgrowsignificantlyfasterthanwith
lightalone.Thecellcycleofthisunicellulargreenalgaecanbesynchronizedbyalternatingperiodsoflight
anddark,aprocesswidelyappliedtostudyregulationofgeneexpressionduringthediurnalcycle(e.g.,Leu
etal.,1990;Breidenbachetal.,1990).
Theattractivenessofthealgaasamodelorganismhasrecentlyincreasedwiththereleaseofseveral
genomic resources (see Chlamydomonas Center, http://www.chlamy.org/), which also administrates
thousandsofmutantstrains,plasmidsandotherresourcesforgeneticandmolecularresearch.Inaddition
togenomicsequencedatathereisalargesupplyofexpressionsequencedataavailableascDNAlibraries
andexpressedsequencetags(ESTs).cDNAlibrariesareavailableonlineandaBAClibrarycanbepurchased
from the Clemson University Genomics Institute. There are also two databases of >50 000 and >160 000
ESTsavailableonline.
ThevariabilityofalgalbiochemicalcompositionhasbeenimpressivelydemonstratedinC.reinhardtiiduring
itsdiurnalgrowthcycle.Duringtheearlylightperiodmostcellularmassisprotein,duringthesecondhalfof
the light period the alga accumulates up to 50% starch. While oil droplets in Chlamydomonas were not
originallyreported,recentstudiesrevealedupto20%TAGundernitrogenstarvation.
BIOTECHNOLOGY
Clamydomonas is not being investigated for mass cultivation and biomass production, though Hu et al.
(2008)haveincludedtwoChlamydomonasspeciesintheircomparativestudyonlipidproductivitywithC.
applantashowinghigherproductivitythanC.reinhardtii.However,thewideknowledgeonitsgeneticsand
cellbiologyhasallowedrevealingnumerousmechanismsofinteresttobiofuelsproduction.
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Analysisofstarchdeficientmutantsrevealedthetightlinkbetweencarbonallocationtostarchorlipid
biosynthesis. Identification of mirna in Chlamydomonas has revealed their usefulness in upregulation of
lipidbiosynthesis(Maor,personalcommunication).
Clamydomonas has the potential to produce large amounts of hydrogen because it can directly split
waterintohydrogenand oxygen usingtheenzyme hydrogenase.Itwas discoveredthatwhen thealga is
deprivedofessentialsulfatesalts,itnolongermaintainstheproteincomplexnecessaryforphotosynthetic
production of oxygen and instead switches to the hydrogenproducing metabolic pathway. However, the
alga can not grow in a sulfur deprived condition for long time before it needs to revert to the oxygen
producing mode (Torzillo et al., 2009; Ghirardi et al., 2010). Detailed understanding of the hydrogen
productionprocessandthehydrogenasecomplexofChlamydomonashaveallowedcreatingmutantswith
enhancedhydrogenproductioncapacity.
Significant efforts have been undertaken to investigate expression of recombinant proteins in
Chlamydomonas reinhardtii (Mayfield et al., 2003). Among the proteins expressesd successfully are
antibodymolecules,bovinemammaryassociatedserumamyloidthatmaybeexpressedsuccessfullyboth
inthecytoplasmorchloroplast.(e.g.Mayfieldetal.,2007).
Culturemedia
C.reinhardiiicanbecultivatedinalargevarietyofsyntheticgrowthmediasuchassagergranick,highsalt
(sueoka),tap,inpresenceorabsenceofacetate(Harris,1989,p31on).Presenceofacetatesignificantly
enhancesthegrowthrateunderdiurnallightregime.
Productionsystem
Possiblebiohydrogenproductionhas beeninvestigatedand modelledinastudytonrelNREL (Wadeand
Amos,2004),thoughnoactualfieldtestsandupscalinghavebeenperformedasactualstudieshavebeen
performedonthelabscalesofar.
For the production of recombinant proteins rather small well controlled photobioreactors are
suggestedratherthanlargeoutdoorscultivationsystems.
Earlygeneticwork,genetictransformationandfullysequencedgenomemakethisalgathefavorite
forexploringthepowerofgeneticengineeringinalgalbiotechnology(Harris,1989).
Majorbreaktroughsarealgaewithincreasedhydrogenproduction,reducedchlorophylletc.Mutants
withincreasedoilcontentduetoimpairedstarchsynthesis(Lietal.,2010),andapplicationofmirna
miRNAtechnologyforenhancedoilproduction(Maor,personalcommunication).
Furthermore the algae has been used to express and produce recombinant medically relevant
peptidesandproteins(Mayfieldetal.,2003,2007).
References
Breidenbach E., Leu S., Michaels A., Boschetti A. (1990) Synthesis of EFTu and distribution of tis mRNA between stroma and
thylakoidsduringthecellcycleofChlamydomonasreinhardii. BiochimicaetBiophysicaActa1048:209216.
GhirardiM.L.,KosourovS.,ManessP.,SmolinskiS.,SeibertM.(2010)AlgalHydrogenproduction.In:FlickingerM.C.,AndersonS.
(eds.) Encyclopedia of Industrial Biotechnology: Bioprocess, Bioseparation, and Cell Technology. John Wiley & Sons, Inc.,
Hoboken,NJ,USA.Vol.1,pp.184198.
Harris E. (1989) The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use.Academic Press, San
Diego.
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HuQ.,SommerfeldM.,JarvisE.,GhirardiM.,PosewitzM.,SeibertM.,DarzinsA.(2008)Microalgaltriacylglycerolsasfeedstocksfor
biofuelsproduction:perspectivesandadvances.ThePlantJournal54:621639.
Leu S., White D., Michaels A. (1990) Cell cycledependent transcriptional and posttranscriptional regulation of chloroplast gene
expressioninChlamydomonasreinhardtii.BiochimicaetBiophysicaActa1049:311317.
LiY.,HanD.,HuG.,DauvilleeD.,SommerfeldM.,BallS.,HuQ.(2010)Chlamydomonasstarchlessmutantdefectivein
ADPglucosepyrophosphorylasehyperaccumulatestriacylglycerol.MetabolicEngineering12:387391.
Mayfield S.P., Manuell A.L., Chen S., Wu J., Tran M., Siefker D., Muto M., MarinNavarro J. (2007) Chlamydomonas
reinhardtiichloroplastsasproteinfactories.CurrentOpinioninBiotechnology18:126133.
MayfieldS.P.,FranklinS.E.,LernerR.A.(2003)Expressionandassemblyofafullyactiveantibodyinalgae.Proceedings
oftheNationalAcademyofScienceoftheUSA100:438442.
Molnar A., Bassett A., Thuenemann E., Schwach F., Karkare S., Ossowski S., Weigel D., Baulcombe D. (2009) Highly
specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii. The Plant
Journal58:165174.
Torzillo G., Scoma A., Faraloni C., Ena A., Johanningmeier U. (2009) Increased hydrogen photoproduction by means of a sulfur
deprivedChlamydomonasreinhardtiiD1proteinmutant.InternationalJournalofHydrogenEnergy34:45294536.
Wade A., Amos A. (2004) Updated cost analysis of photobiological hydrogen production from Chlamydomonas
reinhardtiigreenalgae.Milestonecompletionreport,NREL/mp56035593.
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8.1.5
Haematococcuspluvialis
Figure20GreenandastaxanthincontainingH.pluvialiscells(lightmicroscope).
D.Reinecke,BGU
SYMBOLS:B,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Volvocales
Haematococcaceae
Haematococcus
RelatedSpecies
H. allmanii, H. buetschlii, H. capensis, H. carocellus, H. droebakensis var. fastigatus, H. droebakensis, H.
grevillei, H. insignis, H. lacustris, H. murorum, H. pluvialis, H. salinus, H. sanguineus, H. thermalis, H.
zimbabwiensis.
BIOLOGY
Haematococcus pluvialis is a medium to large unicellular green algae (10 100 m) with a large single
cloroplast,twoflagellae.Youngcellsaremotile,ageingcells(palmelloids)losetheirmobility.Understress
the algae accumulates the carotenoid asthaxanthin in cytoplasmic oil globules and enters a resting stage
(Droop,1954).Thecellwallthickensandaltersitschemicalcompositionduringmaturation.
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Biochemicalcompositionofalgaeandaquaticbiomassmainconstituents
During steady state growth the algae is rich in protein, with less than 10% lipid content, displaying the
typicalgreenalgalfattyacidcompositionwithsignificantamountsofshort(C16,C18)polyunsaturatedfatty
acids mainly in the chloroplast lipids, chlorophylls a and b and photosynthetic carotenoids such as
carotene.
Under stress the alga accumulates initially up to 50% starch, then initiates synthesis of triacylglycerol
(TAG)accumulatingto40%ofcellweightincytoplasmicoilglobules,andupto4%oftheketocarotenoid
astaxanthin(Boussibaetal.,1992,1999).
Growthkineticsandefficiencies
Haematococcusisconsideredaslowgrowingalga.Productivityinoutdoorssystemshasbeenreportedby
HuntleyandRedalje(2007),implyingaphotosyntheticenergyconversionefficiencyof3%.
BIOTECHNOLOGY
CultureMedia
BG11(Rippkaetal.,1979),mBG11orsimilarsyntheticgrowthmedia.
Cultivationmethods
Haematococcusiscultivatedbothinopenpondsorclosedbioreactors.Thetubularphotobioreactorfacility
atQeturaisthelargestoperatingphotobioreactorfacilityformicroalgaeproduction.Largescaleoutdoors
cultivationintwostagemode,photobioreactorforgreencellsandopenpondsforproductionofredcells
wastestedinHawaiiandyieldedalongtermgrowthaverageof38tonsperhectareandyearwith25%oil
content,or422GJha1year1(HuntleyandRedalje,2007).
Productionsystem
Production is managed as semicontinuous or batch cultivation in two stages. The first stage is a nutrient
sufficientgreenstagethatcanbehandledascontinuousorsemicontinuousculture(Boussibaetal.,1997;
Huntley and Redalje, 2007), while the second stage under nutrient limitation for accumulation of
astaxanthin is necessarily a batch cultivation where all the resulting biomass is harvested. One step
cultivationhasbeenproposedbutnotcommerciallydeployed(GarciaMaleaetal.,1999).
Harvestingmethods
Haematococcus,specificallystressedrestingcellssettlespontaneouslyandcaneasilybeharvestedinlarge
funnelsorsedimentationponds.Howevercentrifugationyieldshigherrecoveryofbiomassfaster.
Biomassprocessing
The biomass is dried,dried; cells are broken by bead mills or other suitable technologies. Astaxanthin
containingoilisbeingextracted,e.g.bysupercriticalCO2extraction,andsold.
Contaminations, competitors, cold or heat stress can significantly reduce productivity outdoors, and in
bioreactorsheatingorcoolingarerequiredformaintainingsatisfactoryproductivities.Beingafreshwater
microalgae,Haematococcuscultivationisfrequentlyhamperedbyother,fastgrowingmicroalgaesuchas
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Scenedesmus or Chlorella, zooplankton or even funghi which can drastically reduce the culture
performance.
Astaxanthin production at the tubular bioreactor facility at Algatech Qetura (http://www.algatech.com/,
Boussiba et al., 1997) may be considered the most advanced microalgal production process having
reached full commercialization. This production facility has been running successfully for eight years
uninterrupted under continuous incorporation of innovations for increasing productivity and product
quality.
References
Boussiba S., Bing W., Yuan J.P., Zarka A., Chen F. (1999) Changes in pigments profile in the green alga Haematococcus pluvialis
exposedtoenvironmentalstresses.BiotechnologyLetters21:601604.
BoussibaS.,FanL.,VonshakA.(1992)EnhancementanddeterminationofastaxanthinaccumulationingreenalgaHaematococcus
pluvialis.MethodsinEnzymology.NewYork:AcademicPress,pp.386371.
Boussiba S., Vonshak A., Cohen Z., Richmond A. BenGurion University of the Negev, Israel (1997). A procedure for largescale
productionofastaxanthinfromHaematococcusWO97/28274.
DroopM.(1956a)Haematococcuspluvialisanditsallies.I.TheSphaerellaceae.RevueAlgologique2:5370.
DroopM.(1956b)Haematococcuspluvialisanditsallies.II.NomenclatureinHaematococcus.RevueAlgologique3:182192.
ElliottA.(1934)MorphologyandlifehistoryofHaematococcuspluvialis.ArchivfrProtistenkunde82L.:250272.
GarciaMalea M.C., Acien F.G., Del Rio E., Fernandez J.M., Ceron M.C., Guerrero M.G., MolinaGrima E. (2009) Production of
astaxanthinbyHaematococcuspluvialis:Takingtheonestepsystemoutdoors.BiotechnologyandBioengineering102:651
657.
HepperleD.,NozakiH.,HohenbergerS.,HussV.A.,MoritaE.,KrienitzL..(1998)PhylogeneticpositionofthePhacotaceaewithinthe
Chlamydophyceaeasrevealedbyanalysisof18SrDNAandrbcLsequences.JournalofMolecularEvolution47:42030.
HuntleyM.E.,RedaljeD.J.(2007)CO2 mitigationandrenewableoilfromphotsyntheticmicrobes:Anewappraisal.Mitigationand
AdaptationStrategiesforGlobalChange12:573608.
Kobayashi M., Kurimura Y., Kakizono .T, Nishio N., Tsuji Y. (1997) Morphological changes in the life cycle of the green alga
Haematococcuspluvialis.JournalofFermentationandBioengineering84:9497.
Rippka R., Deruelles J., Waterbury J.B., Herdman M., Stanier R.Y. (1979). Generic assignment, strains histories and properties of
pureculturesofcyanobacteria.JournalofGeneralMicrobiology111:161.
TrikiA.,MaillardP.,GudinC.(1997)GametogenesisinHaematococcuspluvialisFlotow(Volvocales,Chlorophyta).Phycologia36:
190194.
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8.1.6
Dunaliellasp.
Figure21Dunaliellasp.
D.J.Patterson
http://starcentral.mbl.edu/msr/rawdata/viewable/dunaliel
la_1097788928_esmlaw.jpg;Left:
Figure22DunaliellatertiolectaRoscoffCulture
CollectionStrain#6
CNRSStationBiologiquedeRoscoffwww.sb
roscoff.fr/Phyto/gallery/main.php?g2_itemId=371
SYMBOLS:B,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Volvocales
Dunaliellaceae
Dunaliella
RelatedSpecies
D. acidophila, D. assymetica, D. baasbeckingii, D. bardawil, D. bioculata, D. carpatica, D. cordata, D.
euchlora, D. gracilis, D. granulata, D. lateralis, D. maritima, D. media, D. minuta, D. parva, D. peircei, D.
polymorpha, D. primolecta, D. pseudosalina, D. quartolecta, D. ruineniana, D. salina, D. terricola, D.
tertiolecta,D.turcomanica,D.viridisvar.palmelloides,D.viridis,D.viridisf.euchlora.
BIOLOGY
Dunaliellaisagenusofalgae,specificallybelongingtotheDunaliellaceaefamily.Dunaliellasp.aremotile,
unicellular,rodtoovoidshaped(911m)greenalgae,Chlorophyta,whicharecommoninmarinewaters.
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The genus Dunaliella has marine and halophilic representatives. Freshwater species have also been
described.DunaliellaalsohasaverywidepHtolerancerangingfrompH1(D.acidophila)(Gimmleretal.,
1989) to pH 11 (D. salina). In fact, D. salina is one of the most environmentally tolerant eukaryotic
organismsknownandcancopewithasalinityrangefromseawater(=3%NaCl)toNaClsaturation(=31%
NaCl),andatemperaturerangefrom<0Cto>38C(Ginzburg,1987;Borowitzka,1988).
DunaliellaismorphologicallysimilartoChlamydomonas,withthemaindifferencebeingtheabsenceof
acellwallinDunaliella.Ithastwoflagellaofequallengthandasingle,cupshapedchloroplast,whichinthe
marineandhalophilicspecieshasacentralpyrenoid(Borowitzka,1988).D.salinaisahalophilemicroalga
andwasoriginallyidentifiedinseasaltfields.Itisdifferentfromallothergreenalgaecellsbecauseitlacksa
cellwallandiswrappedwithanextremelythinelasticmembrane.Thelackofcellwallallowsthecellto
changeitsvolumewithchangingosmoticpressure.IntheDeadSeaitwasfirstdiscoveredin1941.Another
strainofinteresttobiotechnologyisDunaliellatertiolecta.D.tertiolectaisamarinemicroalgawithacell
sizeof1012m.D.tertiolectaisafastgrowingstraininseawater,reportedtohaveoilyieldofabout
37%,thoughcomparativegrowthratesandlipidproductivitieshavenotbeenreported.
Biochemicalcomposition
In order to survive under such extreme conditions, Dunaliella synthesizes different compounds in a very
highconcentration.Itsresistanceagainsthighsaltconcentrations(35%)islinkedtoitsabilitytosynthesize
andaccumulateglycerolupto10%ofitsdryweight.Duetothisitcanmaintainshapeandfunctionsunder
high osmotic pressures. In order to grow in an environment of high temperature and intense sunlight,
Dunaliella synthesizes betacarotene up to 6 of its dry weight. Studies show that the function of beta
caroteneistoprotectthealgaecellfromdamagescausedbyintenseradiation.
Proteinsaccumulateupto60%ofthedrycellweightandcontainmostaminoacids.Proteinsandamino
acidsareimportantingredientsincosmeticpreparations.Theyareusedinordertocreateacontaminating
layerontheskinsurfacewhilenourishingtheskincells.
The carbohydrates include monosugars (glucose, glactose, mannose, xylose, ribose, rhamnose), di
sugars and 1,4 polysaccharides glucosen and starch. The sugars, especially the polysugars are used as
stabilizers.Theythickenandgivetheproductasmoothandgentletexture.Theyabsorblargequantitiesof
waterandgranttheproductwithmoisturizingtraits.Attachedtothenegativeelectricalchargeofthepoly
sugarsareelectrolytes,releasedinacontrolledwaytotheskincells.Thepresenceofpolysaccharidesinthe
cosmetic product allows this controlled release of the active substances in the product and offers an
efficienttreatmentofskindiseaseswithoutusingsubstanceswhichrisktheuserinsideeffects.
Lipidsaccumulateto618%ofthedrycellweightdependingongrowthconditions.Fattyacidsinclude
palmiticacid,3transacidhexadecanoic,linoleicacidandarachidicacid.Betacarotenecanaccumulateup
to 6% of the cell's dry weight. Glycerol assembles up to 10% of the dry cell weight. Glycerol includes
monogalactoglycerol,digalactoglycerolanddiacylglycerol.
Inadditiontohighlevelofbetacarotene,Dunaliellacontainsthiamine,pyridoxine,riboflavin,nicotinic
acid,biotinandtocopherol(vitaminE).carotene,producedfromDunaliellaiscomposedoftwoisomers:
all trans and cis9, in contrast to betacarotene produced from carrots or synthetic betacarotene.
carotene seems to act as photoprotective sunscreen to protect the chlorophyll and the cell DNA from
thehighirradiancewhichcharacterizesthenormalhabitatofD.salina.Ithasalsobeenproposedthat
carotene also acts as a carbon sink to store the excess carbon produced during photosynthesis under
conditionswheregrowthislimitedbutphotosyntheticcarbonfixationmustcontinue.
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BIOTECHNOLOGY
SinceestablishmentofafirstpilotplantforDunaliellacultivationforcaroteneproductionintheUSSRin
1966, the commercial cultivation of Dunaliella for the production of carotene throughout the world is
nowoneofthesuccessstoriesofhalophilebiotechnology(BenAmotzandAvron,1982).
Although technically the production of glycerol from Dunaliella was shown to be possible, economic
feasibilityislowandnobiotechnologicaloperationpresentlyexists.carotenoidproductionbyDunaliella
is also not competitive with other resources, so that Dunaliella production to date is limited to the
nutraceuticalsmarket.Fortheaccumulationofhighconcentrationsofcaroteneandglycerol,thealgaeis
cultivated at extreme salinity with reported very low productivity of about 2 g m2 day1, which is not
competitiveforbiofuelsproduction.
Productionsystem,harvestingandprocessing
Thelargertheindividualpondsusedtogrowthealgae,thesmallertheproductivityofthesystemseemsto
be. In D. salina the optimum salinity for growth lies between 18 and 22% NaCl whereas the optimum
salinity for carotenoid production is >27% NaCl (Borowitzka et al., 1984). Thus the optimum yield of
caroteneperunitvolumeandtimeisachievedatabout24%NaCl.
Twostrategiescanbeapplied tomaximize theproductionofcarotene.Oneofthesestrategiesis a
twostagegrowthprocessinwhichthealgaearefirstgrownatalowsalinity(=15%NaCl)innutrientrich
mediumtomaximizebiomassproduction,andthentransferredtoahighsalinity,lownutrientmediumin
order to induce carotene production (Borowitzka et al., 1984). A similar, twostage production process
wasproposedbyChenandChi(1981)forglycerolproductionfromDunaliella.
NBT Eilat cultivates Dunaliella in open raceway ponds in hyper saline water obtained after passages
through evaporation ponds and achieves productivities of about 2 g m2 day1. A complete description of
differentproductionprocessesandsites,includingharvesting,processingandeconomicsoftheprocesscan
bedownloadedfrom:
http://www.windseaalgae.org/wsapresentations/day1/Ami%20BenAmotz%20WSA%20April%202009.pdf
Culturemedia
The most commonly used medium for culture of Dunaliella is Modified Johnsons Medium (Borowitzka,
1988). However, these algae can also be grown in a wide range of other media including Guillard's f/2
medium (Guillard and Ryther, 1962), modified ASP medium (McLachlan and Yentsch, 1959) and enriched
seawater(RaoandChauhan,1984).
AtlowsalinitiesprotozoasuchastheciliateFabreasalinaandtheamoebaHeteroamoebasp.caninvade
thecultureandveryrapidlydecimatethealgalproduction.
LowersalinitiesgenerallyfavourthegrowthofthenoncarotenogenicDunaliellaspecies(D.viridis,D.
minutaandD.parva)thatcanovergrowD.salinaanddrasticallyreducethecaroteneproductivityofthe
pond(BurfordandBorowitzka,1987).
A twostage process requires greater capital and running costs and thus may make the process
uneconomic.
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Dunaliella was the first microalga cultivated in large scale for production of a high value product (
carotene)ratherthanbulkbiomass.Itscultivationwasfavouredbythehypersalinemediumwhichreduces
thenumberofpotentialcontaminantsduringlongtermopenpondcultivation.
References
BenAmotzA.,AvronM.(1982)ThepotentialuseofDunaliellafortheproductionofglycerol,caroteneandhighproteinfeed.In:
SanPietroA.(ed.)Biosalineresearch:Alooktothefuture.PlenumPub.Corp.,NewYork,pp.207214.
Borowitzka M.A. (1988) Algal growth media and sources of cultures. In: Borowitzka M.A,. Borowitzka L.J. (eds.) Microalgal
Biotechnology.CambridgeUniversityPress,Cambridge,pp.456465.
Borowitzka L.J., Borowitzka M.A.,MoultonT.P. (1984)ThemasscultureofDunaliellaforfinechemicals:fromlaboratorytopilot
plant.Hydrobiologia116/117:115121.
BurfordM.A.,BorowitzkaL.J.(1987)CompetitionbetweenDunaliellaspeciesathighsalinity.Hydrobiologia151/152:107116.
ChenB.J.,ChiC.H.(1981)Processdevelopmentandevaluationforalgalglycerolproduction.BiotechnologyandBioengineering23:
12671287.
Cifuentes A. (1992) Growth and carotenogenesis in eight strains of Dunaliella salina Teodoresco from Chile. Journal of Applied
Phycology4:111118.
GimmlerH,WeisU.,WeissC.,KugelH.,TreffnyB.(1989)Dunaliellaacidophila(Kalina)Masyukanalgawithapositivemembrane
potential.NewPhytologist113:175184.
GinzburgM.(1987)Dunaliella:agreenalgaadaptedtosalt.AdvancesinBotanicalResearch14:93183.
Guillard R.R.L., Ryther J.H. (1962). Studies of marine planktonic diatoms. I. Cyclotella nana (Hustedt) and Detonula confervacea
Jimenez C., Pick U. (1993) Differential reactivity of carotene isomers from Dunaliella bardawil toward oxygen radicals. Plant
Physiology101:385390.
McLachlanJ.,YentschC.S.(1959)ObservationsonthegrowthofDunaliellaeuchlorainculture.TheBiologicalBulletin116:461471.
Rabbani S., Beyer P., Von Lintig J., Hugueney P., Kleinig H. (1998) Induced betacarotene synthesis driven by triacylglycerol
depositionintheunicellularalgaDunaliellabardawil.PlantPhysiology116:12391248.
RaoP.S.N.,ChauhanV.D.(1984)OnoccurrenceandgrowthofDunaliellafromIndia.I.Enrichedseawaterformasscultureofthe
alga.Phykos23:3337.
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8.1.7
Chlorococcumsp.
Figure23Chlorococcumsp.
UTEX819,D.Reinecke,BGU
SYMBOLS:H,D,E
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Chlorococcales
Chlorococcaceae
Chlorococcum
RelatedSpecies
C. acidum, C. aegyptiacum, C. botryoides, C. choloepodis, C. citriforme, C. costatozygotum, C.
diplobionticum, C. dissectum, C. echinozygotum, C. elbense, C. elkhartiense, C. ellipsoideum, C.
hypnosporum,C.infusionum,C.isabeliense,C.lobatum,C.macrostigmatum,C.minimum,C.minutum,C.
novaeangliae, C. oleofaciens, C. olivaceum, C. pamirum, C. pinguideum, C. polymorphum, C.
pseudodictyosphaerium, C. pyrenoidosum, C. refringens, C. salinum, C. schizochlamys, C. schwarzii, C.
submarinum,C.tatrense,C.vacuolatum.
BIOLOGY
Vegetative cells solitary or in temporary groups of indefinite form, never embedded in gelatin. Cells
ellipsoidaltosphericalandvaryinsize.Cellwallssmooth.Thechloroplastiscupshaped,parietal,withor
withoutaperipheralopeningandhasoneormorepyrenoids.Cellsuniucleate,ormultinucleatejustprior
tozoosporogenesis.Reproductionbyzoospores,aplanospores,orisogametes.Motilecellshavetwoequal
flagellaandremainellipsoidalforatimeaftermotilityceases.
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This freeliving genus is cosmopolitan. Though primarily an edaphic alga, it has been reported from
suchdiversehabitatsashotspringsinCentralAsiaandsoilscollectedinAntarctica.Aquatic,marine,and
aerialisolateshavebeenrecorded.
BIOTECHNOLOGY
SeveralstrainsofChlorococcumhavebeentestedaspotentialsourcesofastaxanthin(ZhangandLee,1997;
LiuandLee,2000;Masojdeketal.,2000;MaandChen,2001).Outdoorculturestoverifyastaxanthinyield
have been performed. An enhanced astaxanthin producing mutant was cultivated in a tubularloop
photobioreactorconsistingoftwoinclinedpanelswith34Pyrexglasstubeseach(eachtubeapproximately
1mlongand1.2cmindiameter),reachingbiomassproductivitiesupto0.3gL1day1andketocarotenoid
productivities up to 3.4 mg L1 day1 (Zhang and Lee, 1999). In a horizontal 50L tubular photobioreactor
Masojdeketal.(2000)obatinedgrowthratesabout0.13h1,fourtimesthatofHaematococcus,butwith
20timeslowersecondarycarotenoidcontent.In10LbubbledtubesTrediciandcoworkers(unpublished
data) obtained productivities of 0.20 g L1 day1 outdoors and 0.27 g L1 day1 indoors under contnuos
illumination,withtemperaturereachingatmiddayvaluesupto45and36C,respectively.
Inaflatplatephotobioreactorunder artificiallightof2000molphotonsm2s1,aultrahighdensity
culture of the marine Chlorocococcum littorale reached a productivity of 380 20 mg l1 h1, with 1cm
lightpathlength,avalue2.4and6.4timeshigherthanthoseobtainedinthe2and4cmreactors.Culture
denisitesashighas84gL1werereachedanddailyCO2fixationratewas16.7gL1(Huetal.,1998).
Chlorococcum has been investigated as hydrogen producer (Schnackenberg et al., 1995; Ueno et al.,
1999), but hydrogen yields are much lower than those obtained with Chlamydomonas and Scenedesmus
spp. (Winkler et al., 2002). Chlorococcum was also proposed for bioethanol production via dark
fermentation of starch (Ueno et al., 1998; Harun and Danquah, 2011; Harun et al., 2011) and was
investigatedasasourceoflipidforbiodieselproduction(Rodolfietal.,2009;Halimetal.,2011).
References
Algaebase:http://www.algaebase.org/search/genus/detail/?genus_id=37477[BIOLOGYsection]
Halim R., Gladman B., Danquah M.K., Webley P.A. (2011) Oil extraction from microalgae for biodiesel production. Bioresource
Technology102:178185.
Harun R., Danquah M.K. (2011) Influence of acid pretreatment on microalgal biomass for bioethanol production. Process
Biochemistry46:304309.
HarunR.,JasonW.S.Y.,CherringtonT.,DanquahM.K.(2011)Exploringalkalinepretreatmentofmicroalgalbiomassforbioethanol
production.AppliedEnergy:doi:10.1016/j.apenergy.2010.10.048.
Hu Q., Kurano N., Kawachi M., Iwasaki I., Miyachi S. (1998) Ultrahighcelldensity culture of a marine green alga Chlorococcum
littoraleinaflatplatephotobioreactor.AppliedMicrobiologyandBiotechnology49:655662.
LiuB.H.,LeeY.K.(2000)SecondarycarotenoidsformationbythegreenalgaChlorococcumsp.JournalofAppliedPhycology12:301
307.
MaR.Y.N.,ChenF.(2001)Enhancedproductionoffreetransastaxanthinbyoxidativestressintheculturesofthegreenmicroalga
Chlorococcumsp.ProcessBiochemistry36:11751179.
Masojdek J., Torzillo G., Kopeck J., Koblek M., Nidiaci L., Komenda J., Lukavsk A., Sacchi A. (2000) Changes in chlorophyll
fluorescencequenchingandpigmentcompositioninthegreenalgaChlorococcumsp.grownundernitrogendeficiencyand
salinitystress.JournalofAppliedPhycology12:417426.
Rodolfi L., Chini Zittelli G., Bassi N., Padovani G., Biondi N., Bonini G., Tredici M.R. (2009) Microalgae for oil: strain selection,
inductionoflipidsynthesis,andoutdoormasscultivationinalowcostphotobioreactor.BiotechnologyandBioengineering
102:100112.
SchnackenbergJ.,IkemotoH.,MiyachiS.(1995)RelationshipbetweenoxygenevolutionandhydrogenevolutioninaChlorococcum
strainwithhighCO2tolerance.JournalofPhotochemistryandPhotobiologyB:Biology28:171174.
UenoY.,KuranoN.,MiyachiS.(1998)Ethanolproductionbydarkfermentationinthemarinegreenalga,Chlorococcumlittorale.
JournalofFermentationandBioengineering86:3843.
UenoY.,KuranoN.,MiyachiS.(1999)Purificationandcharacterizationofhydrogenasefromthemarinegreenalga,Chlorococcum
littorale.FEBSLetters443:144148.
Winkler M., Hemschemeier A., Gotor C., Melis A., Happe T. (2002) [Fe]hydrogenases in green algae: photofermentation and
hydrogenevolutionundersulfurdeprivation.InternationalJournalofHydrogenEnergy27:14311439.
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ZhangD.H.,LeeY.K.,NgM.L.,PhangS.M.(1997)Enhancedaccumulationofsecondarycarotenoidsinamutantofthegreenalga,
Chlorococcumsp.JournalofAppliedPhycology9:147155.
Zhang D.H., Lee Y.K. (1999) Ketocarotenoid production by a mutant of Chlorococcum sp. in an outdoor tubular photobioreactor.
BiotechnologyLetters21:710.
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8.1.8
Neochlorisoleoabundans
Figure24Neochlorisoleoabundans
Picturefromhttp://www.sbs.utexas.edu/utex/algaeDetail.aspx?algaeID=3623
SYMBOLS:D,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Chlorococcales
Chlorococcaceae
Neochloris
RelatedSpecies
BIOLOGY
Neochloris oleoabundans was isolated from the top of a sand dune (2O N; 55O E) in Rub al Khali in Saudi
ArabiaandnamedbyBoldandChantanachat(ChantanachatandBold,1962).Vegetativecellsaredescribed
as being between 6 and 25 m, with a cup shaped parietal chloroplast. Vegetative cells with smaller
diameter(3.5m)howeveraremostcommonwiththestrainUTEX1185,whichoriginatesfromtheoriginal
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BoldandChantnachatisolate.Aftercelldivision,onepyrenoidispresentinthechloroplastandlaterinthe
cellcycleitdividesintotwo.Thecellsareuninucleate.
Sexual reproduction has not been observed (Chantanachat and Bold, 1962); N.oleoabundans
reproducesasexuallythroughformationofzoosporesoraplanospores.Zoosporesarebiflagellate(identical
length of flagella) and between 2 and 3.5 m in width and 3.6 and 4.5 m in length. Zoospores form
vegetative sphaerical cells after a short period of time and are not often observed in laboratory cultures
whiletheyappearwithvaryingfrequencyinoutdoormasscultures.Vegetativecellsmaybeupto22min
diameterandmaycontainvisibleoildropletsinthecytoplasm.
Aplanosporesareformedbyincompleteseparationofdaughtercells:thewallprotoplasts(walllessand
nonmotile cells) are retained within the cell wall of the original mother cell (termed sporangium).
Aplanospores are about the size of zoospores andmay accumulate to considerable numbers before they
areliberatedbyruptureofthecellwallofthesporangium.
BiochemicalcompositioninNeochlorisvarieswithgrowthstagewhich,inamoremodernconceptmaybe
consideredaneffectofgrowthlimitationbyeitherlightornutrientfactors.InTable6itmaybenotedthat
protein content significantly decreases in the stationary phase while lipid and carbohydrate content
increase from exponential to stationary phase. InTable 7, it may be noted that neither total fatty acid,
unsaturated fatty acid or sterol percentage of lipids are affected by growth stage to any appreciable
extent.
Table6Exampleofbiochemicalcompositionthroughgrowthstagesgiveninpercentdryweight.Sourceofgrowthlimitation
2
1
notindicated.Algaewerecultivatedin100mltesttubesatlowirradiation(60mm sec ).AfterGatenbyetal.(2003).
Constituent(%DW) Exponential
Protein
54
Carbohydrate
8
Lipid
Lateexponential
63
10
Stationary
44
18
Latestationary
18
40
22
35
36
19
Table7Exampleoflipidcompositionthroughgrowthstagesgiveninpercentlipid(or,forsterols,inlipid).Conditionsasin
table6.AfterGatenbyetal.(2003).
Constituent(%lipid) Exponential
Fattyacids
32
UnsaturatedFA
89
Sterols(lipid)
Lateexponential
45
85
Stationary
31
88
Latestationary
54
80
9.6
5.1
4.5
5.2
Published studies of the effect of stress conditions are all applying nitrogen limitation as stress factor. A
singlestudy(Pruvostetal.,2011)investigatestheeffectofstressongrosscompositionofNeochloris,other
studiespublishedsofar,focusonlyonlipidproportions.ThefindingsaresummarizedinTable8.
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Table8ChangesingrosscompositionofNeochloristhroughnutrientstarvation.DatafromartificiallyilluminatedPBRs.
System
Unit/fraction
Flatplate
PBR,1and
130L
Notindic.
Light
mol 2
photonsm
s1oras
otherwise
indicated
300500
3klux
1Lbubble
columns
Notindic.
90260mm
bubble
columns
160200
Temp
o
Changeincompositionfrom
nutrientrepletetodepletestage.
Lipid
Protein
Carbo
hydrate
30
20%
3035%
2831
3555%
34
34%
30
12.5%
24.9%
70%
20%
20%
40%
Starvation
method
Reference
4days
nitrate
starvation
To visible
lipiddroplet
accumulation
67days
nitrate
starvation
7dayson
nitrogenfree
medium
(Pruvostet
al.,2011)
(Tornabene
etal.,1983)
(Lietal.,
2008)
(Kawataet
al.,1998)
(Neochloris
sp.)
Photosyntheticefficiencyandproductivity
From data in artificially illuminated (continuous) photobioreactors, Neochloris appears to exhibit slow to
moderategrowthrates(Table9).However,muchhighertransientgrowthrateshavebeenexperiencedin
turbidostatsat30C(unpublisheddata,N.H.Norsker)andmaybeduetomixotrophicgrowthoninternal
lipidpools.
No studies have yet reported figures on photosynthetic efficiency in outdoor cultivation. It has only
been possible to identify one study of cultivation of Neochloris in outdoor systems. The growth rate and
lipidproductivityarereportedinTable10.
Table9GrowthrateandlipidproductivityobtainedwithNeochlorisunderdifferentconditions.Allrefertoartificially
illuminatedsystems.
150
30
replete
1.0
0.495
37.66
1Lflat
plate
reactor
270
25
deplete
replete
0.2
0.41
14.42
126
deplete
0.5
65
50Lplastic
sleeves
1Lbubble
columns
10cm
tubular
horizontal
PBR
90260 mm
bubble
columns
150
25
0.92
0.07
not
indicated
200
30
deplete
2.4
133
23
25
deplete
0.3
16.5
replete
0.9
8.9
replete
0.72
deplete
00.43
160200
30
Reference
Lipid
Product.
(mgL1day1)
Growthrate
(day1)
Biomassrange
(gDWL1)
Nstatus
(repl/depl)
Temp
(C)
Irradiation
(molphotons
m2s1)
System
description
1Lbubble
column.
(Gouveia
etal.,
2009)
(Pruvostet
al.,2009)
(daSilvaet
al.,2009)
(Lietal.,
2008)
(Levineet
al.,2011)
(Kawataet
al.,1998)
(Neochloris
sp.)
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Table10GrowthrateandlipidproductivityofNeochlorisinoutdoorcultivation.
System
description
Irradiation
Biomass Growth
Nstatus
(mol
range 1 rate1
(repl/depl)
photons
m2
(gDWL ) (day )
1
s )
Outdoor
natural
0.02
replete
0.18
raceway pond
daylight
2.78
(batch)
Lipid
productivity
(mgL1day1)
1.64.8
Reference
(daSilva
etal.,
2009)
In all the studies published sofar on Neochloris oleoabundans, the strain UTEX 1185 (the original
isolate)hasbeenapplied.Inonlyonestudyincludedinthepresentmaterial(Kawataetal.1998),thestrain
is not UTEX 1185 but a local strain (Neochloris sp.). There is thus insufficient material to discriminate
betweendifferentlaboratorystrainsintermsofproductivityandgrowthrate.
BIOTECHNOLOGY
CultureMedia
Only photoautotrophic growth of Neochloris oleoabundans has sofar been demonstrated. The culture
mediausedarerepotedinTable11.
Table11MediausedforN.oleoabundansbythecitedreferences.
Referencemedia
Othermedia
BBM
3NBBM
3N3SBBM
Soilextract
B12
Other
Reference
(MD)inorganicbasalmedium
Inorganicbasalmedium
x
x
(modifiedfornutrientstarvation)
(Gatenbyetal.,2003)
(Bandetal.1992)
(Tornabene et al.,
1983)
(Lietal.,2008)
(Archibald and Smith,
1987)
(Pruvostetal.,2009)
(Gouveiaetal.,2009)
(daSilvaetal.,2009)
(Pruvostetal.,2011)
(Levineetal.,2011)
Modified: +boron + vitamins, N

source)
Modified:nonitrogen,+B12
Modified
ModifiedFitzgerald
(Bealetal.,2010)
(Wahal and Viamajala,
2010)
(Kawataetal.,1998)
Bristol
medium
Othermedium
Special
micronutrient
additives
AsingleexperimentwithcultivationofNeochlorisinwastewaterfromanaerobicallydigestedmanureis
reported(Levineetal.,2011):whileNeochlorisinahorizontalchamber,irradiatedat200molphotonsm2
s1onnitrateexhibitedaspecificgrowthrateof0.75day1,asimilarsetupresultedinagrowthrateofabout
0.24day1ondilutedwastewater(growthratesestimatedfrompublishedgraphs).
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Cultivationmethods
OnlyoneexperimentcultivatingNeochlorisinopensystemshasbeenreported.Theracewayareawas2.5
m2,thedepthof15cmanditwasagitatedwithapaddlewheeltocreateaflowvelocityof25cms1.The
culturewasdilutedafter20and36daysofcultivation(daSilvaetal.,2009).Specificgrowthrateduringthe
first20dayswas0.18day1.
Othersystemshavebeenusedforindooruseandamongthese:
bubblecolumnsofvariousdiameterupto26cm(Kawataetal.1998;Lietal.2008;Gouveiaetal.2009)
flatpanelreactors,1130L(Pruvostetal.,2011)
shakenflasks(WahalandViamajala,2010)
plasticbags(daSilvaetal.,2009;Levineetal.,2011)
Productionsystems
At present time (2010), Neochloris has not reached a state of commercial production. Neither has any
informationabouttestingofproductionsystemsinpilotscalebeenpublished.
Harvestingmethods
No studies of harvesting methods for Neochloris have yet been published. Speculating about harvest
methods,itisworthnotingthatthecellsizeofNeochlorisoleoabundansisrathersmall(averagediameter
assumingsphaericalcells3mforvegetativelygrowingcultures)andhighenergyinputforseparationby,
forinstance,centrifugationcouldbeassumed.However,itisgenerallyobservedthatNeochlorissediments
ratherreadilyincultureflasksandmaythereforebesuitableforflocculation.
Flotation or foam separation are other potentially attractive harvesting technologies as Neochloris
culturesarepronetofoaming.
Biomassprocessing
There are no published studies on processing Neochloris biomass for any purpose. For potential use for
biodiesel purposes, it is worth noting that a substantial part of the lipid fraction may be constituted by
triacylglycerols:80%withlipidsconstituting3554%ofthebiomass(Tornabeneetal.,1983).Arecent
studyofoilformationoptimizationdemonstratedaTAGyieldof50%oftotallipidsor18%ofDW(Pruvost
et al., 2009). Relatively high specific TAG productivities by Neochloris is one of the main motives for
focussingonNeochlorisforbiodieselproductionpurposes.
Scalinguplimitation
Production of Neochloris for biodiesel is here considered only for photoautotrophic production methods.
Conversion of carbohydrate sources to biodiesel by heterotrophic production is being considered with
otheralgaespecies,butaheterotrophicgrowthpotentialofNeochlorishasnotbeenestablished.
Due to the lack of experience with outdoor cultivation of Neochloris, it is very difficult to pin point
limitationstoscaleup.
Twostudieshavedealtwithoutdoorproduction.IndaSilvaetal.(2009)productionofNeochlorisina
2.5m2racewaypondresultedinhighbiomassdensities(2.8gDWL1)attheendofa20daysbatchgrowth
runwithspecificgrowthrateof0.18day1.Lipidproductivityatthepeakwas4.8gm2day1(July,Portugal)
whichmustbeconsideredpromisingifsuchratescanbesustainedoverlongerperiods.Itislikelythatthe
lipid productivity can be enhanced by reducing the biomass density in the pond as high average light
intensities are required to obtain high lipid productivity and the other design parameter, culture depth
which determines the average light intensity in the culture, cannot be reduced in raceway ponds for
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practicalreasons.Thiswill,ontheotherhand,addtotheharvestingcostsandmayaddtothevulnerability
oftheculturetocontaminationbyotheralgaeorprotozoansandzooplankton.
In Levine et al. (2011) the use of wastewater from biogas production as fertilizer was studued, but
overgrowthbyotheralgaewasencounteredandclosedreactorsweresuggestedfortheproduction.
For these reasons, the development of photobioreactors with low installation and operating costs is
believedtobethemostpromisingtechnologiesforascaleupstrategy.
References
ArceG.,BoldH.C.(1958)SomeChlorophyceaefromCubansoils.AmericanJournalofBotany45:492503.
Archibald P.A., Smith V.J. (1987) Notes on variation on physiological attributes between aquatic and edaphic species of the
chlorophyceanalgalgenusNeochloris.TransactionsoftheAmericanMicroscopicalSociety106:179182.
Band C.J., Arredondovega B.O., Vazquezduhalt R., Greppin H. (1992) Effect of a saltosmotic upshock on the edaphci microalga
Neochlorisoleoabundans.PlantCellandEnvironment15:129133.
Beal C.M., Webber M.E., Ruoff R.S., Hebner R.E. (2010) Lipid analysis of Neochloris oleaoabundans by liquid state NMR.
BiotechnologyandBioengineering106:573583.
ChantanachatS.,BoldH.C.(1962)PhycologicalStudies.II.Somealgaefromaridsoils.UniversityofTexasPublication(6218).
da Silva T., Reis A., Medeiros R., Oliveira A., Gouveia L. (2009) Oil production towards biofuel from autotrophic microalgae
semicontinuouscultivationsmonitorizedbyflowcytometry.AppliedBiochemistryandBiotechnology159:568578.
GatenbyC.M.,OrcuttD.M.,KreegerD.A.,ParkerB.C.,JonesV.A.,NevesR.J.(2003)Biochemicalcompositionofthreealgalspecies
proposedasfoodforcaptivefreshwatermussels.JournalofAppliedPhycology15:111.
Gouveia L., Marques A., da Silva T., Reis A. (2009) Neochloris oleoabundans UTEX #1185: a suitable renewable lipid source for
biofuelproduction.JournalofIndustrialMicrobiologyandBiotechnology36:821826.
KawataM.,NanbaM.,MatsukawaR.,ChiharaM.,KarubeI.(1998)IsolationandcharacterizationofagreenalgaNeochlorissp.for
CO2fixation.StudiesinSurfaceScienceandCatalysis114:637640.
LevineR.B.,CostanzaRobinsonM.S.,SpataforaG.A.(2011)Neochlorisoleoabundansgrownonanaerobicallydigesteddairymanure
forconcomitantnutrientremovalandbiodieselfeedstockproduction.BiomassandBioenergy35:4049.
LiY.,HorsmanM.,WangB.,WuN.,LanC.(2008)Effectsofnitrogensourcesoncellgrowthandlipidaccumulationofgreenalga
Neochlorisoleoabundans.AppliedMicrobiologyandBiotechnology81:629636.
PruvostJ.,VanVoorenG.,CogneG.,LegrandJ.(2009)InvestigationofbiomassandlipidsproductionwithNeochlorisoleoabundans
inphotobioreactor.BioresourceTechnology100:59885995.
Pruvost J., Van Vooren G., Le Gouic B., CouzinetMossion A., Legrand J. (2011) Systematic investigation of biomass and lipid
productivitybymicroalgaeinphotobioreactorsforbiodieselapplication.BioresourceTechnology102:150158.
Starr R.C. (1955) A comparative study of Chlorococcum meneghini and other spherical, zoosporeproducing genera of the
Chlorococcales.IndianaUniversityPublicationsScience20:1111.
StarrR.C.(1978)CulturecollectionofalgaeattheUniversityofTexasatAustin.JournalofPhycology14:47100.
TornabeneT.G.,HolzerG.,LienS.,BurrisN.(1983)LipidcompositionofthenitrogenstarvedgreenalgaNeochlorisoleoabundans.
EnzymeandMicrobialTechnology5:435440.
Wahal S., Viamajala S. (2010) Maximizing algal growth in batch reactors using sequential change in light intensity. Applied
BiochemistryandBiotechnology161:511522.
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8.1.9
Scenedesmussp.
Figure25Scenedesmussp.LocalisolateSdeBoker,Israel
D.Reinecke,BGU
SYMBOLS:B,D,E,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Sphaeropleales
Scenedesmaceae
Scenedesmus
RelatedSpecies
BIOLOGY
Scenedesmus is a freshwater medium to large size unicellular green alga often appearing in tetrads with
fourto16elongatedcellsconnected(cenobia),althoughcellscanalsoappearasindividualinovalform.
Scenedesmusisversatileinproductionofoilandsecondarymetabolitessuchascarotenoids.Multiple
species produce different carotenoids during logarithmic growth or stress, though cellular levels never
exceed1%ofdryweight.Itisrelevantthatmajorcarotenoidisusuallylutein.Exponentiallygrowingcells
havehighproteincontent(upto50%),lessthan10%lipids,therestvariableamountsofstarchandcellwall
components. Under nutrient stress Scenedesmus species can accumulate high amounts of storage lipids
(TAG) in cytoplasmic oil globules. Fatty acid composition under logarithmic growth includes significant
amountsofshortpolyunsaturatedfattyacidssuchasgammalinolenicandstearidonicacid.Understress,
TAGaccumulatesmostlyC16andC18unsaturatedandsomemonounsaturatedfattyacids.
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Growthkineticsandefficiencies
Scenedesmusisamongthemostvigorouslygrowinggreenalgaeandoutcompetesmostotheralgalspecies
under high nutrient conditions, e. g. in wastewater. It can cause serious contamination problems when
cultivating slower growing algae such as Haematococcus. It is among the faster growing and highest oil
producingstrainstestedbyRodolfietal.(2009)orHuetal.(2008).
Growthratesandproductivityinlaboratorymaingroups
Maximum specific growth rates as higher than 0.12 h1 has been reported for Scenedesmus sp. Lower
specificgrowthrates,of0.04h1,hasbeenreportedforScenedesmusobliquus.Themostrelevantaspectof
thisstrainisitstolerancetohightemperature,including40C,itsmaximumgrowthratebeingobtainedat
temperaturesof3035C.Atlaboratoryscalebiomassproductivitiesof0.9gL1day1hasbeenreportedfor
Scenedesmus almeriensis. High productivity requires the use of high irradiance, but Scenedesmus has
demonstratetoberesistanttoirradianceshigherthan1700molphotonsm2s1withoutphotoinhibition.
Scenedesmusisalsotoleranttoimpulsionusingcentrifugalpumps,inadditiontoaeration,nomechanical
damage being reported by this phenomenon. Concerning pH, Scenedesmus tolerates wide ranges of pH,
from 5 to 10, although optimal pH is in the range of 7.58.0. It is particularly relevant the tolerance of
ScenedesmustoalkalinepHforwastewaterandfluefluegasdepuration.
Photosyntheticefficiencyandproductivityoutdoors
DuetothefactthatScenedesmusishighlyrobustandfastgrowing,ithasbeenproducedoutdoorsusing
bothopensystemsandclosedphotobioreactors.Inopenraceways,biomassproductivitieshigherthan0.5
gL1day1 has been reported, while biomass productivities up to 1.2 gL1day1 have been obtained using
closedphotobioreactors.Overallmeanannualproductivityof0.6gL1day1hasbeenobtainedinpilotscale
tubular photobioreactors (30 m3) with Scenedesmus almeriensis. This value is about 3% solar efficiency.
Solarefficienciesfrom13%hashavebeenreportedforScenedesmus.
Under optimal growth conditions the major component of Scenedesmus biomass are proteins, 4050%
d.wt., next being carbohydrates. The lipid content under adequate growth conditions is lower than 15%,
woithmaximumvaluesof10%correspondingtofattyacids.Itisrelevantthatunderoptimalconditionsthe
carotenoid content, especially lutein, increases up to 1% in some species. Scenedesmus cells can be
stressed by nutrient depletion, nitrogen or phosphorous, which triggers accumulation of lipids. However,
nolipidcontenthigherthan30%areobtainedintheseconditions.
BIOTECHNOLOGY
CultureMedia
Scenedesmus grows well in modified BG11 medium, but also in other growth media, in nutrient rich
wastewateretc,andapparentlymayalsoexploitorganicmoleculesforphotoheterotrophicgrowth.Ithas
been cultivated using commercial fertilizers outdoors, and in brackish water. It is tolerant to the use of
nitrate, ammonia or urea as nitrogen source, thus Scenedesmus is adequate for removal of inorganic
nitrogenfromeffluents.
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Cultivationmethods
Scenedesmuscanbecultivatedineitherdiscontinuous(batch)orcontinuousmode.Indiscontinuousmode
the productivity is lower, thus usually semicontinuous operation being utilized. No large variation in the
composition of the biomass is observed when cultivated in different modes. In continuous mode the
optimaldilutionrateisintherangefrom0.30.4day1.
Productionsystem
Scenedesmusarenotdamagedbyaerationorimpulsioncentrifugalpumps,thusithasbeencultivatedin
most of culture systems developed, including open and closed photobioreactors. Fouling is an important
issue when cultivating this strain. Under adverse conditions, but also under optimal conditions, cells of
Scenedesmusfixonthereactorswalls,reducinglightavailabilityinsidetheculture.Whenclosedreactors
are used it is obligatory to implement selfcleaning systems. This is not necessary when using open
raceways.
Harvestingmethods
AlthoughScenedesmuscellsarelargerthanothermicroalgaecells,theirsettlingvelocitybygravityislow,in
the range of 106 ms1. Thus, Scenedesmus cells cannot be harvested by natural sedimentation. It is
necessary the use of centrifugation or filtration operations. Cells of Scenedesmus are easily harvested by
centrifugation:aslurrycontaining15%d.wtisobtainedundercontinuousoperation,andapastecontaining
30%d.wt.isobtainedunderdiscontinuousoperation.Althoughfiltrationcanbeperformed,itisnotusually
carried out. Natural sedimentation can be performed by previous flocculation. The use of anionic
polyelectrolyteatdosesbelow0.1mg/Lhasbeendemonstratedtobeusefultopreconcentratetheculture
bytwentytimes.
Biomassprocessing
BiomassofScenedesmuscanbeusedasbiomassforfeedinganimalsorfishes,mainlyforitshighprotein
content. The fatty acid profile is not highly valuable because no PUFAs are present. The only especially
valuable components present in the biomass of Scenedesmus are carotenoids. Lutein contents up to 1%
d.wt. has have been reported, it being useful as nutraceutical for human and animals. Lutein can be
extracted using organic solvents, although new processes using supercritical fluids or ethanolwater
mixtureshavebeenproposed.
Scalinguplimitation
NolimitationsaredirectlyrelatedwiththecultureofScenedesmus,differentthantheproductionofother
strains. For scaleup purposes the tolerance of this strain to high temperatures, up to 45 C, is highly
relevant.
Scenedesmus is highly robust and has been used successfully used to treat such problematic waste
effluents as olive mill wastewater, biogas or municipal land fill effluents which are toxic to most
microorganismsincludingbacteria.
AbiotechnologyforproductionandpurificationofluteinfromScenedesmushasbeendevelopedbasedon
cultivation in tubular photobioreactors and extraction and purification of the carotenoid from cellular
lysate,thoughthetechnologyisnotcurrentlycommerciallyapplied.
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References
AbeliovichA.,DikbuckS.(1977)FactorsaffectinginfectionofScenedesmusobliquusbyaChytridiumsp.insewageoxidationponds.
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AllardB.,TemplierJ.(2000)Comparisonofneutrallipidprofileofvarioustrilaminaroutercellwall(TLS)containingmicroalgaewith
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Benemann J.R., Koopman B.L., Murry M., Weissman J.C., Eisenberg D.M., Oswald W.J. (1977) Species control in largescale algal
biomassproduction.Finalreport.UnitedStates.
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degradation.OrganicGeochemistry37:871881.
Buchheim M.A., Michalopulos E.A., Buchheim J.A. (2001) Phylogeny of the Chlorophyceae with special reference to the
Sphaeropleales:astudyof18Sand26SrDNAdata.JournalofPhycology37:819835.
CepakV.,PribylP.(2006)Theeffectofcolourlightonproductionofzooidsin10strainsofthegreenchlorococcalalgaScenedesmus
obliquus.CzechPhycology6:127133.
Cepak V., Pribyl P., Vitova M., Zachleder V. (2007) The nucleocytosolic and chloroplast cycle in the green chlorococcal alga
Scenedesmusobliquus(Chlorophyceae,Chlorococcales)grownundervarioustemperatures.Phycologia46:263269.
CzyganF.(1968)SecondarycarotenoidsingreenalgaeII.Studiesonbiogenesis.ArchivfrMikrobiologie62:209236.
deMoraisM.G.,CostaJ.A.V.(2007)BiofixationofcarbondioxidebySpirulinasp.andScenedesmusobliquuscultivatedinathree
stageserialtubularphotobioreactor.JournalofBiotechnology129:439445.
Dean A.P., Sigee D.C., Estrada B., Pittman J.K. (2010) Using FTIR spectroscopy for rapid determination of lipid accumulation in
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Disch A., Schwender J., Muller C., Lichtenthaler H.K., Rohmer M. (1998) Distribution of the mevalonate and glyceraldehyde
phosphate/pyruvate pathways for isoprenoid biosynthesis in unicellular algae and the cyanobacterium Synechocystis PCC
6714.BiochemicalJournal333:381388.
GonzalezLopezCV,CeronGarciaMdC,FernandezFG,SegoviaBustosC,ChistiY,FernandezSevillaJM.2010.Proteinmeasurements
ofmicroalgalandcyanobacterialbiomass.BioresourceTechnology101(19):75877591.
Greger M., Johansson M. (2004) Aggregation effects due to aluminum adsorption to cell walls of the unicellular green alga
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Grewe C., Menge .S, Griehl C. (2007) Enantioselective separation of allEastaxanthin and its determination in microbial sources.
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GurbuzF.,CiftciH.,AkcilA.(2008)BiodegradationofcyanidecontainingeffluentsbyScenedesmusobliquus.JournalofHazardous
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GutmanJ.,ZarkaA.,BoussibaS.(2009)ThehostrangeofParaphysodermasedebokerensis,achytridthatinfectsHaematococcus
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Hanagata N., Dubinsky Z. (1999) Secondary carotenoid accumulation in Scenedesmus komarekii (Chlorophycea, Chlorophyta).
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HeideH.,KaliszH.M.,FollmannH.(2004)Theoxygenevolvingenhancerprotein1(OEE)ofphotosystemIIingreenalgaeexhibits
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Ho S.H., Chen W.M., Chang J.S. (2010) Scenedesmus obliquus CNWN as a potential candidate for CO2 mitigation and biodiesel
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Kim M.K., Park J.W., Park C.S., Kim S.J., Jeune K.H., Chang M.U., Acreman J. (2006) Enhanced production of Scenedesmus spp.
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8.1.10 Desmodesmussp.
Figure26Desmodemussp.
PicturefromUNIFI
SYMBOLS:H,D,E
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Chlorophyceae
Sphaeropleales
Scenedesmaceae
Desmodesmus
RelatedSpecies
D. communis, D. costatogranulatus, D. bicellularis, D. serratus, D. denticulatus, D. lefevrei, D.
arthrodesmiformis, Desmodesmus sp. Hegewald 198751, D. subspicatus, D. hystrix, D. opoliensis, D.
pannonicus, D. perforatus, D. pirkollei, Desmodesmus sp. CL1, D. maximus, D. tropicus, D. komarekii, D.
multivariabilis,D.pleiomorphus,D.fennicus,D.armatus.
BIOLOGY
Desmodesmus used to be the most speciesrich subgenus of Scenedesmus, but it was given genus status
basedon18SandITS2rDNAphylogenies.Thelargegeneticdistancebetweenthetwosubgeneraandtheir
clear distinct cell wall ultrastructure supported retention of the Scenedesmus Meyen for nonspiny
organismsandformationofagenusDesmodesmus(Chodat)An,FriedletHegewaldforthosewhichcould
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bearspines.Desmodesmusappearsasunicellsorcoenobia24816celled,withlongaxesofcellsparallel,
laterally adjoined, and arranged in a single linear or alternating series. Cells are ellipsoidal to ovoid, and
spines usually are present on the terminal cells and/or medial cells, but may be entirely absent. The cell
wallmayhaveridges,warts,ornets.Thechloroplastisparietal,usuallywithonepyrenoid.
Desmodesmus is an extremely common genus, occasionally abundant, found in the phytoplankton of
ponds and lakes. They are cosmopolitans and able to withstand harsh conditions, such as periods with
stronggrazingpressure.
TherelativelylownumberofstudieswithDesmodesmuscanbeexplainedfrominvestigatorsstillbeing
unaware of the division of the old genus Scenedesmus into the new genera Scenedesmus and
Desmodesmus.
BIOTECHNOLOGY
LittlescientificliteratureisavailableonDesmodesmus,althoughitsapplicationaresimilartothatreported
forScenedesmus.PartoftheliteratureavailableaboutScenedesmusactuallyreferstoDesmodesmus.
VirginiaCoastalEnergyResearchConsortiumcultivatedaDesmodemusstrainforbiodieselproduction
studiesina0.4haopenpondduringaboutayearwithlowandvariablebiomassconcentrationsandquite
lowlipidyields(Stubbins,2009).
References
LrlingM.(2003)PhenotypicplasticityinthegreenalgaeDesmodesmusandScenedesmuswithspecialreferencetotheinduction
ofdefensivemorphology.AnnalesdeLimnologieInternationalJournalofLimnology39:85101.[BIOLOGYsection]
Shubert L.E. (2003) Nonmotile coccoid and colonial green algae. In: Wehr J.D., Sheath R.G. (eds.) Freshwater Algae of North
America:EcologyandClassification.ElsevierScience,pp.253309.[BIOLOGYsection]
StubbinsA.(2009)VirginiaCoastalEnergyResearchConsortiumFinalReport:AlgalBiodieselStudies,July2007toSeptember2009.
Vanormelingen P., Hegewald E., Braband A., Kitschke M., Friedl T., Sabbe K., Vyverman W. (2007) The systematics of a small
spineless Desmodesmus species, D. costatogranulatus (Sphaeropleales, Chlorophyceae), based on its rDNA sequence
analysesandcellwallmorphology.JournalofPhycology43:378396..[BIOLOGYsection]
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8.1.11 Chlorellasp.
Figure27Chlorellaemersonii,leftpicturecontrolculture,rightoictureNstarvedculture
D.Reinecke,BGU
SYMBOLS:B,D,E,H,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Trebouxiophyceae
Chlorellales
Chlorellaceae
Chlorella
RelatedSpecies
Numbersofnamesandspecies:Thereare71species(andinfraspecific)namesinthedatabaseatpresent,
ofwhich32havebeenflaggedascurrentlyacceptedtaxonomically.
C. acuminata, C. angustoellipsoidea, C. anitrata, C. anitrata var. minor, C. antartica, C. aureoviridis, C.
autotrophica, C. botryoides, C. candida, C. capsulata, C. communis, C. conductrix, C. conglomerata, C.
desiccata,C.ellipsoideavar.minor,C.ellipsoidea,C.emersoniivar.rubescens,C.emersoniivar.globosa,C.
emersonii, C. faginea, C. fusca var. rubescens, C. fusca var. vacuolata, C. fusca, C. glucotropha, C.
homosphaera,C.kessleri,C.kolkwitzii,C.lobophora,C.luteoviridisvar.lutescens,C.luteoviridis,C.marina,
C. miniata, C. minor, C. minutissima, C. mirabilis, C. mucosa, C. mutabilis, C. nocturna, C. oocystoides, C.
ovalis, C. parasitica, C. parva, C. peruviana, C. protothecoides, C. protothecoides var. mannophila, C.
pyrenoidosavar.tumidus,C.pyrenoidosa,C.pyrenoidosavar.duplex,C.regularisvar.minima,C.reisiglii,C.
reniformis, C. rugosa, C. saccharophila, C. saccharophila var. ellipsoidea, C. salina, C. sorokiniana, C.
spaerckii, C. sphaerica, C. stigmatophora, C. subsphaerica, C. terricola, C. trebouxioides, C. variabilis, C.
vulgarisf.minuscula,C.vulgarisf.suboblonga,C.vulgarisf.globosa,C.vulgarisvar.viridis,C.vulgarisvar.
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autotrophica,C.vulgarisvar.tertia,C.vulgaris,C.zofingiensis.
BIOLOGY
Chlorella is a large and diverse genus of small unicellular green algae of highest relevance to multiple
aspectsofbiotechnology.
Cells are round or ellipsoidal in shape. Smooth rigid cellulosic cell wall contains contains glucosamine
(chitosan).Nucleusissingleandeccentric,chloroplastsingleandparietal,pyrenoidsingleandcoveredwith
starch envelope. Pyrenoid stroma penetrated with 2 or 3 closely appressed thylakoids. Only asexual
reproductionbyautosporesisknown,autospores(216permothercell)arereleasedbyruptureofparental
cellwall.Chlorella,essentiallycosmopolitan,occursinbothfreshwaterandmarinehabitats.Traditionally,
thegenusChlorellahasbeencomprisedover100species;nevertheless,10wellestablishedspecieswere
recognizedbychemotaxonomicmethods(KesslerandHuus,1992).Basedonmodernpolyphasicapproach,
only four: C. vulgaris Beijerinck, C. lobophora Andreyeva, C. sorokiniana Shihira & Krauss, and C. kessleri
Fott&Novkov(Huusetal.,1999)oreventhree:C.vulgaris,C.lobophoraandC.sorokiniana(Krienitzet
al.,2004)trueChlorellaspeciesarerecognizedrecently.
Strainsofspecielinterest
Chlorella are fast growing freshwater, in some cases marine, water algae, reported to accumulate high
concentrationofoilunderstress(Demirbas,2009).FurthermoreChlorellastrainscanexpressavarietyof
carotenoids, among them astaxanthin. Some species is characterized by a very high growth rate (max =
0.20/h) and tolerance to a high culture temperature (40oC). Its desirable technological properties
(resistancetoshearstress,lowadhesiontoasurfaceofthebioreactor,lowtendencytoformaggregates)
areexpectedtooffersignificantadvantagesforitsuseinlargescaleproductionbioreactors(Douchaand
Lvansk,2009).ItsabilitytogrowsunderhighCO2 permitswithadirectsupplyoffluegascontainingupto
40%(v/v)ofCO2(DouchaandLvansk,2005;Douskovaetal.,2009,2010).
Chlorellavulgarisisafastgrowingspeciesandsomestrainscanaccumulateveryhighconcentrationsof
lipids under stress (Francisco et al., 2010; Hsieh and Wu, 2009; Ly et al., 2010; Liang et al., 2009) while
anotherothersonesaccumulatehighamountofstarch(Doukovetal.,2010).Oilproductionwastested
also in other Chlorella species and strains as termophilic alga Chlorella sorokiana, or heterotrophically
grown Chlorella protothecoides (Xu et al., 2006). Chlorella zofingiensis, can accumulate astaxanthin and
lutein(DelCampoetal.,2004;Liuetal.,2010a,b;Ypetal.,2004).
Biochemicalcompositionandbiomassmainconstituents
The biochemical composition and the essential aminoacid content of Chlorella biomass are repoted in
Tables12and13.
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Table12BasicchemicalbiomasscompositionofaproductionstrainChlorellasp,strainP12.
Item
%algalDW
Moisture
77
Proteins(Nx6.25)
5558
*Lipids
812
Saccharides
1015
Fibre
68
Mineralsubstances
68
Chlorophyll
2.53.5
Nucleicacids
34
*Theproportionofessentialunsaturatedfattyacids(oleic,linoleic,linolenic)inthe
totalfattyacidsunderoptimumgrowthconditionsisintherangeof4060%.
Table13Thepercentproportionofessentialamino
acidsinChlorellaandotherproteinrichsources.
Aminoacid
Chlorella
(dryweight)
Isoleucine
Leucine
Lysine
Methionine
Phenylalanine
Threonine
Tryptophan
Valine
2.01
4.14
3.19
1.04
2.57
2.42
0.80
3.00
Inadditionto23%ofchlorophyll,Chlorellacontainsalsocarotenoids,orangeandyellowdyespigments.
Themostvaluableoftheseisbetacarotene,provitaminA.TheamountofcaroteneinChlorellaisinthe
rangeof0.10to0.25%dryweight
An important component of the Chlorella cells is biologically complexed, and therefore readily
utilisedutilisable, basic minerals (phosphorus, potassium, magnesium, calcium and iron) (Table 14) and
traceelements,whichformpartofenzymecomplexesandvitamins.Theseelementsincludeinparticular
manganese,zinc,molybdenum,copperandcobalt.
Trace elements are often chelated with amino acids. Their concentration and type of binding can be
considerablymodified.Thisoffersusthepossibilitytoobtainalgalbiomasswithadefined,mostlyincreased
content of the desirable elements or their mixture in a natural organic form, which enhances their
biologicalefficiency.
AnothergroupofsubstanceswhichispresentinChlorellainmuchhigherlevelsthaninotherplantsis
vitamins(Table15).StrikingisthehighcontentofvitaminsofgroupB,ascorbicacid(vitaminC),nicotinic
acid(vitaminB3)andtocopherols(vitaminE).
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Table14Proportionofmineralsubstancesand
importanttraceelementsintheChlorelladryweight.
Element
mg/100gDW
Phosphorus
1200
Potassium
879
Sulphur
600
Magnesium
300
Calcium
230
Iron
70
Manganese
14
Zinc
11
Copper
4
Cobalt
0.5
Table15VitamincontentofChlorella.
Vitamins(mgkg1)
Chlorella
(dryweight)
B1thiamine
B2riboflavin
B3nicotinicacid(niacin)
B5pantothenicacid
B6pyridoxine
B12cobalamin
biotin(vitaminH)
folicacid
vitaminE(tocopherol)
vitaminC(ascorbicacid)
carotene(provitaminA)
18
44
219
13
28
0.8
0.3
42
298
655
1050
ChlorellaGrowthFactor(CGF)isawaterextractablecellfractioncontainingfreeaminoacids,peptides,
glycoproteins,polyamines,somevitamins,mineralsandother,asyetnotexactlydefinedcomponents.The
effectsoftheextractarepresentedasstriking,thoughscientificdatatoprovethesestatementsarestillat
laboratoryscale.
ItItpromotestissueregeneration,cellgrowthanddivision.Itstimulatestheproductionofleukocytes
and their phagocytic activity, i.e. the ability to eliminate foreign bacteria and also the production of
lymphocytes responsible for the synthesis of antibodies important factors in the immunity against
infections. It is a suitable dietary supplement during the administration of probiotics, i.e. substances
positively affecting the composition of intestinal microflora. It has been shown that, following an
administration of the algal extract, the organism exhibits a better regeneration of damage caused by
ionising radiation. Chlorella extracts have found their use in topical applications, e.g. in the treatment of
chronicinflammations,eczemas,cruralulcers,burnsandotherbadlyhealingwounds,whicharehealedby
afullyfunctionaltissue.Japaneselaboratorieshaverepeatedlypublisheddataontheantitumouractivity
ofthealgalextractinvitro.
The nutrient solution, in which Chlorella has been cultured, displays also a conspicuous stimulatory
effectwhenusedforwateringfreshlyplantedfruitorforesttreesorvegetables.Thishasbeenattributedto
itsstimulatoryeffectsonplantroottakingandgrowth.Thestimulatoryeffectisascribed,apartfromother
components,tocompoundsofthephytohormonegroupwhichhavebeenidentifiedinthealgalextracts.
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Acomplextreatmentofagriculturalwasteincludingthefollowingmajorsteps:anaerobicfermentationof
suitable waste, cogeneration of the obtained biogas and growth of microalgae consuming the CO2 from
biogasorfluegas,wasverifiedunderfieldconditionsinapilotscalephotobioreactor.Laboratoryanalyses
oftheproducedmicroalgaeconfirmedthatitmeetsthestrictEUcriteriaforrelevantcontaminantslevelin
foodstuff(Doukovetal.,2009;2010a;Katneketal.,2010).
The freshwater alga Chlorella, a highly productive source of starch, might substitute for starchrich
terrestrialplantsinbioethanolproduction.Cheapenhancedstarch biomasscanbeproducedfrom highly
productive Chlorella cultures grown in suitable outdoor photobioreactors in which the photosynthetic
carbon dioxide source is derived from combustion of organic waste, fermentation processes or other
sources (Doucha et al., 2005; Douskova et al., 2009; Mann et al., 2009). This characteristic enhances the
ecological and economic impact of the proposed technology, because of its potential to bioremediate
carbondioxideemissionsfromdifferentCO2sourcesincludingwasteincinerators,powerstations,limekilns,
cogenerationunits,etc.insitu.
BIOTECHNOLOGY
Culturemedia
ChlorellawasoneofthefirstalgaeisolatedasapureculturebyBeijerinckin1890.Sincethehalfofthelast
century,attentionhasbeendrawntowardsitspotentialforautotrophicmasscultivation.Tothispurpose,
manymineralmediaweredevelopedbasedinprincipleonthechemicalcompositionofthealgalcells.The
basicinorganicelementsusedare:N,P,K,Mg,Ca,S,Fe,Cu,Mn,MoandZn(Krauss,1958;OKelly,1968).
ManyformulasareusedforthecultivationofthisgenusseeHamaandMiyachi(1988).Forhighyielded
productionofChlorellaoutdoorsanoptimizedcompositionofnutrientsolutionwasproposedbyDoucha
andLvansk(2006).
ThemineralmediumwasbasedonthemeancontentofP,N,K,Mg,andSinalgalbiomassandhadthe
followinginitialcomposition(mg/L):1100(NH2)2CO,237KH2PO4,204MgSO4.7H2O,40C10H12O8N2NaFe,88
CaCl2, 0.83 H3BO3, 0.95 CuSO4.5H2O, 3.3 MnCl2.4H2O, 0.17(NH4)6Mo7O24.4H2O, 2.7 ZnSO4.7H2O, 0.6
CoSO4.7H2O,0.014NH4VO3indistilledwater.
Cultivation of Chlorella heterotrophically in fermenters is also used (Lee et al., 1997). Heterotrophic
culturemayprovideacosteffective,largescalealternativemethodforcultivationofsomemicroalgaethat
can utilize organic carbon substances as their sole carbon and energy source (Chen and Chen, 2006).
NutrientsolutioncontainingglucoseoracetateasasourceofcarbonforintensivegrowthofChlorella in
fermenter was described by Endo and Shirota (1972) and lately by Doucha and Lvansk (Cz. patent.
288638, 2001). Technology of heterotrophic Chlorella cultivation has been commercially used for
productionofbiomass(DouchaandLvansk,2011)forfoodandfeedaswellasforproductionofbiomass
enrichedbyorganicallyboundselenium(Douchaetal.,2009;,Skivanetal.,2006;Trvneketal.,2008)or
asasourceoflipidsforproductionofbiodiesel(Xiongetal.,2008).
Cultivationmethodsandproductionsystems
Thereisstillanoticeablediscrepancybetweentheextentofcommerciallyoperatedalgalculturesandthe
potential of algae. Since the first experiments with largescale algal cultures in the 1950s (Burlew, 1953)
many types of culture equipment have been developed (Stengel, 1970; Richmond and Becker, 1986;
Tredici,2004).Formostproductsofmicroalgalmasscultivationoutdooropencircularorracewayponds
with a 1530 cm layer of algal suspension are the most used technology for the growth of algae
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(Borowitzka,1999).ThecommercialproductionofChlorellabiomassiscarriedoutexclusivelyintheseopen
systems. Closed reactors (tubular, helical and flat bioreactors, vertical cylinders and sleeves) have been
employedforresearchinsmallfieldinstallations(e.g.Torzillo,1997;PulzandScheibenbogen,1998;Tredici,
2004).TheonlyexceptionisalargescaletubularbioreactorwhichstartedproductionofChlorellabiomass
inCentralGermanyin2000(Pulz,2001)andtubularreactorsusedtoproduceHaematococcusinIsrael.
Open ponds are characterized by simple construction and relatively low building costs. On the other
hand, there are many serious drawbacks of this system: (a) due to thick layer of algal suspension, the
culturemustbegrownatlowdensities(about0.5algalDWl1).Withincreasingdensitiestheproductivity
sharply decreases; (b) low velocity flow (1530 cm s1) of poorly mixed algal suspension leads to low
utilizationoflightenergyandtoaccumulationofoxygendissolvedinthesuspension;(c)theseparationof
low concentrated algae from the nutrient solution at the harvest is highly energy demandinged is the
separationoflowconcentratedalgaefromthenutrientsolutionattheharvest.
The key for reduction of cultivation costs rests in a low area volume of algal suspension. This can be
achievedbydecreasingalgallayertoalowvalueastechnologicallypossible(DouchaandLvansk,2006,
2009).
Technologicalandproductioncharacteristicsofbothsystemsaregivenbelow(Table16).
Table16Characteristicsofculturesinracewaypondsandthinlayerculturesystem.
Culturecharacteristics
culturevolume(lm2)
Racewayponds
150300
Thinlayer
68
culturelayerthickness(mm)
biomassharvestdensity(gl1)
150300
0.51
68
3550
harvest/downstreamprocessing
densitymultiply
surface/volumeratio(m1)
150300
34.3
100
2.54
58
1020
2040
0.050.1
520
25
6070
photosynthetic efficiency (% of
PAR)
arealproductivity(gm2d1)
volumetricproductivity(gl1d1)
efficiencyofCO2utilization(%)
Harvestingmethods
Manymethodsareavailableforharvestingofmicroalgae,consistinginthickeningofalgalbiomassasafirst
step.Theseinclude:centrifugation,electroflotation,andchemicalflocculation,followedbysedimentation
or air flotation, continuous belt filtration, vibrating and stationary screens, sand bed filtration, and
autoflocculation (Richmond, 1986). Speaking about largescale commercial Chlorella cultures, only
centrifugation by means of continuously operating self cleaning centrifuges, is used. The advantage of
centrifugationisitssimplicityandpossibilitytolowerchemicalsandbacterialcontaminationinproduct.On
theotherside,thisprocessisconnectedwithahighenergydemand.Whencommonlyusedracewaypond
culturetechnologyisused(atharvestingdensityabout0.5gl1andthickeningupto150gl1),about30%
ofthetotalcostoftheproductionisaccounted(GudinandThepenier,1986).
Biomassprocessing
The next step after the thickening of algal biomass by centrifugation is disruption of algal cells. The rigid
cellulosiccellwall,oneofthecharacteristicsofunicellularChlorella,causesalowutilizationofcellcontent
by a recipient. The digestibility of ruptured Chlorella cells increases to 80 % (Doucha and Lvansk, 2008)
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compared to 1525 % for unruptured cells (Becker, 1984). To open the cell, methods of freezing, alcalic
alkalineandorganicsolvents,osmoticshocks,sonication,highpressurehomogenizationandbeadmilling
were tested (Molina Grima et al., 2004). For largescale processing of Chlorella cultures disintegration of
cellwallsbybeadmillsismostlyused(Middelberg,1995;DouchaandLvansk,2008).
Spraydryingastheendstepofthedownstreamprocessingprocessisthemostextendedmethodfor
dehydrationofrupturedChlorellacells.Algaebiomassisdispersedintoveryfinedroplets,whosesurface
temperature in the course of several seconds of drying does not surpass 60 oC. Thus, the process is very
considerateandthequalityoftheproductishigh.
Scalingup
ThefirstcommercialproductionofChlorellaand,someyearslater,bluegreenSpirulinaculturesstartedin
JapanandTaiwaninthe1960s.Nowadays,largescaleproductionplantsarelocatedalsointheUSA,China,
India, Thailand, Indonesia, Germany and other countries (Borowitzka, 1996; Lee, 1997; Pulz , 2001;
Borowitzka1996;Tredici,2004;Spolaoreetal.,2006).Theirperyearworldproductionisestimatedtobe
about 8000 tons (Spirulina) and 5000 tons (Chlorella). Most of the products have been used in human
nutrition.
High growth rate, high photosynthetic efficiency, relatively high content of energyrich chemicals on
onesideandexperiencewithlargescalecultureanddownstreamprocessingtechnologiesconcentratein
last decades increasing attention on microalgae as a feedstock for biofuels. Today projects dealing with
algae are focused almost entirely on biodiesel production. Nevertheless, algal strains containing higher
amount of lipids are characterized by low growth rate. Slow growth increases the operational costs and
demandscultivationinclosedbioreactorswhosebuildingisexpensive.
On the other side for economical production of bioethanol, relatively cheap biomass of highyielded
Chlorellacultures,containingenhancedamountofstarchgrowninsuitableopenbioreactorsisperspective
solution.Toproducestarcheconomically,conditionsforculturingstarchenrichedalgaeindensecultures
must be attained. Using a thin layer algal suspension in outdoor cultures, linear growth continues up to
very high biomass concentrations (about 40 g/L) enabling easy and cheap harvesting and processing
(Douchaand Lvansk,2006,2009).However,the contentofstarchin the biomassislow(15%ofDWor
less).
Theconditionsunderwhichstarchcontentincreasesincommerciallyproducedalgalbiomasstoalevel
thatwouldbeviableforbioethanolproductioncanbeachievediftheprocessesandeventsduringwhich
starchisextensivelydegradedaresloweddown,orstoppedcompletely,whilethefactorssupportingstarch
synthesis, namely light intensity, remain sustainable (Figure 27) (Brnyikov et al., 2011). Similarly, the
Chlorella species producing oil as their energy reserves rather than starch can be similarly treated to
markedlyincreasetheiroilcontent(Figure28).
An increase in the production of starch in sulphurlimited culture up to a maximum of 50% starch
contentofalgalbiomass(DW)wasdemonstratedunderfieldconditionsusingtheoutdoorscaleup,thin
layersolarphotobioreactor.
While use of algae with enriched starch content is conventional for bioethanol production, another
attractiveexploitationofstarchfromalgaemightbetheproductionofhydrogen,whichmayberealizedin
thenearfuture(Miuraetal.,1982;Tsygankovetal.,2002;Melisetal.,2004;Chochoisetal.,2009;Meliset
al.2004;Miuraetal.1982;Tsygankovetal.2002).Sulphurlimitationcouldbeoneofthewaystosupport
hydrogenproduction(Melisetal.,2000;Zhangetal.,2002).Ithasbeenshownrecentlythatsomestrains
of Chlorella can produce and accumulate significant volume of hydrogen gas under anaerobic conditions
andsulphurdeprivationsuchasitisreportedinliteratureusingC.reinhardtii(Chaderetal.,2009).
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Figure28Electronmicroscopicphotographsof
daughter(A)andmother(B)cellsofChlorella
grownincompletemineralmedium,inthe
presenceofcycloheximide(1mg/L)(C),andin
sulfurlimitingmedium(D).Nnucleus,Sstarch
granules.Bars:panelsA,B,C=2m;barpanelD
=5m.
Figure29FluorescentmicrophotographyofChlorellacellswith
enrichedcontentofoil(yellowspotsoilstainedbyNileRed
TodecreasepriceofalgalbiomassthefluegasfromvarioussourcescanbeusedasacheapCO2source.
Using pilot outdoor thinlayer bioreactor, built in a livestock farm, flue gas, after utilization of CH4
anaerobically generated in biogas station, was used as a source of CO2 for algal photosynthesis. Besides,
mineralsofliquidconcentrateoftheanaerobicdigestedlivestockexcrementscanbeusedasasourceof
inorganic nutrients for algal growth (Doucha, 2010, unpublished results). The flue gas, after utilization of
biogas produced from distillery stillage (Doukov et al., 2010) or from swine manure (Katnek et al.,
2010)forelectricityandheatproduction,wasalsosuccessfullyappliedasacheapsourceofCO2foralgal
biomassproduction.
Anincreaseintheproductionofstarchinsulfurlimitedcultureuptoamaximumof50%starchcontent
ofalgalbiomass(DW)wasdemonstratedunderfieldconditionsusingtheoutdoorscaleup,thinlayersolar
photobioreactor.DespitetherelativelyunfavourableclimaticconditionsofTrebon(CzechRepublic),atotal
yield of starch calculated per ha over a season of 150 days was 7 tonnes (Doukov et al., 2010). In
optimum locations for photoautotrophic production of algae like Greece, with a season lasting
approximately 250 days, the overall harvest might be increased by a factor of 10 (Doucha and Livansky,
2006).
The remaining parts of the cells, containing largely proteins, can be used as a feed supplement what
furtherdecreasesthecostofstarchproduction.
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HussV.A.R.,FrankC.,HartmannE.C.,HirmerM.,KloboucekA.,SeidelB.M.,WenzelerP.,KesslerE.(1999)Biochemicaltaxonomy
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KatnekF.,abataS.,olcovO.,Malterov.Y,Katnek.P,BrnyikovI.,KuthanK.,ZachlederV.(2010)Infieldexperimental
verificatiom of cultuvation of microalgae Chlorella sp. using the flue gas from a cogeneration unit as a source of carbon
dioxide.WasteManagementandResearch28:961966.
KesslerE.,HussV.A.R.(1992)ComparativephysiologyandbiochemistryandtaxonomicassignmentoftheChlorella(Chlorophyceae)
strainsoftheCultureCollectionoftheUniversityofTexasatAustin.JournalofPhycology28:550553.
KraussR.W.(1958)Physiologyofthefreshwateralgae.AnnualReviewofPlantPhysiology9:207244.
Krienitz L.., Hegewald E.H., Hepperle D., Huss V.A.R., Rohr T., Wolf M. (2004) Phylogenetic relationship of Chlorella and
Parachlorellagen.nov.(Chlorophyta,Trebouxiophyceae).Phycologia43:529542.
Krusteva N.G., Tomova N.G., Georgieva M.A. (1984) Allosteric regulation of NAD(NADP)dependent glyceraldehyde3phosphate
dehydrogenasefromChlorellabyaminoacids,dithiothreitolandATP.FEBSLetters171:137140.
LeeY.K.(1997)CommercialproductionofmicroalgaeintheAsiaPacificrim.JournalofAppliedPhycology9:403411.
LeeY.K.(2001)Microalgalmassculturesystemsandmethods:Theirlimitationandpotential.JournalofAppliedPhycology13:307
315.
Li X., Xu H., Wu Q. (2007) Largescale biodiesel production from microalga Chlorella protothecoides through heterotrophic
cultivationinbioreactors.BiotechnologyandBioengineering98:764771.
Liang Y., Sarkany N., Cui Y. (2009) Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and
mixotrophicgrowthconditions.BiotechnologyLetters31:10431049.
LiuJ.,HuangJ.,FanK.W.,JiangY.,ZhongY.,SunZ.,ChenF.(2010a)ProductionpotentialofChlorellazofingienesisasafeedstockfor
biodiesel.BioresourceTechnology101:86588663.
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Liu J., Zhong Y., Sun Z., Huang J., Sandmann G., Chen F. (2010b) One amino acid substitution in phytoene desaturase makes
Chlorellazofingiensisresistanttonorflurazonandenhancesthebiosynthesisofastaxanthin.Planta232:6167.
MannG.,SchlegelM.,SchumannR.,SakalauskasA.(2009)Biogasconditioningwithmicroalgae.AgronomyResearch7:3338.
MelisA.,SeibertM.,HappeT.(2004)Genomicsofgreenalgalhydrogenresearch.PhotosynthesisResearch82:277288.
Melis A., Zhang L., Forestier M., Ghirardi M.L., Seibert M. (2000) Sustained photobiological hydrogen gas production upon
reversibleinactivationofoxygenevolutioninthegreenalgaChlamydomonasreinhardtii.PlantPhysiology122:127135.
MiddelbergA.P.J.(1995)Processscaledisrruptionofmicroorganisms.BiotechnologyAdvances13:491555.
MiuraY.,YagiK.,ShogaM.,MiyamotoK.(1982)Hydrogenproductionbyagreenalga,Chlamydomonasreinhardtii,inanalternating
light/darkcycle.BiotechnologyandBioengineering24:15551563.
Molina Grima E., Fernndez F.G.A., Medina A.R.(2004) Downstream processingof cellmass and products. In: Richmond A. (ed.)
HandbookofMicroalgalCulture:BiotechnologyandAppliedPhycology.BlackwellScience,pp215251.
OKellyJ.C.(1968)Mineralnutritionofalgae.AnnualReviewofPlantPhysiology19:89112.
Ohhama T., Miyachi S. (1988) Chlorella. In: Borowitzka M.A., Borowitzka L.J. (eds.), Microalgal Biotechnology. Cambridge
UniversityPress,Cambridge,pp.326.
PulzO.(2001)Photobioreactors:Productionsystemsforphototrophicmicroorganisms.AppliedMicrobiologyandBiotechnology57:
287293.
PulzO.,ScheibenbogenK.(1998)Photobioreactors:Designandperformancewithrespecttolightenergyinput.In:ScheperT.(ed.)
AdvancesinBiochemicalEngineering/Biotechnology,SpringerVerlag,Berlin,pp.123152.
RichmondA.(ed.)(1986)CRCHandbookofMicroalgalMassCulture.CRCPrsss,BocaRaton,Florida.
Richmond A., Becker E.W. (1986) Technological aspects of mass cultivation a general outlook. In: Richmond A. (ed.) CRC
HandbookofMicroalgalMassCulture.CRCPress,BocaRaton,Florida,pp245263.
Rodolfi L., Chini Zitteli G., Bassi N., Padovani G., Biondi N., Bonini G., Tredici M.R. (2009) Microalgae for oil: Strain selection,
inductionoflipidsynthesisandoutdoormasscultivationinalowcostphotobioreactor.BiotechnologyandBioengineering
102:100112.
SauerN.,TannerW.(1989)ThehexosecarrierfromChlorella.cDNAcloningofaeukaryoticH+cotransporter.FEBSLetters259:43.
SkivanM.,imnJ.,DlouhG.,DouchaJ.(2006)Effectofdietarysodiumselenite,SeenrichedyeastandSeenrichedChlorellaon
eggSeconcentration,physicalparametersofeggsandlayinghenproduction.CzechJournalofAnimalScience51:163167.
Spolaore P., JoannisCassan C., Duran E., Isambert A. (2006) Commercial application of microalgae. Journal of Bioscience and
Bioengineering101:8796.
Stengel E. (1970) Anlagentype und Verfahren der technischen Algenmassenproduktion. Berichte der Deutschen Botanische
Gesellschaft83:589606.
SyrettP.J.,ThomasE.M.(1973)TheassayofnitratereductaseinwholecellsofChlorella:Straindifferencesandtheeffectofcell
walls.NewPhytologist72:13071310.
Torzillo G. (1997) Tubular bioreactors. In: Vonshak A (ed.) Spirulina platensis (Arthrospira): Physiology, Cell Biology and
Biotechnology.Taylor&Francis,London,pp101115.
TrvnekJ.,RacekJ.,TrefilL.,RodinovH.,KroupovV.,IllekJ.,DouchaJ.,PsekL.(2008)Activityofglutathioneperoxidase(GSH
Px)inthebloodofewesandtheirlambsreceivingtheseleniumenrichedunicellularalgaChlorella.CzechJournalofAnimal
Science53:292298.
Tredici M.R. (2004) Mass Production of Microalgae: Photobioreactors. In: Richmond A. (ed.) Handbook of Microalgal Culture.
BlackwellScienceLtd,Oxford,pp178214.
Tsygankov A., Kosourov S., Seibert M., Ghirardi M.L. (2002) Hydrogen photoproduction under continuous illumination by sulfur
deprived,synchronousChlamydomonasreinhardtiicultures.InternationalJournalofHydrogenEnergy27:12391244.
XiongW.,LiX.,XiangF.,WuQ.(2008)HighdensityfermentationofmicroalgaChlorellaprotothecoidesinbioreactorformicrobio
dieselproduction.AppliedMicrobiologyandBiotechnology78:2936.
XuH.,MiaoX.,WuQ.(2006)HighqualitybiodieselproductionfromamicroalgaChlorellaprotothecoidesbyheterotrophicgrowth
infermenters.JournalofBiotechnology126:499507.
YpJ.M.,ChengL.H.,XuX.H.,ZhangL.,ChenH.L.(2010)EnhancedlipidproductionofChlorellavulgarisbyadjustmentofcultivation
conditions.BioresourceTechnology101:67976804.
YpP.F.,WongK.H.,ChenF.(2004)EnhancedproductionofastaxanthinbythegreenmicroalgaChlorellazofingiensisinmixotrophic
culture.ProcessBiochemistry39:17611766.
Zhang L., Happe T., Melis A. (2002) Biochemical and morphological characterization of sulfurdeprived and H2producing
Chlamydomonasreinhardtii(greenalga).Planta214:552561.
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8.1.12 Parietochlorisincisa
Figure30Parietochlorisincisainbalancedgrowth(left)andnitrogenstarved(right)
I.Khozin,BGU
SYMBOLS:D,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Trebouxiophyceae
Chlorellales
Chlorellaceae
Parietochloris
Parietochlorisincisa
RelatedSpecies
Numbersofnamesandspecies:Thereare6species(andinfraspecific)namesinthedatabaseatpresent.
P.alveolaris,P.bilobata,P.cohaerens,P.incisa,P.ovoidea,P.pseudoalveolaris.
BIOLOGY
Parietochlorisincisaisaunicellular,oleaginous,freshwateralga.Thealgawasisolatedfromtheslopesofa
snowmountaininJapan(Watanabeetal.,1996),analpineenvironmentwhichischaracterizedbyabroad
temperature range, UV radiation and light levels that can be extremely high. Such environments were
foundtobeanaturalhabitatforphototrophicmicroorganismsthataccumulatehighpolyunsaturatedfatty
acids (PUFA) concentrations, including sea ice diatoms, dinoflagellates, and green algae. Indeed P. incisa
wasfoundtoaccumulateunusuallyhighcontentofthelongchainPUFA,arachidonicacid(AA)(Bigognoet
al.,2002a).
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Undernitrogenstarvation,thefattyacidcontentinP.incisaisover35%ofdryweight;AAconstitutesabout
60%oftotalfattyacids,andover90%ofcellAAisdepositedinTAG(KhozinGoldbergetal.,2002).Inthe
lipidlinkedpathwayofitsbiosynthesis,AAismainlyexportedtoTAGandaccumulatedincytoplasmiclipid
bodies (Bigogno et al., 2002b). The pathway of AA biosynthesis involves stepwise desaturation and
elongationofoleicacidviathesocalled6pathway.Thegenesencodingfor12,6,5desaturasesand
6PUFAelongasewereclonedandtheirfunctionswerevalidatedinheterologoussystem(Iskandarovet
al.,2009,2010).
Furthermore,underoptimalgrowthtemperature(25C),AAispartiallyconvertedtothevaluablePUFA
eicosapentaenoic acid (EPA, 20:53). This conversion is more pronounced when alga is exposed to low
temperature (1015oC). The conversion of 6 precursors into 3 fatty acids is catalyzed by a group of
enzymescalled3fattyaciddesaturases(FAD)thatdifferbyfattyacidsubstratepreferenceandcellular
localization. A 5desaturasedeficient mutant isolated at BGU (Cohen et al., 2009) is able to accumulate
thehighvaluePUFADGLA(20:36).
BIOTECHNOLOGY
Growthmedium
mBG11
Growthrate
Growthrateinpanelreactorsandcontinuousilluminationundernitrogenstarvation:
160mg/lperdayaccumulating6%DGLAand18%TAG.
Pilotscalescaleproduction
TheDGLAproductionprocessusingtheP.incisamutanthasbeentestedatBGUatthepilotscaleusing50
200lflatpanelreactorsingreenhouseorclimatecontrolledgrowthrooms.DGLAcontent10%oftotaldry
biomass was achieved after cultivation in Ndeficient medium demonstrating a commercial production
capability.
P.incisaanditsmutantareuniqueamongalgaeinbeingabletoaccumulatehighconcentrationsofPUFA
in TAG in cytoplasmic oil globules. Identification of several desaturase and elongase genes may allow
breakthroughsinengineeringmoreefficientPUFAproducingalgae.
References
BigognoC.,KhozinGoldbergI.,BoussibaS.,VonshakA.,CohenZ.(2002a)Lipidandfattyacidcompositionofthegreenoleaginous
algaParietochlorisincisa,therichestplantsourceofarachidonicacid.Phytochemistry60:497503.
Bigogno C., KhozinGoldberg I., Adlerstein D., Cohen Z. (2002b) Biosynthesis of arachidonic acid in the oleaginous microalga
Parietochlorisincisa(Chlorophyceae):radiolabelingstudies.Lipids37:209216.
CohenZ.,KhozinGoldbergI.,BoussibaS.,VonshakA.;BenGurionUniversityoftheNegevResearchandDevelopmentAuthority,
assignee.200919.02.2009.OverProductionofdihomogammalinolenicacidbyaMutantStrainofParietochlorisincisa.IL.
Iskandarov U., KhozinGoldberg I., Ofir R., Cohen Z. (2009) Cloning and characterization of the 6 polyunsaturated fatty acid
elongasefromthegreenmParietochlorisincisa.Lipids44:545554.
Watanabe S., Hirabayashi S., Boussiba S., Cohen Z., Vonshak A., Richmond A. (1996) Parietochloris incisa comb.nov.
(Trebouxiophyceae,Chlorophyta).PhycologicalResearch44:107108.
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8.1.13 Protothecasp.
Figure31LightmicroscopicviewshowingProtothecamoriformis
Lactophenolcottonbluemountfixation.
Picturefromhttp://content9.eol.org/content/2009/11/25/02/75871_small.jpg
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Trebouxiophyceae
Chlorellales
Chlorellaceae
Prototheca
Relatedspecies
P. blaschkeae, P. chlorelloides, P. ciferrii, P. crieana, P. filamenta, P. hydrocarbonea, P. kruegeri, P.
moriformis, P. moriformis var. betulina, P. portoricensis, P. portoricensis var. trisporus, P. salmonis, P.
segbwema, P. stagnorum, P. trispora, P. ulmea, P. viscosa, P. wickerhamii, P. zopfii var. portoricensis, P.
zopfiivar.hydrocarbonea,P.zopfii.
BIOLOGY
ThegenusProtothecaiscomposedofmicroscopicachlorophyllousorganismswithalifecyclesimilartothat
of the genus Chlorella. The species typically produce thickwalled cells (sporangia) which, at maturity,
dividebyirregularcleavageforming215aplanospores(endospores).Followingruptureofthesporangial
wall,freedaplanosporesenlargeandrepeatthecycle.Onetothreepercentofthesporangiacleavetoform
23thickwalledrestingcells(hypnospores).Nosexualcyclehasbeenobserved.
Protothecacanusedextrose,levulose,galactose,ethanol,nbutanol,isobutanol,isopentanol,hexanol
and glycerol and some species assimilate sucrose, trehalose, npropanol and npentanol. In general,
alcohols are assimilated similarly in both liquid and vapor phases; npentanol is assimilated only in the
vaporphase.SpeciesofProtothecarequirethiamine.
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NumerousstudieshavereportedapathogenicpotentialforP.wickerhamiiandP.zopfii.Thecasesof
human protothecosis are predominantly caused by P. wickerhamii and occur as local (predominantly
cutaneous) and systemic infections mainly in immunecompromised patients, e.g. patients infected with
HIVortreatedwithglucocorticoids.P.blaschkeaewereisolatedfromsomecasesofonychomycosis.Canine
protothecosisiscausedbyP.wickerhamiiandP.zopfii,andischaracterizedbysimilarclinicalsymptomsas
in humans. Worldwide, P. zopfii has been identified to induce a therapyresistant inflammation of the
mammaryglandindairycows.
BIOTECHNOLOGY
P.zopfiiand P.moriformishavebeenproposed,togetherwithChlorellaprotothecoides,asorganismsfor
productionofascorbicacid(Runningetal.,2002).Inalaboratoryscaleexperiment(114Lfermentors)P.
zopfii at 35 C at pH 7 produced 37.8 mg L1 of ascorbic acid, mainly intracellular, with a biomass
preodcutionof27gL1,whileatpH4/5itproduced73mgL1ofextracellularascorbicacidand55gL1of
biomassin29h;similarlyP.moriformisproducedin30hatpH4162mgL1ofextrcellularascorbicacidand
42gL1ofbiomass(Hussetal.,1995;Running,1999).
P.zopfiiisabletoutilisecrudeoilhydrocarbons(aromatics,cyclicandbranchedalkanes)aswellaspure
nalkanes(Uenoetal.,2002,2008).
Solazyme Inc. (San Francisco, USA) has recently patented several processes to obtain oil from
microrganisms including Prototheca, that contains lipids with a higher degree of saturation compared to
other algae and has the advantage of lacking pigments, also through genetic engineering to increase
production(Dillonetal.,2010;Franklinetal.,2010,2011).
References
ArnoldP.,AhearnD.G.(1972)ThesystematicsofthegenusProtothecawithadescriptionofanewspeciesP.filamenta.Mycologia
64:265275.[BIOLOGYsection]
DillonH.F.,ElefantD.,DayA.G.,FranklinS.,WittenbergJ.(2010)Fractionationofoilbearingmicrobialbiomass.WO2010/138620.
FranklinS.,SomanchiA.,EspinaK.,RudenkoG.,ChuaP.(2011)Renewablechemicalproductionfromnovelfattyacidsfeedstocks.
USPatentNo.7,883,882B2.
Franklin S., Somanchi A., Espina K., Rudenko G., Chua P. (2010) Manufactoring of tailored oils in recombinant heterotrophic
microorganisms.WO2010/063031A2.
HussJ.R.,RunningJ.A.,SkatrudT.J.(1995)Lascorbicacidproductioninmicroorganisms.WO95/21933.
RunningJ.A.(1999)ProcessfortheproductionofascorbicacidwithPrototheca.USPatentNo.5,900,370.
RunningJ.A.,SeversonD.K.,SchneiderK.J.(2002)ExtracellularproductionofLascorbicacidbyChlorellaprotothecoides,Prototheca
species, and mutants of P. moriformis during aerobic culturing at low pH. Journal of Industrial Microbiology and
Sudman M.S., Kaplan W. (1973) Identification of the Prototheca Species by immunofluorescence. Applied and Environmental
Microbiology25:981990.[BIOLOGYsection]
Ueno R., Urano N., Wada S., Kimura S. (2002) Optimization of heterotrophic culture conditions for nalkane utilization and
phylogeneticpositionbasedonthe18SrDNAsequenceofathermotolerantProtothecazopfiistrain.JournalofBioscience
andBioengineering94:160165.
UenoR.,WadaS.,UranoN.(2008)Repeatedbatchcultivationofthehydrocarbondegrading,microalgalstrainProtothecazopfii
RND16immobilisedinpolyurethanefoam.CanadianJournalofMicrobiology54:6670.
vonBergenM.,EidnerA.,SchmidtF.,MurugaiyanJ.,WirthH.,BinderH.,MaierT.,RoeslerU.(2009)Identificationofharmlessand
pathogenicalgaeofthegenusProtothecabyMALDIMS.ProteomicsClinicalApplications3:774784.[BIOLOGYsection]
Wolff G., Plante I., Lang B. F., Kck U., Burger G. (1994) Complete sequenceof the mitochondrial DNA of the chlorophyte alga
Prototheca wickerhamii: Gene content and genome organization. Journal of Molecular Biology 237: 7586. [BIOLOGY
section]
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8.2 Rhodophyta
8.2.1
Porphyridiumcruentum
Figure32LightmicroscopicviewshowingPorphyridiumcruentumstrainCCALA415.
Picturefromwww.butbn.cas.cz/ccala/col_images/415.jpg
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Porphyridiophyceae
Porphyridiales
Porphyridiaceae
Porphyridium
Relatedspecies
P.aerugineum,P.cruentum,P.griseum,P.marinum,P.purpureum,P.sordidum,P.violaceum.
BIOLOGY
Porphyridiumiscomposedofsphericaltoovoidunicellswithastellatechloroplastandprominentcentral
pyrenoid.Thecelldiameteris510mintheexponentialphase,716minthestationaryphase.Cellsare
solitary, but often grouped into irregular colonies with an illdefined mucilaginous matrix. Species are
distinguished by chloroplast color. The chloroplasts of freshwater species contain single thylakoids with
phycobilisomes(granulesconsistingoftheaccessorypigments)onbothsides.Thephycobilisomesofblue
coloredspecies,suchasPorphyridiumaerugineum,tendtobehemidiscoidalinshapeandpredominatedby
thebluepigmentphycocyanin.Incontrast,thephycobilisomesoftheredcoloredPorphyridiumpurpureum
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arelargerandhemisphericalandcomposedmostlyoftheredpigmentphycoerythrin.Reproductionoccurs
bycelldivision.Porphyridiumformsgelatinouscoatingsonvarioussurfaces;itiswidespreadinfreshwaters,
brackish environments. Species of Porphyridium can form gelatinous crusts on moist soils and decaying
wood.Inthesehabitats,thesespeciesarereasonablydesiccationresistantandshadetolerant.
BIOTECHNOLOGY
ThemarinemicroalgaPorphyridiumcruentumisofincreasinginterestassourceofvaluablecompoundslike
phycoerythrin, sulfated exopolysaccharides, superoxidedismutase, and polyunsaturated fatty acids with
applicationsinthefood,pharmaceuticalandcosmeticindustries(Fbregasetal.,1998;Dillonetal.,2007).
Phycoerythrin is used as fluorescent dye in immunoassays (Bermejo Romn et al., 2002), sulfated
polysaccharidesareknowninhibitviruses(Huheiheletal.,2002)andshowhypocholesterolemicactivityin
rats(Dviretal.,2000,2009).P.cruentumisalsoconsideredtobeoneofthemostimportantsourcesofthe
polyunsaturated fatty acids eicosapentaenoic acid (20:5 3, EPA) and arachidonic acid (20:46, ARA)
(CohenandCohen1991;GuilGuerreroetal.,2001).
Ina220LailifttubularphotobioreactorP.cruentumreachedaveragebiomassproductivitiesof1.76gL
1
day1 with 2% biomass dry weight of ARA , 1.7% palmitic acid and 1.1% EPA (Rebelloso Fuentes et al.,
1999),similartothoseobtainedbyCamachoRubioetal.(1999).
Porphyridium have been tested in screening for algal strains suitable for biodiesel production or
considered as a potential interesting alga based on its biochemical composition (Griffiths and Harrison,
2009;Rodolfietal.,2009),howeveruptonownoindepthstudiesonthistopichavebeencarriedout.
References
Bermejo Romn R., AlvrezPez J.M., Acin Fernndez F.G., Molina Grima E. (2002) Recovery of pure Bphycoerythrin from the
microalgaPorphyridiumcruentum.JournalofBiotechnology93:7385.
Camacho Rubio F., Acin Fernndez F.G., Snchez Prez J.A., Garca Camacho F., Molina Grima E. (1999) Prediction of dissolved
oxygen and carbon dioxide concentration profiles in tubular photobioreactors for microalgal culture. Biotechnology and
Cohen Z., Cohen S. (1991) Preparation of eicosapentaenoic acid (EPA) concentrate from Porphyridium cruentum. Journal of the
AmericanOilChemists'Society68:1619.
Cohen Z., Vonshak A., Richmond A. (1988) Effect of environmental conditions on fatty acid composition of the red alga
Porphyridiumcruentum:correlationtogrowthrate.JournalofPhycology24:328332..[BIOLOGYsection]
Dillon H.F., Somanchi A., Rao K., Jones. P.J.H. (2007) Nutraceutical compositions form microalgae and related methods of
productionandadministration.WO2007/136428A2.
DvirI.,StarkA.H.,ChayothR,MadarZ.,MalisAradS.(2009)Hypocholesterolemiceffectsofnutraceuticalsproducedfromthered
microalgaPorphyridiumspinrats.Nutrients1:156167.
DvirI.,ChayothR.,SodMoriahU.,ShanyS.,NyskaA.,StarkA.H.,MadarZ.,MalisAradS.(2000)Solublepolysaccharideandbiomass
of red microalga Porphyridium sp. alter intestinal morphology and reduce serum cholesterol in rats. British Journal of
Nutrition84:469476.
Fabregas J., Garcia D., Morales E., Domnguez A., Otero A. (1998) Renewal rate of semicontinuous cultures of the microalga
Porphyridiumcruenturnmodifiesphycoerythrin,exopolysaccharideandfattyacidproductivity.JournalofFermentationand
Griffiths M.J., Harrison S.T.L. (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production.
JournalofAppliedPhycology21:493507.
GuilGuerreroJ.L.,BelarbiE.H., RebollosoFuentesM.M.(2001) Eicosapentaenoicandarachidonicacidspurificationfromthered
microalgaPorphyridiumcruentum.Bioseparation9:299306.
Huheihel M., Ishanu V., Tal J., Malis Arad S. (2002) Activity of Porphyridium sp. polysaccharide against herpes simplex viruses in
vitroandinvivo.JournalofBiochemicalandBiophysicalMethods50:189200.
Rebolloso Fuentes M.M., Garca Snchez J.L., Fernndez Sevilla J.M., Acin Fernndez F.G., Snchez Prez J.A., Molina Grima E.
(1999)OutdoorcontinuouscultureofPorphyridiumcruentuminatubularphotobioreactor:quantitativeanalysisofthedaily
cyclicvariationofcultureparameters.JournalofBiotechnology70:271288.
102:100112.
Sheath R.G. (2003) Red algae. In: Wehr J.D., Sheath R.G. (eds.) Freshwater Algae of North America: Ecology and Classification.
ElsevierScience,pp.197224.[BIOLOGYsection]
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8.3 Bacillariophyta
8.3.1
Benthicdiatoms(Amphora;Amphiprora;Cylindrotheca;Navicula;Nitzschia)
TAXONOMY
Amphorasp.
Figure33Amphora
coffeaeformis.
Figure34Lightmicroscopic
pictureofAmphorasp.
Figure35Lightmicroscopicpictureof
Amphoracoffeaeformis.
AllrightsreservedSeambiotic
008
Copyright19952010Protist
InformationServer
Copyright2010MonashUniversity;
arts.monash.edu.au
SYMBOLS:D,PIV
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Thalassiophysales
Thalassiophysales
Amphora
Relatedspecies
Amphoraisaverylargeandheterogeneousgenus.Thereare1032speciesnamesinthealgaedatabaseat
A. coffeaeformis, A. coffeaeformis punctata, A. coffeaeformis linea, A. coffeaeformis tenuis, A.
coffeaeformistaylori,A.delicatissima,A.delicatissimacapitata.
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Amphiprorahyalina
Figure36LightmicroscopicpictureofAmphiprorasp.
Figure37LightmicroscopicpictureofAmphiprorasp.
Picturefromwww.plingfactory.de/
Picturefromwww.mikroskopieph.de/
SYMBOLS:D,PIV
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Naviculales
Amphipleuraceae
Amphiprora
Amphiprorahyalina
Relatedspecies
Thereare230speciesnamesinthealgaedatabaseatpresent,ofwhich29havebeenflaggedascurrently
acceptedtaxonomically.
Synonym:Amphiprorapaludosavar.hyalina(EulensteinexVanHeurck)Cleve1894.
Cylindrothecasp.
Figure38LightmicroscopicpictureofCylindrotheca
sp.
Figure39Scanningelectronmicroscopicpictureof
Cylindrothecasp.
PicturecourtesyofPAE(UGent)
Picturefromocean.inha.ac.kr/l3.htm
SYMBOLS:D,PIV
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Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Bacillariales
Bacillariaceae
Cylindrotheca
Relatedspecies
There are 6 species names in the algae database at present, of which 2 have been flagged as currently
Potentiallyimportantforbiofuel:
CylindrothecafusiformisReimann&Lewin1964
Cylindrothecaclosterium(BrbissonexKtzing)GrunowinvanHeurck1882
Naviculasp.
Figure40LightmicroscopicpictureofNavicula
gregaria
Figure41LightmicroscopicpictureofNavicula
lanceolata
PicturecourtesyofPAE(UGgent)
PicturecourtesyofPetrZnachor
SYMBOLS:D,PIV
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Naviculales
Naviculaceae
Navicula
Relatedspecies
Naviculaisaverylargeandheterogeneousgenus.Thereare6714speciesnamesinthealgaedatabaseat
Potentiallyimportantforbiofuel
Naviculaacceptata,Naviculasaprophila,Naviculapelliculosa.
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Nitzschiadissipata
Figure42LightmicroscopicpictureofNitzschiadissipata
Picturefromhttp://craticula.ncl.ac.uk/
Figure43Rasterelectronmicroscopicpictureof
Nitzschiasp.
Picturefromhttp://ina.beyerprivat.net/
SYMBOLS:D,PIV
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Bacillariales
Bacillariaceae
Nitzschia
Nitzschiadissipata
Relatedspecies
There are 1024 species names in the algae database at present, of which 321 have been flagged as
Potentiallyimportantforbiofuel:
N.communis,N.frustulum,N.palea.
BIOLOGY
Benthicdiatomsareallraphid,pennatediatoms.
Amphorasp.
The benthic Amphora species appear epiphytic, epilithic or epipelic (in or attached to sediments). The
foulingA.coffeaeformisisacommonmarinespecies.
CellsofAmphoraaresolitary,sometimessessilebutusuallymotile,almostalwayslyingingirdleview
andthenappearingellipticalorlanceolate,withtruncateends(Roundetal.,1990).Amphoraspecieshave
typicalasymmetricalvalvemorphology.Itsdorsiventralfrustuleresemblesathirdofanorange(Hendey,
1964)withbothraphesystemsonthesame(ventral)sideofthecell.Cellsusuallyhaving1or2,sometimes
many,plastidswhichareextremelydiverseinposition,shapeandstructure(Roundetal.,1990).Celllength
and width varies with species, roughly ranging from 14 55 mand 2.5 9 m respectively (Sala et al.,
1998).
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Amphiprorahyalina
Amphiprora (currently placed in the genus Entomoneis) is a benthic genus, epipelic of brackish marine
sediments;occasionallyinfreshwater.A.hyalinaisamarinespecies.
Amphiprora/Entomoneis cells are solitary and twisted about the apical axis (twisted frustules) usually
lying in girdle view and then appearing bilobate. This torsion of the cell means that valves or whole
frustulescanpresentagreatvarietyofaspectsdependingonexactlyhowtheylierelativetotheobserver.
Cells contain one platelike plastid or two plastids, one on each side of the median transapical plane. A
varietyofporeandraphestructuresisfoundinEntomoneis.Therapheismedian,sigmoid,onaraisedkeel
formingawing(Roundetal.,1990).
Cylindrothecasp.
Cyclindrothecaisbenthicandwidelydistributedintheepipelon(livingon(orin)finesediments)ofmarine
habitats;rarelyoccurringinfreshwater.Thegenusisveryabundantincoastalwatersworldwide(Roundet
al.,1990).
Cylindrothecacellsaresolitary,longandnarrow,needlelikeandonlyweaklysilicified.Thefrustulesare
twisted about the apical axis, consequently the cells rotate as they move through the sediment. Cells
containtwoormanychloroplaststhatareplatelikeordiscoid(Roundetal.,1990).
Naviculasp.
AllNaviculaarebenthicdiatoms.CellsofNaviculasensulatodiatomshavenaviculoid(boatshaped)cells
that may exist singly or in ribbons. Navicula is Latin for "small ship". The valves are symmetrical both
apically and transapically, and may have rounded, acute, or capitate ends. The central area is often
distinctlyexpanded.Theycontaintwogirdleappressedplastids,oneoneithersideoftheapicalplane.The
raphidsystemiswelldevelopedwitharapheoneachvalvewhichmakescellshighlymotile.
Nitzschiadissipata
Nitzschiacomprisesbothplanktonicandbenthicspecies.N.dissipataisabenthicspeciesandwasrecorded
infreshwater(e.g.Aboaletal.,2003;Dayetal.,1995;KrammerandLangeBertalot,1988;Robertsetal.;
2004)aswellasincoastalwaters(TadrosandJohansen,1988).Nitzschiaisoneofthemostdifficultgenera
for species identification and many features are only seen by electron microscopy (e.g. Mann, 1986;
Trobajo et al., 2004, 2006). Nitzschia cells are usually linear to lanceolate and may be solitary or colony
forming.Mostspecieshavetwochloroplasts,oneineachendofthecell.Eachvalvepossessesaraphethat
is more or less eccentric and supported by fibulae (=bridges of silica between portions of the valve on
either side of the raphe, giving a ladderlike appearance). The two raphes of a frustule are positioned
diagonallyopposite(nitzschioid).Valvestriae(=lineswithsmallholes)areusuallyuniseriate.
Biofilmformation
Benthic diatoms are the most common benthic microalgae, which are abundant in many softsediment
aquatichabitats(estuaries,shallowsubtidalseas,coralreefflats,lakes,andrivers)andcancontributeupto
50% of the total autotrophic production in some ecosystems. They form biofilms, a matrix of cells,
sedimentsandextracellularpolymericsubstances(EPS)(UnderwoodandPaterson,2000).Itisknownthat
diatommucilagesarerichinpolysaccharidesandproteoglycansandaresecretedthroughthechannelsand
pores in the diatom frustule. In some benthic species, these compounds produce structures (tubes, pads
andstalks)thatareusedforattachmenttosurfacesandoftencontributetobiofoulingproblems.Epipelic
diatomsdonotproducepermanentstructuresbutsecretelargequantitiesofextracellularmucilagesthat
are involved in motility (Underwood and Paterson, 2000). Motility is an essential adaptation for
photosynthetic organisms in these environments, allowing cells to migrate into the illuminated (photic)
zoneofsedimentnearthesurfaceafterperiodsofsedimentmixingordeposition.Indiatoms,thismotility
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is generated by the production of extracellular polymeric substances (EPS), primarily polysaccharides

(Underwoodetal.,2004).Themechanismofmovementindiatomsisuniqueformicrobialcellsandrelies
ontheextrusionofmucilagethroughaslitinthesurfaceofthesilicafrustule(cellwall).Thisslit,knownas
theraphe,maybepresentononlyasinglevalveofthefrustule(monoraphid)oronbothvalves(biraphid)
(Underwoos and Paterson, 2000).The production of EPS in the aquatic environment is ecologically
significant because EPS and other carbohydraterich exudates can be used by bacteria, meiofauna, and
macrofaunaasacarbonsourceandEPScanincreasethestabilityofsediments(Underwoodetal.,2004).
The production of extracellular carbohydrates shows some dependency on environmental conditions, for
example,irradianceandnutrientconditions.Theproductionofsomeextracellularcarbohydratesceasesin
darkness,butstudiesonaxenicculturesofbenthicdiatomsandnaturalsedimentassemblageshaveshown
production of EPS in dark as well as under illuminated conditions. Continued EPS production in darkness
indicates that EPS production is not directly coupled to the photosynthetic production of carbohydrates
(Underwood et al., 2004). Nutrient limitation can also increase extracellular carbohydrate production in
benthic diatoms: this production of extracellular carbohydrates is assumed to be due to unbalanced
metabolism,withcellsreleasingfixedcarboninexcessoftheirenergeticrequirements,becauseofgrowth
beingpreventedbynutrient(N,P)limitation(Underwoodetal.,2004).DiatomEPSconsistsofarelatively
undefined complex mixture of proteins, proteoglycans and carbohydrates (Underwood and Paterson,
2000).There is evidence that benthic diatoms produce a number of different types of EPS, which vary in
structure and sugar composition, and that the production of these EPS depends on environmental
conditionsandthenutrientstatusofthecells(Underwoodetal.,2004).Thebiosyntheticpathwayforthese
carbohydratesandthemechanismscausingchangesinEPScompositionareyettobeelucidated.Glucans
either maybethe precursorofEPSormayactasthephotoassimilate carbonstore,providingenergy for
EPSsynthesisduringperiodsofdarkness.Thislatter hypothesisissupportedbythesignificantcorrelation
found between glucan catabolism and EPS production in the dark in a number of benthic species. This
provides indirect evidence that glucans are involved in the production of EPS (Underwood et al., 2004).
Microphytobenthic biofilms can have high rates of photosynthesis and a significant proportion of their
photoassimilatedcarbonisreleasedintotheenvironmentasextracellularcarbohydrates(Underwoodand
Paterson,2000).
Biochemicalcompositionunderoptimalandstressedconditions
Amphora sp.: The biochemical composition vary among Amphora species as indicated by the diverse
valuesreportedbydifferentstudies(delaPea,2007;Gordonetal.,2006;Khatoonetal.,2009;Sheehan
et al., 1998). In addition, different culture conditions result in significant variations in growth and the
biochemicalcompositionofthecellsofthesamestrainasshownbydelaPea(2007).Heshowedthatthe
proximatechemicalcomposition(protein,carbohydrates,fattyacidcontent,chlorophylla)ofAmphorasp.
is highly dependent on light intensity, the culture location (indooroutdoor) and the type of enrichment
used(delaPea,2007).AhigherproteinandcarbohydratecontentofAmphorasp.wasnotedincultures
located inside the laboratory compared to cultures grown outside (probably due to the more regulated
cultural conditions inside like constant irradiance and temperature). Lipid content ranged from 26 81%
depending on culture site and nutrients used (de la Pea, 2007). Amphora is rich in total lipids and fatty
acids with a high amount of polyunsaturated fatty acids (PUFAs) especially EPA and a high amount of
essentialaminoacids(Gordonetal.,2006;Khatoonetal.,2009).GriffithsandHarrison(2009)calculated
theaveragetotallipidcontentforAmphorasp.fromavailableliteraturedata.
51%cdw(celldryweight)undernutrientrepletelaboratoryconditions
40%cdwinoutdoorponds
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Amphiprorahyalina:A.paludosavar.hyalinacontainsahighamountofEPA(28%)(CorreaReyesetal.,
2009). Proximate composition (dry weight percentage) in A. paludosa var. hyalina (CorreaReyes et al.,
2009):
11.33%0.36protein
20.96%0.94nitrogenfreeextracts
8.10%0.49lipids
59.61%1.01ash
The average total lipid content for Amphiprora hyalina calculated from available literature data by
GriffithsandHarrison(2009)is:
22%cdwundernutrientrepletelaboratoryconditions
28%cdwunderNdeficientlaboratoryconditions
37%cdwunderSideficientconditions
Cylindrotheca sp.: The average total lipid content for Cylindrotheca sp. calculated from literature by
GriffithsandHarrison(2009)is:
27%cdwundernutrientrepleteconditions
27%cdwunderNdeficientconditions
Navicula sp.: The average total lipid content for N. acceptata calculated from literature by Griffiths and
Harrison(2009)is:
N. saprophila contains large quantities of EPA and is considered a potential source of this important
fatty acid (Kitano et al., 1997). It was shown that EPA production was enhanced under mixotrophic
conditions in CO2 enriched (about 2%) atmosphere in the presence of acetate as compared with
photoautotrophicconditions(Kitanoetal.,1997;1998).ThebiomassofafreshwaterN.saprophilastrain
hasthefollowingcomposition(Pilny,2009):
Protein
Lipid
Crudefiber
Ash
growninWCmedium
46.03%
28.57%
<15.4%
11%
mediumwithhalfstrengthphosphorusandhalf
strengthnitrogen(ureaasasource)
38.82%
18.24%
<11.86%
11.86%
Saline strains of N. saprophila have been found to produce significant amounts of carbohydrate in some
media(Barclayetal.,1986).TheaveragetotallipidcontentforN.saprophilacalculatedfromliteratureby
GriffithsandHarrison(2009):
Chelf(1990)showedthatnitrogenconcentrationwasthevariablewiththegreatesteffectonneutral
lipidaccumulationinN.saprophila.Nitrogendeficiencyledtoahighersynthesisoflipids.
Nitzschiadissipata:VariousNitzschiaspecieshavehighoilcontent(Chisti,2007;GriffithsandHarrison,
2009; Mata et al., 2010) and have been suggested for production of EPA (Spolaore et al., 2006). EPA
production in Nitzschia sp. was enhanced under mixotrophic conditions in the presence of acetate as
compared with photoautotrophic conditions (Kitano et al., 1997). Some Nitzschia species are cultured
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heterotrophically (e.g. Barclay et al., 1994; Wen and Chen, 2003). In the ASP program different Nitzschia
species were isolated (a.o. N. dissipata, N. palea, N. communis, N. pusilla, etc.) (Sheehan et al., 1998).
Preliminary screening experiments indicated that N. dissipata had the best growth rates and lipid
accumulation potential (Sheehan et al., 1998). The average total lipid content for N. dissipata calculated
fromliteraturebyGriffithsandHarrison(2009):
BIOTECHNOLOGY
Benthicdiatoms(Amphora,Amphiprora,Cylindrotheca,Navicula,Nitzschia)arecommonlyusedinabalone
culture, where they act as inductors for larval settlement and as food for the early juvenile stages. They
excreteextracellularpolymericsubstancesthatplayanimportantroleinabalonelarvalsettlement(Brown
etal.,1997;PulzandGross,2004;CarbajalMirandaetal.,2005;seerefsindelaPea,2007;CorreaReyes
etal.,2009).
SpeciesofthegenusNitzschiaoccurinnearlyeverydiatomassemblageinfresh,brackish,andmarine
habitats. Many of them are considered to be important indicators of organic pollution and high nutrient
loads,makingthemimportantforwaterqualitystudiesandbiomonitoring(LangeBertalot,1979;VanDam
etal.,1994).
For their high lipid content benthic diatoms have been considered as sources for biodiesel, though
experiments were carried only at laboratory scale (mainly in flasks), obtaining quite low growth and
productivity (Griffith and Harrison, 2009; Rodolfi et al., 2009). Navicula saprophila was genetically
transformedtooptimizelipidproduction(Dunahayetal.,1995).
CultureMedia
For marine and brackish strains the most common medium used is medium F or f/2 added with Silicon
(GuillardandRyther,1962)andadjustedinsalinityaccordingtonecessity.Forfreshwaterstrainsmedialike
Chu(Gerloggetal.,1950)orBG11(Rippkaetal.,1979)addedwithSiliconcanbeused.
Cultivationmethods
The benthic diatoms, in general, have to be grown in photobioreactors purposely designed, because of
theirtendencytoformbiofilms.Thesimplestsolutionisrepresentedbythebenthicalgaegrowthchamber
(BAGC),whichwasusedtogrowamixedbenthicalgaecommunityondairymanurebyWilkieandMulbry
(2002). The culture medium (220 L) was recirculated from a tank to the chamber by a submersible
recirculationpumpwhilealgalgrowthwassupportedbyscreensof96:5x96:5cmwith3x4mmmesh,for
atotalgrowingareaof0.93m2.
AnotherpossiblesolutionwasthatbyLebeauetal.(2000),whoproposedaphotobioreactordesignfor
immobilised microalgal cells. The aim of this photobioreactor was to cultivate microalgae, especially
benthicdiatomsthatsufferfrombioturbation.Inthesephotobioreactors,thesurface/volumeratioofthe
matrix (agar or alginate) was maximised to offer the maximal contact between microalgae and
nutrient/light.Theimmobilizedcellphotbioreactorconsistedof10Lglassandstainlesssteelcylinders.The
inoculum was entrapped in a tubular agar gel layer, then iserted into the cylinder containing the liquid
culturemedium.ThereactorwastestedwithHasleaostreariafortheproductionofthepigmentmarennine
(Rossignoletal.,2000).
Another solution is to multiply the adhesion surface inside the reactor. This has been achieved in
different ways. The simplest is that proposed by de la Pea (2007) that cultivated Amphora in a 8 L
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rectangularacrylicglassaquariacontaining5Lofculturemedium.Tenpiecesof121212cm2areaacrylic
glassplateswereinstalledineachaquariumtoserveasdiatomsettlementplates.Theplateswereprovided
with orange polyvinyl stand pipes to keep them in an upright and slanting position. A twopoint mild
aerationwasprovidedbyglasswoolfilteredairtoprovideaneffectivegasexchangeandtoallowthealga
to settle onto the acrylic glass plates. A more complex device was proposed by AvendaoHerrera and
Riquelme(2007),thatusedanewsystemcalledtheTanakaphotobioreactor,thatconsistsofaborosilicate
tubemeasuring40x4cm,witha4.5x1cmsidearmeffluenttubeatthebottomandaPVCstopperatthe
topwhichhasanopeningtoallowtheentryofa5mmdiameterglassrod.Theinteriorofthetubecontains
anarrayofabout350sterilizedpolyethylenebristlesinabottlebrusharrangementmeasuring30x4cm,
providinganextensivesurfaceareaforadhesion.TheyusedthereactortocultivateNaviculavenetawith
associated bacteria.This configuration was slightly modified by SilvaAciares and Riquelme (2008) with a
reactor called bristles photobioreactor (PBB). PBB is constructed of transparent acrylic plastic 1 m in
lengthwithaninsidediameterof18cmandoutsidediameterof19cm.Thetopofthetubeisclosedbya
PVCcapwhithaconnectorfortheintroductionoftheairline.Thebaseofthetubeisclosed.Withinthe
bodyofthePBB,throughouttheentirelengthofthetube,thereisastructureresemblingalargebottle
brushbearingPVCbristlesmeasuring17.5cm.Thebristlesandthestructureweredesignedassupports
forthegrowthofthe(adherent)benthicdiatoms.PVCbristleswerethebestmaterialforthispurposein
comparisonwithanumberofothertypesofplasticmaterialstested.Thecirculationofthecellsuspension
throughout the photobioreactor is obtained using an airlift system. They cultivated Nitzschia, Amphora,
NaviculaandCylindrothecaclosterium obtainingvolumetricproductivitiesin therange0.120.28gL1d1,
withNitzschiabeingthebestandoneNaviculatheworseperformer.
Onanindustrialscale,SBAEIndustriesnv(Belgium)hasdevelopedandpatentedanoutdoortechnology
called DiaForce for production of benthic diatoms. DiaForce pumps water through channels filled with
artificial carriers on which the diatoms attach. DiaForce technology is used for the production of diatom
polycultures.
Somespecies,likesomeCylindrotheca,canbegrownalsoinbubbledphotobioreactors.
Harvestingmethods
Benthic(attached)freshwateralgaemayhaveanadvantageoverplanktonic(suspended)algaeintheease
of separation and recovery of algal biomass from an aqueous stream. AvendaoHerrera and Riquelme
(2007) report that harvesting of the N. veneta cultures was carried out at the end of each culture cycle
(each 7 days). As much as possible of the diatom (>85%) was dislodged from the bristles within the
photobioreactorusinga1minagitationoftheglassrod,drainingthereactorthroughthesidearm.Toavoid
handlingofthephotobioreactor,theremainingdiatombacterialbiomass(approximately15%ofthetotal)
wasusedtostartanewculturecycle. MicroalgaewereharvestedmanuallyfromthePBB,dislodgingthe
diatomsbyvigorousagitationofallthePVCbristlesinthephotobioreactorcolumnforaperiodof2min
(SilvaAciaresandRiquelme,2008).SBAEstrianglecarriersfacilitatetheharvestingprocessasalgalbiomass
isdirectlycollected,byliftingthemfromthewater.Thisconcentratesthebiomassx100andreducesthe
costcomparedtocentrifugationofthealgaeoutofthewater.
Allthesystemsproposed,exceptSBAEsone,havebeendevelopedinrelationtoaquaculturenecessities,
so they are of rather difficult scalingup. To exploit benthic diatoms as source of biodiesel, it will be
necessarytodesignnovelphotobioreactors,specificfortheircultivation,thatcanbescaledup.
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(Bacillariophyta),inrelationitsuseasabioindicator.NovaHedwigia79:433445.
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(Bacillariophyta).JournalofPhycology42:13531372.
Underwood G.J.C., Paterson D.M. (2000) The importance of extracellular carbohydrate production by marine epipelic diatoms.
AdvancesinBotanicalResearch40:183240.
UnderwoodG.J.C.,BoulcottM.,RainesC.A.,WaldronK.(2004)Environmentaleffectsonexopolymerproductionbymarinebenthic
diatoms:dynamics,changesincomposition,andpathwaysofproduction.JournLofPhycology40:293304.
Van Dam H., Mertens A., Sinkeldam J. (1994) A coded checklist and ecological indicator values of freshwater diatoms from the
Netherlands.NetherlandsJournalofAquaticEcology28:117133.
WenZ.Y.,JiangY.,ChenF.(2002)HighcelldensitycultureofthediatomNitzschialaevisforeicosapentaenoicacidproduction:fed
batchdevelopment.ProcessBiochemistry37:14471453.
WenZ.Y.,ChenF.(2003)Heterotrophicproductionofeicosapentaenoicacidbymicroalgae.BiotechnologyAdvances21:273294.
WilkieA.C.,MulbryW.W.(2002)Recoveryofdairymanurenutrientsbybenthicfreshwateralgae.BioresourceTechnology84:81
91.
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8.3.2
Phaeodactylumtricornutum
Figure44Lightmicrographsof
Figure45Scanningelectronmicroscopyimagesof
thefusiform.
(a)thethreemorphotypes:left,fusiform;
(a)triradiate(b)andovoid(c)morphotypesofP.
topright,triradiate;bottomright,oval.(b)a
tricornutum
smallclusterofcells.Eachcellis
approximately15minlength
(Franciusetal.,2008)
ImagescourtesyofAlessandraDeMartino
(Vardietal.,2008)
SYMBOLS:D,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Bacillariophyceae
Naviculales
Phaeodactylaceae
Phaeodactylum
Relatedspecies
PhaeodactylumtricornutumistheonlyspeciesinthegenusPhaeodactylum.
BIOLOGY
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Generaldescription
Phaeodactylumbelongstothepennatediatoms(Bacillariophyceae).Itisamemberoftheraphidpennate
clade(MedlinandKaczmarska,2004).ExaminationofintraspeciesgeneticdiversitybasedonrDNAinternal
transcribed spacer 2 (ITS2) sequences and amplified fragment length polymorphism (AFLP) analyses
indicatefourdifferentgenotypeswithinthespecies(DeMartinoetal.,2007).
Phaeodactylum tricornutum is fast growing and has emerged as a model system for physiological,
biochemical, and molecular studies mainly because of its ease of culture and the ability to be routinely
geneticallytransformed(Scalaetal.,2002,Montsantetal.,2005).
It was chosen as the second diatom for whole genome sequencing after the centric diatom
Thalassiosira pseudonana. The completed P. tricornutum genome, sequenced by the Joint Genome
Institute(JGI),isapproximately27.4megabases(Mb)insizeandispredictedtocontainfewergenesthanT.
pseudonana(Bowleretal.,2008).P.tricornutumshares57%ofitsgeneswithT.pseudonana.Aremarkably
high number of P. tricornutum predicted genes appears to have been transferred between diatoms and
bacteriaandarelikelytoprovidenovelmetaboliccapacities(Bowleretal.,2008).
Phaeodactylumtricornutumisunicellularorformssmallcellclusters.Cellsarecirca3mwideand8to20
mlong(Lewinetal.,1958),andcontainasingleplastid(Roundetal.,1990).Unlikeotherdiatomsitcan
exist in different morphotypes (i.e. fusiform, triradiate or oval) (Borowitzka and Volcani, 1978). This
plasticityisrelatedtotheatypicalnatureofthecellwall,whichisonlypoorlysilicifiedcomparedwithother
diatoms.Inaddition,Phaeodactylumdoesnotexhibitthesizereductionrestitutioncyclethatissounique
for the diatoms. The species appears to be unique in that it does not have an obligate requirement for
silicicacid.Theovoidformistheonlymorphotypewhichisabletosynthesizetruesinglesilicavalveswhen
growninthepresenceofsilicicacid.Siliceousfrustuleswereneverobservedinfusiformortriradiatecells,
or in oval cells grown in the absence of silicic acid (De Martino et al., 2007). Fusiform and triradiate
morphotypesarecharacterizedbycellwallspossessingalmostexclusivelyorganiccomponents(DeMartino
etal.,2007;Franciusetal.,2008).
Physiologicalcharacteristics
Although not considered to be of great ecological significance, P. tricornutum has been found in several
locationsaroundtheworld,typicallyincoastalareaswithwidefluctuationsinsalinityaswellasininland
waters(Rushforthetal.,1988).
The ability of the species to adapt to changing environmental conditions could be related to the
pleiomorphiccharacterofthecellsandthedifferentmorphotypesarethoughttobeadaptedforsurvivalin
different habitats. It is shown that the three morphotypes are physiologically different. The oval
morphotype appears to be induced as an acclimation response to suboptimal growth conditions (De
Martino et al., 2007). The oval and triradiate morphotypes may represent distinct ecophenotypes each
one specifically adapted to growth in particular conditions (De Martino et al., 2007). In contrast to oval
cells,triradiatecellsappeartobemuchmoresensitivetostressandrapidlydisappear(orconverttooval
fusiformcells)whengrowthconditionsaresuboptimal(DeMartinoetal.,2007).
Fusiform and triradiate forms are common when grown in liquid media. Fusiform cells tend to
transformintotheovoidmorphotypewhentheyaretransferredonasolidmedium(agar)whilethereverse
transformation occurs upon transfer in liquid medium (Lewin et al., 1958). It has also been noted that
fusiformandtriradiatecellsaremorebuoyantthanovalcells(Lewinetal.,1958)andsowouldbeexpected
to be better adapted to a planktonic lifestyle than oval cells. De Martino et al. (2007) suggested that P.
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tricornutum should predominantly occur in benthic communities because oval and round cells tend to
aggregateandsinkandcanadherestronglytosurfaces.Verylittleisknownonthenatureandbiological
roleofthetriradiatefusiformtransition(Franciusetal.,2008).
P. tricornutum is extensively used as a food source for the aquaculture industry because of its ease of
cultivationanditsrichoilcontent.However,different culture conditionsresultinsignificantvariationsin
thebiochemicalcompositionofthecellsand,therefore,intheirnutritiousvalue.
Onaverage,thebiomassofP.tricornutumcontains(RebollosoFuentesetal.,2001):
36,4%crudeprotein,
26,1%availablecarbohydrates,
18,0%lipids,
15,9%ash.
The biomass produced at low external irradiance was richer in protein and eicosapentaenoic acid
(RebollosoFuentesetal.,2001).
Thetotallipidcontentandthefattyacidandlipidclass(polarmembranelipidsvsneutralstoragelipids)
composition are dependent on different chemical (nutrient starvation, salinity, growthmedium pH) and
physicalstimuli(temperatureandlightintensity).Inadditiongrowthphaseand/oragingoftheculturehas
an effect on oil content and composition (Hu et al., 2008). This hampers rigorous comparison across
experimentsunderdifferentconditions(GriffithsandHarrison,2009).
GriffithsandHarrison(2009)calculatedtheaveragetotallipidcontentforP.tricornutumfromavailable
literaturedata:
21%cdw(celldryweight)undernutrientrepleteconditions,
26%cdwunderNdeficientconditions.
Otherliteraturementionvaluesfortotaloilcontentof:
31%dw(Sheehanetal.,1998),
18.7%biomassdw(Rodolfietal.,2009),
1857%dw(Mataetal.,2010).
ThefattyacidcompositionandthepartitioningintostoragelipidsinP.tricornutumisintenselystudied,
e.g. (Alonso et al., 1998, 2000; RebollosoFuentes et al., 2001; Tonon et al., 2002; Meiser et al., 2004;
Alonsoetal.,1998;Liangetal.,2006;Meiseretal.,2004;Ohseetal.,2009;RebollosoFuentesetal.,2001;
Tononetal.,2002;Yuetal.,2009).
In Phaeodactylum, the verylongchain fatty acids (vlc PUFA) arachidonic acid (AA) (C20:46),
eicosapentaenoicacid(EPA)(C20:5x3)anddocosahexaenoicacid(DHA)(C22:63)arethemajorfattyacid
speciesaccountingforapproximately30%ofthetotalfattyacidcontent(Huetal.,2008).
P.tricornutumisabletoproducethenutritionallyrelevantEPA(3.95%CDW)inhighproportiontothe
totalfattyacidcontentandhasimportantadvantagesasapotentialcommercialproducerofEPAbecauseit
isfastgrowingwithlowamountsofotherPUFAs,suchasDHAandAA,whichhasimportantadvantagesin
simplifying recovery. (Lebeau and Robert, 2003; Meiser et al., 2004). Unlike in most algal species/strains
examinedwhereTAGsarecomposedprimarilyofC14C18fattyacids(saturatedormonounsaturated)in
P.tricornutum,partitioningofverylongchain(>C20)intoTAGshavebeenobserved(Huetal.,2008).The
TAGyieldfromP.tricornutumisabout14%oftotaldryweight(Yuetal.,2009).
Phosphorus limitation result in increased lipid content, mainly TAG, in P. tricornutum (Reitan et al.,
1994).Phosphorusdeprivationwasfoundtoresultinahigherrelativecontentof16:0and18:1andalower
relativecontentof18:4x3,20:5x3and22:6x3(Reitanetal.,1994).
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Adecreaseofthenitrogenconcentrationcausedthegalactolipid(GL)fractiontodecreasefrom21to
12%.Conversely,bothneutrallipids(NLs)andphospolipids(PLs)increasedfromabout73to79%andfrom
6 to 8%, respectively. TAG was the lipid class with the highest increase, from 69 to 75% (Alonso et al.,
2000).
Unlikeinsomeotherdiatoms,cultureagehadalmostnoinfluenceonthetotalfattyacidcontentinP.
tricornutumthatremainedaround11%ofdryweight(Alonsoetal.,2000).Conversely,cultureagehada
greater impact on lipid classes, producing changes in amounts of triacylglycerols (TAG) which ranged
between 43% and 69%, and galactolipids (GLs) that oscillated between 20% and 40%. In general, the
contentofpolarlipidsofthebiomassdecreasedwithcultureage(Alonsoetal.,2000)..
CarotenoidsinPhaeodactylumtricornutumarelow,rangingfrom0.115to0.45g/100gdrybiomass.In
P.tricornutum,chlorophyllsarethemainpigments(1.172.87g)(RebollosoFuentesetal.,2001).
Growthkineticsandproductivity
Growthrateandbiomassproductivityareinfluencedbyenvironmentalconditions,availableresourcesand
choiceofculturesystem.
Ingeneral,P.tricornutumshowsgoodgrowthattemperaturesbetween15and25C.Formostisolates,
growthceasesattemperaturesabove30C.Inrelationtotheappropriateconditionsforthecultures,some
researchershavefoundinthisspeciesareductioninthephotosynthesisactivityofnearly75%atpHvalues
above9.0andunder5.5.Ontheotherhand,ithasalsobeeninformedthat,forrestrictivelightconditions,
the specific growth rate decreases at a pH above 9.0. The influence of illumination in microalgae growth
ratesisoneofthemostcontroversialfactors.Accordingtosomeinvestigators,P.tricornutumusesaphoto
adaptation strategy, developing an increase of the cellular content of chlorophyll under limiting light
conditions.Thiseffectcausesthatthephotosyntheticefficiency,andconsequentlythespecificgrowthrate,
changesverylittleforawideirradiancerange(reviewedinPerezetal.,2008).
TheinfluenceofseveralcultureconditionsonthespecificgrowthrateofP.tricornutumwasstudiedin
batchculturesbyPerezetal.(2008).TheycalculatedanoptimumpHof7,8andaspecificgrowthrateof
0.064h1wasachievedforcertainnitrateconditionsandillumination.However,forpHvaluesbetween6
and9growthwasfavorableanddidnotshowgreatvariation.Thetemperatureoptimumwasachievedat
20.4Cinaeratedculturesandat22.3Cinnonaeratedcultures.Betteradaptationtolowtemperatures
thanhighoneshasbeenobtained.Theexperimentscarriedoutwithdifferentirradiancesdrivetoasimple
Monodsequationfortheirradianceinfluenceongrowth,withsemisaturationirradianceof10.2and6.8
molphotonsm2s1foraeratedandnonaeratedcultures,respectively.Furthermore,aerationaffectsto
thegrowthofP.tricornutum,withhighergrowthratesunderaeratedconditions(Perezetal.,2008).
Analysisofbiomassproductivity,lipidcontentandtheircombinationtoyieldlipidproductivityhasbeen
done across literature data by Griffiths and Harrison (2009). Lipid productivity is a critical variable for
evaluatingalgalspeciesforbiodieselproduction,becauselipidcontentdoesnotcorrelatedirectlywithlipid
productivity.
Average literature data for P. tricornutum grown in laboratory under nutrient replete conditions
Laboratory averages under nutrient replete conditions from literature for P. tricornutum (Griffiths and
Harrison,2009):
biomassgrowth(averagedoublingtime):25h
biomassproductivityonavolumetricbasis:0.34gL1day1
biomassproductivityonasurfaceareabasis:20gm2day1
lipidproductivitycalculatedfromlaboratorybiomassproductivity(ingramsperlitreperday)and
nutrientrepletelipidcontentinnutrientrepleteconditions:72mgL1day1
lipidproductivityreporteddirectlyintheliterature:45mgL1day1
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Rodolfietal.(2009)showedabiomassproductivityof0.24gL1day1andalipidproductivityof44.8mg
L1day1forP.tricornutumF&MM40straincultivatedin250mLflasks.
BIOTECHNOLOGY
Paheodactylum tricornutum is widely used in aquaculture as feed for penaid shrimp larve, freshwater
prawnlarvae,bivalvelarvaeandpostlarvaeandmarinezooplankton(Tredicietal.,2009).Phaeodactylum
hasbeenproposedasasourceofeicosapentaenoicacid(EPA,20:53)(Velosoetal.,1991;MolinaGrimaet
al.,1994).Itisaspeciesofinterestforbiodieselproduction.Phaeodactylumextractsareusedincosmetics
(Nizard et al., 2007; http://www.cosmeticsdatabase.com/). Fatty acids from Phaeodactylum have shown
antibacterialactivity(Desboisetal.,2009)andtheywerefoundinhigheramountsinthefusiformthanin
the oval cell form (Desbois et al., 2010). Antibacterial and antiinflammatory activities were found in
polysaccharides(GuzmanMurilloandAscencio,2000;Guzmanetal.,2003),whilegalactolipidswerefound
toinduceapoptosisinmammalianengineeredcelllines(Andrianasoloetal.,2008).
Growthmedium
P.tricornutumisusuallygrowninMannandMyers(1968)medium.
Cultivationsystemsandmethods
In Spain, biomass productivity in an outdoor 200L airlift tubular photobioreactor operated in
continuouswas1.21,.9gL1day1(1932gm2day1),dependingondilutionrateandsuperficialliquid
velocity (Molina et al., 2001). In the same location, in a 75L helical tubular photobioreactor biomass
productivies up to 1.3 g L1 day1 with a photosynthetic efficiency up to 15% were obtained (Hall et al.,
2003).InGermany,inanairliftflatpanelsystemwith33Lmodules,abiomassproductivyof0.530.73gL1
day1wasobtainedinmidyearmonths,whileonanannualbase,anaverageof27mgL1day1ofEPAwas
achieved (http://www.igb.fraunhofer.de/www/gf/umwelt/algen/en/Eicosapentaensaeure_en.html). In
Portugal,in2.2m2ponds,averageproductivitiesof4g(ashfreedryweight)m2day1wereachieved,with
anEPAproductivityof0.15gm2day1(Velosoetal.,1991).EPAproductivitiesupto47.8mgL1day1were
achieved in the airlift tubular photobioreactor, with biomass productivity up to 2.57 g L1 day1 (Molina
Grimaetal.,1994).
Phaeodactylumcanalsogrowmixotrophicallyondifferentcarbonsources,amongwhichglycerol(the
byproduct of biodiesel production) yielding higher biomass and EPA productions than those obtained in
photoautotrophicconditions(CronGarciaetal.,2005).Inthree60Lreactors,abubblecolumnandtwo
airlift photobioreactors, outdoors an increase in biomass and EPA productivity was achived by adding
glycerol(FernndezSevillaetal.,2004).
Harvestingmethods
Phaeodactylumtricornutmcanbeharvestedbycentrifugation,e.g.atat1800g(Ibez Gonzleset al.,
1998). Flocculation was studied by Veloso et al. (1991) and calcium hydroxide was found to be the best
flocculantatconcentrationsof30100mgL1.Theadditionofchitosandidnotimprovedtheefficiencyof
flocculation.
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8.3.3
Chaetocerosmuelleri
Figure46Lightmicroscopicpicture
ofChaetocerosmuelleri.
InstantAlgae
Figure47Scanningelectron
microscopicpictureofChaetoceros
sp.
Figure48Scanningelectron
microscopicpictureofaresting
sporeofChaetocerossp.
PictureformSDSUCenterForInland
Water
PictureformSDSUCenterFor
InlandWaters
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Mediophyceae
Chaetocerotales
Chaetocerotaceae
Chaetoceros
Chaetocerosmuelleri
Relatedspecies
Chaetoceros muelleri var. subsalsum Johansen & Rushforth 1985. The taxonomic relationships are not
adequatelyresolvedinChaetoceros(see1.5)andseveralcrypticspecieswithinC.muelleriareconceivable
(Johansenetal.,1990).
Potentiallyimportantforbiofuel:Chaetoceroscalcitrans
BIOLOGY
ThediatomgenusChaetocerosisamongthelargestgeneraofdiatomsandcomprisesseveralhundredsof
species.Thegenusbelongstothebiormultipolarcentricclass(Mediophyceae),withChaetoceroshavinga
bipolarvalveoutline.Thecellsoccurinchainsheldtogetherbylongspinescalledsetaethatextendfrom
thecornersofthecells.Manyspeciesformrestingspores,whichareoftenabundantinthefossilrecord.
Therefore,thisgenusisimportantwithinthedisciplinesofmarinebiology,marinegeology,oceanography,
and aquaculture. Chaetoceros is usually divided in 2 subgenera which are further divided in numerous
sections:
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subgenus Phaeoceros with species characterized by numerous chloroplasts throughout the cell and
robustspinysetaecontainingchloroplasts.
subgenus Hyalochaete consisting of species having many parietal plastids and thin setae lacking
chloroplasts.
TheevolutionaryhistoryofthegenusChaetocerosispoorlyknown,andtheclassificationlargelybased
onobservationsofthemorphologyandultrastructureofsomespecies.Moleculardatawillbenecessaryto
unravelphylogeneticrelationships.
CellsofChaetocerosarejoinedinchainsthatarecoiled,curvedorstraight.Occasionallythecellsare
solitary.Cellsarenarrowlytobroadlyellipticalinvalveviewandrectangularingirdleviewandareunited
by fusion or interlocking of setae produced from the valve. They contain one or more small platelike
plastids. Cell width (apical axis) varies with species, roughly ranges from <10 m to 50 m. Vegetative
cells/colonies are weakly silicified. It can be difficult to identify Chaetoceros taxa because they are
notoriouslyvariableinmorphology.C.muelleriisalsoavariabletaxon,bothinvegetativecellsandresting
spores and it has been reported under more than 10 different specific names. Most populations of non
marine, noncolonial Chaetoceros species have been reported as C. muelleri. Two different forms of C.
muellerihavebeendescribed:thenominateformC.muellerivar.muelleriischaracterizedbythefrequent
presenceofasmallprocessonthevalvefaceassociatedwithchainformation(chainsofonly24cells)and
smoothlycurvedsetae,whileC.muellerivar.subsalsumlacksvalvarprocessesandconsequentlydoesnot
formcolonies.Bothformshavenoornamentationontheirrestingspores.
Proximate composition (dry weight percentage) in C. muelleri is 43.1% crude protein, 17.1 %
carbohydrates,21.5%lipidsand18.3%ash.
Chaetoceros is an important marine planktonic genus and some species are major bloom formers in
both oceanic and coastal habitats. Only a few species have been recorded in fresh water. Chaetoceros
muellerihas beenreportedmainlyfrombrackishwatersofvaryingsalinities andtemperaturesinEurope
and North America. The species is only rarely reported from freshwater habitats and has never been
recordedfromtruemarinesystems.
BIOTECHNOLOGY
Chaetocerosmuelleriisusedintheaquacultureindustry,primarilyasfeedforbivalveshellfishandshrimp
(Brown et al., 1997). This is due to its fatty acid profile, comprising 520% eicosapentaenoic acid (EPA,
C20:53),0.21%docosahexaenoicacid(DHA,C22:63)and<0.2%arachidonicacid(AA,C20:46)ofthe
totalfattyacidcontentanditsappropriatesizeforfeedingofthelarvae(e.g.58m)(Brownetal.,1997).
Chaetocershasbeenevaluatedasasourceforbiodieselproduction,althoughonlyatlaboratoryscale.
The average total lipid content for C. muelleri is about 19% cell dry weight under nutrient replete
conditions, 27% under N deficient conditions, 36% under Si deficient conditions (Griffiths and Harrison,
2009).Thehighlipidcontentofthealgaeofthisgenus(upto40%)iscounterbalancedbyalowbiomass
productivity(70mgL1day1)thatmakestheexploitationofthisalgadifficult(Rodolfietal.,2009).
Outdoor mass cultivation of Chaetoceros is usually carried out in the hatcheries, often indoors, in
cylinders and tanks that do not allow high productivities (Tredici et al., 2009). In 3000L tanks, biomass
productivity of C. muelleri ranged from 22.5 to 45.7 mg L1day1 indoors, while outdoors the mean
productivityvariedfrom29.0to69.7mgL1day1inwinterandspring,respectively(LpezElasetal.,2005).
Ina350LFlatPlatephotobioreactoroutdoors,ZhangandRichmond(2003)obtainedwithC.muellerivar.
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subsalsumproductivitiesashighas0.15gL1day1withadailyharvestof10%andalightpathlengthof20
cm.
References
Blinn D.W. (1984) Growthresponses to variations in temperature and specific conductance by Chaetoceros muelleri
(Bacillariophyceae).BritishPhycologicalJournal19:3135.[BIOLOGYsection]
315331.
GranH.H.(1897)Botanik.Prophyta:Diatomaceae,SilicoflagellataogCilioflagellata.DenNorskeNordhavsExpedition18761878.7,
pp.136.[BIOLOGYsection]
Hasle G.R., Syvertsen E.E. (1997) Marine diatoms. In: Tomas C.R. (ed.) Identifying Marine Phytoplankton, Academic Press, San
Diego,pp.5385.[BIOLOGYsection]
JaimeCeballos B.J., HernndezLlamas A., GarciaGalano T., Villarreal H. (2006) Substitution of Chaetoceros muelleri by Spirulina
platensismealindietsforLitopenaeusschmittilarvae.Aquaculture260:215220.[BIOLOGYsection]
Johansen J., Rushforth S. (1985) A contribution to the taxonomy of Chaetoceros muelleri Lemmermann (Bacillariophyceae) and
relatedtaxa.Phycologia24:437447.[BIOLOGYsection]
LpezElas J.A., Voltolina D., EnrquezOcaa F., GallegosSimental G. (2005) Indoor and outdoor mass production of the diatom
Chaetocerosmuelleriinamexicancommercialhatchery.AquaculturalEngineering33:181191.
Medlin L.K.,Kaczmarska I. (2004) Evolution of the diatoms: V. Morphological and cytological support for the major clades and a
taxonomicrevision.Phycologia43:245270.[BIOLOGYsection]
Ostenfeld C.H. (1903) Plankton from the sea around the Fres. In: Warming, E. (ed.) Botany of the Fres, Nordisk Forlag,
Copenhagen,pp.588611.[BIOLOGYsection]
102:100112.
RoundF.E.,CrawfordR.M.,MannD.G.(1990) Thediatomsbiologyandmorphologyofthegenera,CambridgeUniversityPress,
Cambridge,UK.[BIOLOGYsection]
uses. In: Burnell G., Allan G. (eds.) New technologies in aquaculture: Improving production efficiency, quality and
environmentalmanagement,WoodheadPublishing,Cambridge,pp.610676.
Zhang C.W., Richmond A. (2003) Sustainable, highyielding outdoor mass cultures of Chaetoceros muelleri var. subsalsum and
Isochrysisgalbanainverticalplatereactors.MarineBiotechnology5:302310.
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8.3.4
Cyclotellacryptica
Figure49Lightmicroscopicviewofa
chainoflivingcellsfromstrain
CCMP332inCCMPofCyclotella
cryptica.
Figure50SEMofthevalveofC.
cryptica.
Figure51SEMofthevalveofC.
cryptica.
Overallviewofthedistalvalve
surface.Arrowdenotesthelocation
ofafultoportulaoffsetfromthe
valvecenter
Proximalviewofthevalve.Wide
ribsarepredominantanda
fultoportulaislocatedbyan
arrow.
Picturefrom(Tesson,Hildebrand,
2010)
B.Beszteri,AWI,Bremerhaven
Picturefrom(Tesson,Hildebrand,
2010)
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Mediophyceae
Thalassiosirales
Stephanodiscaceae
Cyclotella
Cyclotellacryptica
Relatedspecies
Agenusofaround100species.Thereare230speciesnamesinthealgaedatabaseatpresent,ofwhich108
havebeenflaggedascurrentlyacceptedtaxonomically.
BIOLOGY
Cyclotella is a radial centric genus of around 100 species. The genus is part of the Mediophyceae class
comprising bi and multipolar centrics and the radial Thalassiosirales, the order including Cyclotella. The
Thalassiosirales are a sistergroup of the pennates. Strutted processes (fultoportula) through which chitin
threadsaresecretedforchainformationandflotationarerestrictedtotheThalassiosiralesandaresimple
tubeswhichpenetratesthesilicaframeworkwithadjacentpores.
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Cyclotellacellsareshort,drumshaped,freelivingorformingfilaments,chainsorrarelyclusters,united
by mucilage. They have numerous discoid plastids. C. cryptica exhibits very stiff, thin, crystalline, and
chitinous fibril appendages that attach to the pores of its valves. The valves are almost flat to slightly
concentricallyundulate.ThediameterofC.crypticarangesfrom525m.C.crypticahasbeenshownto
display extreme morphological plasticity depending on salinity. Its frustules show the morphology
consideredcharacteristicofC.meneghiniana,acloselyrelatedspecies,whengrowninlowsalinitymedium,
whereas they produced the typical C. cryptica morphology at higher salinities. C. cryptica can produce
auxosporesfromvegetativecells.
C.crypticaisaplanktonicspeciesoccurringinmarineandbrackishenvironments.ThedistributionofC.
cryptica is widespread. In marine and brackish environments, C. cryptica requires NO3 as its source of
nitrogen.Itiscapableofheterotrophicgrowthinthedarkonglucose.Thissuggestsitcansurviveinbottom
waters or muds with high glucose content for extended periods of time. It probably recommences
photoautotrophic growth higher in the water column when environmental conditions improve. It is also
knowntogrowphotoheterotrophically.
BIOTECHNOLOGY
Cyclotellaspeciesareculturedforaquaculture(brineshrimp,Artemia)(LebeauandRobert,2003),mainly
due to its ability to prodcuce EPA. Wood et al. (1999) compared the growth of C. cryptica on different
organic carbon substrates and on medium without organic carbon sources: growth was stimulated by
supplementationwithglucoseandglycerolandwasinhibitedbyacetate,andgrewbetteronnitratethan
onammonia.Thefattyacidcompositionseemedtobeaffectedprincipallyinthedegreeofunsaturation,
that was lower with organic carbon. Under heterotrophic conditions in 19L carboys Pahl et al. (2010)
reached2.1gL1after12daysofcultivation.
The first successful transformation of a microalgal strain with potential for biodiesel production was
achievedin1994,withsuccessfultransformationofthediatomsCyclotellacrypticaandNaviculasaprophila
(Dunahayetal.,1995).Thisopenedwaysforgeneticengineeringforenhancedlipidproduction.
In C. cryptica a dramatic increase in the lipid content of the cultures was seen under Ndeficient
conditionsincellsgrown at30C:the totallipids,asapercentageofAFDW, increasedfrom15%to44%,
and the increase in total lipids was due to increases in both the neutral lipid and polar lipid fractions
(Sheehanetal.,1998).ACyclotellaspecies,exhibitedanincreaseoflipidcontentofmorethan40%ofdry
weightuponSilimitation.However,lipidproductivity(9gm2day1),wasnotsignificantlydifferentbetween
Sideficient and the Sisufficient controls, because of the high productivity of the Si sufficient culture
(Sheehanetal.,1998).
BiomassproductivityforC.crypticagrowninoutdoorpondsinCaliforniawasabout30gm2day1with
aphotosyntheticefficiencyof7.0%(Sheehanetal.,1998).
References
DunahayT.G.,JarvisE.E.,RoesslerP.G.(1995)GenetictransformationofthediatomsCyclotellacrypticaandNaviculasaprophila.
HerthW.,ZugenmaierP.(1977)UltrastructureofthechitinfibrilsofthecentricdiatomCyclotellacryptica.JournalofUltrastructure
Research61:230239.[BIOLOGYsection]
Lebeau T., Robert J.M. (2003) Diatom cultivation and biotechnologically relevant products. Part I: Cultivation at various scales.
Medlin LK., Kaczmarska I. (2004) Evolution of the diatoms: V. Morphological and cytological support for the major clades and a
taxonomicrevision.Phycologia43:245270.[BIOLOGYsection]
PahlS.L.,LewisD.M.,ChenF.,KingK.D.(2010)Growthdynamicsandtheproximatebiochemicalcompositionandfattyacidprofile
oftheheterotrophicallygrowndiatomCyclotellacryptica.JournalofAppliedPhycology22:165171.
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SchultzM.E.(1971)SalinityrelatedpolymorphisminthebrackishwaterdiatomCyclotellacryptica.CanadianJournalofBotany49:
12851289.[BIOLOGYsection]
biodieselfromalgae.
Tesson B., Hildebrand M. (2010) Dynamics of silica cell wall morphogenesis in the diatom Cyclotella cryptica: Substructure
formationandtheroleofmicrofilaments.JournalofStructuralBiology169:6274.[BIOLOGYsection]
WhiteA.W.(1974)Growthoftwofacultativelyheterotrophicmarinecentricdiatoms.JournalofPhycology10:292300.[BIOLOGY
section]
Wood B.J.B., Grimson P.H.K., German J.B., Turner M. (1999) Photoheterotrophy in the production of phytoplankton organisms.
JournalofBiotechnology70:175183.
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8.3.5
Odontellaaurita
Figure52LightmicroscopicviewshowingOdontellaaurita
PicturefromAlgaebase.com(KarlBruun)
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Mediophyceae
Triceratiales
Triceratiaceae
Odontella
Odontellaaurita
Relatedspecies
Synonymandrelatedspecies:
Odontellaauritavar.minima(Grunow)DeToni
OdontellasubaequaKtzing
BIOLOGY
Odontella aurita cells are oblong in girdle view, with long 'spines' and raised apical elevations. It often
formschainslinkedbytheprocesses,withthetubularspinescrossingeachother.Cellshavemanyplastids,
small and discoid. The diameter varies from 10 to 100 m. Valves are elliptical or lanceolate, with no
separation into face and mantle. Valve face is plain or with fine granules, spinules or spines, sometimes
withtworidges(whichmaybefibriate)runningoneithersidedelimitinganellipticalareainthecentre.At
eachendthereisanelevation,sometimeslowandblunt,elsewherehornlike,whichbearsanocellus.Wall
isloculate,withfineexternalporesandroundinternalforamina.Theedgeofthevalvemantleissometimes
recurved so that a groove runs around just above the free edge. The spines, which are very variable in
length,areactuallytheexittubesoftherimoportulae,andareplacedinthecentreofthevalvesorcloseto
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thebasesoftheelevations,diagonallyoppositeeachother;theycanhavesmallapicalspinules.Internally
the rimoportulae are sessile and lie in slight depressions. Copulae are split, with ligulae and clustered
fimbriaealongtheadvalvaredge;areolaearesimple,inverticalrows.Thevalvocopulaismodifiedtofitthe
'sculptured'edgeofthevalvemantle.
O.auritaisamarinealga,planktonicorepiphytic.Itisveryabundantthroughouttheoceans.
BIOTECHNOLOGY
O.auritacontains22%ofeicosapentaenoicacid(EPA,20:53)and0.91.7%ofdocosahexaenoicacid(DHA,
22:63)withrespecttotoalfattyacidcontent.Ithasbeenapprovedforhumanconsumptionin2002by
theEuropeanCommunityandismainlysoldinformofdriedbiomassasadietarysupplement(Trediciet
al.,2009).
O.auritaiscultivatedinopenpondsintheVandeRegioninFrance.ItdevelopsfromtheendofApril
until half October. With stable growing conditions the average doubling time is 2.6 days and the specific
growthrateisof31%day1.O.auritaisharvestedmechanicallybysedimentationorbyfiltration,followed
bycentrifugation.Thealgalpasteisthenfrozen,andislyophilised(freezedried)beforemarketing.
O.auritaisarichsourceofthecarotenoidfucoxanthin(Moreauetal.,2006).Thismoleculeexhibited
cytostatic activity and this effect could have important implications for the application of this kind of
microalgae in food manufacturing and the formulation of ocular implant products used in cataract
treatment(Moreauetal.,2006).
Odontellaauritaisalsousedincosmeticindustry(Patt,2008;RedziniakandDonguy,2009).
Althoughthisalgaiscommercially producedandit isreportedtohavelipid contentsofabout713%
under nutrient replete conditions, no studies have been carried out to assess its biodiesel production
potential.
References
MoreauD.,TomasoniC.,JacquotC.,KaasR.,LeGuedesR.,CadoretJ.P.,MullerFeugaA.,KontizaI.,VagiasC.,RoussisV.,Roussakis
C.(2006)CultivatedmicroalgaeandthecarotenoidfucoxanthinfromOdontellaauritaaspotentantiproliferativeagentsin
bronchopulmonaryandepithelialcelllines.EnvironmentalToxicologyandPharmacology22:97103.
PattL.M.(2008)Methodsandcompositionsforpreventingandtreatingagingorphotodamagedskin.USPatentNo.7,384,916.
Redziniak G., Donguy D. (2009) Use of fatsoluble extract of Odontella aurita for restructuring skin, composition for use and
cosmeticmethodusingtheextract.JapanesePatentJP2009029806.
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8.3.6
Skeletonemasp.
Figure53LightmicroscopicviewshowingSkeletonemacostatum
PicturefromAlgaebase.com(KarlBruun)
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Mediophyceae
Thalassiosirales
Skeletonemaceae
Skeletonema
Relatedspecies
BIOLOGY
Cellsaregenerallycolonialandcontainoneortwocupshapedchloroplasts.Cellsjoinedbylongmarginal
processestoformfilaments,whichappearinthelightmicroscopelikeshortbeadsjoinedbynumerousfine
threads.Valvescircular;valvefaceconvextoflat;mantlesdeep.Valveswithaprominentnetworkofcostae
externally,becomingpseudoloculatenearthemargin.Internallywithdistinctcribraontheflatorslightly
ribbedsurface.Asingleringofprocessesoccursaroundthetopofthemantle.Thesearecloselyassociated
with a ring of fultoportulae, the external openings of which are short tubes hidden in the bases of the
processes. The processes are semicircular in crosssection and expand at their apices to form 'knuckles'
whichinterlockwiththeprocessesfromtheadjacentcell;theyaresometimesmuchlongerthanthecell
and can interlock with either one or two processes of the sibling valve. Apical axis is 221 m while
pervalvar axis is 261 m. Discrimination among species of the Skeletonema costatum group is very
difficult.
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Skeletonema occurs in coastal waters throughout the world where it can be an extremely common
diatom. Several species are reported in various oceanic regions but all these need reinvestigation. In
additionseveralfossilspecieshavebeendescribed.
BIOTECHNOLOGY
Skeletonema is widely used in aquaculture as bivalve larvae andpostlarvae and for penaeid shrimp larve
(Tredicietal.,2009).Grosscompositionisabout40%protein,20%carbohydratesand20%lipidand,among
fatty acids, about 6% eicosapentaenoic acid (EPA, 20:55) and 2% docosahexaenpic acid (DHA, 22:63)
(Brownetal.,1989;Rodolfietal.,2009).
Skeletonemaisworldwidecultivatedinhatcheriesusinglowproductivitysystemssucastanks,sleeves
andcylinders(Tredicietal.,2009).Biomassproductivitiesareusuallyverylow(intheorderof104cellsml1
day1,correspondingtotensofmilligramsdryweightperliterperday)(Laing,1991;Popovichetal.,2011).
Outdoorsin1000m3cultivationvolume,madeof20unitsof50m34mdepthand28m2surfacearea,12
tonsofdrybiomassareproducedyearly(Kittoetal.,1999).Weiss(2008)reportsabiomassproductivityof
about 20 g m2 day1 cultivating S. costatum in ponds with waste CO2 and turbine cooling seawter.
Skeletonemacanbeharvestedthroughautoflocculationandsedimentation(Weiss,2008).
AnextractfromthepeptidicfractionofSkeletonemacostatumcontainingtheactiveprinciplenamed
grevillineisusedincosmeticsasaskintreatmentforerythema(RocquetandReynaud,2008).
Skeletonemahasbeenlistedamongpossibleinterestingstrainsforbiodieselproduction(Rodolfietal.,
2009;Popovichetal.,2011),althoughatpresentnodataaboutpilotscalecultivationforthispurposeare
available.
References
Algaebase:http://www.algaebase.org/search/genus/detail/?genus_id= 43754[BIOLOGYsection]
Brown M.R., Jeffrey S.W., Garland C.D. (1989) Nutritional aspects of microalgae used in mariculture; a literature review. CSIRO
MarineLaboratories,Report205.
Kitto M.R., Regunathan C., Rodrigues A. (1999) An industrial photosynthetic system for Skeletonema costatum in arid regions.
LaingI.(1991)Cultivationofmarine,unicellularalgae.LaboratoryLeafletNo.67,MinistryofAgriculture,FisheroesandFood,UK.
Popovich C.A., Damiani C., Constenla D., Leonardi P.I. (2011) Lipid quality of the diatoms Skeletonema costatum and Navicula
gregaria from the South Atlantic Coast (Argentina): evaluation of its suitability as biodiesel feedstock. Journal of Applied
Phycology:DOI10.1007/s108110109639y.
RocquetC.,ReynaudR.(2008)Anaturalwaytorelievetheskinfromerythema:grevilline.CosmeticScienceTechnology2008:129
136.
102:100112.
StazioneZoologicadiNapoli[BIOLOGYsection]:
http://www.szn.it/SZNWeb/cmd/ShowArchiveItem?TYPE_ID=SPECIE&ITEM_ID=6883&LANGUAGE_ID=1&_languageId_=2
WeissH.(2008)Methodforgrowingphotosyntheticorganisms.PatentWO2008/107896A2.
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8.3.7
Thalassiosirapseudonana
Figure54Lightmicroscopicviewshowing
Thalassiosirapseudonanaculture
Figure55Scanningelectronmicrographshowing
theintricatecellwallpatternofT.pseudonana
PicturecourtesyofSBAE
PhotobyNilsKrger,GeorgiaInstituteofTechnology
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Bacillariophyta
Mediophyceae
Thalassiosirales
Thalassiosiraceae
Thalassiosira
Relatedspecies
Thereare186speciesnamesinthealgaedatabaseatpresent,ofwhich82havebeenflaggedascurrently
Potentiallyimportantforbiofuel:T.weissflogii
BIOLOGY
Thalassiosira is a radial centric genus including more than 100 species. The genus is part of the
Mediophyceae class comprising bi and multipolar centrics and the radial Thalassiosirales to which
Thalassiosira belongs. Fultoportula or strutted processes are the defining character of the order
Thalassiosirales. A molecular phylogeny showed the the paraphyletic nature of the families
Thalassiosiraceae, Skeletonemaceae, and Stephanodiscaceae within the order and of the genus
Thalassiosira,asdefinedpresently.SomespeciesofThalassiosiramaybecloserrelativesofStephanodiscus,
whereasothersaremorecloselyrelatedtosomespeciespresentlyassignedtothegenusCyclotella(both
fromthefamilyStephanodiscaceae),ascomparedwithotherspeciesofthegenusThalassiosira.
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T. pseudonana was chosen as the first eukaryotic marine phytoplankton for whole genome
sequencing. T. pseudonana is a model for diatom physiology studies, belongs to a genus widely
distributed throughout the world's oceans, and has a relatively small genome (34 mega base
pairs).Inspiteofthefactthatthecentricandpennatelineageshaveonlybeendivergingfor90
million years, their genome structures are dramatically different and a substantial fraction of
genes(40%)arenotsharedbytheserepresentativesofthetwolineages.T.pseudonanacanalso
begeneticallytransformed.
GenerallyThalassiosiracellscontainnumerouschloroplasts;cellsarediscordrumshaped,occurringas
singlecells,aschains(connectedbyacentralorseveralorganicthreads),orinmucilagecolonies.Organic
threadsusuallyextrudefromstruttedprocesses.Thecircular,areolatedvalvespossessoneorsometimes
severalcentralandalwaysmarginalstruttedprocesses.Theycanalsobedistributedoverthewholevalve.
Restingsporesknownforsomespecies.ThemorphologicalidentificationofThalassiosiraspeciesismainly
basedonultrastructuredetailslikethenumberandlocationoftherimoportulae(tubesonvalveconnecting
thediatomprotoplastwiththeoutside)andfultoportulaeprocessesonthevalve.Colonyconfigurationand
theformofthecellsalsoprovidecharactersforspeciesidentificationunderthelightmicroscope.However,
for identification of most species examination of the frustules in the electron microscope is essential. T.
pseudonanaisarelativelysmallspecies(celldiameter215m)withvariablevalvemorphology.Thecell
diameter is larger than the pervalvar axis. T. pseudonana showed no evidence of the size reduction
restitutioncyclethatissouniqueforthediatoms.
T.pseudonanaisacosmopolitanspecieslivinginbothfreshandcoastalwaters.
BIOTECHNOLOGY
Thalassiosiraspeciesareculturedinaquaculturebecausetheirhighnutritionalvalue(LebeauandRobert,
2003).T.pseudonanaisusedasfeedforbivalvemolluskslarvae(Brownetal.,1997;Duerretal.,1998).The
biomassofT.pseudonanacontains3046%protein,636%carbohydrates,2131%lipidsdependingon
harveststageandonlightphotoperiodandirradiancelevel(Brownetal.,1996).
ThalassiosirageneshavealreadycontributedtoresearchandcommercialeffortstoproducevlcPUFAs
intransgeniccropplants(Tononetal.,2004,2005).
Thalassiosirahasbeenevaluatedasapotentialbiodieselproduceronlyatlaboratortscale.Theaverage
totallipidcontentforT.pseudonanacalculatedfromliteraturebyGriffithsandHarrison(2009)isof16%
celldryweightundernutrientrepleteconditionsand26%underNdeficientconditions.Cellsinlogarithmic
phasehavehighproportionsofpolarlipids(79to89%oftotallipid)andlowtriacylglycerol(10%oftotal
lipid).Cellsinstationaryphasecontainlesspolarlipid(48to57%oftotallipid)andmoretriacylglycerol(22
to 45% of total lipid), with an increase in saturated and monounsaturated fatty acids and a decrease in
PUFAs (Brown et al., 1996; Zhukova, 2004). Yu et al. (2009) showed a TAG yield from T. pseudonana of
about 1418% of total dry weight. Silicatestarved cells accumulated an average of 24% more TAGs than
thosestarvedfornitrate;however,thechemotypesoftheTAGsproducedweregenerallysimilarregardless
of the starvation condition employed. Under laboratory conditions, Rodolfi et al. (2009) found for T.
pseudonanalowbiomass(80mgL1day1)andlipid(17.4mgL1day1)productivities.
References
ArmbrustE.V.,BergesJ.A.,BowlerC.,etal.(2004)ThegenomeofthediatomThalassiosirapseudonana:Ecology,evolution,and
metabolism.Science306:7986.[BIOLOGYsection]
BowlerC.,AllenA.E.,BadgerJ.H.,etal.(2008)ThePhaeodactylumgenomerevealstheevolutionaryhistoryofdiatomgenomes.
Nature456:239244.[BIOLOGYsection]
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BrownM.R.,DunstanG.A.,NorwoodS.J.,MillerK.A.(1996)Effectsofharveststageandlightonthebiochemicalcompositionofthe
diatomThalassiosirapseudonana.JournalofPhycology32:6473.
315331.
DuerrE.O.,MolnarA.,SatoV.(1998)Culturedmicroalgaeasaquaculturefeeds.JournalofMarineBiotechnology7:6570.
Hasle G.R. (1976). Examination of diatom type material: Nitzschia delicatissima Cleve, Thalassiosira minuseula Kraaske, and
CyclotellananaHustedt.BritishPhycologicalJournal11:101110.[BIOLOGYsection]
Hasle G.R., Heimdal B.R. (1970) Some species of the centric diatom genus Thalassiosira studied in the light and electron
microscopes.NovaHedwigia31:559597.[BIOLOGYsection]
Hasle G.R., Syvertsen E.E. (1997) Marine diatoms. In: Tomas C.R. (ed.) Identifying Marine Phytoplankton, Academic Press, San
Diego,pp.5385.[BIOLOGYsection]
Hoppenrath M., Beszteri B., Drebes G., Halliger H., Van Beusekom J.E.E., Janisch S., Wiltshire K.H. (2007) Thalassiosira species
(Bacillariophyceae,Thalassiosirales)intheNorthSeaatHelgoland(Germanbight)andsylt(NorthFrisianWaddenSea)A
firstapproachtoassessingdiversity.EuropeanJournalofPhycology42:271288.[BIOLOGYsection]
KaczmarskaI.,BeatonM.,BenoitA.C.,MedlinL.K.(2005)MolecularphylogenyofselectedmembersoftheorderThalassiosirales
(Bacillariophyta)andevolutionofthefultoportula.JournalofPhycology42:121138.[BIOLOGYsection]
Lebeau T., Robert J.M. (2003) Diatom cultivation and biotechnologically relevant products. Part I: Cultivation at various scales.
Poulsen N., Chesley P., Krger N. (2006) Molecular genetic manipulation of the diatom Thalassiosira pseudonana
(Bacillariophyceae).JournalofPhycology42:10591065.[BIOLOGYsection]
102:100112.
TononT.,HarveyD.,QingR,LiY.,LarsonT.R.,GrahamI.A.(2004)Identificationofafattyacid11desaturasefromthemicroalga
Thalassiosirapseudonana.FEBSLetters563:2834.
TononT.,QingR.W.,HarveyD.,LiY.,LarsonT.R.,GrahamI.A.(2005)Identificationofalongchainpolyunsaturatedfattyacidacyl
coenzymeasynthetasefromthediatomThalassiosirapseudonana.PlantPhysiology138:402408.
YuE.T.,ZendejasF.J.,LaneP.D.,GaucherS.,SimmonsB.A.,LaneT.W.(2009)Triacylglycerolaccumulationandprofilinginthemodel
diatomsThalassiosirapseudonanaandPhaeodactylumtricornutum(Baccilariophyceae)duringstarvation.JournalofApplied
Phycology21:669681.
Zhukova N.V. (2004) Changes in the lipid composition of Thalassiosira pseudonana during its life cycle. Russian Journal of Plant
Physiology51:702707.
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8.4 Eustigmatophyceae(phylumHeterokontophyta)
8.4.1
Monodussubterraneus
Figure56LightmicroscopicviewshowingMonodussp.
PicturefromUNIFI
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Xanthophyceae
Mischococcales
Pleurochloridaceae
Monodus
Monodussubterraneus
Heterokontophyta
Eustigmatophyceae
Monodopsidaceae
Monodospis
Monodospissubterranea
Relatedspecies
This genus is of uncertain collocation. Most of the algae collection display Monodus under the
Eustigmatophyceae,butseveraltaxonomiesstillreportthisgenusundertheXanthophyceae.Pigmentsare
similar to Eustigmatophyceae while its cell structure is different from the standard eustimatophyte
structure.Thereare17species(andinfraspecific)namesinthedatabaseatpresent.
Synonym:
Monodussubterraneus=Monodopsissubterranea
Monodusovalis=Pseudocharaciopsisovalis
BIOLOGY
Monodushasbeenpreviouslyclassifiedasaxanthophyte,thenithasbeenmovedtoeustigmatophytes,but
ithasintermediatecharacters.Cellsaresolitary,510mlongandlessthan10mindiameter.Cellsare
freefloating, spherical, ovoid, elliptical or cylindrical in shape. Chloroplast is terminal, yellowgreen, with
embeddedpyrenoid.Reproductionoccursbyautosporesandzoospores;aplanosporesandcystshavealso
beennoted.InMonodussubterraneus(syn.Monodopsissubterranea)zoosporesarenotpresent.Monodus
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is euplanktonic and metaphytic in dystrophic, mesotrophic, and eutrophic ponds, pools, seeps, and lakes
ofteninassociationwithSphagnum,sedges,and/orTypha.
BIOTECHNOLOGY
Monodus subterraneus has been cultivated at pilot plant for its ability to produce high amounts of
eicosapentaenoicacid(EPA,20:55),whichsynthesisinthisalgahasbeenwidelystudied(KhozinGoldberg
etal.,2002;LiuandLin,2005;KhozinGoldbergandCohen,2006).Atpresent,onlyoneditarysupplement
conatianingMonodusforEPAiscommercialised.
Under artificial illumination, in a cultivation system consisting of a helical tubular photobioreactor
(Biocoil)constructedfromtransparentPVCtubing tocontainatotalvolumeof4.5liter,Luetal.(2001)
foundthatbiomassandEPAproductivitieswere1.31.7gL1day1and4456mgL1day1,respectively.
Outdoors,M.subterraneuswascultivatedinaflat,inclinedmodularphotobioreactor(FIMP),consisting
ofaseriesoffourindividual14Lglassreactorsmeasuring70cmhigh,90cmlongand2.8cmwide,Qiang
etal.(1997)obtainedamaximalEPAproductivityof58.9mgL1day1andabiomassproductivityof1.55gL
1
day1 at the optimal cell density of 4 g L1. Lu et al. (2002) cultivated M. subterraneus in a 75L helical
reactorandina57Lbubblecolumnobtaininglowbiomassproductivityinbothphotobioreactors,although
productivity was higher in the helical reactor than in the bubble column regardless of the operational
conditions,withmaximumvaluesof0.54and0.16gL1d1,respectively,andamaximumEPAproductivity
of9mgL1d1.
Monodus has been tested in laboratory as a potential strain for biodiesel production (Rodolfi et al.,
2009).
References
Algaebase:http://www.algaebase.org/search/genus/detail/?genus_id= 45848[BIOLOGYsection]
KhozinGoldbergI.,CohenZ.(2006)Theeffectofphosphatestarvationonthelipidandfattyacidcompositionofthefreshwater
eustigmatophyteMonodussubterraneus.Phytochemistry67:696701.
KhozinGoldbergI.,DidiCohenS.,ShayakhmetovaI.,CohenZ.(2002)Biosynthesisofeicosapentaenoicacid(EPA)inthefreshwater
eustigmatophuteMonodussubterraneus(Eustigmatophyceae).JournalofPhycology38:745756.
Liu C.P., Lin L.P. (2005) Morphology andeicosapentaenoic acid production by Monodussubterraneus UTEX 151. Micron 36: 545
550.
Lu C., Rao K., Hall D., Vonshak A. (2001) Production of eicosapentaenoic acid (EPA) in Monodus subterraneus grown in a helical
tubularphotobioreactorasaffectedbycelldensityandlightintensity.JournalofAppliedPhycology13:517522.
LuC.,AcinFernndezF.G.,CaizaresGuerreroE.,HallD.O.,MolinaGrimaE.(2002)OverallassessmentofMonodussubterraneus
cultivationandEPAproductioninoutdoorhelicalandbubblecolumnreactors.JournalofAppliedPhycology14:331342.
Ott D.W., OldhamOtt C.K. (2003) Eustigmatophyte, Raphidophyte, and Tribophyte Algae. In: Wehr J.D., Sheath R.G. (eds.)
FreshwaterAlgaeofNorthAmerica:EcologyandClassification,Elsevier,Rotterdam,pp.423469.[BIOLOGYsection]
Qiang H., Zhengyu H., Cohen Z., Richmond A. (1997) Enhancement of eicosapentaenoic acid (EPA) and linolenic acid (GLA)
production by manipulating algal density of outdoor cultures of Monodus subterraneus (Eustigmatophyta) and Spirulina
platensis(Cyanobacteria).EuropeanJournalofPhycology32:8186.
102:100112.
Whittle S.J, Casselton P.J. (1975) The chloroplast pigments of the algal classes Eustigmatophyceae and Xanthophyceae. I.
Eustigmatophyceae.EuropeanJournalofPhycology10:179191.[BIOLOGYsection]
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8.4.2
Nannochloropsissp.
Figure57LightmicroscopicviewshowingNannochloropsissalina
Picturefromwww.sbroscoff.fr/Phyto/gallery/main.php?g2_itemId=323
SYMBOLS:D,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Eustigmatophyceae
Eustigmatales
Monodopsidaceae
Nannochloropsis
Relatedspecies
Thereare6species(andinfraspecific)namesinthedatabaseatpresent,alltaxonomicallyaccepted.
Speciesofinterestforbiofuel:
N.gaditana,N.granulata,N.limnetica,N.oceanica,N.oculata,N.salina.
BIOLOGY
Nannochloropsiscells(Karlsonetal.,1996;FawleyandFawley,2007)arenonmotile,sphericaltoovoid,2
4 m in diameter, with a single chloroplast lacking a pyrenoid and containing chlorophyll a. Major
carotenoidsareviolaxanthin,vaucheraxanthinandneoxanthin,besidescarotene.Amongothers,algaeof
thisgenuscansynthesizelowamountsofcanthaxanthinandastaxanthin.Zoosporesarenotpresent,then
reproductionisbyautospores(2to6).Speciesattributionismainlycarriedoutbygeneticanalysis(rbcLand
18SrDNA).Cellspresentthesocalledredbody,aprimarycharacteristicofthevegetativecellsofmany
eustigmatophytes, that is an eyespot located extraplastidially and not surrounded by membranes. The
composition of the cell wall is still unclear. Small refractile bodies can also be observed within the
cytoplasm and can be both mobile (Brownian motion) or immobile. The genus Nannochloropsis includes
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both marine and freshwater species, though the biotechnology of this alga is at present limited to the
marinespecies.
Underartificiallightinabubblemixed29Lphotobioreactor,RebellosoFuentesetal.(2001)havefoundan
average dry biomass composition of: 29% protein, 36% carbohydrate, 18% lipid, 9% ash, and plamitic,
palmitoleic, oleic and eicosapentaenoic acid (on average 2.2%) as major fatty acids. Mineral composition
was on average (mg/100g d.wt): 659 Na, 533 K., 972 Ca, 316 Mg, 136 Fe, 103 Zn, 35 Cu, 3.4 Mn, 529 S.
Brown et al. (1998) report a different composition for Nannochloropsis CS246 tested for oyster rearing:
17%protein,23%carbohydrate,26%lipid,16%ash.
Nannochloropsis is widely cultivated for its high content of eicosapentaenoic acid (EPA; 20:5 3),
presentinamountsupto45%ofthebiomass,andbecauseofitssmallcells(23mdiameter)asfeedfor
rotifersinthegreenwatertechnique.Modulationoffattyacidcompositionbycultureparameterssuch
as light intensity, lightdark cycles, temperature, salinity and nutrients has been widely investigated and
reviewedbySukenik(1999).Patternsofvariationinthelipidclassandfattyacidcompositionduringbatch
cultivation of N. oculata have also been investigated (Hodgson, et al. 1991). Generally, the higher the
biomassproductivity,thehigherisEPAproductivity(ChiniZittellietal.1999).
Nannochloropsishasbeenalsoproposedassourceoflipidforbiodieselproductionbecauseitisableto
reachover60%lipidsafternitrogenstarvation(ShifrinandChrisholm,1980;Rodolfietal.,2009)mainlyas
TAGscontainingsaturatedandmonounsaturatedfattyacids(Bondiolietal.,2010).
BIOTECHNOLOGY
CultureMedia
Nannochloropsisgrowsinseawater,withmaximalgrowthrateinthesalinityrangefrom25to30g/L,butit
can tolerate salinities between 10 and 35 g/L (Renaud and Parry, 1994). Seawater sources can be both
natural and artificial seawater; the last one made either from single salts or from a mixture of seawater
saltsavailableonthemarket.F/2orFmedium(GuillardandRyther,1962)iswidelyusedforcultivationof
Nannochloropsis.Inindustrialscalemicroalgaeplants,whereculturevolumesandthusamountsofculture
medium to be prepared and treated before discharge are considerable and highly expensive, particularly
whenartificialseawaterisused,thepossibilityofreutilizingtheexhaustgrowthmediumafterbiomass
separation would be of high relevance. However, Richmond and Zou (1999) report the presence of
inhibitorysubstances,releasedfromthecellsintotoculturemediumwhichmayreducegrowthandlimit
the degree of recycling. Moreover it was observed (Rodolfi et al., 2003) that the accumulation of
metabolitesproducedbythemicroalga,togetherwithparticulatematerialsuchascellwallsreleasedafter
celldivisionandbacteria,causesgrowthcessationaftertwoweeks.Reutilizationofculturemediumseams
possibleonlyaftertreatmentwithactivatedcharcoalfilters.
Cultivationmethodsandproductionsystems
Inhatcheries,Nannochloropsisismainlycultivatedindoors,inbatchandsemibatch,inpolyethylenebags,
cylinders (Fulks and Main, 1991) and annular columns. Experiments with artificial or mixed (artificial and
natural) light sources aiming to evaluate biomass productivity in different culture conditions have been
carried out in alveolar and glass panels, annular columns, biofence systems and a new devised helical
tubular reactor with continuous harvesting regimen (Chini Zittelli et al., 2000, 2003; Zou et al., 2000;
Sandnesetal.,2005;Briassoulisetal.,2010).
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Cultivationinlargescaleoutdoorracewaypondsupto3000m2(20cmdepth)hasbeencarriedoutin
Eilat,Israel(Sukeniketal.,1993)bydailyharvesting1050%ofpondvolume.Averageproductivityof7and
22 g m2 were recorded in winter and in summer, respectively (Sukenik, 1991). The authors reported a
culture collapse after 2 months of cultivation because of grazers and contaminating algal species. At
presentinIsraelSeambioticcultivatedNannochloropsisinracewayspondusingfluegastoproducebiomass
asfeedstockforbiodiesel.InJapanNannochloropsisoculataisextensivelycultivatedintankskeptoutdoors
withseawaterandchemicalfertilizers(Okauchi,1991).Decreaseincelldensitywasrecordedintherainy
season and the phenomenon was mainly attributed to predation by protozoa, partially solved by
chlorinationandapplicationofterramycin.Boussibaetal.(1987)foundeffectivetheuseofureaandNH4+
as nitrogen source (instead of NO3) at a concentration of 25 mM against contaminating diatoms and
predators.
ChiniZittellietal.(1999)cultivatedNannochloropsisoutdoorsinanairmixednearhorizontaltubular
reactorwithdimensionsofupto30m2(600Lofculture)keptinsemicontinuousregimen.Withareactorof
this type but of a smaller size, a productivity of about 15 g m2 day1 was obtained in June. Outdoor
Nannochloropsiscultivationwasalsoperformedbyusinga500Lglassplatereactor(Zhangetal.,2001)and
GreenWallPanels.Inthefirstreactorproductivitiesof10and14.2gm2illuminatedsurfaceareaperday
wereobtainedinwinterandinsummer,respectively.Inthesecondreactorproductivitiesfrom9to20gm2
per ground area per day, depending on cultivation season and on panel orientations, were achieved
(Tredici,unpublisheddata).
Nannochloropsis has been cultivated in a 2800 liters tubular photobioreactor (50 m2 overall surface
area) at the Estacin Experimental de Cajamar Las Palmerillas(Spain). In February, with a daily dilution
rateofabout35%(fivedaysperweek),theaveragebiomassconcentrationwas1.2gL1andthevolumetric
productivityabout0.42gL1day1(Prof.Acienpersonalcommunication).
Nannochloropsishasalsobeenproposedassourceofoilforbiodieselproduction.Rodolfietal.(2009)
havesuggestedatwophasestrategy(afirststagecultivationofthemicroalgainthepresenceofnitrogen
carried out in closed photobioreactors, followed by a second stage of nitrogen starvation, preferably
carried out in open ponds), and have estimated a productivity of 20 ton of lipids per ha per year in the
Mediterranean region. Lipid content increase after nitrogen starvation was not observed in N. salina
(Boussibaetal.,1987),thustheselectionoftherightstrainisofcrucialimportanceforthisapplication.
Harvestingmethods
DuetoNannochloropsissmallcellsize,themostadequatesystemforbiomassrecoveryiscentrifugation,
that gives a product (paste) with 2530% dry material. To reduce the volume to be centrifuged,
autoflocculationcanbeused.The pHincrease,consequent totheinterruptionofCO2inletintheculture
exposed tosunlight,can causecellaggregationandthussedimentation,whichallowsto concentratethe
culture 1020 times. The same occurs by addition of NaOH. Flocculation of Nannochloropsis by using
cationicstarchwasnoteffective(Vandammeetal.,2010).
From experiments carried out at lab scale with two algae, Nannochloropsis and Scenedesmus, a
potentialreductionofharvestingcostsof20%wasproposedasaresultofpretreatmenttocentrifugation
withsubmergiblemicrofilatrationorultrafiltrationmembranes(Gorietal.,2010).
Biomassprocessing
Umduetal.(2009)foundthatlipidtransesterificationwithheterogeneouscatalysts(Al2O3supportedCaO
and MgO catalyst) was more active than with pure CaO and MgO in the production of biodiesel from N.
oculata.
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Patil et al. (2011) demonstrated a onestep process for direct liquefaction and conversion of wet
Nannochloropsis biomass (50% lipid) to biodiesel under supercritical methanol conditions, a process
allowingthesimultaneousextractionandtransesterificationofthebiomass.Theoptimalconditionsforthe
processwere:ratiobetweenwetalgaeandmethanolaround1:9,255Cand25minreactiontime.These
milder conditions compared to those necessary for pyrolisis seem to prevent formation of byproducts.
Other processes to obtain oil and biodiesel from Nannochloropsis biomass, such as catalytic Mcgyan
process,hydrothermalliquefactionandgasificationofthebiomass,directpyrolisisandcatalyticpyrolysisof
Nannochloropsisresiduehavebeendeveloped(Krohnetal.2011,Browetal.,2010,BillerandRoss,2011;
Panetal.,2010)
Scalinguplimitation
1) Low tolerance to temperatures above 30C (Sukenik et al., 2009; Sandnes et al., 2005).
Temperatures above 30 C can cause reduction in productivity and formation of cell aggregates
which favour contamination by bacteria and grazers (Rodolfi et al., 2003) and further depress
productivity.Accurate(andexpensive)coolingofthecultureisthusnecessary.
2) Small cell size allows less energy consumption for mixing, but more energy is required for
separatingthecellsfromtheculturemedium.
3) Thick cell wall, that allows mixing of the culture even with centrifugal pumps, but increases
difficultyofcellbreakage,whichisnecessaryforoilextraction.
Nannochloropsisisabletoaccumulatelipidsinamountshigherthan50%ofdrybiomassafter
nitrogenstarvation
NannochloropsisisabletoaccumulatelipidsmainlyasTAGandsaturatedandmonounsaturated
fattyacids,moresuitableforbiodieselproduction
AfteroilextractiontheresidueisavaluablePUFArichfractionthatcanbeusedasfeedorfood
ingredient
References
Biller P., Ross A.B. (2011) Potential yields and properties of oil from the hydrothermal liquefaction of microalgae with different
biochemicalcontent.BioresourceTechnology102:215225.
BondioliP.,DellaBellaL.,RivoltaG.,CasiniD.,PrussiM.,ChiaramontiD.,ChiniZittelliG.,BassiN.,RodolfiL.,TrediciM.(2010)Oil
productionbythemarinemicroalgaNannochloropsissp.F&MM24.Proceedingsofthe18thEuropeanBiomassConference
andExhibition,37May,Lyon,France.
Boussiba S., Vonshak A., Cohen Z., Avissar Y., Richmond A. (1987) Lipid and biomass production by the halotolerant microalga
Nannochloropsissalina.Biomass12:3747.
BriassoulisD.,PanagakigP.,ChionidisM.,TzenosD.,LalosA.,TsinosC.,BerberidisK.,JacobsenA.(2010)Anexperimentalhelical
tubularphotobioreactorforcontinuousproductionofNannochloropsissp.BioresourceTechnology101:67686777.
Brown M.R., McCausland M. A., Kowalsi K. (1998) The nutritional value of fourAustralian microalgal strains fed to Pacific oyster
Crassostreagigasspat.Aquaculture165:281293.
BrownT.M.,DuanP.,SavageP.E.(2010)HydrothermalliquefactionandgasificationofNannochloropsissp.EnergyFuels24:3639
3646.
Chini Zittelli G., Lavista F., Bastianini A., Rodolfi L., Vincenzini M., Tredici M.R. (1999) Production of eicosapentaenoic by
Nannochloropsis.culturesinoutdoortubularphotobioreactors.JournalofBiotechnology70:299317.
Chini Zittelli G., Pastorelli R., Tredici M.R. (2000) A modular flat panel photobioreactor (MFPP) for indoor mass cultivation of
Nannochloropsissp.underartificialillumination.JournalAppliedPhycology12:521526.
Chini Zittelli G., Rodolfi L., Tredici M.R. (2003) Mass cultivation of Nannochloropsis sp. in annular reactors. Journal Applied
Phycology15:107114.
FawleyK.P.,FawleyM.W.(2007)ObservationsonthediversityandecologyoffreshwaterNannochloropsis(Eustigmatophyceae),
withdescriptionsofnewtaxa.Protist158:325336.
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FulksM.,MainK.L.(eds)(1991)Rotifersandmicroalgaeculturesystems.ProceedingsofaUSAsiaWorkshop.TheOceanicInstitute,
Honolulu.
Gori R., Munz G., Lubello C., Daddi D., Biondi N., Tredici M.R. (2010) Energy demand and economic evaluation of solidliquid
separation systems used for harvesting and concentration of cultivated microalgae. Venice 2010, Third International
SymposiumonEnergyfromBiomassandWasteProceedings.
Guillard R.R.L., Ryther J.H. (1962) Studies of marine planktonic diatoms. I. Cyclotella nana (Hustedt) and Detonula confervacea
HodgsonP.A.,HendersonR.J.,SargentJ.R.,LeftleyJ.W.(1991)Patternsofvariationinthelipidclassandfattyacidcompositionof
Nannochloropsisoculata(Eustigmatophyceae)durtingbatchculture.IThegrowthcycle.JournalAppliedPhycology3:169
181.
KarlsonB.,PotterD.,KuylenstiernaM.,DersenM.A.(1996)Ultrastructure,pigmentcomposition,and18SrRNAgenesequencefor
Nannochloropsis granulata sp. nov. (Monodopsidaceae, Eustigmatophyceae), a marine ultraplankter isolated from the
Skagerrak,northeastAtlanticOcean.Phycologia35:253260.
Krohn B.J.K., McNeff C.V., Yan B., Nowlan D. (2011) Production of algaebased biodiesel using the continuous catalytic Mcgyan
process.BioresourceTechnology102:94100.
Okauki M. (1991) The status of phytoplankton production in Japan. In: Fulks M., Main K.L. (eds) Rotifers and microalgae culture
systems.ProceedingsofaUSAsiaWorkshop.TheOceanicInstitute,Honolulu.
Pan P., Hu C., Yang W., Li Y., Dong L., Zhu L., Tong D., Qing R., Fan Y. (2010) The direct pyrolysis and catalytic pyrolysis of
Nannochloropsissp.residueforrenewablebiooils.BioresourceTechnology101:45934599.
PatilP.D.,GudeV.G.,MannarswamyA.,DengS.,CookeP.,MunsunMcGeeS.,RhodesI.,LammersP.,NirmalakhandanN.(2011)
Optimizationofdirectconversionofwetalgaetobiodieselundersupercriticalmethanolconditions.BioresourceTechnology
102:118122.
RebellosoFuentesM.M.,NavarroPrezA.,GarcaCamachoF.,RamosMirasJ.J.,GuilGuerreroJ.L.(2001)Biomassnutrientprofiles
ofthemicroalgaNannochloropsis.JournalofAgricultureandFoodChemistry49:29662972.
RenaudS.M.,ParryD.L.(1994)Microalgaeforuseintropicalaquaculture.IIEffectofsalinityongrowth,grosschemicalcomposition
andfattyacidcompositionofthreespeciesofmarinemicroalgae.JournalofAppliedPhycology6:347356.
Richmond A., Zou N. (1999) Efficient utilisation of high photon irradiance for mass production of photoautotrophic micro
organisms.JournalofAppliedPhycology11:123127
RodolfiL.,ChiniZittelliG.,BarsantiL.,RosatiG.,TrediciM.R.(2003)GrowthmediumrecyclinginNannochloropsismasscultivation.
BiomolecularEngineering:243248.
Rodolfi L., Chini Zittelli G., Bassi N., Padovani N., Biondi N., Bonini G, Tredici MR. (2009) Microalgae for oil: strain selection,
102:100112.
Sandnes J.M., Kllqvist T., Wenner D., Gislerd H.R. (2005) Combined influence of light and temperature on growth rates of
Nannochlorpsisoceanica:linkingcellularresponsestolargescalebiomassproduction.JournalofAppliedPhycology17:515
525.
Shifrin M.S., Chrisholm S.W. (1980) Phytoplankton lipids: environmental influences on production and possible commercial
applications.In:ShelefG.,SoederC.J.(eds.)AlgaeBiomass,ElsevierNorthHollandBiomedicalPress,Amsterdam,pp.624
645.
Sukenik A. (1999) Production of eicosapentaenoic acid by the marine eustigmatophyte Nannochloropsis. In: Cohen Z. (ed.)
ChemicalsfromMicroalgae,Taylor&FrancisLtd,London,pp.4156.
SukenikA.,ZmoraO.,CarmeliY.(1993)Biochemicalqualityofmasrineunicellularalgaewithspecialenphasisonlipidcomposition.
II,Nannochloropsissp.Aquaculture117:313326
Sukenik A., Beardall J., Kromkamp J.C., Kopeck J., Masojdek J., van Bergeik S., Gabai S., Shaaham E., Yamshon A. (2009)
Photosyntetic performance of outdoor Nannochloropsis mass cultures under a wide range of environmental conditions.
AquaticMicrobialEcology56:297308.
Umdu E. S., Tuncer M., Seker E. (2009) Transestrification of Nannochloropsis oculata microalgas lipid to biodiesel on Al2O3
supportedCaOandMgOcatalysts.BioresourceTechnology100:28282831.
VandammeD.,FoubertI.,MeesschaertB.,MuylaertK.(2010)Flocculationofmicroalgaeusingcationicstarch.JournalofApplied
Phycology22:525530.
ZhangC.W.,ZmoraO.,KopelR.,RichmondA.(2001)AnindustrialflatplategallsreactorformassproductionofNannochloropsissp.
(Eustigmatophyceae).Aquaculture195:3549.
ZouN.,ZhangC.W.,CohenZ,RichmondA.(2000)Productionofcellmassandeicosapentaenoicacid(EPA)inultrahighcelldensity
culturesofNannochloropsissp.(Eustigmatophyceae).EuropeanJournalofPhycology35:127133.
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8.5 Haptophyta
8.5.1
Isochrysissp.
Figure58LightmicroscopicviewshowingIsochrysissp.
PicturefromUNIFI
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Haptophyta
Prymnesiophyceae
Isochrysidales
Isochrysidaceae
Isochrysis
Relatedspecies
Thereare3speciesnamesinthedatabaseatpresent,alltaxonomicallyaccepted.
Isochrysisgalbana
BIOLOGY
Isochrysis presents slightly elongated motile cells with a capacity to change shape, a stigma, and two
smoothapicalflagellaoncetotwicethelengthofthecells.ParkehasmentionedforstrainsofI.galbana,
the typespecies, the possibility of an important benthic phase, and, furthermore, the presence of
haptophycean scales on the cellbody of the motile cells. These scales are very difficult to detect, being
embedded in a thick mucilage matrix. There is also a very reduced haptonema. I. galbana cell has two
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chloroplasts found in two sides of the cell, and osmiophilic granules with median electron density in the
chloroplasts.Thepyrenoidissituatedinthechromatophore.
TwoIsochrysysspeciesaremainlybenthic:I.maritimaandI.litoralis.Isochrysismaritimacellscanbe
seentobehemispherical,3x6m,formingregularcubicmassesoffourtoeightcells(typicalaspectofthe
young culture). The cellwalls are pectic. A small red stigma is present and the swarmers have a positive
phototactic reaction. Each cell contains one or two parietal yellowgreen chloroplasts without any
detectable pyrenoids. Preliminary thinsectioning though, has revealed an intraplastidial slightly bulging
pyrenoid. Each cell contains a chrysolaminarin vacuole, fat globules, and peripheral muciferous bodies
whicharevisibleonlywhenstained.Inoldercultures,thecellsbecomegloboseandsurroundedbyvarious
concentric mucilage envelopes. Asexual reproduction occurs in this species by division of the nonmotile
individualsandalsobyproductionofswarmers.Thelatterareliberatedfromthickwalledsporangia.The
swarmersareelongated(3x4.55m)andshowacapacitytochangeshape.Twoflagellariseanteriorly;
they are slightly unequal, 6 and 8 m in length. The method of swimming of this species is highly
characteristic:theflagellabehavehomodynamically,movementoftheindividuals,generallyinabackward
direction, being rapid, with rotation of the body round the long axis. In I. litoralis young cells are non
motile,hemispherical(46x68m)orround(5m)whileoldercellsaresphericalvaryinginsize(515
m). It has a thick pectic envelope. There are two parietal goldenyellow chromatophores with one
intraplastidialpyrenoidperchromatophore.Asexualreproductionoccursbydivisionofthenonmotilecells
and by production of elongated swarmers (4 x 7 m) with two apical subequal (78 and 89 m)
homodynamicflagella.SwimmingmovementsaresimilartothoseofIsochrysismaritima.Cellsarecovered
withonelayerofovalplatescales(0.3x0.18m)withathickenedmarginandradiatingridges.
BIOTECHNOLOGY
Isochrysisiscommonlyusedintheaquacultureindustry(Tredicietal.,2009).Foritshighdocosahexaenoic
acid(DHA,22:63)contentitisoftenusedtoenrichzooplanktonsuchasrotifersorArtemia.Itisaprimary
algaeusedinshellfishhatcheries(oysters,clams,mussels,andscallops)andinsomeshrimphatcheries.
IsochrysishasbeenproposedasasouceofDHAfornutraceuticalsanddietarysupplements(Liuand
Lin,2001;PoissonandErgan,2001;)althoughatpresentonlyonecommercialproductcontaingIsochrysis
isinthemarket.Isochrysissp.extractshavebeenatentedforuseincosmetics(Herrmannetal.,2010).
ChiniZittellietal.(2004)outdoorsin120LannularcolumnsobtainedwithIsochrysissp.avolumetric
productivity of 0.31 g L1 day1, while in a fullscale arrangement the overall areal productivity (including
both the area occupied by the columns and the space between two rows of columns, as described by
Tredici,2004)wasof32.1gm2day1.Ina2.5m2pondtheproductionrateofIsochrysisgalbanabiomass
was29.5gm2day1andthatoflipids6.5gm2day1,while,ina100m2pond,theaverageproductivitiesof
biomass and of lipids were 23.5 g m2 day1 and 5.6 g m2 day1, respectively (Boussiba et al., 1988).
HarvestingcanbeachievedbymeansofflocculationwithFeCl3anddissolvedairflotation(Boussibaetal.,
1988).
Isochrysishasbeentestedatlaboratoryscaleforbiodieselproduction(Rodolfietal.,2009);however
theculturecarriedoutbyBoussibaetal.(1988),althoughnotintendedforbiodieselproduction,evidenced
thesuitabilityofthisalgaasbiodieselfeedstock.
References
BillardC.,GayralP.(1972)TwonewspeciesofIsochrysiswithremarksonthegenusRuttnera.EuropeanJournalofPhycology7:289
BoussibaS.,SandbankE.,ShelefG.,CohenZ.,VonshakA.,BenAmotzA.,AradS.,RichmondA.Outdoorcultivationofthemarine
microalgaIsochrysisgalbanainopenreactors.Aquaculture72:247253.
Chini Zittelli G., Somigli S., Rodolfi L., Tredici M.R. (2004) Outdoor mass cultivation of Isochrysis sp. in annular columns. 1er
CongresoLatinoamericanosobreBiotecnologiaAlgal,Argentina2004.ISBNNo.9871130325,pp.4548.
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HerrmannM.,JoppeH.,PertileP.,ZanellaL.(2010)ExtractsofIsochrysissp.PatentApplicationNo.US2010/0080761A1.
LiuC.P.,LinL.P.(2001)UltrastructuralstudyandlipidformationofIsochrysissp.CCMP1324.BotanicalBulletinofAcademiaSinica
42:207214.
PoissonL.,ErganF.(2001)DocosahexaenoicacidethylestersfromIsochrysisgalbana.JournalofBiotechnology91:7581.
102:100112.
Tredici M.R. (2004) Mass production of microalgae: photobioreactors. In: Richmond A. (ed.) Handbook of Microalgal Cultures,
BiotechnologyandAppliedPhycology,Blackwell,Oxford,pp.178214.
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8.5.2
Pavlovasp.
Figure59LightmicroscopicviewshowingPavlovalutheri
Picturefromhttp://www.eol.org/
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Haptophyta
Pavlovophyceae
Pavlovales
Pavlovaceae
Pavlova
Relatedspecies
Thereare12speciesnamesinthedatabaseatpresent,11ofwhicharetaxonomicallyaccepted.
P.gyrans,P.lutheri,P.pinguis,P.salina.
BIOLOGY
All species are known to produce motile swarmers though in a number of species they do not always
constitute the dominant phase (P. ennorea, P. granifera, P. noctivaga, P. virescens), which is instead the
nonmotilepalmelloidstage.
Themotilecellsareelongate(P.gyrans,P.granifera,P.pinguis,P.salina,P.virescens)thoughothers
maybetypicallybroadlyoval,rhombic,cordateor,indeed,almostcircular(P.helicata,P.Iutheri).However,
considerable variation can be observed in the shape of all species and some (e.g., P. gyrans) can display
markedmetaboly.Theappendagesarealwaysinsertedlaterallyoranteriolaterallyononeoftheflattened
facesofthecellwhichbyconventionisreferredtoastheventralface.Inallspeciestheflagellaarisefroma
depressionofvaryingdepth.ThelonganteriorlydirectedflagellumbeatswithacharacteristicsinusoidalS
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shapedwaveactioneitherinaflatplaneordescribingahelicalpathasin,forexample,P.helicata.Inlength
itvariesfromspeciestospeciesrangingfrom610m(P.calceolata)to1120m(P.helicata).Itcarriesan
investiture of long fine hairs and small electrondense bodies the latter arranged in a regular helix.
Variationsarefoundfromspecies tospeciesinthepresenceor absenceof thebodies, theirmorphology
and their arrangement. The bodies are reffered to as scales (knobscales) by analogy with the Golgi
producedscalesofothermembersofthePrymnesiophyceae.Comparedwiththelongflagellum,theshort
orposteriorflagellumisrelativelyuniforminlengthandexternalmorphology.Therangeoflengthrecorded
isbetween2.5and6.0manddistallythereisataperedtip.Theshortflagellumusuallybeatswitharather
stiff, inflexible action and is often directed towards the cell posterior and held close to the cellbody. P.
salinaandP.helicatadifferfromotherspeciesofPavlovainthattheshortflagellumisreducedtoastump
about 0.2 m long. The haptonema is always short and difficult to see in the living cell. It does not coil
neither does it contract. In some species there may be a long terminal "filopodium" but even without
filopodia the haptonema is frequently attenuated distally. Normally the surface of the haptonema is
naked but in the case of P. lutheri there is a superficial covering of small knobscales. All species are
"naked"inthatthereisnocellwallalthoughthenonmotilecellsmaybeinvestedwithlayersofmucilage.
Although some species have been recorded as possessing two chloroplasts, there is usually only one
consistingoftwolaterallobesextendingroundthecellbothventrallyanddorsallyapproximatelyforminga
VorHshape.SomemembersofthePavlovalesareunusualamongstthePrymnesiophyceaeinthatthey
have stigmata. In P. gyrans, P. granifera and P. pinguis the conspicuous orangered stigma consists of a
concavelayeroflipidglobules,650innumberandsituatedadjacenttothechloroplastmembranecloseto
the flagellar insertion. Within the Pavlovales, conspicuous pyrenoids are found only in P. gyrans, P.
granifera, P. pinguis. The cells of all species usually contain two discrete colourless bodies which are
presumedtoconstitutethereservemetabolite,a13glucan.
The available records show that the Pavlovales is, perhaps, a very successful goup having
representatives in oceanic, coastal, brackish and fresh water habitats. Some species are markedly
euryhaline which is clearly of advantage in brackish habitats. Individual species may also be widely
distributedgeographically.
BIOTECHNOLOGY
Pavlovaisanimportantalgainaquaculture,whereitisusedtofeedbivalvelarvaeandpostlarvae,penaeid
shrimp larvae, freshwater prawn larvae and marine zooplankton (Tredici et al., 2009). It contains high
amountsofbotheicosapentaenoic(EPA,20:53)anddocosahexaenoic(DHA,22:63)acids.
It is usually cultivated in low efficient photobioreactors in the hatcheries (Tredici et al., 2009). A
comparisonstudycarriedoutbyPonisetal.(2006)usingstandardhatcherytechniques(a10Lcarboy)and
a 4L Flat Alveolar Panel, both under continuous artificial illumination. The volumetric productivity of P.
lutheriintheFAPwasonaverage0.25gdryweightL1day1duringthebatchgrowthphaseandincreased
to 0.53 g L1 day1 when a semicontinuous regimen was adopted, while in the carboys the average
volumetricproductivitywas0.10gL1day1.
Nopilotscalestudiesconcenringbiodieselproductionhasbeen uptonowcarriedout.At laboratory
scale,bothP.salinaandP.lutherishowedaquitehighlipidcontent(31and35%,respectively)(Rodolfiet
al.,2009),similartothatobtainedintheFlatAlveolarPanel(Ponisetal.,2006).
References
GreenJ.C.(1980)ThefinestructureofPavlovapinguisGreenandapreliminarysurveyoftheorderPavlovales(Prymnesiophyceae).
EuropeanJournalofPhycology15:151191.[BIOLOGYsection]
PonisE.,ParisiG.,LeCozJ.R.,RobertR.,ChiniZittelliG.,TrediciM.R.(2006)Effectoftheculturesystemandculturetechniqueon
biochemicalcharacteristicsofPavlovalutherianditsnutritionalvalueforCrassostreagigaslarvae.AquacultureNutrition12:
322329.
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102:100112.
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8.6 Dinophyta
8.6.1
Crypthecodiniumcohnii
Figure60LightmicroscopicviewshowingCrypthecodiniumcohnii.
Picturefromhttp://researchpages.net/media/resources/2009/07/26/Crypthe2.jpg
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Dinophyta
Dinophyceae
Gonyaulacales
Crypthecodiniaceae
Crypthecodinium
Relatedspecies
ThereisonlyspeciesCryptechodiniumcohnii,synonymCryptechodiniumsetense.
BIOLOGY
Crypthecodinium cohnii was first described as Glenodinium cohnii, then transferred to the genus
Gyrodinium,andfinallyCrypthecodiniumcohniiwasintroduced,thatapparentlyrepresentsasupraspecies
encompassingseveralbiologicalspecies.
Two forms of C. cohnii have been reported, swimming cells and cysts with different dimensions. The
swimmingcellsshowvelocitiesoftheorderof1km.year1.Themotileswimmingcellshavetwounequal
flagella.Thecingulumsometimesalmostcompletelygirdlesthecellandsometimesismuchdisplacedand
does not form a complete loop around the body. C. cohnii has a very delicate theca (1520 m) whose
plates are mainly composed of cellulose and are barely visible, unless using a special staining method.
Duringcelllocomotion,thethecadeforms.Thethecaiscontainedinrelativelyfewalveoliwithapattern
thatcan be determined (thecal platetabulation)andusedfortaxonomicpurposes.Thecystsaresolitary
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andovoidinshapeandcanstayinadormant/survivalstageorstartdividing(vegetativecysts).Thenon
motile cysts contain greater levels of lipids, including docosahexaenoic acid (DHA, 22:63), than the
swimmerformofthemicroalga.C.cohniihasbothasexualandsexualreproduction.Ithasbeenreported
thatthelatterisinducedbynutrientdepletionandhasbeenobservedindense,rapidlygrowingcultures.
The gametes form groups of 38 cells which move around each other until two cells, anisogametes or
isogametes, establish contact with their ventral sides and begin to fuse. Then one of the two transverse
flagellaisshed,andthedevelopingzygote,withtwolongitudinalflagella,swimsaway.Sometimelaterthe
zygote encysts. The cysts divide and release normally 2, but sometimes 48, dinoflagellates. The new
dinoflagellates are motile as soon as they leave the cyst. C. cohnii is an obligatory heterotrophic. Carbon
andothernutrientsareobtainedfromlivingordecomposingseaweeds.Itmighthaveparasiticcapabilities
in or on macrophytes, as motile cells appear to have a peduncle, an organelle employed by other
dinoflagellatesonphagotrophy.Somestrainsarephagotrophic:thecellcontentofthepreyisingestedbya
feedingtube,leavingthepreyscellcoatinthemedium.Itgrowswellonorganiccarbonsubstratessuchas
glucose,dextrose,ethanol,aceticacid,sodiumacetate,carobpulpsyrup.
Thismicroalgaisbrackish,littoralandneritic.Itisoftenpresentamongmacrophytes,particularlyFucus
spp.Crypthecodiniumcohniilikedinoflagellateshavebeenobtainedfromvarioushabitatsincludingopen
ocean beaches, polluted brackish bays and estuaries, steaming mangrove swamps and frozen harbors.
Nevertheless, the strains of essentially globally dispersed dinoflagellates resembling C. cohnii are not
membersofthesamegenepool.
BIOTECHNOLOGY
C.cohniicanaccumulaterelativelyhighamountsoflipids(upto20%byweight)withDHAcontentsupto
3050%ofthetotalfattyacidsandnootherpolyunsaturatedfattyacidspresentabove1%(Coutoetal.,
2010).ForDHAproduction,thelimitingnutrientshouldbenitrogensincecellgrowthanddivisionishalted
due to the lack of this nutrient for de novo protein and nucleotide synthesis, and the supplied carbon is
convertedintostoragelipids(triacylglycerols)richinDHA.Therefore,industrialC.cohniifermentationsare
usuallyacarbonfedbatchandprogressesintwostages:thefirstistheactivegrowthphaseduringwhich
the lipid content of the biomass is low; once the nitrogen source is depleted, carbon is continuously
suppliedtothefermenter(Coutoetal.,2010).Inmostofthecommercialcultivationprocesses,glucoseis
used as the carbon source and energy source, as it represents an easily accessible feedstock for many
industrialfermentationprocesses(Coutoetal.,2010).ThemaximumoverallvolumetricproductivityofDHA
reported on glucose is 19 mg L1 h1, while in a fedbatch cultivation with pure ethanol as feed, 83 g dry
biomass L1, 35 g total lipid L1 and 11.7 g DHA L1 were produced in 220 h, with an overall volumetric
productivityofDHAwas53mgL1h1;withaceticacidascarbonsource,DHAproductivitiesofupto45mg
L1 h1 were achieved (de Swaaf et al., 2003). DHA from C. cohnii is added tomany commercial products,
includinginfantformulas(Tredicietal.,2009).About240tonnesofDHAoilperyearareproducedinthUSA
atapriceog43g1(BrennanandOwende,2010).
C.cohniihasbeenconsideredalsoasabiodieselfeedstock(GriffithsandHarrison,2009).Thefattyacid
composition of the remaining lipid fraction from the biomass leftover with lower content in
polyunsaturatedfattyacidscouldbeadequateforfurtherusesasfeedstockforbiodiesel,contributingto
theeconomyoftheoverallprocesssuggestinganintegratedbiorefineryapproach(Coutoetal.,2010).
References
BrennanL.,OwendeP.(2010)BiofuelsfrommicroalgaeAreviewoftechnologiesforproduction,processing,andextractionsof
biofuelsandcoproducts.RenewableandSustainableEnergyReviews14:557577.
Couto R.M., Calado Simes P., Reis A., Lopes Da Silva T., Martins V.H., SnchezVicente Y. (2010) Supercritical fluid extraction of
lipidsfromtheheterotrophicmicroalgaCrypthecodiniumcohnii.EngineeringinLifeSciences10:158164.
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de Swaaf M.E., Pronk J.T., Sijtsma L. (2003) Fedbatch cultivation of the docosahexaenoicacidproducing marine alga
Crypthecodiniumcohniionethanol.AppliedMicrobiologyandBiotechnology61:4043.
MendesA.,ReisA.,VasconcelosR.,GuerraP.,LopesdaSilvaT.(2009)CrypthecodiniumcohniiwithemphasisonDHAproduction:a
review.JournalofAppliedPhycology21:199214.[BIOLOGYsection]
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8.7 Labyrinthulomycetes
8.7.1
Schizochytriumsp.
Figure61LightmicroscopicviewshowingSchizochytriumaggregatumATCC28209.
Zoosporangiumcontainingnumerouszoospores(upperlargecell).
Picturefrom
http://syst.bio.konanu.ac.jp/labybase/images/Schizochytrium_aggregatum_sporangium_400.jpg
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokonta
Labyrinthulomycetes
Thraustochytriales
Thraustochytriaceae
Schizochytrium
Relatedspecies
Thereareatleastfivespeciesreported.
S.aggregatum,S.limacinum,S.mangrovei,S.minutum,S.octosporum.
BIOLOGY
Schizochytrium is a genus of marine thraustochytrid protists (related to heterokont algae) with mono
centric thalli. It possesses the following characteristic features: a multilayered wall composed
predominatelyofLgalactose,anorganelletermedasagenogenetosomefromwhichtheectoplasmicnets
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ariseandbiflagellateheterokontzoosporesinmanyofdescribedgenera.Schizochytriumischaracerisedby
successive bipartition of a vegetative cell, resulting in a cluster of cells, each of which develops into a
zoosporangiumorzoospore.Itisanosmoheterotroph.Theyareubiquitouslydistributedandcanbefound
onthealgalsurface,inestuarinehabitats,seawaterandsalinesoil.
BIOTECHNOLOGY
DHAAlgalOilisproducedviaheterotrophicfermentationusingSchizochytrium.DHAAlgalOilisintendedto
be used as a direct food ingredient to increase dietary intake of the long chain omega3 fatty acid DHA.
Schizochytrium sp. has been utilized in aquaculture applications, including enrichment of DHA in Artemia
and rotifers used to feed larval fish and shrimp. A product for aquaculture applications has been
successfullyutilizedforover7yearsasanexcellent,stabledietarysourceofDHAinshrimplarvacultureand
finfish (red seabream, Japanese flounder) culture with no adverse effects (Hammond et al., 2001). Dried
Schizochytriumsp.microalgaehasalsobeendeterminedtobegenerallyrecognizedassafe(GRAS)foruse
as a DHArich ingredient in broiler chicken and laying hen feed at levels up to 2.8 and 4.3%, respectively
and, since 1997, DHAenriched eggs from hens fed a diet containing approximately 1% DRM are now
commercially marketed in the United States, Mexico, Germany, Spain, Portugal, Benalux countries, Italy,
Norway,andIsrael(Hammondetal.,2001).
Under glucose and nitrogen fedbatch culture conditions and with control of glucose, pH and oxygen
levels, some Schizochytrium strains have been shown to grow to biomass densities of 200 g L1 in short
fermentationcyclesof90100h,accumulating4045gL1DHA(Chietal.,2009).
The oleaginous S. limacinum has capability of producing significant amounts of total lipid and DHA
whengrowinginavarietyofcarbonsourcessuchasglucose,fructose,aswellascomplexcarbonsources
suchassweetsorghumjuice(Ethieretal.,2011).Thetotallipidcontainedinthebiomasscanbeusedasa
sourceofbiodieselproduction.S.limacinumhasbeenidentifiedascapableofutilizingglycerolasacarbon
source, which makes it possible to convert waste glycerol from the biodiesel industry to DHA (Chi et al.,
2009).Thehighestbiomassproductivityof3.88gL1day1wasobtainedbyfeedingwith60gL1ofglycerol,
whilethehighestDHAproductivityof0.52gL1day1wasobtainedat90gL1ofglycerolduetothehigher
DHAcontent(Ethieretal.,2011).InadditiontoDHA,S.limacinumalsocontainsahighleveloftotalfatty
acid (ca 50% of dry biomass) and is a suitable feedstock for producing biodiesel via the direct
transesterification method (Johnson and Wen, 2009). The biodiesel prepared was subjected to ASTM
standardtests:parameterssuchasfreeglycerol,totalglycerol,acidnumber,soapcontent,corrosivenessto
copper, flash point, viscosity, and particulate matter met the ASTM standards, while the water and
sediment content, as well as the sulphur content did not pass the standard and require further
investigations(JohnsonandWen,2009).
References
ChiZ.,LiuY.,FrearC.,ChenS.(2009)StudyofatwostagegrowthofDHAproducingmarinealgaeSchizochytriumlimacinumSR21
withshiftingdissolvedoxygenlevel.AppliedMicrobiologyandBiotechnology81:11411148.
Ethier S., Woisard K., Vaughan D., Wen Z. (2011) Continuous culture of the microalgae Schizochytrium limacinum on biodiesel
derivedcrudeglycerolforproducingdocosahexaenoicacid.BioresourceTechnology102:8893.
HammondB.G.,MayhewD.A.,HolsonJ.F.,NemecM.D.,MastR.W.,SanderW.J.(2001)SafetyAssessmentofDHARichMicroalgae
fromSchizochytriumsp.II.DevelopmentalToxicityEvaluationinRatsandRabbits.RegulatoryToxicologyandPharmacology
33:205217.
JohnsonM.B.,WenZ.(2009)ProductionofbiodieselfuelfromthemicroalgaSchizochytriumlimacinumbydirecttransesterification
ofalgalbiomass.EnergyandFuels23:51795183.
Kamlangdee N., Fan K.W. (2003) Polyunsaturated fatty acids production by Schizochytrium sp. isolated from mangrove.
SongklanakarinJournalofScienceandTechnology25:643650.[BIOLOGYsection]
Raghukumar S. (2002) Ecology of the marine protists, the Labyrinthulomycetes (Thraustochytrids and Labyrinthulids). European
JournalofProtistology38:127145.[BIOLOGYsection]
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8.7.2
Thraustochytriumsp.
Figure62SEMimageshowingTharustochytrium.
Picturefromtolweb.org/tree/ToLimages/thrastosem2.png
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokonta
Labyrinthulomycetes
Thraustochytriales
Thraustochytriaceae
Thraustochytrium
Relatedspecies
Thereareatleast17specieswithinthisgenus.
T. aggregatum, T. antarcticum, T. arudimentale, T. aureum, T. benthicola, T. caudivorum, T. gaertneri, T.
globosum,T.indicum,T.kerguelense,T.kinnei,T.motivum,T.pachydermum,T.proliferum,T.roseum,T.
rossii,T.striatum.
BIOLOGY
Thraustochytrium is a genus of marine thraustochytrid protists (heterotrophic organisms related to
heterokont algae). Vegetative stages of thraustochytrids consist of single cells which are globose to
subglobose, measuring 4 to 20 m in diameter, mostly growing epibiontically on various substrata. Most
thraustochytrids reproduce by means of zoospores which possess a long anterior flagellum and a short
posterior flagellum. In Thraustochytrium the cytoplasmic contents of the mature cell, the sporangium,
dividedirectlyintozoospores.Itlivesmainlyondetritus.
BIOTECHNOLOGY
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Biotechnology of Thraustochytrium is similar to that of Schizochytrium, although no strain of this genus

have been at present commercially produced. DHA production rate is similar to that of many
Schizochytriumstrains(Raghukumar,2008).
AlloilsproducedbyThraustochytriumsp.are>99%inthetriacylglycerolformand<0.05%offreefatty
acids(Scottetal.,2011).Usingrawglycerolascarbonsourceproducedcomparableresultsforbiomass,oil
andDHAtothoseobtainedwhenglucosewasused(Scottetal.,2011).
References
Raghukumar S. (2008) Thraustochytrid marine protists: Production of PUFAs and other emerging technologies. Marine
ScottS.D.,ArmentaR.E.,BerrymanK.T.,Norman.A.W.(2011)Useofrawglyceroltoproduceoilrichinpolyunsaturatedfattyacids
byathraustochytrid.EnzymeandMicrobialTechnology48:267272.
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8.7.3
Ulkeniasp.
SYMBOLS:D
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokonta
Labyrinthulomycetes
Thraustochytriales
Thraustochytriaceae
Ulkenia
Relatedspecies
Thereareatleast6specieswithinthisgenus.
U.amoeboidea,U.minuta,U.profunda,U.radiata,U.sarkariana,U.visurgensis.
BIOLOGY
Ulkeniaisagenusofmarinethraustochytridprotists(heterotrophicorganismsrelatedtoheterokontalgae).
InUlkeniathesporangiumisformedfromthematurethallusbytheescapeoftheprotoplastfromthecell
wall s one amoeboid mass, and thesucessive cleavage of the naked protoplast into zoospores. Ulkenia
amoeboidea produces spores that assume the shape of an amoeba and move with a slow sinusoidal
movement, then these cells turn into cells which produce zoospores. It is found on decaying biomass of
macroalagaeandinmarineanimalguts.
BIOTECHNOLOGY
DHArichoilfromUlkeniasp.,containingtypically45%DHA,isobtainedthroughhetrotrophicfermentation
by a German Company and is used as a food ingredient in such foods as breads, cakes and biscuits,
breakfast cereals, cream cheese, modified milk and milk products, beverages, functional drinks (Food
StandardsAustraliaandNewZealand,2005).
Ulkenia,aswellasotherthraustochytrids,hasbeenproposedasabiodieselfeedstockduetoitshigh
lipidcontent(Fisheretal.,2008).
References
Raghukumar S. (1996) Morphology, taxonomy and ecology of Thraustochytrids and Labyrinthulids, the marine counterparts of
zoosporic fungi. In: Dayal R. (ed.) Advances in zoosporic fungi, MD Publications PVT Ltd, New Delhi, pp. 3558. [BIOLOGY
section]
FisherL.,NichollsD.,SandersonK.(2008)Productionofbiodiesel.WO2008/067605A1.
FoodStandardsAustraliaandNewZealand(2005)Finalassessmentreport.ApplicationA522.DHArichmicroalgaloilfromUlkenia
sp.asanovelfood.
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9 Macroalgae
9.1 Chlorophyta
9.1.1
Caulerpasp.
TAXONOMY
ThegenusCaulerpacurrentlycontains86recognizedspecies.
Caulerparacemosa
Figure63Caulerparacemosa(Forsskl)J.Agardh
Spain
CarolinaPenaMartn
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Bryopsidophyceae
Bryopsidales
Caulerpaceae
Caulerpa
Caulerparacemosa
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Caulerpataxifolia
Figure64Caulerpataxifolia(M.Vahl)C.Agardh
CalaD'Or,Mallorca,BalearicIslands,Spain;takenatadepthof7monadeadrhizomeofPosidoniaoceanica.
EduardoInfantesOanes
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Bryopsidophyceae
Bryopsidales
Caulerpaceae
Caulerpa
Caulerpataxifolia
BIOLOGY
Thalluscomposedofhorizontalstolonanchoredbycolorlessrhizoids,bearingerectphotosyntheticfronds
(assimilators) of extremely diverse morphology including threadlike, bladelike, pinnate, spongy and
vesicularstructures.Radialbranchingregardedasprimitive,bilateralasmorerecent,interpretationswhich
aresupportedbyultrastructureofchloroplasts.Reductionoflightalsoresultsinchangeoferectportions
fromradialtobilateralsymmetry.InspeciessuchasC.racemosamorphologyhasbeenshowntovarywith
habitat,resultingindescriptionofnumerousvarietiesorforms(orevenspecies)whichprobablyshouldbe
regarded as ecads. Growth apical and indeterminate. Extent of development of stolons and rhizoids
depends on substratum; stolons can reach to 12 m.m2, but generally are more ramified in epilithic
specimens; entire alga can reach to 1m. in length. Thallus composed of coenocytic filaments or siphons;
walls principally of 1,3 xylan, with numerous trabeculae (branching cylindrical ingrowths of the wall)
traversingthelumen.Whenwounded,asbygrazingfish,bladeorrhizomeexudeayellowishstickymass
whichhardenstoawoundplugofcarbohydrateinafewminutes.Genusheteroplastic.Largechloroplasts
with pyrenoid and starch grains seem to be more primitive, being replaced by small chloroplasts lacking
them.Amyloplastsmixedwithchloroplasts.Isolatedchloroplastssurviveanddivideinvitroformorethan2
weeks. Reproduction probably primarily by fragmentation of stolon. Holocarpic anisogamous sexual
reproduction described for some species. Meiosis, where examined, occurs at gametogenesis with
formation of gametes in unmodified areas of the thallus, without separation by cross walls; gametes
liberated in gelatinous extrusions through superficial papillae. Some species of Caulerpa produce the
poisoncaulerpicin,makingtheiruseasafoodresourcehazardous.
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Geographic distribution is global tropical to subtropical, with some species extending into the
Mediterranean Sea and temperate regions of Australia. Highest species diversity occurs in S. Australia.
Vertical range includes the intertidal to at least 50m; substrata include unconsolidated sand of seagrass
meadowsandhardsurfacessuchascoralrock.
BIOTECHNOLOGY
Caulerpa racemosa is used in Bangladesh, Japan, South Pacfic Islands, Vietnam as food and in the
Philippinesasmedicine(ZemkeWhiteandOhno,1999).Inthislattercountry,C.taxifoliaisalsoused,both
asfoodandmedicine.HeretheannualproductionofCaulerpais810tdryweight,allformculturedalga
(ZemkeWhiteandOhno,1999).
References
Algaebase:http://www.algaebase.org/search/genus/detail/?genus_id=32944&session=abv4:96D919560808f1B19AmHOH2E91A9
BeachK.S.,BorgeasH.B.,SmithC.M.(2006)Ecophysiologicalimplicationsofthemeasurementoftransmittanceandreflectanceof
tropicalmacroalgae.Phycologia45:450457.
Braune W. (2008) Meeresalgen. Ein Farbbildfhrer zu den verbreiteten benthischen Grn Braun und Rotalgen der Weltmeere,
A.R.G.GantnerVerlag.Ruggell,pp.1596,266pls.
Falco C., Menezes de Szchy M.T. (2005) Changes in shallow phytobenthic assemblages in southeastern Brazil, following the
replacementofSargassumvulgare(Phaeophyta)byCaulerpascalpelliformis(Chlorophyta).BotanicaMarina48:208217.
Guiry M.D., Guiry, G.M. (2010) AlgaeBase. Worldwide electronic publication, National University of Ireland, Galway.
http://www.algaebase.org;searchedon31March2010.
HodgsonL.M.,PhamHuuT.,LewmanomontK.,McDermidK.J.(2004)AnnotatedchecklistofspeciesofCaulerpaandCaulerpella
(Bryopsidales, Caulerpaceae) from Vietnam, Thailand and the Hawaiian Islands. In: Abbott I.A., McDermid K.J. (eds.)
TaxonomyofEconomicSeaweedswithreferencetothePacificandotherlocations,VolumeIX.,pp.2138.
KraftG.T.(2007)AlgaeofAustralia.MarinebenthicalgaeofLordHoweIslandandthesouthernGreatBarrierReef,1.Greenalgae,
AustralianBiologicalResourcesStudy&CSIROPublishing,.Canberra&Melbourne,pp.ivi,1347,110textfigs;11pls.
Lam D.W., Zechman F.W. (2006) Phylogenetic analyses of the Bryopsidales (Ulvophyceae, Chlorophyta) based on RUBISCO large
subunitgenesequences.JournalofPhycology42:669678.
LittlerD.S.,LittlerM.M.(1997)AnillustratedfloraofthePelicanCays,Belize.BulletinoftheBiologicalSocietyofWashington9:1
149.
Nuber N., Gornik O., Lauc G., Bauer N., Zuljevic A., Papes D., Zoldos V. (2007) Genetic evidence for the identity of Caulerpa
racemosa(Forsskal)J.Agardh(Caulerpales,Chlorophyta)intheAdriaticSea.EuropeanJournalofPhycology42:113120.
PedrocheF.F.,SilvaP.C.,AguilarRosasL.E.,DreckmannK.M.,AguilarRosasR.(2005)Catlogodelasalgasmarinasbentnicasdel
PacficodeMxico.I.Chlorophycota.pp.iviii,17146.Ensenada,Mxico:UniversidadAutnomadeBajaCalifornia.
Phillips J.A. (2009) Reproductive ecology of Caulerpa taxifolia (Caulerpaceae, Bryopsidales) in subtropical eastern Australia.
EuropeanJournalofPhycology44:8188.
Robledo D., FreilePelegrn Y. (2005) Seasonal variation in photosynthesis and biochemical composition of Caulerpa spp.
(Bryopsidales,Chlorophyta)fromtheGulfofMexico.Phycologia44:312319.
Serio D., Alongi G., Catra M., Cormaci M., Furnari G. (2006) Changes in the benthic algal flora of Linosa Island (Straits of Sicily,
MediterraneanSea).BotanicaMarina49:135144.
SkeltonP.A.,SouthG.R.(2004)NewrecordsandnotesonmarinebenthicalgaeofAmericanSamoaChlorophyta&Phaeophyta.
CryptogamieAlgologie25:291312.
Stam W.T., Olsen J.L., Zaleski S.F., Murray S.N., Brown K.R., Walters L.J. (2006) A forensic and phylogenetic survey of Caulerpa
species(Caulerpales,Chlorophyta)fromtheFloridacoast,localaquariumshops,andecommerce:establishingaproactive
baselineforearlydetection.JournalofPhycology42:11131124.
TerradosJ.,MarbaN.(2006)Isthevegetativedevelopmentoftheinvasivechlorophycean,Caulerpataxifolia,favoredinsediments
withahighcontentoforganicmatter?BotanicaMarina49:331338.
VarelaAlvarez E., Andreakis N., LagoLeston A., Pearson G.A., Serrao E.A., Procaccini G., Duarte C.M., Marba N. (2006) Genomic
DNAisolationfromgreenandbrownalgae(CaulerpalesandFucales)formicrosatellitelibraryconstruction(Note).Journalof
Phycology42:741745.
ZemkeWhiteW.L.,OhnoM.(1999)Worldseaweedutilisation:anendofcenturysummary.JournalofAppliedPhycology11:369
376.
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9.1.2
Ulvasp.
TAXONOMY
ThegenusUlvacontains97relatedspecies.
Ulvalactuca
Figure65UlvalactucaLinnaeus
Spiddal,Co.Galway,Ireland;lowershorerockpools
AnnaSolerVila
SYMBOLS:B,E,PIV
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Ulvophyceae
Ulvales
Ulvaceae
Ulva
Ulvalactuca
Ulvarigida
Figure66UlvarigidaC.Agardh
BlackHd.,Co.Clare,Ireland;lowershorerockpools
M.D.Guiry
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SYMBOLS:B,E,PIV
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Ulvophyceae
Ulvales
Ulvaceae
Ulva
Ulvarigida
BIOLOGY
Generaldescription
Thisisasmallgenusofmarineandbrackishwatergreenalgae.Itisedibleandisoftencalled'SeaLettuce'.
Specieswithhollow,onelayeredthalliwereformerlyincludedinEnteromorpha,butitiswidelyaccepted
nowthatsuchspeciesshouldbeincludedinUlva.
The thallus of ulvoid species is flat and bladelike and is composed of two layers of cells. There is no
differentiationintotissues;allthecellsoftheplantaremoreorlessalikeexceptforthebasalcells,which
areelongatedtoformattachmentrhizoids.Eachcellcontainsonenucleusandhasacupshapedchoroplast
withasinglepyrenoid.
Ulvaundergoesaverydefinitealternationofgenerations.Biflagellateisogametesareformedbycertain
cellsofthehaploid,gametangialplant.Theseareliberatedandfuseinpairstoformadiploidzygotewhich
germinatestoformaseparatediploidplantcalledthesporophyte;thisresemblesthehaploidgametangial
plant in outward appearance. Certain cells of the sporophyte undergo meiosis and form zoospores in
sporangia;thesezoosporesarequitedifferenttothegametesinthattheyformquadriflagellatezoospores
(with 4 flagella). These are released, swim around for a time, settle and germinate to form the haploid
gametangialthallus.Notethatthehaploidgametesarecapableofsettlingandgerminatingwithoutfusion
to form a haploid thallus directly; most Ulva populations reproduce by this form of parthenogenesis and
sexualreproductionisnotverycommon.(Algaebase.2010)
Thebiochemicalcompositionofwildmacroalgaeissubjecttogreatspatialandseasonalvariations.InTable
17compositionofUlvasp.isreported.
Table17BiochemicalcompositionofUlvasp.inEurope,exceptforpigmentvalues.Compositionexpressedas%
dryweightexceptforWaterandash.FW=Freshweight.1Ortizetal.(2006).2ChakrabortyandSantra(2008).3
BriandandMorand(1997).
Ulvasp.
Water(%FW)
88.4165to833
Ash
11117to353
61.5141to613
Total
Carbohydrate
Uronicacids
7.711.53
Glucose
3.99.33
Mannose
0.71.23
Rhamnose
16.623.43
Xylose
2.95.33
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Protein
Lipid
Iodine
K
Na
Ca
Mg
S
P
Vitamin(mg.kg1)
Pigments
Ulvasp.
27.2110.3to17.23
0.311.8to3.53
182203
0.95.93
1.45.63
2.02.73
2.843
0.130.243
A
C:38023.1to7893
B1:0.423.983
B12:0.0400.0643
E:8.737.53
(Chla:2.06mg.g1/Chlb:1.35
mg.g1/carotenoids:19.55g.g1)2
BIOTECHNOLOGY
Culturemedia/Cultivationmethods/Growthkineticsandproductivity
Ulvasp.hasbeencultivatedworldwideinanintegratedpolyculture,inlinewitheithersewagetreatment
plants or aquaculture farms on land. Ulva sp. is also cultivated on Nori nets for food purposes in Japan.
Table18representsthevariouscultivationsystemsandtheirproductivityaroundtheworld.
Species
Table18.Ulvaproductivityinvariouscultivationsystemsaroundtheworld.
Location/Seas Production
Keywords
Reference
on (or culture
duration)
Tank/Pond/RacewayCultivation
Ulvalactuca
USA(Florida)/ 19gDWm2day1
8months
(averageover8months)
Outdoorcultivation
9,4gFWm2day1
Associatedwithsewage
(8 days only) in shallow treatmentplant.Low
tanks
salinity,notaimedat
biomassproduction.
Tank/Pond/RacewayCultivation:IntegratedMultitrophicaquaculture
Ulvalactuca
Greece
(Aegeansea)
Ulvalactuca
Israel
upto376gFWm2day1
Sparusaurata
Biofiltration;fishpond
effluents;yield;chemical
analysis
Biofiltration;stocking
Spain
density;yield;biomass
(Andalucia) /
Ulvarotundata
33.6 g DW m2 day1 and
evolution;nitrogen;
Spring
GR7.5%day1
phosphorus
Gracilariopsis
10.2 g DW m2 day1 and
GR6.5%day1
longissima
Sparusaurata
Israel
/
Biofiltration;recirculation;
Haliotis discus WinterSpring FCR= 2.6625 with Ulva ammoniatoxicity
hannai
lactuca
DeBusketal.,
1986
Tsagkamiliset
al.,2010
Msuyaand
Neori,2008
Hernandezet
al.,2005
Schuenhoffet
al.,2003
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Species
Paracentrotus
lividus
Ulvalactuca
Sparusaurata
Dicentrarchus
labrax
Ulvarotundata
Sparusaurata
Location/Seas Production
on (or culture
duration)
Tapes
philippinarum
Ulvalactuca
Gracilariaspp.
Sparusaurata
Ulvalactuca
Sparusaurata
Ulvarigida
Oncorhynchus
mykiss
Gracilaria
tenuistipitata
Sparusaurata
Ulvalactuca
Ulvalactuca
Reference
Biofiltration;raceway
cultivation;yield
Mataand
Santos,2001
Portugal
Spring
94117gFWm2day1
/
29gDWm2day1andGR
21.8%day1
Israel(1year)
0.67%GRday1and28kg Biofiltration;nutrient
m2year1
budget;seaweedyield;
0.9% GR day1 (juveniles) abalonegrowthrate
0.34% GR day1 (young
adults)
78kgFWm2year1
pooryield
Neorietal.,
2000
Israel
0.26%bodyweightday1
Neorietal.,
1998
Haliotis discus
hannai
Ulvalactuca
Gracilaria
conferta
Haliotis
tuberculata
Ulvalactuca
Gracilaria
conferta
Sparusaurata
Haliotis
tuberculata
Keywords
230gFWm2day1
highlyerraticgrowth
Israel / year
round
0.3%GRday1FCR=20
25gFWseaweedper
gramofabalone
produced
Israel
250gFWm2day1
Spain (Canary
Islands)
40gDWm2day1
Sweden
(56months)
49%GRday1
Israel
Israel
55gDWm2day1
Biofiltration;nitrogen
recycling;pilotscale,
modular,landbased
system;yield;abalone
growthrate
Theoreticalyieldand
revenueprojections;
systemdesign
Shpigeland
Neori,1996
Biofiltration;recirculation;
waterquality;effluent
Neorietal.,
1996
Biofiltration;yield;flow
rate
Growthrate;biomass;
nutrientuptake
Jimenezdel
Rioetal.,1996
Haglundand
Pedersen,
1993
Yield;growthrate;marine
fishponds
Neorietal.,
1991
Biofiltration; growth rate; Vandermeulen

nitrogencontent
and Gordin,
1990
GR = growth rate (% of the fresh weight), FW= fresh weight, DW= dry weight, pm= per month, FCR= Food
conversionratio;inred:Finfish,inBlue:Shellfish,inGreen:Macroalgaecultivated
Productionsystems
There is a wide range of production systems on land or at sea, each of them are applied for a specific
purposes:
the production of Ulva sp. has been performed using Tank / Pond / Raceway cultivation in either
integratedmultitrophicaquaculture(IMTA)systems(asreferencedintable1)orinsewagetreatment
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plantasbiofilters(Giacconeetal.,1976;GuistandHumm,1976;Rytheretal.,1979;Tsagkamilisetal.,
2010),hencereducingtheimpactoffishfarmingonrisksofeutrophication.Thecultivatedmaterial
originatesfromthevegetativegrowthofwildpopulations.
Ulvasp.isalsocultivatedonsuspendednets(similartoPorphyra,Nori,cultivation)forfoodpurposes
in Japan. Nets are spread on wild population of Ulva sp. and used as collectors. Besides, research
programs have been developed to ensure reliable and consistent crop (Critchley in Werner et al.,
2004). Ulva intestinalis (previously known as Enteromorpha intestinalis) cultivation has also been
establishedfromprotoplasts(RusigandCosson,2001).Theauthorsclaimthattheplantsregenerated
fromprotoplastsmayalsobeusedasseedstocktofacilitatepropagationformacroalgalculture.
Nevertheless, most biomass is harvested from the wild after blooming events, therefore no
productionsystemisrequired.
Harvestingmethods
Tank/Pond/RacewayCultivation
Eitherwithnetsormanually.
Netscultivation
Although Ulva and Porphyra exhibit rather similar morphological characteristics (flat and bladelike
shape)andarecultivatedonsimilarnets,harvestingmethodsdiffers.Ulvasp.isharvestedmanually
leavingabout2cmofthebladeonthenettoallowregenerationandamonthlygrowth/harvesting
cycle(Perez,1997).
Blooms
Ulva sp. can also be mechanically harvested from the wild, mostly during blooms (green tides)
where the material is collected on sandy beaches in large quantities (several thousands of tons). In
thatcasebulldozer,rakingmachines,siftingmachines,scrapingmachinesandbaler(Haypress)have
beenusedaccordingtothescaleandthicknessofthedepositonthebeach(upto1mhigh)(Briandin
GuiryandBlunden,1991).TheexistenceofasubtidalandfloatingbloomingpopulationsofUlvasp.
hasalsobeencollectedfromboatequippedwithconveyorbeltcollectorinfranceandfromspecial
reapingmachinesintheOdensefjordinDenmark.MorandandMerceron(1999)reportedarecurrent
collectionofover50000m3peryearforBrittany(France)alone,whichrepresentsabout2000truck
arrivals and departures with associated hydrological and ecological impact (sand removal, erosion,
etc).
Biomassprocessing
Once harvested, the biomass is processed according to the end product. Different levels of quality are
requiredwhetherthematerialwillbeusedforfeedandfoodpurposes,fertilizerorenergypurposes.Paper
productionwasalsooneoftheanswerstoresolveMacroalgalbloomsofUlvasp.
Energy
Macroalgaearestoredinatankwheretheacidogenicfermentationoccurs.Theliquefactionjuicesfromthe
storage of Ulva sp. are collected by draining and pressing the biomass. The juice is then transferred to
another bioreactor for methanogenic fermentation (anaerobic digestion) (Morand and Merceron, 2006).
The cake resulting from the pressing process can be used as organic enriching or fertilizing agent in
agriculture.
Matsui et al. (2006) describe their field test plant process as divided in four parts: Pretreatment,
fermentation, biogas storage and generation. In pretreatment part, seaweeds are smashed and diluted
withwatertosuppresstheeffectofsaltandmakeappropriateslurry.CollectedUlvasp.containedforeign
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bodiessuchassand,whichdonotaffectfermentationdirectlybutdecreaseavailablevolumeoftanks.Thus
theyarewashedbywaterandforeignbodieswereremovedbeforetheywereusedforfermentationtests.
Infermentationpart,therearetwoprocesses(prefermentationandmethanefermentation)forahigher
efficiencyoffermentation.Theseaweedslurryistreatedbyprefermentation(acidproduction)anduseof
methane fermentation as substrate. Capacity of a prefermentation tank is 5 kL. In the methane
fermentationprocessbiogasisproduced.Capacityofamethanefermentationtankis30kL.Themethane
fermentationtankcontainsporousmatrixinsideforinmobilizingbacterialcells.Biogasisrefined(desulfur)
and stored in a gasholder (30 kl). Residue of methane fermentation is dehydrated and used as fertilizer.
Biogasismixedwithcitygas(naturalgas)andfedtothegasenginegenerator(acogenerationsystem).A
gasmixeroftheenginewasimprovedtomixwithbiogasandcitygas.Electricity(10kW)generatedbythe
gasengineisusedforelectricequipmentsoftheplant.Heat(23kW)fromtheengineisusedforheatingup
energyforfermentationtanks.Anequipmentofdeodorizationusingmicroorganismsisset.
Fertilizer
Theharvestedbiomasscanbedirectlyspreadonfieldsassoiladditive,butresultsinasubsequentstream
andaquiferpollution.Besides,thepossibilityofspreadingislimitedbythefieldavailabilitysufficientlyclose
tothesea.
Compostingalgaewithanothersubstrateallowsincreasingtheareaofutilization,butnotsufficiently
becauseofthetransportationcostandthelowvalueofcompost(MorandandMerceron,2004).
Foodandfeedpurposes
Thebiomasscomingfrombloomsisdriedandmilledforuseinpremixedpoultryfeeds(Braultetal.,1983).
However,animalfeedingonlyusessmallamountsofalgae,interestingasfoodsupplement(i.e.carotene)
(Morand and Merceron, 2004). Human food requires highly clean material, which is often incompatible
withharvestingmethodsofblooms,thereforeonlyUlvasp.cultivatedonnetsinJapanarecommercially
usedforfoodpurposesafterdrying,millingandconditioning.
Scalinguplimitations
Ulva sp. are relatively robust species with great tolerance to low salinity and temperature changes. The
mainscalinguplimitationsconcernsthecultivationintanks,pondsandracewayswherelandavailabilityis
scarce.
OtherthanthatHanisak(1987)mentionsthatUlvasp.exhibitshighyieldsbutwerenotsustainablefor
significantperiodsasthespecieswouldbecomereproductiveandshedspores.Aseachcellinthethallus
can become reproductive, it was not unusual for an entire culture to sporulate and be lost overnight,
leaving only empty cells. A search was initiated for sterile strains, as Ulva in particular lends itself to
digestionwithhighmethaneyieldsbecauseofitsfavourablecarbohydrateandproteincontent.
References
BlidingC.(1969"1968")AcriticalsurveyofEuropeantaxainUlvales,PartII.Ulva,Ulvaria, Monostroma,Kornmannia.Botaniska
Notiser121:535629,47figs.
BraultD.,BriandX.,GolvenP.(1983)Lesmareesvertes:premierbilanconcernantlesessaisdevalorisation.In:Basesbiologiques
delaquaculture,Ifremer,Montpellier,pp.3342.
Braune W. (2008) Meeresalgen. Ein Farbbildfhrer zu den verbreiteten benthischen Grn Braun und Rotalgen der Weltmeer,
A.R.G.GantnerVerlag,Ruggell,pp.1596,266pls.
BriandX,MorandP.(1997)AnaerobicdigestionofUlvasp.1.relationshipbetweenUlvacompositionandmethanisation.Journalof
AppliedPhycology9:511524.
BrodieJ.,MaggsC.A.,JohnD.M.(2007)GreenseaweedsofBritainandIreland,BritishPhycologicalSociety,London,pp.ixii,1242,
101figs.
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Burrows E.M. (1991) Seaweeds of the British Isles. Volume 2. Chlorophyta, Natural History Museum Publications, London, pp. xi
238,60figs,9plates.
Chakraborty S., Santra S.C. (2008) Biochemical composition of eight benthic algae collected from Sunderban. Indian Journal of
MarineSciences37:329332.
DawesC.J.,MathiesonA.C.(2008)TheseaweedsofFlorida,UniversityPressofFlorida,Gainesville,Florida,pp.iviii,1592,plsILI.
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liquamiurbanieindustriali.Ingeneriaambientale5:572582.
GuistG.G.,HummH.J.(1976)EffectsofsewageeffluentongrowthofUlvalactuca.FloridaScientist39:26771.
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http://www.algaebase.org;searchedon20October2010.
GuiryM.D.,BlundenG.(1991)SeaweedresourcesinEurope:usesandpotential,JohnWiley&Sons,Chichester;NewYork,pp.xi,
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HanisakD.M.(1987)CultivationofGracilariaandothermacroalgaeinFloridaforenergyproduction.In:BirdK.T.,BensonP.H.(eds.)
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HaydenH.S.,WaalandJ.R.(2004)AmolecularsystematicstudyofUlva(Ulvaceae,Ulvales)fromthenortheastPacific.Phycologia
43:364382.
HernandezI.,FernandezEngoM.A.,PerezLlorensJ.L.,VergaraJ.J.(2005)Integratedoutdoorcultureoftwoestuarinemacroalgae
asbiofiltersfordissolvednutrientsfromSparusauratawastewaters.JournalofAppliedPhycology17:557567.
JimenezdelRioM.,RamazanovZ.,GarciaReinaG.(1996)Ulvarigida(Ulvales,Chlorophyta)tankcultureasbiofiltersfordissolved
inorganicnitrogenfromfishpondeffluents.Hydrobiologia326/327:6166.
KraftG.T.(2007)AlgaeofAustralia.MarinebenthicalgaeofLordHoweIslandandthesouthernGreatBarrierReef,1.Greenalgae,
AustralianBiologicalResourcesStudy&CSIROPublishing,Canberra&Melbourne,pp.iivi,1347,110textfigs;11pls.
LoiseauxdeGorS.,NoaillesM.C.(2008)AlguesdeRoscoff,EditionsdelaStationBiologiquedeRoscoff,Roscoff,pp.1215.
LoughnaneC.J.,McIvorL.M.,RindiF.,StengelD.B.,GuiryM.D.(2008)Morphology,rbcLphylogenyanddistributionofdistromatic
Ulva(Ulvophyceae,Chlorophyta)inIrelandandsouthernBritain.Phycologia47:416429.
MataL.,SantosR.(2001)CultivationofUlvarotundata(Ulvales,Chlorophyta)inraceways,usingsemiintensivefishpondeffluents:
yieldandbiofiltration.Proceedingsofthe17thInternationalSeaweedSymposium.
Matsui T., Amano T., Koike Y., Saiganji A., Saito H. (2006) Sustainable nonfuel products/production systems from biomass
resources.Paperpresentedat2006AmericanInstituteofChemicalEngineers(AIChE)Conference,SanFrancisco,California
(www.aiche.org)Session412http://aiche.confex.com/aiche/2006/techprogram/S3516.HTM.
MorandP.,BriandX.,CharlierR.H.(2006)AnaerobicdigestionofUlvasp.3.Liquefactionjuicesextractionbypressingandtechnico
economicbudget.JournalofAppliedPhycology18:741755.
MorandP.,MerceronM.(2004)Coastaleutrophicationandexcessivegrowthofmacroalgae.In:PandalaiS.G.(ed.)RecentResearch
DevelopmentsinEnvironmentalBiology1.ResearchSignpost,Trivandrum,India,pp.395449.
Morand,P.,Merceron,M.(2005)Macroalgalpopulationandsustainability.JournalofCoastalResearch21:10091020.
MsuyaF.E.,NeoriA.(2008)Effectofwateraerationandnutrientloadlevelonbiomassyield,Nuptakeandproteincontentofthe
seaweedUlvalactucaculturedinseawatertanks.JournalofAppliedPhycology20:10211031.
NeoriA.,RaggN.L.C.,ShpigelM.(1998)Theintegratedcultureofseaweed,abalone,fishandclamsinmodularintensivelandbased
systems:II.Performanceandnitrogenpartitioningwithinanabalone(Haliotistuberculata)andmacroalgaeculturesystem.
AquaculturalEngineering17:215239.
NeoriA.,ShpigelM.,BenEzraD.(2000)Asustainableintegratedsystemforcultureoffish,seaweedandabalone.Aquaculture186:
279291.
NorrisJ.N.(2010)MarinealgaeoftheNorthernGulfofCalifornia:ChlorophytaandPhaeophyceae,SmithsonianContributionsto
Botany94:ix,1276.
OrtizJ.,RomeroN.,etal.(2006)Dietaryfiber,aminoacid,fattyacidandtocopherolcontentsoftheedibleseaweedsUlvalactuca
andDurvillaeaAntarctica.FoodChemistry99:98104.
PacficodeMxico.I.Chlorophycota,UniversidadAutnomadeBajaCalifornia,Ensenada,Mxico,pp.iviii,17146.
PerezR(1997)Cesalguesquinousentourent:conceptionactuelle,rledanslabiosphre,utilisations,culture,IFREMERPlouzan,
France,pp.xi272.
RytherJ.H.,DeBoerJ.A.,LapointeB.E.(1979)Cultivationofseaweedsforhydrocolloids,wastetreatmentandbiomassforenergy
conversion.JensenA.,SteinJ.R.(eds.)ProceedingsoftheIXthInternationalseaweedsymposium,SciencePress,Princeton,
pp.116.
Rusig A.M., Cosson J. (2001) Plant regeneration from protoplasts of Enteromorpha intestinalis (Chlorophyta, Ulvophyceae) as
seedstockformacroagalculture.JournalofAppliedPhycology13:103108.
ScagelR.F.(1957)AnannotatedlistofthemarinealgaeofBritishColumbiaandnorthernWashington(includingkeystogenera).
Bulletin,NationalMuseumofCanada150:vi289.
Shhuenhoff A., Shpigel M., Lupatsch I., Ashkenazi A., Msuya F.E., Neori A. (2003) A semirecirculating, integrated system for the
cultureoffishandseaweed.Aquaculture221:167181.
ShpigelM.,NeoriA.(1996)Theintegratedcultureofseaweed,abalone,fishandclamsinmodularintensivelandbasedsystems:1.
Proportionsofsizeandprojectedrevenues.AquaculturalEngineering15:313326.
SmithG.M.(1944)MarinealgaeoftheMontereyPeninsula,StanfordUniversityPress,Stanford,pp.ix,622,98pls.
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TsagkamilisP.,DanielidisD.,DringM.,KatsarosC.(2010)RemovalofphosphatebythegreenseaweedUlvalactucainasmallscale
sewagetreatmentplant(IosIsland,AegeanSea,Greece).JournalofAppliedPhycology22:331339.
VandermeulenH.,GordinH.(1990)AmmoniumuptakeusingUlva(Chlorophyta)inintensivefishpondsystems:masscultureand
treatmentofeffluent.JournalofAppliedPhycology2:363374.
WernerA.,ClarkeC.,KraanS.(2004)StrategicreviewofthefeasibilityofseaweedaquacultureinIreland.NDPMarineRTDIDesk
StudySeries,DK/01/008.2004
376.
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9.1.3 Cladophorasp.
Figure67CladophoraglomerataFabioRindi(left)andCladophorarupestris(Linnaeus)KtzingMichaelD.
Guiry(right).Ireland
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Ulvophyceae
Cladophorales
Cladophoraceae
Cladophora
Cladophoraglomerata/rupestris
Relatedspecies
ThegenusCladophoracontains180relatedspecies.
BIOLOGY
Thalli of uniseriate branched filaments with apical and/or intercalary growth. Branches sparse to profuse
with branches inserted laterally below apex of cell or apically on cell (a pseudodichotomy). If attached,
branchingrhizoidsarisefrombasalcellandothercellsinbasalregion,orsimplediscoidholdfastproduced.
Chloroplasts parietal, either densely packed discoid and/or united in a reticulum. Pyrenoids in many
chloroplasts, bilenticular, flanked by two bowlshaped starch bodies. Cells multinucleate, nuclei dividing
more or less synchronously with nuclear membrane remaining intact. Cell wall polysaccharide mainly
crystalline cellulose, forming numerous lamellae of microfibrils in crossed fibrillar pattern. Asexual
reproduction by biflagellate or quadriflagellate zoospores only method of reproduction in some species,
some species only reproducing by thallus fragmentation. Akinetes produced by most species under
unfavourableconditions;theseareswollen,thickwalled,filledwithstarch.Thickwalledrhizoidsandbasal
portionsofmainaxesalsoperennatingdevices.Sexualreproductionbyregularalternationofgenerations
producing biflagellate isogametes and quadriflagellate zoospores. Meiosis precedes spore production.
Zoidangiawith13pores,cruciatezooidsreleasedunderpressure.ZoidsofC.rupestriswithwelldeveloped
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layerofgranulesoutsidetheplasmalemmaandinsidethe'fibrouslayer'whichmaybeinvolvedincellwall
synthesis.
Cladophora is cosmopolitan in temperate and tropical regions, occurring in freshwater, brackish and
marineconditions.Thevariousspeciesarefoundintertidallyinwaveexposedtoveryshelteredhabitats,in
brackishpools,lagoonsandmudflats,andinmoreorlesseutrophicfreshwaterstreamsandlakeswithpH>
7. Certain unattached species can be aggregated into spherical mats, 210 cm in diameter, called
"Cladophora Balls". Cladophora glomerata can be a rapid colonizer of rocky stream beds and at
temperatures>15Creadilyproduceszoosporesandnewvegetativegrowthfollowingwinterdormancyor
severefloods.CellwallsofthefreshwaterspeciesC.glomerataincludesilica,anessentialnutrientforthis
species. Calcium is also an essential element and CaCO3 depositions occur in older cells. In seawater,
CladophoranormallyabsorbsbicarbonateratherthanCO2forphotosynthesis.
BIOTECHNOLOGY
In Thailand, Cladophora edible freshwater alga, is known as Kai. It is abundant in the Nan and Mekong
rivers in the Northern part of Thailand. The local people around these rivers collect it for domestic
consumption and it is sold in the markets and is now cultured by using wastewater from fish rearing
(KhuantrairongandTraichaiyaporn,2011).
Cladophoraglomerataisgenerallyconsideredasagoodbioindicatorofheavymetalsinaquaticbodies
(Whittonetal.,1989).
References
AlgaeBase:http://www.algaebase.org/search/genus/detail/?genus_id=37
http://www.algaebase.org;searchedon02July2010.
JohnD.M.(2002)OrderCladophorales(=Siphonocladales).In:JohnD.M.,WhittonB.A.,Brook,A.J.(eds.)TheFreshwaterAlgalFlora
oftheBritishIsles.Anidentificationguidetofreshwaterandterrestrialalgae,CambridgeUniversityPress,Cambridge,pp.
468470.
Khuantrairong T., Traichaiyaporn S. (2011) The nutritional value of edible freshwater alga Cladophora sp. (Chlorophyta) grown
underdifferentphosphorusconcentrations.InternationalJournalofAgriculture&Biology13:297300.
Thomas D.N., Collins J.C., Russell G. (1988) Interactive effcts of temperature and salinity upon net photosynthesis of Cladophora
glomerata(L.)Ktz.C.rupestris(L.)Ktz.BotanicaMarina31:7377.
WhittonB.A.,BurrowsI.G.,KellyM.G.(1989)UseofCladophoraglomeratatomonitorheavymetalsinrivers.JournalofApplied
Phycology1:293299.
Wiencke C., Davenport J. (1987) Respiration and photosynthesis in the intertidal alga Cladophora rupestris (L.) Ktz. under
fluctuatingsalinityregimes.JournalofExperimentalMarineBiologyandEcology114:183197.
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9.1.4 Codiumsp.
TAXONOMY
ThegenusCodiumcurrentlycontains127recognizedspecies.
Codiumfragile
Figure68Codiumfragile(Suringar)Hariot.
Kommetjie,CapePeninsula,SouthAfrica
MichaelD.Guiry
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Bryopsidophyceae
Bryopsidales
Codiaceae
Codium
Codiumfragile
Codiumparvulum
Figure69Codiumparvulum(BoryexAudouin)
P.C.Silva,Atlit,IsraelDr.AlvaroIsrael
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SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Chlorophyta
Bryopsidophyceae
Bryopsidales
Codiaceae
Codium
Codiumparvulum
BIOLOGY
Thallusspongy,anchoredtorocksorshellsbyaweftofrhizoids,varyinginsizefrom1cmto10mlong.
Habit varying widely applanate, pulvinate, digitaliform, globular, petaloid, membraniform, or
dichotomously branched; erect or repent; branches wholly terete or variously flattened, at times
anastomosing. Internal structure composed of a colorless medulla of densely intertwined siphons and a
green palisadelike layer of vesicles called utricles. Organelles, including innumerable nuclei and discoid
chloroplasts(butnoamyloplasts)areconfinedtoalayerofcytoplasmappressedtoawallofwhichmannan
is an important constituent. Chloroplasts lack pyrenoids; carotenoid pigments include siphaxanthin and
siphonein. Incomplete septa (plugs) formed by centripetal deposition of wall material. Utricles arise
primarily by enlargement of sympodial branches of medullary siphons, secondarily by budding or by
productionofadditionalutricleformingmedullarysiphonsfrombasalportionofexistingutricles.Mature
utricles cylindrical or clavate, the apical wall usually thickened and often ornamented in a pattern
characteristicofparticularspecies.Rhizoidalsiphons,whichbecomeburiedinthemedulla,alsoproduced
from basal portion of utricles. Colorless hairs, each with a basal plug, produced by utricles shortly below
theirapices,caducousattheplug,whichremainsasaprominentscar.Gametangiaproducedlaterallyby
utricles, each with a basal plug above a short pedicel; fusiform to ovoid, the contents cleaving into
biflagellate gametes following meiosis. Gametes extruded in a gelatinous mass through apical rupture.
Malegametescontainonlyoneortwochloroplasts,femalegametesseveraltimeslarger,withnumerous
chloroplasts,thetwo typesproducedonthesamethallus(monoecious)ormoreoftenondifferent thalli
(dioecious).Zygotedevelopsintoamorphousprostratevesiclethatproduceserectelongatevesicles;these
in turn initiate primary utricleproducing siphons which eventually consolidate into a multiaxial thallus.
Asexual reproduction by parthenogenesis, fragmentation, or the cutting off of modified aborted
gametangia.DespitetheubiquityofCodiumverylittleisknowaboutitsbiology.Thelifehistoryoutlined
aboveisgeneralizedfromfragmentarystudies.Codiumparvulumhasbeenrecentlydescribedasblooming
inNothernshoresofIsrael.
BIOTECHNOLOGY
CodiumfragileisusedasfoodinKoreaandthePhilippines,inthelatterofwhichotherspeciesofCodium
are also used. These are used also in other countries such as Argentina, Hawaii, Israel and Indonesia
(ZemkeWhite and Ohno, 1999). Codium in Korea is produced in an amount of 150 kg dry weight year1
References
AlgaeBase:http://www.algaebase.org/search/genus/detail/?genus_id=39&sk=0
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IsraelA.,EinavR.,SilvaP.C.,PazG.,ChacanaM.E.,DouekJ.(2010)FirstreportoftheseaweedCodiumparvulum(Chlorophyta)in
Mediterraneanwaters:recentbloomsonthenorthernshoresofIsrael.Phycologia49:107112.
KusakinaJ.,SnyderM.,KristieD.N.,DadswellM.J.(2006)MorphologicalandmolecularevidenceformultipleinvasionsofCodium
fragileinAtlanticCanada.BotanicaMarina49:19.
Lam D.W., Zechman F.W. (2006) Phylogenetic analyses of the Bryopsidales (Ulvophyceae, Chlorophyta) based on RUBISCO large
subunitgenesequences.JournalofPhycology42:669678.
Mondragon J., Mondragon J. (2003) Seaweeds of the Pacific Coast. Common marine algae from Alaska to Baja California, Sea
Challengers,Monterey,California,pp.iv,597.
PacficodeMxico.I.Chlorophycota,UniversidadAutnomadeBajaCalifornia,Ensenada,Mxico,pp.iviii,17146.
Provan J., Booth D., Todd N.P., Beatty G.E., Maggs C.A. (2008) Tracking biological invasions in space and time: elucidating the
invasive history of the green alga Codium fragile using old DNA. Diversity and Distributions A Journal of Conservation
Biogeography14:343354.
ScagelR.F.(1957)AnannotatedlistofthemarinealgaeofBritishColumbiaandnorthernWashington(includingkeystogenera).
Bulletin,NationalMuseumofCanada150:vi289.
Schorie,D.,SeligU.,SchubertH.(2009)SpeciesandsynonymlistoftheGermanmarinemacroalgaebasedonhistoricalandrecent
records.Rostock.MeeresbiologieBeitraeg21:7135.
SilvaP.C.(1955)ThedichotomousspeciesofCodiuminBritain.JournaloftheMarineBiologicalAssociationoftheUnitedKingdom
34:565577.
SmithG.M.(1944)MarinealgaeoftheMontereyPeninsula,StanfordUniversityPress,Stanford,pp.ix,622,98pls..
376.
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9.2 Rhodophyta
9.2.1
Chondruscrispus
Figure70Chondruscrispus(Stackhouse)
NewQuay,Co.Clare,Ireland;plantto60mmlong;onrock
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Florideophyceae
Gigartinales
Gigartinaceae
Chondrus
Chondruscrispus
Relatedspecies
C.armatus,C.canaliculatus,C.elatus,C.giganteus,C.giganteusf.flabellatus,C.nipponicus,C.ocellatusf.
crispus,C.ocellatusf.parvus,C.ocellatus,C.ocellatusf.aequalis,C.ocellatusf.crispoides,C.pinnulatusf.
conglobatus,C.pinnulatus,C.verrucosus,C.yendoif.subdichotomus,C.yendoi,C.yendoif.fimbriatus,C.
crispusvar.lonchophorus,C.crispusvar.filiformis.
BIOLOGY
Cartilaginous,darkpurplishred,red,yellowishorgreenishfrondsto150mmhigh,gametophyteplantsare
ofteniridescentunderwaterwheningoodcondition.Stipecompressed,narrow,expandinggraduallytoa
flat,repeatedlydichotomouslybranchedfrond,intuftsfromadiscoidholdfast.Axilsrounded,apicesblunt
or subacute, frond thicker in centre than margins. Very variable in breadth of segments. Very variable in
branching, colour and thickness. Cystocarps are embedded and aggregated distally or confined to
proliferations. Cystocarps lack a surrounding filamentous hull, the major generic defining character of
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Chondrus.Maleplantsareoftensaidtobeextremelyrare,butarereportedtooccurinnormallyexpected
proportions in at least some populations of the most commerically valuable species, C. crispus.
Tetrasporangiaoccurinbranchedchainsformedlaterallyoncellsoftheinnercortex.
C.crispusisalmostexclusivelyrestrictedtorockandrangesfromintertidalto24mdepths,beingalso
tolerantofloweredsalinities.ItoccursinthenorthAtlanticfromNewJerseytoSpainandpossiblyMorocco
andtheCapeVerdeIslands,aswellasJapan.
BIOTECHNOLOGY
Thisgenusiseconomicallyimportant,currentlyineasternCanadaandFrance,andformerlyinBritainand
Ireland.Muchemphasishasbeendevotedtoitscultivationinoutdoortanksasrawmaterialforcommercial
utilization(ChenandTaylor,1978;Simpsonetal.,1978;SimpsonandShacklock,1979;ShacklockandCroft,
1981;Bidwelletal.,1985).Bidwelletal.(1984,1985)devisedandtestedoutdoortankmethodsgenerally
applicabletoChondrusculturebothwithinandoutsideofitsnativerange,butconcludethatworldprices
donotalloweconomicallyviable cultivationineasternCanada wherethelargestnaturalpopulations are
found. C. crispus is used for carrageenan production in France, Spain and US, and as food in Ireland and
France,whileinJapanC.ocellatusisusedasfood(ZemkeWhiteandOhno,1999).Canadaproduces10,000
tdryweightperyearofChondrus,France1,260,Japan500,Spain300,US120,Portugal30andIreland3
References
Bates C.R., Saunders G.W., Chopin T. (2005) An assessment of two taxonomic distinctness indices for detecting seaweed
assemblageresponsestoenvironmentalstress.BotanicaMarina48:231243.
Bidwell R.G.S., Lloyd N.D.H., McLachlan J. (1984) The performance of Chondrus crispus (Irish moss) in laboratory simulations of
environmentsindifferentlocations.ProceedingsoftheInternationalSeaweedSymposium6:292294.
BidwellR.G.S.,McLachlanJ.,LloydN.D.H.(1985)TankcultivationofIrishmoss,ChondruscrispusStackh.BotanicaMarina28:8797.
ChenL.C.M.,TaylorA.R.A.(1978)MedullarytissuecultureoftheredalgaChondruscrispus.CanadianJournalofBotany56:883
886.
DixonP.S.,IrvineL.M.(1977)SeaweedsoftheBritishIsles.Volume1.Rhodophyta.Part1.Introduction,Nemaliales,Gigartinales,.
BritishMuseum(NaturalHistory),London,pp.xi252,90figs.
Hommersand M.H., Guiry M.D., Fredericq S., Leister G.L. (1993) New perspectives in the taxonomy of the Gigartinaceae
(Gigartinales,Rhodophyta).ProceedingsoftheInternationalSeaweedSymposium14:105120,41figs.
MikamiH.(1965)AsystematicstudyofthePhyllophoraceaeandGigartinaceaefromJapananditsvicinity.MemoirsoftheFaculty
ofFisheriesHokkaidoUniversity5:181285.
ShacklockP.F.,CroftG.B.(1981)EffectofgrazersonChondruscrispusinculture.Aquaculture22:331342.
Silva P.C., Basson P.W., Moe R.L. (1996) Catalogue of the benthic marine algae of the Indian Ocean. University of California
PublicationsinBotany79:11259.
Simpson F.J., Shacklock P.F. (1979) The cultivation of Chondrus crispus. Effect of temperature on growth and carrageenan
production.BotanicaMarina22:295298.
Simpson F.J., Neish A.C., Shacklock P.F., Robson D.R. (1978) The cultivation of Chondrus crispus. Effect of pH on growth and
productionofcarrageenan.BotanicaMarina21:229235.
376.
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9.2.2
Mastocarpusstellatus
Figure71Mastocarpusstellatus(Stackhouse)
GuiryMuighInis,Co.Galway,Ireland;onrock
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Florideophyceae
Gigartinales
Phyllophoraceae
Mastocarpus
Mastocarpusstellatus
Relatedspecies
M.jardinii,M.pacificus,M.papillatus,M.yendoi.
BIOLOGY
Cartilaginous,purplishbrownfronds,oftenindensetufts,arisingfromadiscoidholdfast,to200mmhigh.
Narrowcompressedstipeexpandsintostraplikeblade,usuallyinrolledtoformachannel,withthickened
margins.Repeatedlydichotomouslybranched,axilsacute.Upperpartoffrondwithpapillaeto10mmor
more long on surfaces and margins on female plants. Male plants lack papillae and are generally rare.
Tetrasporophyteapurplishblackcrust(Petrocelisphase).
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M. stellatus occurs on rocks in lower intertidal, often in large continuous mats, widespread and
abundant. Both the gametophyte and tetrasporophytic stages are characteristic of high to mid intertidal
rocksonopencoasts,althoughitalsooccurssubtidally.
M. stellatus occurs in the north Atlantic Ocean from Russia to Portugal and from Morocco to
MauritaniaandfromRhodeIslandtoNewfoundland.
BIOTECHNOLOGY
M. stellatus is used as a source of carrageenan in Spain and Portugal and as food in Ireland. Annual
productionisof600tdryweightinSpain,70inPortugaland5inIreland(ZemkeWhiteandOhno,1999).
References
AlgaeBase:http://www.algaebase.org/search/species/detail/?species_id=24&sk=0&from=results
http://www.algaebase.org/search/genus/detail/?genus_id=20
Bates C.R., Saunders G.W., Chopin T. (2005) An assessment of two taxonomic distinctness indices for detecting seaweed
assemblageresponsestoenvironmentalstress.BotanicaMarina48:231243.
GuiryM.D.,WestJ.A.(1983)LifehistoryandhybridizationstudiesonGigartinastellataandPetroceliscruenta(Rhodophyta)inthe
NorthAtlantic.JournalofPhycology19:474494.
Silva P.C. (1952) A review of nomenclatural conservation in the algae from the point of view of the type method. University of
CaliforniaPublicationsinBotany25:241323.
376.
Zuccarello G.C., Schidlo A., McIvor L., Guiry M.D. (2006) A molecular reexamination of speciation in the intertidal red alga
Mastocarpusstellatus(Gigartinales,Rhodophyta)inEurope.EuropeanJournalofPhycology40:337344.
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9.2.3
Grateloupiaturuturu
Figure72GrateloupiaturuturuYamadawithBifurcariabifurcata
Finisterre,Galicia,Spain,2008
IgnacioBrbara
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Florideophyceae
Halymeniales
Halymeniaceae
Grateloupia
Grateloupiaturuturu
Relatedspecies
ThegenusGrateloupiacontains87relatedspecies
BIOLOGY
Grateloupia is compressed to foliose, linear to lanceolate, rarely unbranched, usually branched
proliferously, to one or more orders, in one or more planes. Stipitate. Holdfast a basal disc. Lubricous to
leathery. Medulla filamentous, with rhizoids. Inner cortex of anastomosing stellate cells, outer cortex of
anticlinalfilesofmoreorlessisodiametriccells,progressivelysmallertowardcuticle.Femalereproductive
structures in ampullae, sparingly branched usually to only two orders. Carpogonia terminating a 2celled
branchontheprimaryampullarfilament.Connectingfilamentscanfusewithasuccessionofauxiliarycells
in separate ampullae. A single, outwardly directed gonimoblast initial produces a compact, embedded
cystocarpwithmanysmallcarposporangia.Smallauxiliaryfusioncellformed.Carpostomeusuallypresent.
Spermatangia superficial, sometimes in nemathecial sori. Tetrasporophyte isomorphic. Tetrasporangia
cruciate,attachedsubbasallytointermediatecorticallayer.
Grateloupiaisdistributedinwarmtemperate(totropical)watersthroughouttheworld.G.turuturuisa
marinespecies.
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BIOTECHNOLOGY
G.filicinaisusedasfoodinIndonesiaandJapan(ZemkeWhiteandOhno,1999).
References
BarreiroR.,QuintelaM.,BrbaraI.,CremadesJ.(2006)RAPDdifferentiationofGrateloupialanceolaandtheinvasiveGrateloupia
turuturu(Gigartinales,Rhodophyta)intheIberianPeninsula.Phycologia45:213217.
D'Archino R., Nelson W.A., Zuccarello G.C. (2007) Invasive marine red alga introduced to New Zealand waters: first record of
Grateloupiaturuturu(Halymeniaceae,Rhodophyta).NewZealandJournalofMarineandFreshwaterResearch41:3542.
De Clerck O., Gavio B., Fredericq S., Brbara I., Coppejans E. (2005) Systematics of Grateloupia filicina (Halymeniaceae,
Rhodophyta),basedonrbclsequenceanalysesandmorphologicalevidence,includingthereinstatementofG.minimaand
thedescriptionofG.capensissp.nov.JournalofPhycology41:391410.
Figueroa F.L., Korbee N., de Clerck O., Brbara I., Gall E.A.R. (2007) Characterization of Grateloupia lanceola (Halymeniales,
Rhodophyta), an obscure foliose Grateloupia from the Iberian Peninsula, based on morphology, comparative sequence
analysisandmycosporinelikeaminoacidcomposition.EuropeanJournalofPhycology42:231242.
GavioB.,FredericqS.(2002)Grateloupiaturuturu(Halymeniaceae,Rhodophyta)isthecorrectnameofthenonnativespeciesin
theAtlanticknownasGrateloupiadoryphora.EuropeanJournalofPhycology37:349360.
Verlaque M., Brannock P.M., Komatsu T., VillalardBohnsack M., Marston M. (2005) The genus Grateloupia C. Agardh
(Halymeniaceae, Rhodophyta) in the Thau Lagoon (France, Mediterranean): a case study of marine plurispecific
introductions.Phycologia44:477496.
Xia B.M. (2004) Flora algarum marinarum sinicarum Tomus II Rhodophyta No. III Gelidiales Cryptonemiales Hildenbrandiales,
SciencePress,Beijing,pp.ixxi,1203,plsIXIII.
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9.2.4
Palmariapalmata
Figure73Palmariapalmata(Linnaeus)Kuntze
Spiddal,Co.Galway,Ireland;lowerintertidal
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Florideophyceae
Palmariales
Palmariaceae
Palmaria
Palmariapalmata
Relatedspecies
P. callophylloides, P. decipiens, P. georgica, P. hecatensis, P. marginicrassa, P. mollis, P. moniliformis, P.
stenogona.
BIOLOGY
Thalluswitherectstipitatefrondsarisingfromabasaldisc,bladesflattened,becomingcartilaginouswith
age, simple, dichotomously or palmately divided, frequently with marginal, palmately divided
proliferations; construction multiaxial, cortex of 25 layers of small pigmented cells increasing in number
ratherthansizeasthethallusmaturestoformameristoderm,medullacompactwith25layersoflarge,
rounded,looselycoherent,almostcolourlesscellswithstringsofbeadlikechloroplasts.Gametangialplants
dioecious;spermatangiaformedinlarge,irregular,tortuoussoriovermostofthesurfaceoferect,frondose
blades similar in morphology to the tetrasporangial plants; carpogonia occurring as single cells on young
plantletsonly;tetrasporangialplantdevelopingdirectlyfromthefertilizedcarpogoniumandovergrowing
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thecarpogonialplant,carposporophytelacking.Tetrasporangialarge,inextensive,irregular,tortuoussori
generally covering much of the surface of the young frond when mature, formed in a terminal position
fromcorticalcells,interspersedwithmodified,pigmented,sterilefilaments,regeneratingrepeatedlyfrom
abasalgenerativestalkcell.Sporesregularlycruciatelyordecussatelyarranged.
Marine. On rock, mussels and epiphytic on several algae, intertidal (at all levels but particularly near
lowwater)andshallowsubtidal,especiallyonupperpartofLaminariahyperboreastipes(toadepthof5
m),widelydistributed,abundant.
BIOTECHNOLOGY
PalmariapalmataisafooditemofancientusageintheNorthAtlantic.Thisspeciesisvariouslyknownas
dulse,dillisk,duilleasgorduillisg(IrishGaelic),thenarrowerformsbeingcalledcreannach(IrishGaelic),soll
(Icelandic), sousll, sl, blm (Norwegian), gomon vaches (French), and darusu (Japanese). Dulse is a
goodsourceofvitaminsandminerals,particularlywhencomparedtohigherplantvegetables(Morganet
al.,1980),andisstilleateninScotland,Ireland,Iceland,BrittanyandNorway,although,inmanyofthese
areas,itsuseisonlyoccasional.Dulsecontainslargeamountofseveralunusualcarbohydratesincludingan
unusualshortchainedone,floridoside,andthiscanformupto30%ofthedryweight.Scotland,Norway,
IcelandandeasternCanadaallproducesmallamountsofdulseforhumanconsumption.InIrelandabout
20 dry tonnes are sold each year, while in Canada 100 t are produced yearly (ZemkeWhite and Ohno,
1999).
References
http://www.algaebase.org/search/species/detail/?species_id=1
Irvine L.M., Guiry M.D. (1983) Palmariales. In: Irvine L. (ed.) Seaweeds of the British Isles. Volume 1. Rhodophyta, Part 2A
Cryptonememiales(sensustricto),Palmariales,Rhodymeniales,Vol.1(2A),BritishMuseum,London,pp.6598.
MorganK.C.,ShacklockP.F.,SimpsonF.J.(1980)SomeaspectsofthecultureofPalmariapalmataingreenhousetanks.Botanica
Marina23:765770.
MorganK.C.,WrightJ.L.C.,SimpsonF.J.(1980)ReviewofchemicalconstituentsoftheredalgaPalmariapalmata(dulse).Economic
Botany34:2750.
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9.2.5 Solieriachordalis
Figure74Solieriachordalis(C.Agardh)J.Agardh
StephanieBondu(UBOBrest,France)
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Rhodophyta
Florideophyceae
Gigartinales
Solieriaceae
Solieria
Solieriachordalis
Relatedspecies
S.anastomosa,S.dichotoma,S.dura,S.filiformis,S.jaasundii,S.pacifica,S.robusta,S.tenuis.
BIOLOGY
Plantsofthelargestspeciesreach30cminlength.Thalliareerectorrecumbentfromacrustoseorfibrous
holdfast, terete to compressed, and irregularly to subdichotomously branched. Apices are multiaxial, the
successive axial cells producing single periaxial derivatives either in orthostichous rows or rotated 120
degrees in a zig zag fashion. The medulla is broad and laxly filamentous with abundant rhizoids, and is
surroundedbyacortexofprogressivelysmallersubisodiametriccells.Carpogonialbranchesare3celled,at
times bearing sterile cells on the basal cell, and emit a single nonseptate connecting filament from
fertilized carpogonia. Auxiliary cells are prominently situated in an "auxiliary cell complex" prior to
diploidization. Cystocarps are deeply embedded in the axes, often in clusters, and consist of an ostiolate
pericarp with a filamentous inner hull surrounding a carposporophyte in which a large central fusion cell
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gives rise peripherally to short chains of gonimoblast cells bearing single, terminal carposporangia.
Tetrasporangiaarelaterallypitconnectedtobearingcells.
S.chordalisisamarinespecies.
BIOTECHNOLOGY
AlgaeofthegenusSolieriaareusedasfoodinMyanmar(ZemkeWhiteandOhno,1999).
References
Bondu S., Cocquempot B., Deslandes E., Morin P. (2008) Effects of salt and light stress on the release of volatile halogenated
organiccompoundsbySolieriachordalis:alaboratoryincubationstudy.BotanicaMarina51:485492.
DeMasiF.,GargiuloG.M.(1982)Solieriachordalis(C.Ag.)J.Ag.(Rhodophyta,Gigartinales)enMditerrane.Allionia25:109111.
Deslandes E., Floc'h J.Y., BodeauBellion C., Brault D., Braud, J.P. (1985) Evidence for (iota)carrageenans in Solieria chordalis
(Solieriaceae)andCalliblepharisjubata,Calliblepharispurpureum(Rhodophyllidaceae).BotanicaMarina28:317318.
http://www.algaebase.org;searchedon30June2010.
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9.3 Phaeophyceae(phylumHeterokontophyta)
9.3.1
Alariaesculenta
Figure75Alariaesculenta(Linnaeus)Greville
Spiddal,Co.Galway,Ireland.Frondsexposedatlowwater;16mmlens
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Alariaceae
Alaria
Alariaesculenta
Relatedspecies
A. angusta, A. crassifolia, A. crispa, A. elliptica, A. esculenta f. latifolia, A. esculenta f. angustifolia, A.
fragilis,A.grandifolia,A.marginata,A.ochotensis,A.paradisea,A.praelonga,A.pylaiei.
BIOLOGY
Plantswitholiveoryellowbrownfrondsto4mlongand25cmwide.Attachedbyarootlikeholdfastat
the base from which a narrow flexible stipe arises which continues into the leafy part of the plant as a
distinct midrib. The reproductive structures, apparent as darkbrown areas, are confined to unbranched
leafy appendages borne on the stipe, usually in two rows. This is the only kelplike plant in Ireland and
Britainwithadistinctmidribandistheonlyonewithsporangiaborneatthebaseofthefrondinspecial
leafletscalledsporophylls.
Generallygrowingonrockinveryexposedplaces,oftenformingabandatlowwaterandintheshallow
subtidal,butalsooccurringintidalpoolsinthelowershore.
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BIOTECHNOLOGY
A. esculenta, as well as other Alaria species, is used as food in European, North American and Asian
countries(ZemkeWhiteandOhno,1999).ForfurtherinformationonAlariabiotechnologyseeparagraph
BiotechnologyforLaminaria,SaccharinaandSaccorhiza.
References
AlgaeBase:http://www.algaebase.org/search/species/detail/?species_id=82
Lane C.E., Saunders G.W. (2005) Molecular investigation reveals epi/endophytic extrageneric kelp (Laminariales, Phaeophyceae)
gametophytescolonizingLessoniopsislittoralisthalli.BotanicaMarina48:426436.
LoiseauxdeGorS.,NoaillesM.C.(2008)AlguesdeRoscoff,EditionsdelaStationBiologiquedeRoscoff,Roscoff,pp.[1]215.
WiddowsonT.B.(1971)AtaxonomicrevisionofthegenusAlariaGreville.Syesis4:1149.
376.
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9.3.2 Undariapinnatifida
Figure76Undariapinnatifida(Harvey)Suringar
Venice,Italy;onchainsofvaporettostop
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Alariaceae
Undaria
Undariapinnatifida
Relatedspecies
U.crenata,U.undarioides
BIOLOGY
Life history diplohaplontic with alternation of large sporophyte bearing unilocular meiosporangia with
paraphyses(sori)andmicroscopicdioeciousandoogamous,heteromorphousgametophytes(fordetail,see
Laminaria). Haploid chromosome number is 30 in U. pinnatifida. Sporophyte annual, appearing in winter
anddisintegratingthefollowingautumn.Sporophytecomposedofholdfastwithhaptera,stipe,andblade.
Meristematic region situated at transition zone between stipe and blade. Stipe compressed at base to
flattened above, with wings (greatly expanded to narrow) which are more or less undulatoplicated and
with sori or sterile. Blade linear to rounded or with pinnate lobes, with midrib or thickened fascia.
Cryptostomata and dotlike mucilage glands present. Sori develop in summer on both surfaces of wings
(sporophylls),oronbothsidesofmidriborfascia,oratthesametimeonsporophyllsandblade.Inculture,
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sori of U. pinnatifida are formed in longday conditions, probably induced by high water temperatures.
Zoospores of U. pinnatifida germinate between 13 and 24C gametophytes grow well at 1523C (Saito
1956).Survivalrangeofgamtophytesis1to28C.OptimaltemperaturesforformationandgrowthofU.
pinnatifidasporophytesis1020Cbutat5and25Csporophytesarestillformed.Structureofsporophyte
asinLaminariacomposedofphotosyntheticmeristoderm,parenchymaticcortexandcentralmedulla.InU.
pinnatifidaparthenosporophyteswithnormalmorphologymaydevelopatlowratesreleasingonlyfemale
determinedzoosporeswhenmaturing.
GenusoriginallyendemictothenorthwesternPacific,growingsubtidallyonrocksinwarmtemperate
waters of Japan, Korea and China. Recently, U. pinnatifida was introduced to France, New Zealand and
Tasmania,probablyviaoysterculturesandships.Sincethenthespeciesisexpandingitsdistributionrange.
BIOTECHNOLOGY
Genus of great economic importance as food source in Japan and Korea, especially U. pinnatifida (trade
name: Wakame). Total annual yield from natural harvest and cultivation sites of Undaria spp. was about
130,000tonsfreshweightin1967.Longlineculturestartedintheearly1960'sandmeanwhileproduction
increased to about 30% of the wild harvest in Japan. In recent years, first progress in tissue culture,
protoplast isolation and cryopreservation of U. pinnatifida was achieved. Annual productions in t dry
weightare: Australia6,China20,000,Japan18,310(allfromculture)andKorea83,398(allfromculture)
(ZemkeWhite and Ohno, 1999). For further information on Undaria biotechnology see pargraph on
BiotechnologyforLaminaria,SaccharinaandSaccorhiza.
References
AdamsN.M.(1997)CommonseaweedsofNewZealand,CanterburyUniversityPress,Christchurch,pp.148,48pls.
Lane C.E., Mayes C., Druehl L.D., Saunders G.W. (2006) A multigene molecular investigation of the kelp (Laminariales,
Phaeophyceae)supportssubstantialtaxonomicreorganization.JournalofPhycology42:493512.
PedrocheP.F.,SilvaP.C.,AguilarRosasL.E.,DreckmannK.M.,AguilarRosasR.(2008)CatlogodelasalgasbenthnicasdelPacfico
de Mxico II. Phaeophycota, Universidad Autnoma Metropolitana and University of California Berkeley, Mexicali &
Berkeley,pp.ivi,15146.
Sliwa C., Johnson C.R., Hewitt C.L. (2006) Mesoscale dispersal of the introduced kelp Undaria pinnatifida attached to unstable
substrata.BotanicaMarina49:396405.
YoshidaT.(1998)MarinealgaeofJapan.UchidaRokakuhoPublishing,Tokyo,pp.251222.
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9.3.3 Ascophyllumnodosum
Figure77Ascophyllumnodosum(Linnaeus)LeJolis
CibhantSruthin,anCheathrRua,Co.naGaillimhe;10.5mmlens
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Fucaceae
Ascophyllum
Ascophyllumnodosum
Relatedspecies
The genus Ascophyllum only includes Ascophyllum nodosum. A free floating variety is known as
Ascophyllumnodosumvar.mackayi(Turner)Cotton.
BIOLOGY
This is a brown seaweed that is closely related to Fucus. It forms a single bladders centrally in long,
flattened straplike fronds. The fronds hang downwards, draping sheltered intertidal rocks. Many fronds
growfromthebaseandtheplantgenerallyregeneratesnewfrondsfromthebasewhenoneofthelarger
fronds are damaged. There is evidence that clumps can be over 400 years old and may be even older.
AscophyllumiscurrentlyconfinedtotheNorthAtlanticbasin,butplantshavebeenfoundgrowinginSan
FranciscoBay,butthespeciesdoesnotpersistthere.Theplantsareusedaspackingforshellfishfromthe
NorthAtlanticandwhendiscardedmaytakehold.
BIOTECHNOLOGY
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Ascophyllum nodosum is used in agriculture as a fertilizer and plant growth promoter (France, Canada,
China,Iceland,US)andforalginateproduction (Ireland,Norway,UK) (ZemkeWhiteand Ohno,1999).All
theannualproductionisharvestedfromthewild.Annualproductionis:Ireland8,999tdryweight,Norway
6,632, Iceland 4,400, UK 3,500, China 3,000, Canada 2,500, France 1,700 and US 280 (ZemkeWhite and
Ohno,1999).
References
BaardsethE.(1970)SynopsisofbiologicaldataonknobbedwrackAscophyllumnodosum(Linnaeus)LeJolis.FAOFisheriesSynopsis
38(Rev.1).
ChoG.Y,RousseauF.,ReviersB.De,BooS.M.(2006)PhylogeneticrelationshipswithintheFucales(Phaeophyceae)assessedbythe
photosystemIcodingpsaAsequences.Phycologia45:512519.
McHughD.J.(2003)Aguidetotheseaweedindustry.FAOFisheriesTechnicalPaper441.
376.
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9.3.4
Fucussp.
TAXONOMY
Relatedspecies
F. ceranoides, F. cottonii, F. distichus, F. evanescens, F. gardneri, F. lagasca, F. parksii, F. serratus, F.
setaceus,F.spataeformis,F.spiralis,F.vesiculosus,F.virsoides.
Fucusserratus
Figure78FucusserratusLinnaeus
Spiddal,Co.Galway,Ireland;lowerintertidal
M.D.Guiry
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Fucaceae
Fucus
Fucusserratus
Fucusspiralis
Figure79FucusspiralisLinnaeus
Flaggyshore,Finavarra,Co.Clare,Ireland;receptacleswithsterileedge
M.D.Guiry
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SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Fucaceae
Fucus
Fucusspiralis
Fucusvesiculosus
Figure80FucusvesiculosusLinnaeus
TrnaReilige,AnCheathrRua,Co.Galway,Ireland
M.D.Guiry
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Fucaceae
Fucus
Fucusvesiculosus
BIOLOGY
FrondsarespirallytwistedwithnovesiclesandevenmargininF.spiralis,flatwithvesiclesandevenmargin
in F. vesiculosus and flat with no vesicles and serrated margin in F. serratus. F. spiralis is hermaphrodite,
with rounded inflated receptacles with no dichotomies and a sterile rim; F. vesiculosus is dioecious, with
ellipsoidalelongate pointed inflated receptacles with 12 dichotomies and no definite rim; F. serratus is
dioeciouswithnoinflatedreceptacleswithextendedgrowthandseveraldichotomiesandnodefiniterim.
Crossfertilizationwithhybridformationbetweenpairsofthesespecieshavebeenobserved.
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F.spiralisisnormallyfoundinthehighestpositionontheshore,whereitmaybeleftexposedforseveral
daysduringneaptides,F.vesiculosusoccupiesthemidtidezoneincompetitionwithAscophyllumnodosum
and is alternatively exposedsubmerged, while F. serratus occupies the lowest level, that is more rarely
exposed.
BIOTECHNOLOGY
SeveralFucusspeciesareusedasfood(France,Portugal,Alaska),foragricultureuses(France,Canada),and
as alginate source (Ireland). Annual production of Fucus is of 80 t dry weight in Ireland, 2 in France and
0.04inPortugal(ZemkeWhiteandOhno,1999).
References
BillardE.,DaguinC.,PearsonG.,SerraoE.,EngelC.,ValeroM.(2005)Geneticisolationbetweenthreecloselyrelatedtaxa:Fucus
vesiculosus,F.spiralisandF.ceranoides(Phaeophyceae).JournalofPhycology41:900905.
BillardE.,SerraoE.A.,PearsonG.A.,EngelC.R.,DestombeC.,ValeroM.(2006)Analysisofsexualphenotypeandprezygoticfertility
in natural populations of Fucus spiralis, F. vesiculosus (Fucaceae, Phaeophyceae) and their putative hybrids. European
BrawleyS.H.,CoyerJ.A.,BlakesleeA.M.,HoarauG.,JohnsonL.E.,ByersJ.E.,StamW.T.,OlsenJ.L.(2009)Historicalinvasionsofthe
intertidal zone of Atlantic North America associated with distinctive patterns of trade andemigration. Proceedings of the
NationalAcademyofSciencesoftheUnitedStatesofAmerica106:82398244.
BurrowsE.M.,LodgeS.(1951)AutoecologyandthespeciesprobleminFucus.JournaloftheMarineBiologicalAssociationofthe
UnitedKingdom30:161176.
CerantolaS.,BretonF.,ArGallE.,DeslandesE.(2006)Cooccurrenceandantioxidantactivitiesoffucolandfucophloretholclasses
ofpolymericphenolsinFucusspiralis.BotanicaMarina49:347351.
ColemanM.A.,BrawleyS.H.(2005)Arelifehistorycharacteristicsgoodpredictorsofgeneticdiversityandstructure?Acasestudyof
theintertidalalgaFucusspiralis(Heterokontophyta,Phaeophyceae).JournalofPhycology41:753762.
Gabrielson P.W., Widdowson T.B., Lindstrom S.C. (2004) Keys to the seaweeds and seagrasses of Oregon and California.
PhycologicalContribution6:iv181.
Garbary D.J., Brackenbury A., McLean A.M., Morrison D. (2006) Structure and development of air bladders in Fucus and
Ascophyllum(Fucales,Phaeophyceae).Phycologia45:557566.
Larsen A., SandJensen K. (2005) Salt tolerance and distribution of estuarine benthic macroalgae in the KattegatBaltic Sea area.
Phycologia45:1323.
Mathieson A.C., Dawes C.J., Wallace A.L., Klein A.S. (2006) Distribution, morphology, and genetic affinities of dwarf embedded
FucuspopulationsfromtheNorthwestAtlanticOcean.BotanicaMarina49:283303.
Moss B.L. (1950) Studies in the genus Fucus. II. The anatomical structure and chemical composition of receptacles of Fucus
vesiculosusfromthreecontrastinghabitats.AnnalsofBotany14:395410.
MossB.L.(1950)StudiesinthegenusFucusIII.StructureanddevelopmentoftheattachingdiscsofFucusvesiculosus.Annals of
Botany14:411419.
Ngeli C. (1849) Gattungen einzelliger Algen, physiologisch und systematisch bearbeitet. Neue Denkschriften der Allg.
SchweizerischenGesellschaftfrdieGesammtenNaturwissenschaften10(7):iviii,1139,plsIVIII.
PearsonG.,LagoLestonA.,ValenteM.,SerraoE.(2006)SimpleandrapidRNAextractionfromfreezedriedtissueofbrownalgae
andseagrasses.EuropeanJournalofPhycology41:97104.
PetersA.F.,MarieD.,ScornetD.,Kloareg,B.,CockJ.M.(2004)ProposalofEctocarpussiliculosus(Ectocarpales,Phaeophyceae)asa
modelorganismforbrownalgalgeneticsandgenomics.JournalofPhycology40:10791088.
PerrinC.,DaguinC.,VandeVlietM.,EngelC.R.,PearsonG.A.,SerroE.A.(2007)Implicationsofmatingsystemforgeneticdiversity
of sister algal species: Fucus spiralis and Fucus vesiculosus (Heterokontophyta, Phaeophyceae). European Journal of
Phycology42:219230.
PowellH.T.(1960)ThetypificationofFucusspiralisL.BritishPhycologicalBulletin2:17.
SerroE.A.,AliceL.A.,BrawleyS.H.(1999)EvolutionoftheFucaceae(Phaeophyceae)inferredfromnrDNAITS.JournalofPhycology
35:382394.
Wallace A.L., Klein A.S., Mathieson A.C. (2004) Determining the affinities of salt marsh fucoids using microsatellite markers:
evidence of hybridization and introgression between two species of Fucus (Phaeophyta) in a Maine estuary. Journal of
Phycology40:10131027.
376.
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9.3.5
Himanthaliaelongata
Figure81Himanthaliaelongata(Linnaeus)S.F.Gray
LesGlnanislands,Finistre,Bretagne,France.
BenoitQueguineur
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Himanthaliaceae
Himanthalia
Himanthaliaelongata
Relatedspecies
ThegenusHimanthaliaonlyincludesthespecieHimanthaliaelongata.
BIOLOGY
ThegrowthformofHimanthaliadiffersfromthatofanyothermemberoftheFucales.Theyoungplants
arefirstrecognizableontheshoreassmallvesicles,palegreenishbrownincolour.Thesevesiclesflatten
out and after I2 years growth they form the small leathery discshaped structure which is commonly
referredtoasthe'button'stage.Fromthese'buttons'thelongthongsarise,andinsomehabitatstheymay
reachalengthofabout2mormore.The'button'isgenerallyinterpretedasequivalenttothevegetative
thallusoftheotherFucales,whilethethongscorrespondtothefertilereceptacles.Gametesareshedover
alongperiod,theyarereleasedfromlateJuly,onthroughthewinter.InHimanthaliaonlyonevegetative
meristem is ever produced. The single apical cell of this divides many times; each new daughter cell
becomingtheapicalcellofareproductiveapex.Eachofthesehasdeterminategrowth.Bythetimethat
the gametes are shed the identity of the apical meristem has been lost and the receptacle subsequently
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disintegrates.Thereisnomeristemleftontheplanttocontinueevegetativegrowth,andonceanapical
cellislosttheplantisunabletoregenerateanewone.
Himanthaliagrowsat,ornearlowwaterlevel,sothatinsomehabitatstheplantsareuncoveredatlow
springtides,whereasinotherlocalitiestheymayneverbeexposed.
BIOTECHNOLOGY
Himnathaliaelongataisusedasfood(MacArtainetal.,2007).
References
GallardoGarciT.,PrezRuzafaI.M.(2001)HimanthaliaLyngb.In:GmezGarreta,A.(ed.)FloraphycologicaibericaVol.1Fucales.,
UniversidaddeMurcia,Murcia,pp.6971.
MossB.(1969)ApicalmeristemsandgrowthcontrolinHimanthaliaelongata(S.F.Gray).NewPhytologist68:387397.
MacArtain P., Gill C.I.R., Brooks M., Campbell R., Rowland I.R.(2007) Nutritional value of edible seaweeds. Nutrition Reviews 65:
535543.
SetchellW.A.(1931)Someearlyalgalconfusions.UniversityofCaliforniaPublicationsinBotany16:351366.
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9.3.6
Cystoseirasp.
TAXONOMY
Relatedspecies
C. abiesmarina, C. algeriensis, C. amentacea, C. baccata, C. barbata, C. barbatula, C. brachycarpa, C.
compressa, C. corniculata, C. crassipes, C. crinita, C. crinitophylla, C. dubia, C. elegans, C. foeniculacea, C.
funkii, C. geminata, C. hakodatensis, C. helvetica, C. humilis, C. hyblaea, C. indica, C. jabukae, C.
mauritanica, C. mediterranea, C. montagnei, C. myrica, C. neglecta, C. nodicaulis, C. occidentalis, C
osmundacea, C. pelagosae, C. planiramea, C. platyclada, C. sauvageauana, C. schiffneri, C. sedoides, C.
setchellii, C. sonderi, C. spinosa, C. squarrosa, C. susanensis, C. tamariscifolia, C. trinodis, C. usneoides, C.
wildpretii,C.zosteroides.
Cystoseirabaccata
Figure82Cystoseirabaccata(S.G.Gmelin)P.C.Silva
Finavarra,Co.Clare,Ireland;MLWNlagoon
M.D.Guiry
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Sargassaceae
Cystoseira
Cystoseirabaccata
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Cystoseiratamariscifolia
Figure83Cystoseiratamariscifolia(Hudson)Papenfuss
LesGlnanislands,Finistre,Bretagne,France.
BenoitQueguineur
SYMBOLS:B
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Sargassaceae
Cystoseira
Cystoseiratamariscifolia
BIOLOGY
C.baccataplantsareusuallysolitary,1mormoreinlength,attachedbyathick,conicalattachingdisc.Axis
simpleorbranched,upto1minlength,flattened,about1x0.4cmintransversesection;apexsmoothand
surroundedduringperiodsofactivegrowthbyincurredyounglaterals.Lateralbranchsystemsdistichous,
alternate, radially symmetrical, profusely branched in a repeatedly pinnate fashion and bearing sparse,
filiform,occasionallybifurcateappendagesonthebranchesofhigherorders;deciduous,leavingdecurrent
baseswhichgiveanirregular,zigzagoutlinetotheaxis.Cryptostomatalacking.Aerocystspresentinaxesof
branchesofhigherorder,sometimesinchains;seasonal,particularlynumerousinautumn.Receptacles15
cmlong,formedfromaxesofultimateramuli,irregularlynodoseandbearingsimple,filiformappendages.
BIOTECHNOLOGY
Nobiotechnologicaluseknown,exceptatresearchlevel(contaminantbiosorption,productionofbioactive
metabolites).
References
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Snchez I., Fernndez C., Arrontes J. (2005) Longterm changes in the structure of intertidal assemblages following invasion by
Sargassummuticum(Phaeophyta).JournalofPhycology41:942949.
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9.3.7
Halidryssiliquosa
Figure84Halidryssiliquosa(Linnaeus)Lyngbye
SpanishPoint,Co.Clare,Ireland;lowerintertidalpools
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Sargassaceae
Halidrys
Halidryssiliquosa
Relatedspecies
H.dioica,H.murmanica
BIOLOGY
Thallusto2mlong,compressedtoughandleatherly,attachedbystrongholdfast.Branchesdistichousand
regularly alternate. Air bladders, up to 5 cms long, are oblong and stalked towards the ends of the
branches.Theairbladdersaredividedbycrosswallsvisiblewhensectionedlongitudinally.Common.Low
littoral rock pools and upper sublittoral. Widespread around the British Isles. Europe: Portugal, Atlantic
coastsofSpainandFrance,Netherlands,BalticSea,NorwayandtheFaroes.
BIOTECHNOLOGY
Nobiotechnologicaluseknown,exceptatresearchlevel(productionofbioactivemetabolites).
References
BrauneW.(2008).Meeresalgen.EinFarbbildfhrerzudenverbreitetenbenthischenGrnBraunundRotalgenderWeltmeere.,
Morton O., Picton B.E. (2010) Halidrys siliquosa. In: Encyclopedia of Marine Life of Britain and Ireland
http://www.habitas.org.uk/marinelife/species.asp?item=ZR7160
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9.3.8
Sargassummuticum
Figure854Sargassummuticum(Yendo)Fensholt
Finavarra,Co.Clare,Ireland;
M.D.Guiry
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Fucales
Sargassaceae
Sargassum
Sargassummuticum
Relatedspecies
ThegenusSargassumcontains345relatedspecies.
BIOLOGY
Thallusto10200cmormoreinlength,withonetoafewsimple,teretetocompressed,stipes120cmlong
arisingfromadiscoidconicalholdfast.Stipesbearingradiallyordistichouslyborne,longprimarybranches,
producedseasonallyfromthestipeapicesandsubsequentlydeciduous,leavingscarsorotherresidueson
thestipe.Primarybranches10cmto200cmormorelong,distichously,tristichouslyorradiallybranched
withaterete,angular,compressedorthreesidesaxes;basallateralssimpleorbranched,compressedand
relatively narrow to (in most species) leaflike, (1)315(25) mm broad, entire ot with dentate margins;
upperlateralsusualybranched,withslender,compressedtoterete,ramuli.Airbladders(vesicles)normally
present, subspherical to ovoid, petiolate, mutic or apiculate, replacing ramuli or axilliary to the laterals.
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Growthfroma3sidedcellinanapicaldepression.Structureofacentralmedullaofelonagatecellsinthe
stipeandbranchaxes,withacoreofisodiametriccellsandasurfaceofphaeoplasticmeristoderm,activein
the larger branched. Reproduction. Thallus monoecious or dioecious. Receptacles unisexual or bisexual,
developed in axils of laterals or ramuli, simple or usually in branched clusters, terete or compressed,
smooth,verrucoseorspinous,withscatteredconceptaclesandostioles,growingapicallywithconceptacles
maturingbelow;conceptaclesunisexualorbisexual.
BIOTECHNOLOGY
SargassumisusedasfoodinmanycountriesinAsiaaswellasinHawaii.Itisalsousedfortheproductionof
alginateand,infewcountries,foragriculturalpurposes.Annualproduction,allfromwildharvesting,isof
5,000tdryweightinthePhilippines,2,249inIndiaand400inVietnam(ZemkeWhiteandOhno,1999).
References
A.R.G.GantnerVerlag.,Ruggell,pp.1596,266pls.
ConnanS.,DelisleF.,DeslandesE.,GallE.A.(2006).IntrathallusphlorotannincontentandantioxidantactivityinPhaeophyceaeof
temperatewaters.BotanicaMarina49:3946.
FarnhamW.F.,FletcherR.L.,IrvineL.M.(1973)AttachedSargassumfoundinBritain.Nature243:231232.
KilarJ.A.,HanisakM.D.,YoshidaT.(1992)Ontheexpressionofphenotypicvariability:whyisSargassumsotaxonomicallydifficult?.
In: Abbott I.A. (ed.) Taxonomy of Economic Seaweeds with reference to Pacific and Western Atlantic species, Vol.3,
CaliforniaSeaGrantCollege,LaJolla,California,pp.95117.
LeeK.,YooS.A.(1992)KoreanspeciesofSargassumsubgenusBactrophycusJ.Agardh(Sargassaceae,FucalesIn:AbbottI.A.(ed.)
TaxonomyofEconomicSeaweedswithreferencetoPacificandWesternAtlanticspecies,Vol.3,CaliforniaSeaGrantCollege,
LaJolla,California,pp.139147
PedrocheP.F.,SilvaP.C.,AguilarRosasL.E.,DreckmannK.M.,AguilarRosasR.(2008)CatlogodelasalgasbenthnicasdelPacfico
de Mxico II. Phaeophycota, Universidad Autnoma Metropolitana and University of California Berkeley, Mexicali &
Berkeley,pp.ivi,15146.
PhillipsN.(1995)BiogeographyofSargassum(Phaeophyta)inthePacificbasin.FucalesIn:AbbottI.A.(ed.)TaxonomyofEconomic
SeaweedswithreferencetoPacificandWesternAtlanticspecies,Vol.5,CaliforniaSeaGrantCollege,LaJolla,California,pp.
107145.
Stiger V., Horiguchi T., Yoshida T., Coleman A.W., Masuda M. (2003) Phylogenetic relationships within the genus Sargassum
(Fucales,Phaeophyceae),inferredfromitITSnrDNA,withanemphasisonthetaxonomicrevisionofthegenus.Phycological
Research51:110.
Tseng C.K., Yoshida T., Chiang Y.M. (1985) East Asiatic species of Sargassum subgenus Bactrophycus J.Agardh (Sargassaceae,
Fucales), with keys to the sections and species. In: Abbott I.A., Norris J.N. (eds.) Taxonomy of Economic Seaweeds with
referencetosomePacificandCaribbeanspecies,Vol.I,CaliforniaSeaGrantCollege,LaJolla,California,pp.114.
VarelaAlvarez E., Andreakis N., LagoLeston A., Pearson G.A., Serrao E.A., Procaccini G., Duarte C.M., Marba N. (2006) Genomic
DNAisolationfromgreenandbrownalgae(CaulerpalesandFucales)formicrosatellitelibraryconstruction(Note).Journalof
Phycology42:741745.
YoshidaT.,StigerV.,HoriguchiT.(2000)Sargassumborealesp.nov.(Fucales,Phaeophyceae)fromHokkaido,Japan.Phycological
Research48:125132.
376.
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9.3.9
Laminaria,Saccharina,Saccorhiza
TAXONOMY
Laminariasp.
Relatedspecies
L. abyssalis, L. agardhii, L. angustata, L. appressirhiza, L. bongardiana f. taeniata, L. bongardiana f. sub
simplex, L. brasiliensis, L. bulbosa var. nodipes, L. bulbosa var. gymnopoda, L. bulbosa var. brevipes, L.
bulbosavar.linearis,L.bullata,L.complanata,L.cordata,L.digitata,L.digitatavar.ligulata,L.digitatavar.
lyrata, L. digitata var. pseudosaccharina, L. digitata var. elliptica, L. digitata var. bifida, L. ephemera, L.
farlowii,L.hyperboreaf.cucullata,L.hyperborea,L.inclinatorhiza,L.japonicaf.membranacea,L.japonica
f. longipes, L. longipes, L. multiplicata, L. nigripes, L. ochroleuca, L. pallida, L. platymeris, L. rodriguezii, L.
ruprechtii,L.saccharinavar.vividissima,L.saccharinavar.vittata,L.saccharinavar.vividissima,L.setchellii,
L.sinclairii,L.solidungula,L.yezoensis.
Laminariadigitata
Figure86Laminariadigitata(Hudson)J.V.Lamouroux
Spiddal,Co.Galway,Ireland;plantsonrocksatMLWS
M.D.Guiry
SYMBOLS:B,PIV
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Laminariales
Laminariaceae
Laminaria
Laminariadigitata
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Varieties
L.digitatavar.ligulata,L.digitatavar.lyrata,L.digitatavar.pseudosaccharina,L.digitatavar.elliptica,L.
digitatavar.bifida.
Laminariahyperborea
Figure87Laminariahyperborea(Gunnerus)Foslie
Dog'sBay,Roundstone,Co.Galway,Ireland;plantsatlowwater;10.5mmlens
M.D.Guiry
SYMBOLS:B,PIV
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Laminariales
Laminariaceae
Laminaria
Laminariahyperborea
Varieties
Laminariahyperboreaf.cucullata
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Laminariaochroleuca
Figure88LaminariaochroleucaBachelotdelaPylaie
Spain,Galicia,ACorua,RadeACorua,2004,lowerintertidal
IgnacioBrbara
SYMBOLS:B,PIV
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Laminariales
Laminariaceae
Laminaria
Laminariaochroleuca
Saccharinalatissima
Figure89Saccharinalatissima(Linnaeus)C.E.Lane,C.Mayes,L.D.Druehl&G.W.Saunders
NewQuay,Co.Clare,Ireland;plantsatMLWS
M.D.Guiry
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SYMBOLS:B,PIV
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Laminariales
Laminariaceae
Saccharina
Saccharinalatissima
Relatedspecies
S. angustata, S. angustata subsp. siberica, S. bongardiana f. taeniata, S. bongardiana f. subsimplex, S.
bongardiana f. subsessilis, S. bongardiana f. bifurcata, S. bongardiana, S. cichorioides f. sikotanensis, S.
cichorioidesf.sachalinensis,S.cichorioidesf.coriacea,S.cichorioides,S.cichorioidesf.sinuicola,S.coriacea,
S.crassifolia,S.dentigera,S.groenlandica,S.gurjanovaef.lanciformis,S.gurjanovae,S.gyrata,S.japonica,
S.japonicaf.diabolica,S.japonicaf.longipes,S.kurilensis,S.longicruris,S.longipedalis,S.longissima,S.
ochotensis,S.plana,S.religiosa,S.sculpera,S.sessilis,S.yendoana.
Saccorhizapolyschides
Figure90Saccorhizapolyschides(Lightfoot)Batters
CoolivaQuay,Co.Clare,Ireland;plantsonsandyrocksatlowwater
M.D.Guiry
SYMBOL:B,PIV
Phylum
Class
Order
Family
Genus
Species
Heterokontophyta
Phaeophyceae
Tilopteridales
Phyllariaceae
Saccorhiza
Saccorhizapolyschides
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Relatedspecies
S.dermatodea
BIOLOGY
GeneralDescriptionofMajordivisionsandclasses
LaminariaandSaccharinaaretwosmallgeneraoflargebrownseaweedscommonlycalledkelps.Theorder
of the Laminariales share an almost identical life history: The sporophyte is the dominant phase and the
gametophyte is a microscopic phase. There is therefore an alternation of sporophyte and gametophyte.
Sporophytesofbothgeneraaredifferentiatedintoholdfast(below),stipeandlamina;thegametophyteis
undifferentiatedandisfilamentousandcreeping.
Sexualreproductionisoogamous;antherozoids(malegametes)areproducedbyamalegametophyte
andeggsareproducedinoogoniabythefemalegametophyte.Thefemaleliberatesapheromonecalled
lamoxirene,alowmolecularweight(C11),volatilehydrocarbonwhich,inadditiontoattractingthesperm,
initiatestheirreleasefromtheantheridiaofthemalegametophyte.Fertilizationtakesplacewiththeegg
partially extruded from the oogonium and the zygote develops in situ to form a sporophyte (the
macroscopickelp).Thesporophytedevelopssporangiaonthesurfaceofthebladesinunilocularsporangia
(calledsuchbecausetheyarenotdividedbycrosswallsorlocules);thecontentsofthesesporangiadivide
meioticallyandformnumerousflagellatedzoosporeswhicharehaploid.Thesezoosporesswimawayand
eventuallysettleanddevelopintothegametangialthalli.Thelifehistoryisthereforeheteromorphic;the
gametophytesandthesporophytehaveadifferentmorphology.
LaminariaandSaccharinaspeciesarefoundonrockyshoresatlowtideandinthesubtidaltodepthsof
830minthenorthAtlanticandnorthPacific;somespeciesoccuratdepthsofupto120m(Mediterranean
andBrazil),butthisrequiresextraordinarywaterclarity.
Saccorhiza polyschides sporophytes are annual, a single stipe attached initially by a small disc,
sometimeswithafewshorthaptera,sooncoveredbyadiscoidcircumferentialswelling(rhizogen)ofthe
stipe, which forms downwardgrowing hapteroid protuberances that surround the initial holdfast,
succeededbyoneormorewhorlsofhaptera;allhapteraunbranched.Stipeflattened,broadeningtoforma
single lanceolate or expanded and split blade. Hairpits on young blades, deciduous with age; costae
lacking. Medulla containing long, thickwalled longitudinal conducting cells (solenocysts) and lateral
connectingcells(allelocysts)ratherthansievetubes.Mucilageductsabsent.Sporangiasoridiscontinuous
patches at base of blades, paraphyses lacking a hyaline appendage, zoospores with a red eyespot.
Gametophytes of S. polyschides dioecious and sexually dimorphic. Marine, littoral to 19 m, distributed
either in the western Mediterranean and the Atlantic from the Tropic of Cancer to southern Norway
(Algaebase,2010).
Table5BiochemicalcompositionofPhaeophyceaerelevantforbiotechnology.
Water(%FW)
Ash
TotalCarbohydrate
Laminaria
hyperborea
(lamina+stipe)
Laminaria
digitata
(lamina)
Saccorhiza
polyschides
(lamina)
Saccharina
latissima
7789
7390
90.993.0
1637
13.837.6
26.58
61
References
Baardseth&Haug,1953;Indergaard&Minsaas,
1991;Jensenetal.,1985
Jensen&Haug,1956;Ruperez,2002;Adamset
al.,2011;SanchezMachadoetal.,2004
Kuppers&Kremer,1978
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Saccorhiza
polyschides
(lamina)
16.223.4
Alginicacid
Laminaria
hyperborea
(lamina+stipe)
1734
Laminaria
digitata
(lamina)
2045
Laminaran
030
024.6
Mannitol
425
532.1
2.511.1
Fucoidan
24
24
10.4
6.2
5.510.3
traces
1.2
Protein
414
8.15
9.414.4
6.11
Lipid
0.63
0.55.96
0.50.9
0.5
0.83.9
0.41.3
0.82.4
0.74
0.31.1
0.040.09
Fiber
Othercarbohydrate
Phlorotannins
References
Saccharina
latissima
2430
Jensen & Haug, 1956; Indergaard & Minsaas,

1991; Jensen et al., 1985; Kuppers & Kremer,
1978
Jensen&Haug,1956;Adamsetal.,2011
Jensen & Haug, 1956; Jensen et al., 1985;
Adamsetal.,2011;Kuppers&Kremer,1978
Indergaard&Minsaas,1991;Indergaard,1983
Jensen&Haug,1956;Jensenetal.,1985;Haug
&Jensen,1954;MacArtainetal.,2007
Indergaard&Minsaas,1991;Indergaard,1983
1991; Jensen et al., 1985; Kuppers & Kremer,
1978
Jensen & Haug, 1956; Jensen et al., 1985;
Kuppers&Kremer,1978;Haug&Jensen,1954;
Marshametal.,2007;Schaaletal.,2010
Pedersen,1980
Ca
1.43.0
1.005
Mg
0.60.7
0.50.8

1991;Jensenetal.,1985;Haug&Jensen,1954
Indergaard & Minsaas, 1991; Jensen et al.,
1985;Hanssenetal.,1987;Jensen,1954
Indergaard & Minsaas, 1991; Jensen et al.,
1985;Hanssenetal.,1987;Jensen,1954
Ruperez, 2002; Hanssen et al., 1987; Jensen,
1954
Indergaard & Minsaas, 1991; Hanssen et al.,
1987;Jensen,1954
Hanssenetal.,1987
Hanssenetal.,1987
Iodine
6.311
1.33.8
14.7
Na
1.63.0
0.92.2
4.6
1.21.3
0.2
BIOTECHNOLOGY
Cultivationmethods/Productivity
Table20Kelpcultivationtrials,methodsandproductivityinEurope.Extrapolatedfiguresobtainedafterthedesign
of Tseng (1987) (t FW ha1 year1) and for a single crop of 8 Months, 5324 m of linear cultivated rope ha1. The
extrapolated figures are displayed in order to compare values of productivity but do not reflect a scaledup
situation.
Location/Season
(orculture
duration)
Production
(aspublished)
Tank/Pond/RacewayCultivation:Integrated
Salmon
Canada
Saccharina
6.59%GRday1
latissima
SeaCultivation
Laminaria
saccharina
Laminaria
saccharina
Laminaria
saccharina
Saccorhiza
polyschides
Canada
(8months)
UK
(6months)
IsleofMan/UK
38 kg FW m1
ropepm
4.228.4 kgFW m
1
ropepm
Production
extrapolated
afterTseng
(1987)design
(tFWha1year1)
Keywords
Reference
Nutrientuptake;
growthrate;
biomass;flow
rate;kelpdensity
Subandaret
al.,1993
Yield;cultivation
methodology
Druehletal.,
1988
Kain,1991
detachedfrom
ropesinstrong
watermovement
127.8340.7
178.91209.6
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Alariaesculenta
5.6kgm1 rope
year1(FW)
(extrapolated
fromKain&
Dawes,1987)
144plantsm1
ropein8months
29.8
Laminaria
digitata
Ouessant/France
Alariaesculenta
Ireland
15.6kgm1in5
months
132.9
Laminaria
saccharina
Kiel/Germany
0.5kgFWm1
after3months
8iftriplethe
durationtomake
upforayear
Laminaria
saccharina
Helgoland/Germa
ny
4kgFWm1in6
months
28.4
Alariaesculenta
Ireland
35kgFW/mrope
after45months
SeaCultivation:Integrated
Salmosalar
Scotland
Laminaria
saccharina
Laminaria
hyperborea
Saccorhiza
polyschides
Mytilusedulis
Galicia/Spain
Laminaria
saccharina
2
1
10kgm year
(FW)
2kgm2year1
(FW)
17kgm2 year1
(FW)
6.2kgm1ropein
4months
Cleansmall
plantsforfood
purposes
hybridisation
experiments
Perez,1997
freefloating
culturesofsmall
plantsattached
toropesresulted
inlargerplants
thanseeded
ropes.Ropes
overseeded.
Positiveeffectsof
L.saccharina
cultureinthe
BalticSea.
Ringstructurefor
offshore
cultivation
reproducible
procedurefrom
fertilisationto
offshore
cultivation
CRMtrials
citedin
Werneretal.,
2004
53.2
10.6
90.5
Sanderson,
2006
Unpublished
data(Kelly&
Dworjanyn,
2008)
31.953.2
Kraan,2000
Buck&
Buchholz,
2004
Arbona,1997
66
Peteiro et al.,
2006
decreasein
harvestable
biomassafter13
monthsdueto
lossofplants.
GR=growthrate(%ofthefreshweight),FW=freshweight,DW=dryweight,pm=permonth,FCR=Foodconversion
ratio
Productionsystems
Worldaquaticplantproductionbyaquaculturewas15.1milliontonnes(US$7.2billion)in2006.Japanese
kelp (Saccharina japonica 4.9 million tonnes) showed the highest production, followed by Wakame
(Undariapinnatifida2.4milliontonnes)(FAO,2009).Manyproductionsystemsarecurrentlyinusebut
theyallarisefromthesametechniques:Fragmentsofadultplants,juvenileplants,sporelingsorsporesare
seededontoropesorothersubstrataandtheplantsgrowntomaturityinthesea.Toachievethis,intimate
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knowledgeofboththebiologyandlifehistoryoftheplantsiscritical.Forexample,kelpscannotbegrown
fromfragmentsasthereisahighlevelofspecializationandfragmentsofsporophytesdonotregenerate.
InAsia
Cultureofsporelings
The first step consists in the collection of the zoospores onto the seeding cords. To do this, fronds with
maturesporangialsoriaresubjectedtopartialdryingintheairandthenplacedinasmallcontainerwith
seawater.Theliberatedzoosporesreadilyattachthemselvestotheseedingcords.Thegametophytesand
earlysporophytesareculturedinwaterof810Cinglasshouseforaboutthreemonths,bywhichtimethe
juvenilesporophytesare23cminlength.
Transplantation
When the water temperature has dropped to about 20 C, sporeling cords are removed from the
glasshouseandonfloatingrafts.Inamonthorso,thesporelingswillhavegrowntojuvenilesof1015cm
orlonger.Thesejuvenilesporophyteswillbeeventuallybroughttothetransplantingroomandplacedin
tanksfilledwithseawater.Duetotheirfastgrowthonthecordstheyarethinned.Plantsareremovedfrom
theoriginalsporelingcordandinsertedinthetwistsofthekelpropes,atadensityofabout30juvenilesto
eachropeof2m.
Culturemethods
Therearetwobasicfloatingraftkelpcultivationmethods.Oneisthehangingkelprope(alsocalledsingle
raft)cultivationmethod.Theotheristhehorizontalkelprope(alsocalleddoubleraft)cultivationmethod.
Thefloatingline,isabout60meterslongfloatedatthesurfacebybuoysgenerallymadeofglassorplastics
andanchoredterminallybyanchoringlinestowoodenpegsdrivenintotheseabottom.Eachkelpropehas
about30plantstwistedinitandisabout2minlength.Inthesingleraftmethod,thekelpropesarehung
downfromfloatinglineandweighteddownbyasmallpieceofstone.Inthedoubleraftmethod,thetwo
kelpropesarelinkedortiedtogetheratoneendandtheotherendstiedtofloatinglines.Thehangingkelp
ropemethodhastheadvantageofbetterwatermovementbuthasthedefectofunevengrowthofkelps.
Thehorizontalkelpropemethodgivesanevengrowthofkelp.However,ithasthedisadvantageofbeing
more resistant to water motion. Generally, the singleraft method is better adapted to comparatively
clearerwaterregions,andthedoubleraftmethodtoturbidregionswithlowerwatertransparency,suchas
theZhejiangcoast(Feijiu,1988).Theuseoffertilizerisstillinplacebutnowlimitedtothefirstmonthsof
development,theassociationwithfish/shellfishfarmingallowstogreatlyreducetheadjunctionofnutrient.
Severaltypeofassociationareapplied:
Saccharinajaponica+Undariapinnatifida
Saccharinajaponica+Mytilussp.
Saccharinajaponica+Haliotis(Abalone)
Saccharinajaponica+Undariapinnatifida+Mytilussp.
Saccharinajaponica+Undariapinnatifida+Haliotis
Saccharinajaponica+Undariapinnatifida+Haliotis+Mytilussp.
The productivity increases by 45% and the income rises by 15 to 20% when compared to monoculture
(Perez,1997).
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InEurope
ProductionsystemshavebeeninvestigatedforresearchpurposesinvariouspartsofEuropesincethelate
1970s. The technique of freeliving culture of the microscopic gametophytes has been developed in
Ifremerin1984.Thistechniqueallowsthefarmertoseedtheropesondemand,andafterashortenedpre
cultivationperiodinthehatchery,theropeswithyoungsporelingscanbetransferredatsea.Thedesignof
hollowcylindricalcollectorshavealsosimplifiedandhencereducedthetimeneededtosetthecultureat
sea.ProductionatacommercialscaleissetupforUndariapinnatifidainFrancewhereitpeakedat100t
DWin1995(FAO,2002),andreachesasaturatedmarketforfoodpurposes.
InIreland,cultivationtrialshaveusedhybridizationofNorthAtlanticstrainsofAlariaesculentainorder
toimproveproductivityaswellasproteincontenttotargetthefoodandfeedmarket(Kraanetal.,2000)
Trialswithringsstructurescomparedtolonglineshavealsotakenplaceto proposeadesignforoffshore
seaweedaquaculture(BuckandBuchholz,2004).
The use of polyculture (Integrated MultiTrophic Aquaculture), has been performed for some time,
howeverrecentresearchhasfocusedontheuseofseaweedasbiofilterstotakeadvantageofnutrientload
fromfishfarming.
Harvestingmethods
InAsia
Harvestingtakesplacewhenthefrondsaremature.Thetimeforharvestingisimportanttokelpfarmers.
SinceSaccharinaissoldonthemarketonthebasisofdryweight,andsincethewetweighttodryweight
ratiochangesfrommonthtomonth,thecriterionforselectingharvesttimemusttakeintoconsideration
thehighestperunitareaproductionrateplusthelowestwettodryratio.
Harvestingisdonemanually,thekelpropesaredetachedfromthefloatingline,andcollectedinsmall
boats,manyofwhicharetowedinalonglinebyamotorboattotheshores.
InEurope
Manual harvesting of longlines of kelp is not economically viable in a highly competitive area due to the
highcostoftheworkforceinWesternEurope.Thereforemechanizationandminoradaptationofmussels
harvestingboatstokelpslonglineisnecessary.
Biomassprocessing
InAsia
When the boats reach the wharf or shore, the plants are transported to land and dried under the sun.
Formerly the kelp was sold on the market only in its crude dried form, but recently small package of
shreddedandseasonedformswithdifferentflavourshavebeenintroducedtothemarketandhavebeen
verywellreveivedbytheChinesepeople.
InEurope
Insomecases,thecropwasusedforthefoodmarket.Hencethecropwasdried,sometimesflavouredand
sold as a snack or seavegetables. (Laminaria saccharina and Alaria esculenta in the Island of Man, UK,
UndariapinnatifidainBrittany,France)(Werneretal,2004).
Biomassresultingfromcultivationtrialsforresearchpurposeshasmostlyleadtoknowledge,skillsand
data production on seaweed aquaculture. However, some European projects currently uses the biomass
producedforanaerobicdigestion.
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Therearenoserioustechnologicalbarriers(Kain,1991)toscalingupseaweedaquaculture,asithasalready
been demonstrated spectacularly in Asia. However, limitations in a European context rather lie in the
selection of adequate sites (both regarding the requirement of the targeted seaweed species and the
competitionwithothercoastalresourceusers),thedevelopmentofmechanicalharvestingtechniquesand
assessment of the environmental impact of largescale aquaculture. Fei (2004) advocates for largescale
seaweed cultivation as a way of solving eutrophication under the following conditions: (a) Largescale
cultivation could be conducted within the region experiencing eutrophication; (b) Fundamental scientific
andtechnologicalproblemsforcultivationshouldhavebeensolved;(c)Cultivationshouldnotimposeany
harmfulecologicaleffects;(d)Cultivationmustbeeconomicallyfeasibleandprofitable.
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chemical composition of the bioenergy feedstock Laminaria digitata for thermochemical conversion. Bioresource
Technology102:226234.(SpecialIssue:BiofuelsII:AlgalBiofuelsandMicrobialFuelCells)
ArbonaJ.F.(1997)AroutinemethodformasscultivationofAlariaesculenta(Greville1830).M.Sc.Aquaculture,NationalUniversity
ofIreland,Galway.
BaardsethE.,Haug,A.(1953)Individualvariationofsomeconstituentsinbrownalgae,andreliabilityofanalyticalresults.Reports
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BuckB.H.,BuchholzC.M.(2004)Theoffshorering:Anewsystemdesignfortheopenoceanaquacultureofmacroalgae.Journalof
AppliedPhycology16:355368.
ConnanS.,DelisleF.,DeslandesE.,GallE.A.(2006)IntrathallusphlorotannincontentandantioxidantactivityinPhaeophyceaeof
temperatewaters.BotanicaMarina49:3946.
Dieck I.T., Oliveira Filho E.C. de (1993) The section Digitatae of the genus Laminaria (Phaeophyta) in the northern and southern
Atlantic:crossingexperimentsandtemperatureresponse.MarineBiology115:151160.
Druehl L.D., Baird R., Lindwall A., Lloyd K.E., Pakula S. (1988) Longline cultivation of some Laminariaceae in British Columbia,
Canada.AquacultureandFishManagement19:253263.
FAO (2009) The State of World Fisheries and Aquaculture, FAO Fisheries and Aquaculture Department, Food And Agriculture
OrganizationoftheUnitedNations.Rome.http://www.fao.org/docrep/011/i0250e/i0250e00.htm
Fei,X.(2004)Solvingthecoastaleutrophicationproblembylargescaleseaweedcultivation.Hydrobiologia512:145151.
GuiryM.D.(2010)Seaweed.ie.Worldwideelectronicpublication,NationalUniversityofIreland,Galway.http://www.seaweed.ie;
searchedon19November2010.
Hanssen J.F., Indergaard M., stgaard K., Bvre O.A., Pedersen T.A., Jensen A. (1987) Anaerobic digestion of Laminaria spp. and
Ascophyllumnodosumandapplicationofendproducts.Biomass14:113.
Haug A., Jensen A. (1954) Seasonal variations on the chemical composition of Alaria esculenta, Laminaria saccharina, Laminaria
hyperboreaandLaminariadigitatafromNorthernNorway.ReportsoftheNorwegianInstituteofSeaweedResearch4.
IndergaardM.,MinsaasJ.(1991)Animalandhumannutrition.In:GuiryM.D.,BlundenG.(eds.)SeaweedResourcesinEurope:Uses
andPotential,JohnWiley&Sons,Chichester,pp.2164.
JensenA.(1954)Omtangogtareihusdyrf6ret.MedlemsbladfordenNorskeVeterinaerforening6:123.
JensenA.,HaugA.(1956)GeographicalandseasonalvariationinthechemicalcompositionofLaminariahyperboreaandLaminaria
digitatafromtheNorwegiancoast..ReportsoftheNorwegianInstituteofSeaweedResearch14:118.
JensenA.,IndergaardM.,HoltT.J.(1985)SeasonalvariationinthechemicalcompositionofSaccorhizapolyschides(Laminariales,
Phaeophyceae).BotanicaMarina28:375381.
Kain(Jones)J.M.(1971)SynopsisofbiologicaldataonLaminariahyperborea.FAOFisheriesSynopsisNo87(Rev.1):74pp..
Kain J.M. (1991) Cultivation of attached seaweeds. In: Guiry M.D., Blunden G. (eds.) Seaweed Resources in Europe: Uses and
Potential,JohnWiley&Sons,Chichester,pp.309377.
KellyM.,DworjanynS.(2008)ThePotentialofMarineBiomassforAnaerobicBiogasProduction,TheCrownEstate,London.
Kraan S., Tramullas A.V., Guiry M.D. (2000) The edible brown seaweed Alaria esculenta (Phaeophyceae, Laminariales):
hybridization,growthandgeneticcomparisonsofsixIrishpopulations.JournalofAppliedPhycology12:577583.
Kraan S (2000) The genus Alaria (Alariaceae, Phaeophyceae) explored: phylogenetics, hybridization and aquaculture. PhD thesis.
NationalUniversityofIreland,Galway.
KuppersU.,KremerB.P.(1978)Longitudinalprofilesofcarbondioxidefixationcapacitiesinmarinemacroalgae.PlantPhysiology
62:4953
Lane C.E., Saunders,G.W. (2005) Molecular investigation reveals epi/endophytic extrageneric kelp (Laminariales, Phaeophyceae)
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Lane C.E., Mayes C., Druehl L.D., Saunders G.W. (2006) A multigene molecular investigation of the kelp (Laminariales,
Phaeophyceae)supportssubstantialtaxonomicreorganization.JournalofPhycology42:493512.
MacArtainP.,GillC.I.R.,BrooksM.,CampbellR.,RowlandI.R.(2007)Nutritionalvalueofedibleseaweeds.NutritionReviews 65:
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MarshamS.,ScottG.W.,TobinM.L.(2007)Comparisonofnutritivechemistryofarangeoftemperateseaweeds.FoodChemistry
100:13311336
McHughD.J.(2003)Aguidetotheseaweedindustry.FAOFisheriesTechnicalPaper441
MorrisseyJ.,KraanS.,GuiryM.D.(2001)AguidetocommerciallyimportantseaweedsontheIrishCoast,.BordIascaighMhara,Dun
Laoghaire.
NortonT.A.(1970)SynopsisofbiologicaldataonSacchorizapolyschides.FAOFisheriesSynopsis83:1193.
PapenfussG.F.(1950)ReviewofthegeneraofalgaedescribedbyStackhouse.Hydrobiologia2:181208.
PedersenA.(1980)Fenolinnholdibrunalger(Phaeophyceae)somfunksjonavvekstypeogsalinitet.Ph.D.thesis.Inst.Marinbiol.,
Univ.Bergen.
PerezR.(1997)Cesalguesquinousentourent:conceptionactuelle,rledanslabiosphre,utilisations,culture,EditionIFREMER
Plouzan,France.
PeteiroC.,SalinasJ.M.,Freire.,FuertesC.(2006)CultivationoftheautoctonousseaweedLaminariasaccharinaoffthegalician
coast(NW):productionandfeaturesofthesporophytesforanannualandbiennialharvest.Thalassas22:4552.
RoederV.,CollnJ.,RousvoalS.,CorreE.,LeblancC.,BoyenC.(2005)IdentificationofstressgenetranscriptsinLaminariadigitata
(Phaeophyceae)protoplastculturesbyexpressedsequencetaganalysis.JournalofPhycology41:12271235.
RuperezP.(2002)Mineralcontentofediblemarineseaweeds.FoodChemistry79:2326.
IndergaardM.(1983)Theaquaticresource.I.Thewildmarineplants:aglobalbioresource.In:CoteW.A.(ed.)Biomassutilization,
PlenumPublishingCorporation,pp.137168.
Snchez I., Fernndez C., Arrontes J. (2005) Longterm changes in the structure of intertidal assemblages following invasion by
Sargassummuticum(Phaeophyta).JournalofPhycology41:942949.
SanchezMachado D.I., LopezCervantes J., LopezHernandez J., PaseiroLosada P. (2004) Fatty acids, total lipid, protein and ash
contentsofprocessededibleseaweeds.FoodChemistry85:439444.
SchaalG.,RieraP.,LerouxC.(2010)TrophicecologyinaNorthernBrittany(BatzIsland,France)kelp(Laminariadigitata)forest,as
investigatedthroughstableisotopesandchemicalassays.JournalofSeaResearch63:2435.
Subandar A., Petrell R.J., Harrison P.J. (1993) Laminaria culture for reduction of dissolved inorganic nitrogen in salmon farm
effluent.JournalofAppliedPhycology5:455463.
FeijiuW.(1988)MarinephytocultureinChina.http://www.fao.org/docrep/field/003/AB719E/AB719E02.htm.
WernerA.,ClarkeC.,KraanS.(2004)StrategicreviewofthefeasibilityofseaweedaquacultureinIreland.NDPMarineRTDIDesk
StudySeries,DK/01/008.2004.
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10 Otheraquaticbiomass
10.1 Egeriadensa
Figure91EgeriadensaPlanch.
JeffSchardtoftheFloridaFishandWildlifeConservationCommission
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Magnoliophyta
Liliopsida
Alismatales
Hydrocharitaceae
Egeria
Egeriadensa
BIOLOGY
Egeriadensaisasubmersedplant,likealargeElodea(uptothreetimesthesizeofElodea).Egeriagrows
eitherrootedinthebottomorfreefloating.Stemsareupto2mormorelongand3mmthickandmaybe
highlybranched.Whereverbranchesoccur,twonodes(stemjoints)arecompactedcloselytogether,giving
theimpressionthatthenodeisdoubled.Fine,unbranchedwhiterootsformatthedoublenodes,andonly
fragmentscontainingthisdoublenodecangrowintonewplants.Leaves,likethoseofElodea,areinwhorls,
buttypicallywhorlsof4or5,alongstems.Theleavesarenotstronglyrecurvedandare1030mmlong,
andupto4mmwide;0.5mmbelowtheleaftipthewidthis0.51mm.Flowersareabout2cm.acrossand
have3petals.Theyoccuronashortstalkaboutaninchabovethewaterandareproducedprimarilyinthe
springthroughearlysummer,butoccasionallyappearlaterinthegrowingseason.Thisplantonlyflowersin
warmedwater(nativeareas).Elsewherereproductionisvegetative.
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Itgrowsinponds,reservoirs,gravelpits,ditches,swampsandcanals.Itismostcommonineutrophic
water, and will persist in driedup water bodies. Occasionally it is found on completely dry ground. It is
knowntohavesomefrosttolerance.
Egeria is native to South America. It was introduced by the aquarium trade and currently is the top
selling aquatic plant for use in aquaria as "oxygenators". It has been sold under several names including
"oxygenatingplant",elodea,andAnacharis.
Egeriadensahas22%ash,20%protein,29%cellulose.
BIOTECHNOLOGY
Egeriadensahasbeentestedforremovalofnutrientsandheavymetalsfromwatersandasrawmaterialto
producecheapandgoodqualityorganicfertilizers.
References
Caro Lara I., Romero Otalora Z., Lora Silva R. (2009) Production of organic fertilizers with elodea (Egeria densa) present on the
Fuquenelagoon.RevistaU.D.C.AActualidad&DivulgacinCientfica12:91100.[BIOTECHNOLOGYsection]
Champion P.D., Tanner C.C. (2000) Seasonality of macrophytes and interaction with flow in a New Zealand lowland stream.
Hydrobiologia441:112.[BIOLOGYsection]
DaddsN.,BellS.(n.a.)Invasivenonnativeplantsassociatedwithfreshwaters.AGuidetotheiridentification.Preparedonbehalfof
Plantlife, Royal Botanic Garden Edinburgh, Scottish Natural Heritage, Scottish Environment Protection Agency, Scottish
Water.[BIOLOGYsection]
Mdenes A.N., Pietrobelli J.M.T. de A., EspinozaQuiones F.R. (2009) Cadmium biosorption by nonliving aquatic macrophytes
Egeriadensa.WaterScienceandTechnology60:293300.[BIOTECHNOLOGYsection]
FeijoC.,GarcaM.E.,MomoF.,TojaJ.(2002)NutrientabsorptionbythesubmergedmacrophyteEgeriadensaPlanch.:effectof
ammonium and phosphorus availability in the water column on growth and nutrient uptake. Lirnnetica 21: 93104.
[BIOTECHNOLOGYsection]
FBIS
Freshwater
Biodata
Information
System
New
Zealand
(2005)
[BIOLOGY
section]:
https://secure.niwa.co.nz/fbis/validate.do?search=common[Accessed5August2005]
Florence J., Chevillotte H., Ollier C., Meyer J.Y. (2007) Egeria densa. Base de donnes botaniques Nadeaud de l'Herbier de la
Polynsie franaise (PAP). Available from: http://www.herbiertahiti.pf/Selection_Taxonomie.php?id_tax=5052 [Accessed
26March2008][BIOLOGYsection]
GetsingerK.D.,DillonC.R.(1984)Quiescence,growthandsenescenceofEgeriadensainLakeMarion.AquaticBotany20:329338.
[BIOLOGYsection]
HaramotoaT.,IkusimaI.(1988)LifecycleofEgeriadensaPlanch.,anaquaticplantnaturalizedinJapan.AquaticBotany30:389
ITIS
(Integrated
Taxonomic
Information
System)
(2002)
Online
Database.
Egeria
densa.
http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=38972[AccessedMarch2005][BIOLOGY
section]
KayS.,HoyleS.(1999)Aquaticweedfactsheet.BrazilianelodeaorEgeria,Egeriadensa.NCStateUniversity,CollegeofAgriculture
andLifeScience,CropSceinceDepartment.[BIOLOGYsection]
LittleE.C.S.(1979)Handbookofutilizationofaquaticplants.FAOFisheriesTechnicalPaperNo.187,Rome.[BIOLOGYsection]
MacdonaldI.A.W.,ReaserJ.K.,BrightC.,NevilleL.E.,HowardG.W.,MurphyS.J.,PrestonG.(eds.)(2003)Invasivealienspeciesin
southern Africa: national reports & directory of resources. Global Invasive Species Programme, Cape Town, South Africa.
[BIOLOGYsection]
RoyB.,PopayI.,ChampionP.,JamesT.,RahmanA.(2004)AnIllustratedGuidetoCommonWeedsofNewZealand,Egeriadensa,
2ndEdition,NewZealandPlantProtectionSociety.[BIOLOGYsection]
USDAGRIN (Germplasm Resources Information Network) (2003) Egeria densa. National Genetic Resources Program [Online
Database]NationalGermplasmResourcesLaboratory,Beltsville,Maryland.[BIOLOGYsection]
USDANRCS (Natural Resource Conservation Service) (2002) Egeria densa. The PLANTS Database Version 3.5 [Online Database]
NationalPlantDataCenter,BatonRouge,LA.[BIOLOGYsection]
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10.2 Eichhorniacrassipes
Figure92Eichhorniacrassipes(Mart.)Solms(1803).
IllustrationbyOhze
SYMBOLS:B,E
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Magnoliophyta
Liliopsida
Commelinales/Liliales
Pontederiaceae
Eichhornia
Eichhorniacrassipes
Relatedspecies
E.azurea,E.diversifolia,E.heterosperma,E.paniculata,E.paradoxa
BIOLOGY
Eichhorniacrassipesisaperennialaquaticherb;rhizomeandstemsarenormallyfloating,withrootingat
thenodes,withlongblackpendantroots.Leavesareusuallywithinflatedspongypetioles,theleafblades
are circular to reniform, 412 cm wide. Inflorescence is a contracted panicle, 415 cm long, with several
flowers; perianth is lilac, bluish purple, or white, the upper lobe bearing a violet blotch with a yellow
center. Stamens are 6; stalk of the inflorescence soon becomes goosenecked, forcing the dead flowers
under the water; capsule is dehiscent, surrounded by the perianth, membraneous, manyseeded. In
general,seedproductionintemperatepopulationswasfoundtobehalfthatoftropicalpopulations,mainly
duetodifferencesinlevelsofinsectvisitation.Seedsare1114,winged,1.12.10.60.9mm.Thediploid
numberofchrmomosomeis32.
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Waterhyacinthisreportedtotolerateannualprecipitationof8.2to27.0dm(meanof8cases=15.8),
annualtemperatureof21.1to27.2C(meanof5cases=24.9),andestimatedpHof5.0to7.5.Leavesare
killedbyfrost,andplantscannottoleratewatertemperatures>34C.Waterhyacinthisreportedtotolerate
grazing.
NativetoBrazil,nowgrowinginmosttropicalandsubtropicalcountries.Morethan50countriesinwhich
water hyacinth is weed have been listed. It grows in ponds, ditches, canals, calm waters of rivers at an
elevationof0300mandadepthrangeof2m.EngineershaveestimatedthatthePanamaCanalwouldbe
impassablewithinthreeyearswithoutcontinuousaquaticweedcontrolmeasures.
Fresh plant contains 95.5% moisture. On a dry matter basis, it is 75.8% organic matter, 1.5% N, and
24.2%ash.Theashcontains28.7%K2O,1.8%Na2O,12.8%CaO,21.0%Cl,and7.0%P2O5.Proteinisbout12
18%ofdrymatter.Proteincontains,per100g,0.72gmethionine,4.72gphenylalanine,4.32gthreonine,
5.34glysine,4.32gisoleucine,0.27gvaline,and7.2gleucine.2328%ofdryweightiscellulose.
Onadrymatterbasis,standingcropis12.8t ha1,maximum productivity14.6gm2day1,maximum
yield54.7gm2day1.
BIOTECHNOLOGY
Water hyacinth would be ideally suited for nutrient removal systems. Aquatic plants, when growing in
watercontainingamplequantitiesofN,PandK,willexploitthesituationbyluxuryconsumptionofthese
elements,farinexcessofwhattheyneedforhealthygrowth.Asitfloatsonthesurfaceandisnotrooted,
harvesting is facilitated. By continuous harvesting the population could be kept in a rapidly expanding
phase during which uptake rates of nutrients are at their highest. Waters beneath dense stands are
anaerobicsoadditionalNwouldbelostbydenitrification.Therewouldbeconsiderablemicrobialactivity
beneath the hyacinths and nutrients would be absorbed by these organisms. However, considerable
organic matter would reach the water by the loss of root fragments that probably have a fairly high
biologicaloxygendemand(BOD)anditmightprovenecessarytouseconventionalsewageholdingpondsto
reducetheBODpriortofinalrelease.Waterhyacinthrootsnaturallyabsorbpollutants,includingsuchtoxic
chemicalsaslead,mercury,andstrontium90,aswellassomeorganicpollutant,inconcentrations10,000
timesthatinthesurroundingwater.
WaterhyacinthshavebeenusedashumanfoodinthePhilippinesinwartime.Thesoftwhitebudof
theplantwaseateneitherraw,orasasalad,orasaningredientinvegetabledishes.Itwascalledrepollo
(cauliflower). Under conditions of relative food abundance it is considered unlikely that it would still be
usedasfood.
Dried and cleansed plants, can be used as fertilizer, poultry feed, additives to cattlefeed, and plant
mulch.Waterhyacinthproteinshowsdeficientlevelsinonlytwooftheessentialaminoacids,valineand
methionine,ascomparedtotheFAOreferencepattern.Adietcontaininganadequateproteinlevelwillbe
balanced in lysine if it contains 4.2% of that amino acid in its protein content. Corn is deficient in lysine,
containingonly0.8%.Howeverwaterhyacinthcontains5.3%lysine,whilemilkcontains7.8%.Itisevident
thatwaterhyacinthcouldservetoimprovethelysinecontentofacorndiet.Leafproteinwhichcontains
6.3% lysine has been reported to be an effective supplement for barley in pig rations. As a feed for
ruminants,waterhyacinthshouldbeadministeredasacombinationofthelamina,wheretheproteinand
fibre mainly are, and the petiole where the carbohydrate are mainly located. Animals eating fresh water
hyacinth as sole ingredient of the diet could not ingest sufficient dry matter, quite apart from the
imbalance of the nutrients. On a dry matter basis water hyacinth is better than straw but a little low in
proteintocomparewithhay.Thephysicalstructureoftheplantisnotsuitableforhayorsilagemakingand
theproductwouldnothavemuchnutritionalvalue.Thenutritionalvaluecouldbeincreasedbymixingwith
molasses. The hyacinth is rich in minerals, but it would be simple to incorporate any necessary food
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additives in the processing. Pig farming using water hyacinth grown in ponds fertilized with human and
animalwasteiswellestablishedinMalaysia.Thehyacinthcanbefedfreshafterremovaloftheroots.Some
peopleboiltheplant.Pigsreadilyeatthehyacinthandthriveonit.Somefarmershavemachinesforslicing
the plants into small pieces. The high vitamin content (A, B and C) and easy availability of the plant are
important advantages. But the high moisture content and the possibility of the material being
contaminated with pathogens seem to deter the extensive practice of fresh plant feeding. Consequently
mostfarmers(7080%)prefertocooktheslicedhyacinthwithotherfeedingredientsfor5to6hoursand
feedtheboiledmaterial.Silageisalsofedtothepigs.Goodcompostcanbemadefromhyacinth.
Attempts have been made to utilize the plant as a raw material for paper, plastics and other
commercial products, but so far no industry based on water hyacinth appears to have been established.
Thefibrousstem,constitutingabout40%ofthewholeplant,issuitableforpapermaking.Theadditionof
juteorcottonfibrestotheextentof810%ontheweightofthepulpisconsiderednecessaryasthepaper
prepared from the stems alone is translucent. A plastic material suitable for the production of moulded
articles and boards has been prepared from water hyacinth. In Asia, the plants are collected at the
beginningofthecoldweather,lefttodryandthedrymaterialusedalongwithjutesticksasfuel.Theashes
aresubsequentlyusedasmanure.Thepossibilityofusingthedriedweedfortheproductionofpowergas
andpoweralcoholhasbeenconsidered.Threemethodshavebeensuggested,viz.saccharificationbyacid
digestion and subsequent fermentation, gasification by air and steam with recovery of ammonia, and
bacterialfermentationandutilizationoftheevolvedgasforpowerproduction.Potassiumchloride(0.1ton
ofKClpertonofdryhyacinth)isrecoveredinalltheprocesses.Startingfrom1tonofdriedwaterhyacinth,
59Lofethylalcoholand0.2tonsofresidualfibre(8.1MJ)areobtainedbythefirstprocess.Gasificationby
airandsteamgives,pertonofdriedmaterial,3753kgofammoniumsulphateand1133m3ofgas(0.16
MJ) containing hydrogen, 16.6%; methane, 4.8%; carbon monoxide, 21.7%; carbon dioxide, 4.1%; and
nitrogen, 52.8%. Bacterial fermentation gives per ton of material 750 m3 of gas (0.63 MJ) containing:
methane,51.6%;hydrogen,25.4%;carbondioxide,22.1%;andoxygen,1.2%.Thecommercialpossibilities
oftheprocesseshavenotbeenproved.
References
Anonymous(1976)Waterhyacinthssoakuppollution.BioScience26:234.[BIOTECHNOLOGYsection]
BaileyL.H.(1949)ManualofCultivatedPlants.Macmillan,NewYork.
Duke J. Handbook of energy crops. http://www.hort.purdue.edu/newcrop/duke_energy/dukeindex.html [BIOTECHNOLOGY
section]
GohlB.(1981)Tropicalfeeds.Feedinformationsummariesandnutritivevalues.FAOAnimalProductionandHealthSeries12.FAO,
Rome.[BIOLOGYsection]
Little E.C.S. (1979) Handbook of utilization of aquatic plants. FAO Fisheries Technical Paper No. 187, Rome. [BIOTECHNOLOGY
section]
HammerR.L.(1996)Eichhorniacrassipes.InRandallJ.M.,MarinelliJ.(eds.)InvasivePlants:WeedsoftheGlobalGarden.Brooklyn
BotanicGardenInc.,NewYork,p.99.[BIOLOGYsection]
Holm L.G., Plunknett D.L., Pancho J.V., Herberger J.P. (1977) The world's worst weeds. University Press of Hawaii, Honolulu.
[BIOLOGYsection]
HolmL.G.,PanchoJ.V.,HerbergerJ.P.,Plucknett,D.L.(1979)Ageographicalatlasofworldweeds.JohnWiley&Sons,NewYork.
[BIOLOGYsection]
Integrated Taxonomic Information System (ITIS) catalogue of life 2010 http://www.catalogueoflife.org/annual
checklist/2010/search/all/key/eichhornia/match/1(accessedonthe10thofJune2010)[BIOLOGYsection]
MataiS.,BagchiD.K.(1980)Waterhyacinth:aplantwithprolificbioproductivityandphotosynthesis.In:GnanamA.,Krishnaswamy
S.,KahnJ.S.(eds.)ProceedingsoftheInternationalSymposiumonBiologicalApplicationsofSolarEnergy.MacMillanCo.of
India,Madras,pp.144148.[BIOLOGYsection]
PenfoundW.T.,EarleT.T.(1948)Thebiologyofthewaterhyacinth.EcologicalMonographs18:449472.[BIOLOGYsection]
ReedC.F.(1970)SelectedweedsoftheUnitedStates.AgricultureHandbook366.USDA,Washington,DC.[BIOLOGYsection]
USDA,NRCS(2001)ThePLANTSDatabase,Version3.1.(http://plants.usda.gov).NationalPlantDataCenter,BatonRouge,LA
708744490USA.
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10.3 Elodeacanadensis
Figure93ElodeacanadensisMichaux(1803).
IllustrationbyC.A.M.Lindman
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Magnoliophyta
Liliopsida
Hydrocharitales
Hydrocharitaceae
Elodea
Elodeacanadensis
Relatedspecies
E.bifoliata,E.callitrichoides,E.granatensis,E.nuttallii,E.potamogeton,E.spinosa.
BIOLOGY
Elodeacanadensisisasubmergedaquaticperennialfreshwaterherb,usuallyfirmlyrootedtothebottom
mud and producing a thick green mat below the water surface. Short thread like stolons giving rise to
slender vertical stems to 3 m long. Leaves are dark green oblonglinear, formed at intervals of 325 mm
along the stem, in groups of three, each 612 mm long and about 15 mm wide, usually with forward
pointingteethonthemargins.Rootsarefilamentous,risingfromthenodesalongthestolons.Flowersare
whiteorpalepurplewiththreesepalsandpetals.Theyaresolitary,formingintheaxilsoftheleavesand
growingtowardsthesurfaceonthreadlikestalksabout30cmlong..Fruitsarecapsuleslessthan1cmin
length.Itisdispersedbyseedsandfragmentsviawatercurrents.ItisadioeciousplantfloweringfromJune
toAugust.Pollinationoccursnearthewatersurfaceandpollenisdistributedbywindandwatercurrents.
Vegetative reproduction by fragments is very common. Mass development has been reported multiple
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times in the last century. It is preyed upon by a high number of freshwater organisms including fish and
birds.
It is diffuse in surface standing waters, and surface running waters, like shallow lakes, ponds, pools,
ditchesandstreamswithslowmovingwater.Itgrowsupto3mwaterdepthand,inexceptionalcases,up
to16mindepth.IttoleratespHvaluesfrom6.0to7.5andtemperaturesfrom1to25C.
ItoriginatesfromNorthAmericaninlandwaters.ThefirstEuropeanrecordwasreportedfromIreland
in 1836. It subsequently became widespread in north and central European countries. After a rapid
colonizationofnorthernEuropethepopulationsdeclinedduetotheintroductionofElodeanuttallii.Today
thepopulationisstable.Asitcanbeverydominant,itcompetesfornutrientsandspacewithotherplants.
Itcanbioaccumulatenutrientsandmodifythehabitatbyreducingwatermovement.Thespeciesisknown
to outcompete other plants. During dense blooms, impairs boating, fishing, swimming, and water skiing.
Cloggingofwaterintakepipesofpowerplantsandotherindustrieswerereported.
Elodea is anchored to the bottom deposits of a lake or river by adventitious roots, but nutrients are
probablyabsorbedmainlybyleavesandstemsincontactwiththefreewater.Althoughtheconcentration
ofdissolvedsaltsinthesurroundingwaterwilllargelycontrolthedensityandcompositionoftheplant,the
evidenceindicatedadifferenceincompositionbetweenspecies,eveninthesamelake,aswellasseasonal
changes.Itfollowsthatifthiswaterweedistobeexploitedcommercially,preliminaryplantanalysesare
neededtodeterminethebestspeciesandseasonforharvesting.
Crudeproteinisabout12%,ash28%andfiber16%.
BIOTECHNOLOGY
Toobtain1tonofdryElodeacanadensis14tonsofwetmaterialwouldhavetobeprocessed.E.canadensis
is an excellent food for cattle and pigs, when fed to sheep it was found to be unpalatable but it was
acceptedwhenmixedwithpasture(1:5drymatterbasis).Itappearstocontainallthevitaminsinatleastas
highanamountasalfalfa.OfmoreinterestisthefactthatthebiologicalvalueoftheproteininElodeais
about70%thatofalfalfaandthedigestibilityisapparentlybetter.Cystineseemstobethelimitingamino
acidinElodeaaswellasinalfalfa.E.canadensiscanbeusedasanonexpensivesupplementalfoodinorder
toincreasegrowthandsurvivalinsummerlingnoblecrayfish,A.astacus,thathasthepotentialtoconsume
thismacrophyteinnature.
Itshowsthetypicalproductivityofsubmergedmacrophytesintemperatezonesas27tonsdryweight
ha1year1and1759tonsfreshweightha1year1.
References
Bowmer H., Kathleen S.W., Jacobs L., Sainty G.R. (1995) Identification, biology and management of Elodea canadensis,
Hydrocharitaceae.JournalofAquaticPlantManagement33:1319.[BIOLOGYsection]
Champion P.D., Hofstra D.E., Clayton J.S. (2007) Border control for potential aquatic weeds. Stage 3. Weed risk management.
Science&TechnicalPublishingNewZealandDepartmentofConservation.[BIOLOGYsection]
DAgaroE.,RenaiB.,GherardiF.(2004)EvaluationoftheAmericanwaterweed(ElodeacanadensisMichx.)assupplementalfoodfor
thenoblecrayfish,Astacusastacus.BulletinFranaisdelaPcheetdelaPisciculture372373:439445.[BIOTECHNOLOGY
section]
Department of Agriculture and Food, Australia: http://agspsrv95.agric.wa.gov.au/dps/version02/01_plantview.asp? [BIOLOGY
section]
Gollasch S. (2006) Elodea canadensis. DAISIEDelivering Alien Invasive Species Inventories for Europe. http://www.europe
th
aliens.org/pdf/Elodea_canadensis.pdf(accessedonthe10 ofJune2010)[BIOLOGYsection]
Integrated Taxonomic Information System (ITIS) catalogue of life 2010 http://www.catalogueoflife.org/annual
checklist/2010/search/all/key/elodea/match/1(accessedonthe10thofJune2010)[BIOLOGYsection]
th
KewsWorldChecklistofSelectedPlantFamilieshttp://apps.kew.org/(accessedonthe10 ofJune2010)[BIOLOGYsection]
section]
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10.4 Lagarosiphonmajor
Figure93Lagarosiphonmajor(Ridley)Moss
ControlofaquaticinvasivespeciesinIreland(CAISIELife+project)
SYMBOLS:B
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Magnoliophyta
Liliopsida
Alismatales
Hydrocharitaceae
Lagarosiphon
Lagarosiphonmajor
Relatedspecies
L. cordofanus, L. hydrilloides, L. ilicifolius, L. madagascariensis, L. muscoides, L. rubellus, L. steudneri, L.
verticillifolius.
BIOLOGY
L. major is a rhizomatous, perennial, submerged aquatic plant. It reaches its maximum growth in clear
wateruptoadepthof6.5m,butmayonlygrowto1minmurkywater.Ithasnumerousthreadlikeroots,
which are adventitious and, along with rhizomes, anchor it to the bottom. Stems, which can reach the
surface,arebrittleandsparselybranched,35mmindiameterandcurvedtowardsthebase(Jshaped).The
leavesare520mmlongand23mmwide,andoccurinalternatespiralsalongthestem.Theygenerally
have tapered tips curving downwards towards the stem, except in low alkalinity water where they are
straight. The threepetalled female flowers are very small, clearwhite on the surface, and grow on very
thin white to almost translucent filamentlike stalks. Neither the male flower, which floats freely to the
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surface, nor fruit or seeds have been recorded outside of its native range. Since the species is dioecious
(sexes on different plants) both must be present for sexual reproduction. Only female plants are known
outside of the native range of this species. All reproduction in introduced regions is therefore asexual
primarilybyfragmentationorlocalgrowthbyrhizomatousspread.
L.majorgrowsbestinclear,stillorslowmovingfreshwaterwithsiltyorsandybottoms.Itprefersthe
cooler waters of the temperate zone, with optimum temperatures of 2023 C and a maximum
temperatureofaround25C.Itcanliveinhighandlownutrientlevelsandgrowsbestunderconditionsof
high light intensity. It also tolerates relatively high pH (i.e. alkaline conditions). Growth of L. major is
greatestinshelteredareasprotectedfromwind,wavesandcurrents.
ItisnativeofsouthernAfrica,isfoundinhighmountainstreamsandponds.Ithasspreadthroughout
the world as an aquarium plant and is also known as an 'oxygen plant'. However, dense infestations can
actuallyconsumemoreoxygenthantheyproduce,andreducewaterqualityandavailableoxygen.
BIOTECHNOLOGY
L. major and other aquatic species grown in small outdoor tanks can be used successfully to assess the
effects of cropprotection products on nontarget aquatic flora. The possible utilization by harvesting for
stockfeedinNewZealandlakeswasevaluated;however,theuseoftheplantsasfodderwasthoughttobe
unsuitablebecauseofthecontentofarsenicaccumulatedbytheplantsfromthethermalwatersthatenter
thelakes.
References
AiroS.,SconfiettiR.(1995)Insituexperimentsonproductivity ofaquaticmacrophytesinapond.RivistadiIdrobiologia34:147
156.
CAISIElife+project(2010)controlofaquaticinvasivespeciesinIreland.www.caisie.ie.Accessedonthe06thofOctober2010.
Champion P.D., Tanner C.C. (2000) Seasonality of macrophytes and interaction with flow in a New Zealand lowland stream.
Hydrobiologia441:112.
Coffey B.T., Clayton J.S. (1987) Submerged macrophytes of Lake Pupuke Takapuna New Zealand. NewZealand Journal of Marine
andFreshwaterResearch21:193198.
CONABIO(2008)SistemadeinformacinsobreespeciesinvasorasenMxico.EspeciesinvasorasPlantas.ComisinNacionalpara
el
Conocimiento
y
Uso
de
la
Biodiversidad.
Fecha
de
acceso.
www.conabio.gob.mx/invasoras/index.php/Especies_invasoras__Plantas
ConservatoireBotaniqueNationalDeMascarin(BOULLETV.coord.)(2007)Lagarosiphonmajor.Indexdelaflorevasculairedela
Runion(Trachophytes):statuts,menacesetprotections.Version2007.1(misejour12juin2007).
CookC.D.K.(2004)AquaticandWetlandPlantsofSouthernAfrica.BackhuysPublishers,TheNetherlands.
deCarvalhoR.F.,BromilowR.H.,GreenwoodR.(2007)UptakeofpesticidesfromwaterbycurlywaterweedLagarosiphonmajorand
lesserduckweedLemnaminor.PestManagementScience63:789797.[BIOTECHNOLOGYsection]
EgloffF.(1975)NewandnoteworthyspeciesofSwissflora.BulletindelaSocieteBotaniqueSuisse84:333342.
GlobalInvasiveSpeciesDatabase:http://www.issg.org/database/species/ecology.asp?si=403&fr=1&sts=sss[BIOLOGYsection]and
[BIOTECHNOLOGYsection]
ITIS
(Integrated
Taxonomic
Information
System)
(2005)
Online
Database
Lagarosiphon
major.
http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=565981
JamesC.S.,EatonJ.W.,HardwickK.(1999)Competitionbetweenthreesubmergedmacrophytes,ElodeacanadensisMichx,Elodea
nuttallii(Planch.)StJohnandLagarosiphonmajor(Ridl.)Moss.Hydrobiologia415:3540.
section]
RattrayM.R.(1995)TherelationshipbetweenP,FeandMnuptakesbysubmersedrootedangiosperms.Hydrobiologia308:117
120.
RattrayM.R.,HowardWilliamsC.,BrownJ.M.(1994)RatesofearlygrowthofpropagulesofLagarosiphonmajorandMyriophyllum
triphylluminlakesofdifferingtrophicstatus.NewZealandJournalofMarineandFreshwaterResearch28:235241.
RiisT.,BiggsB.J.,FlanaganM.(2003)SeasonalchangesinmacrophytebiomassinSouthIslandlowlandstreams,NewZealand.New
ZealandJournalofMarineandFreshwaterResearch37:381388.
Roy B., Popay I., Champion P. James T., Rahman A. (2004) An Illustrated Guide to Common Weeds of New Zealand 2nd Edition.
Lagarosiphonmajoroxygenweed.NewZealandPlantProtectionSociety.
State of Queensland (2004) Lagarosiphon major description. The State of Queensland (Department of Natural Resources and
Mines).
StricklandR.,HardingJ.,ShearerL.(2000)TheBiologyofLakeDunstan.CawthronReportNo.563;ContactEnergyLimited.
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SymoensL.,TriestS.(1983)MonographoftheAfricangenusLagarosiphon.BullettinduJardinBotaniqueNationaldelaBelgique53:
441488.
UniversityofFlorida(2001)Lagarosiphonmajor(Ridley)Moss.NonNativeInvasiveAquaticPlantsintheUnitedStates,Centerfor
AquaticandInvasivePlants,UniversityofFloridaandSeaGrant.
Wells R.D., DeWinton M.D., Clayton J.S. (1997) Successive macrophyte invasions within the submerged flora of Lake Tarawera,
centralNorthIsland,NewZealand.NewZealandJournalofMarineandFreshwaterResearch31:449459.
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10.5 Lemnaminor
Figure95LemnaminorLinnaeus
RobertH.Mohlenbrock@USDANRCSPLANTSDatabase/USDANRCS.1995.Northeastwetlandflora:Fieldoffice
guidetoplantspecies.NortheastNationalTechnicalCenter,Chester
SYMBOLS:B,E,PIV
TAXONOMY
Phylum
Class
Order
Family
Genus
Species
Magnoliophyta
Liliopsida
Arales
Lemnaceae
Lemna
Lemnaminor
Relatedspecies
L.aequinoctialis,L.disperma,L.gibba,L.japonica,L.minuta,L.obscura,L.perpusilla,L.tenera,L.trisulca,
L.turionifera,L.mnavaldiviana,L.yungensis.
BIOLOGY
GeneralDescriptionofMajordivisionsandclasses:
TheLemnaceaeisamonocotyledonousfamilyof4genera:Spirodela,Lemna,WolffiaandWolfiella,and37
species.AllLemnaceaespeciesaresmallaquaticplants,commonlycalledduckweeds(LemnaandSpirodela
species)andwatermeals(Wolffiaspecies).Themajorityofresearchinvolvingtheseplantshasbeendone
withonlyafewspecies;primarilyL.gibbaandL.minor,SpirodelapolyrrhizaandSpirodelapunctata,andto
alesserextent,Wolffiaglobosa.Forthemajorityofspecies,littleisknownoftheirbiologyandtheabilityto
extrapolateanyofthetechnologicalapplicationsdevelopedforthebetterstudiedspecies(Stomp,2005).
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Physiologicalcharacteristics
Theplantsareperennials,growinganywherethereiswaterandsun.Theseplantsrequirelittlemechanical
supportorvasculartissue(thesmallestmembersofthefamilycompletelylackxylemandphloem)somost
cellsresemblematuringleavesexpendingverylittlephotosyntheticenergyonplantstructures.Asaresult,
vegetativepropagationoftheseplantshashighbiomassyieldpotential.
MembersofthefamilyLemnaceaearetinyaquaticmonocotsthatrangeinsizefrom1.5cmlong(Spirodela
polyrhiza)tolessthanamillimeter(Wolffiaglobosa).
All members of the Lemnaceae are small, freefloating, freshwater plants whose geographical range
spanstheentireglobe(Landolt,1986).MembersoftheLemnaceaearethemostmorphologicallyreduced
plantsknown.PlantsofSpirodelaandLemnaspeciesconsistofafrond,arootorroots(thenumberofroots
are species specific) and, when present, a flower. The morphology of Wolffia species is further reduced,
with plants consisting of tiny fronds devoid of roots and producing single flowers. Wolfiella species are
morevariedinmorphology.
Table 21 Biochemical composition of Duckweed. Composition
expressedas%dryweightexceptforWaterandash.FromLandolt
andKandeler(1987a),exceptforstarch(ChengandStomp,2009).
Water(%FW)
Ash
TotalCarbohydrate
Crudefiber
Starch
Protein
Lipid
K
Na
Ca
Mg
Cu
Si
S
P
Duckweed(Lemnaceae)
8697
12.027.6
14.143.6
5.716.2
675(somestrainsinlabcondition)
6.845.0
1.89.2
0.037.0
0.031.3
0.184.5
0.042.8
0.21033.2
0.415.35
0.337.0
0.032.8
Theproteincontentofduckweedsisoneofthehighestintheplantkingdom,butitisdependentongrowth
conditions. Typically duckweeds are rich in leucine, threonine, valine, isoleucine and phenylalanine. They
tendtobelowincysteine,methionine,andtyrosine.
Duckweed growth can be optimized to produce high levels of protein or high levels of starch. The
plant'sdryweightaccumulationvariesbyspeciesandgrowthconditionsandrangesfrom6to20%offresh
weight (Landolt and Kandeler, 1987b; Tillberg, 1979). Protein content of a number of duckweed species
grown under varying conditions has been reported to range from 15 to 45% dry weight (Chang, 1977;
Porath,1979;Appenroth,1982).Thesevaluesplacetheproteincontentofdryduckweedbiomassbetween
alfalfameal(20%)andsoybeanmeal(41.7%)(Hillman,1961).Weroutinelygrowduckweedondiluteswine
wastewaterandget30to35%proteinofdryduckweed.Duckweedstarchcontentisdependentongrowth
conditions, e.g., pH, phosphate concentration (TasseronDeJong, 1971; McLaren and Smith, 1976) and
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developmental states con trolled by the plant hormones, cytokinin (TasseronDeJong, 1971; McCombs
and Ralph, 1972) and abscissic acid (Landolt and Kandeler, 1987b; McLaren and Smith, 1976). Starch
contentsrangingfrom3to75%havebeenreported(LandoltandKandeler,1987b;ReidandBieleski,1970).
Aduckweedstarchcontentof75%iscomparabletocorn,whosestarchcontentrangesfrom65to75%(Lin
andTanaka,2006).
Growthkineticsandproductivity
Doublingtimesvarybyspeciesandenvironmentalconditionsandareasshortas20to24hoursandmany
species have doubling times of 2 to 3 days (Chang et al, 2003; Venkararaman et al, 1970). Intensive
laboratorycultureofduckweedhasachievedhighratesofbiomassaccumulationperunittimeatculture
densitiesof12kgm2(LandoltandKandeler,1987b).Inwastewatertreatmentresearch,agrowthrateof
0.2kgdryweightm2week1hasbeenachieved(Chengetal.,2002a).Toachievethesegrowthrates,only
lowconcentrationsofnutrientsarerequired.Oronandcoworkers(1988)achievedoptimalgrowthratesat
20ppmnitrogenutilizingmunicipalwastewater.Ourresearchwithwastewaterindicatesthathighgrowth
ratescanbeachievedatnitrogenlevelslessthan10ppm(Chengetal.,2002a,b).
BIOTECHNOLOGY
Culturemedia
Stomp(2005)andothershavedemonstratedduckweedgrowthonavarietyofnutrientsolutionslistedin
Table 22. Although differences in growth rates have been observed, generally, duckweed will grow on
almost any dilute, inorganic salt solution that supplies essential macro and micronutrients. The plants
toleratearangeofpH,formostspeciesrangingbetweenpH4.5and7.2.Anumberoforganicbuffers,e.g.
EDTA, citrate, tartaric acid, MES, MOPS, and compounds which stabilize proteins, such as PVP, can be
added to the growth medium without significantly affecting growth rates. This is an important
considerationifrecombinantproteinsaresecretedandaretoberecoveredfromplantgrowthmedium.If
theplantsaregrownunderlightlevelsinsufficienttosupportrobustphotosyntheticgrowth(<300molm2
s1),sucrosecanbeaddedtothemediumtosustainluxuriantgrowthatconcentrationsof0.53%,with1%
being preferable. Other carbon sources have been investigated for their ability to support frond growth
(Frick,1991).
Table22Mediautilizedforcallusinduction,establishmentandfrondregenerationofselectLemnaceaespecies
(Stomp,2005).
Species
Process
BasalMedium
L.aequinoctialis
Callusinduction
MS+3%sucrose
Callusmaintenance
MS
Frondlikestructures MS
L.gibba
Callusinduction
Callusmaintenance
Frondregeneration
MS
PlantGrowthRegulators
Reference
2,4D01mg/L;2iP10mg/L
same
same
Chang&Hsing,1978
2,4D45M;2iP4.9M
Chang&Chui,1976;1978
L.gibba
Callusinduction
MS
2,4DorIAALala
Slovin&Cohen,1985
L.minor
Callusinduction
Callusmaintenance
FNO+2%sucrose
FNO
2,4D5mg/L;2iP0.5mg/L
2,4D2mg/L;2iP0.2mg/L
Frick,1991
Frondregeneration
FNO
2iP0.2mg/L
L.gibba
Callusinduction
Callusmaintenance
Frondregeneration
MS+3%sucrose
MS
MS
2,4D2050M;BA2M
2,4D120M;BA2M
BA10M
Moon&Stomp,1997
L.minor
Callusinduction
Callusmaintenance
Frondregeneration
MS+3%sucrose
MS
SH
2,4D2050M;BA2M
2,4D120M;BA2M
BA10M
Stomp,unpublished
L.gibba
Callusinduction
Callusmaintenance
Frondregeneration
B5+1%sucrose
B5
B5
dicamba50mg/L;BA2mg/L
PCA10mg/L;Picloram2mg/L;2iP2mg/L
TDZ1mg/L
Lietal.,2004
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Species
Process
BasalMedium
PlantGrowthRegulators
Reference
S.oligorrhiza
Callusinduction
Callusmaintenance
Frondregeneration
WP+1.5%galactose
WP+2%sorbitol
WP
WP+0.5%sucrose
dicamba50mg/Lpretreatment
PCA5mg/L;Picloram2mg/L;2iP2mg/L
PCA10mg/L;2iP2mg/L
TDZ1mg/L
Lietal.,2004
S.punctata
Callusinduction
Callusmaintenance
Frondregeneration
MS+1%sorbitol
WP+2%sorbitol
WP+0.5%sucrose
2,4D3.5mg/L;dicamba15mg/L;2iP2mg/L
2,4D1mg/L;NAA5mg/L;TDZ0.5mg/L
2iP1mg/L
Lietal.,2004
Cultivationmethods
Culturingmethodssimilartothoseusedformicroorganismsare easilyadaptedtogrowduckweedinthe
laboratory.
Productionsystems
Wetlandsandpondsarethemostcommonsitestofindduckweeds,butotherquietbodiesofwatermay
harborthem.Duckweedsmayalsobefoundonthefringesoflargerlakesandinquietbackwatersand
sloughscutofffrommightyrivers.
Duckweedareroutinelygroundonswinewastewater.Duckweedsofeverygenus,Spirodela,Landoltia,
Lemna,WolffiaandWolffiellaarefoundworldwide,althougheachspecieshascertainareasandclimates
whereitisparticularlywelladapted.
Harvestingmethods
Duckweedscanbeharvestedwithmechanicalskimmersorwithfinenets(whichislabourintensive).Some
oftheplantswillbemissedbytheskimmer,sointimethepopulationwillregrowtotheoriginalsize.This
isnotadisadvantageifyouhaveagoodusefortheplantsthatyouwillharvest(Cross,2010).
Biomassprocessingforbiofuelpurposes
Enzymatichydrolysisoftheduckweedbiomasswithamylaseshasbeenusedtoyieldahydrolysatewitha
reducingsugarcontentcorrespondingto50.9%oftheoriginaldryduckweedbiomass.Fermentationofthe
hydrolysateusingyeastgaveanethanolyieldof25.8%oftheoriginaldryduckweedbiomass.Theseresults
indicate that the duckweed biomass can produce significant quantities of starch that can be readily
convertedintoethanol.Duckweedbiomasswouldrequirelittleornomechanicalgrindingbecauseofthe
smallsizeoftheplantsandbecauseitisagreen,hydratedbiomass.Thelackofamillingsteptoprepare
biomassforfermentationtranslatesintoasubstantialsavingsinenergy,oneofthemajorcostsinthecorn
toethanolprocess(ChengandStomp,2009).
SinceLemnaceaegrowonfreshwatermedium(orwastewater)andavailabilityofwatercanberestrictedin
somepartsofEurope,thecultivationofLemnaceaeshouldremainlocatedwherewatersupplyisabundant.
Besides,Lemnaceaearefastgrowingorganismsandmaybedisseminatedbymigratorybirds,severalwater
bodiesmightbeunwillinglyinoculated.
References
Alaerts G.J., Mahbubar R., Kelderman P. (1996) Performance analysis of a fullscale duckweedcovered sewage lagoon. Water
Research30:843852.
AppenrothK.J.,AugstenH.,LiebermannB.,FeistH.(1982)EffectsoflightqualityonaminoacidcompositionofproteinsinWolffia
arrhiza(L.)Wimm.usingaspeciallymodifiedBradfordmethod.BiochemieundPhysiologiederPflanzen177:251258.
ChangW.C.,ChiuP.L.(1976)Inductionofcallusfromfrondsofduckweed(LemnagibbaL.).BotanicalBulletinofAcademiaSinica
17:106109.
ChangW.C.,ChiuP.L.(1978)RegenerationofLemnagibbaG3throughcallusculture.ZeitschriftfurPflanzenphysiologie89:9194.
Chang W.C.,Hsing Y.I. (1978) Callus formation and regeneration of frondlike structures in Lemna perpusilla 6746 on a defined
medium.PlantScienceLetters13:133136.
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Chang S.M., Yang C.C., Sung S.C. (1977) The cultivation and the nutritional value of Lemnaceae. Bullettin of the Institute of
Chemistry,AcademiaSinica24:1930.
ChengJ.J.,StompA.M.(2009)Growingduckweedtorecovernutrientsfromwastewatersandforproductionoffuelethanoland
animalfeed.CLEANSoil,Air,Water37:1726.
ChengJ.,BergmannB.A.,ClassenJ.J.,StompA.M.,HowardJ.W.(2002a)NutrientrecoveryfromswinelagoonwaterbySpirodela
punctata.BioresourceTechnology81:8185.
Cheng J., Landesman L., Bergmann B.A., Classen J.J., Howard J.W., Yamamoto Y.T. (2002b) Nutrient removal from swine lagoon
liquidbyLemnaminor8627.TransactionsoftheASAE45:10031010.
CrossJ.W.(2010)TheCharmsofDuckweed.http://www.mobot.org/jwcross/duckweed.htm(accessed10Nov.2010).
DenHartogC.(1975)Thoughtsaboutthetaxonomicalrelationshipswithinthelemnaceae.AquaticBotany1:407416.
FrickH.(1991)CallogenesisandcarbohydrateutilizationinLemnaminor.JournalofPlantPhysiology137:397401.
HillmanW.S.(1961)TheLemnaceaeorduckweeds.Areviewofthedescriptiveandexperimentalliterature.BotanicalReview27,:
221287.
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Landolt E., Kandeler R. (1987a) Biosystematic investigations in the family of duckweeds (Lemnaceae). Veroff. Geobot. Inst. ETH,
Zurich.vol.2,pp.4243.
LandoltEandKandeler,R.(1987b).ThefamilyofLemnaceaeamonographicstudy.Volume2.Biosystematicinvestigationsinthe
familyofduckweeds(Lemnaceae)(vol.4).Veroff.Geobot.Inst.ETH,Zurich.
LiJ.,JaniM.,VunshR.,VishnevetskyJ.,HannaniaU.,FlaishmanM.,PerlA.,EdelmanM.(2004)Callusinductionandregenerationin
SpirodelaandLemna.PlantCellReports22:457464.
Lin Y., Tanaka S. (2006) Ethanol fermentation from biomass resources: current state and prospects. Applied Microbiology and
McCombsP.J.A.,RalphR.K.(1972)Protein,nucleicacidandstarchmetabolismintheduckweed,Spirodelaoligorrhiza,treatedwith
cytokinins.BiochemistryJournal129:403417.
McLaren J.S., Smith H. (1976) The effect of abscisic acid on growth, photosynthetic rate and carbohydrate metabolism in Lemna
minor.NewPhytologist76:1120.
Moon H.K., Stomp A.M. (1997) Effects of medium components and light on callus induction, growth and frond regeneration in
Lemnagibba(duckweed).InVitroCellularandDevelepmentalBiologyPlant33:2025.
OronG.,deVegtA.,PorathD.,(1988)Nitrogenremovalandconversionbyduckweedgrownonwastewater.WaterResearch22:
179184.
PorathD.,HepherB.,KotonA.(1979)Duckweedasanaquaticcrop:evaluationofclonesforaquaculture.AquaticBotany7:273
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SlovinJ.P.CohenJ.D.(1985)GenerationofvariantlinesofLemnagibbaG3viatissueculture,foruseinselectingauxinmetabolism
mutants.PlantPhysiology77(4suppl.):11.
StompA.M.(2005)Theduckweeds:Avaluableplantforbiomanufacturing.In:ElGewelyM.R.(ed.)BiotechnologyAnnualReview,
Elsevier,Rotterdam,pp.6999.
TasseronDeJongJ.,VeldstraH.(1971)Investigationsoncytokinins.Part2:InteractionoflightandcytokininsasstudiedinLemna
minor.PhysiologiaPlantarum24:239241.
Tillberg E., Holmvall M., Ericsson T. (1979) Growth cycles in Lemna gibba cultures and their effects on growth rate and
ultrastructure.PhysiologiaPlantarum46:512.
Venkararaman R., Seth P.N., Maheshwari S.C. (1970) Studies on the growth and flowering of a shortday plant, Wolffia
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11 Concludingremarks
Algaehavebeenrecognizedasvaluableresourceandexploitedbyhumansforthousandsofyears.Thelast
century has seen intensive research efforts to exploit the huge potential of micro and macro algae and
otheraquaticplantsforfood,fodder,energyandbiomaterials,aswellasenvironmentalapplicationssuch
aswastewatertreatmentandbioremediationthataresummarizedinthedocumentabove.
While a significant industry based on exploitation of macroalgae biomass has developed, microalgal
biomass production remains restricted to a few high value nutraceutical products such as pigments (
carotene from Dunaliella and astaxanthin from Haematococcus) or dried microalgal biomass (Chlorella,
Spirulina).
Thehugeeffortsinvestedrecentlyininvestigatingthepotentialofmicroalgaeforbiofuelsproduction
have significantly advanced our understanding in algal biology, cultivation, harvesting and biomass
processing,butnocommerciallyviablesustainableproductionpathwayexistsandinconsequencenoalgal
biofuelsproductionfacilityisonlinetoday.
Difficultiestobeovercomearemostlyinreducingproductionandprocessingcostsofalgalbiomass,by
means of progress in engineering and culture management, and by selecting and improving strains for
maximalproductivityatminimalcosts.Dozensofalgalspeciesandstrainsholdpromiseinthisrespect,and
nostrain maybesingled outforextraordinarysuitabilityunder thewidevarietyof cultivationconditions
imaginable. Many algae may besides fuel provide high value products or protein for improving the
economic balance of the production process. Recent advances in genomics and genetic engineering,
demonstratedinChlamydomonas,Phaeodactylumandothers,holdspromisefortargetedimprovementsof
algal strains to adapt them specifically for enhanced growth and productivity under selected conditions.
Alternatively great opportunities may reside in utilization of algal biomass cultivation for environmental
applications such as waste water treatment or remediation, under recovery of valuable nutrients and
biomassproduction.
All possible schemes proposed have been demonstrated in the small scale, but intensive research
efforts are still necessary to further our understanding on algal biology and production processes. Any
significant advance in biofuels production is dependent on massive funding for the establishment of
multiple,differentanddiversealgalbiomassproductionfacilitiesofsufficientsizeforallowinggainingthe
necessaryinsightsanddatatowardsreachingsustainableandprofitableproductionprocesses.
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ANNEXI
AVAILABILITYofALGAE
ThemainCultureCollectionsofalageandcyanobacteriaarelistedinthetablebelow:
Collection+website
CCAP(CultureCollectionofAlgaeandProtozoa)
http://www.ccap.ac.uk/cultures/cultures.htm
SAG(SammlungvonAlgenkulturenderUniversittGttingen)
http://sagdb.unigoettingen.de/
CCMP(ProvasoliGuillardNationalCenterforCultureofMarinePhytoplankton)
https://ccmp.bigelow.org/node/56
UTEX(TheCultureCollectionofAlgaeattheUniversityofTexasAustin)
http://web.biosci.utexas.edu/utex/Search.aspx
CCALA(CultureCollectionofAutotrophicOrganisms)
http://www.butbn.cas.cz/ccala/index.php
PCC(ThePlymouthCultureCollectionofMarineAlgae)
http://www.mba.ac.uk/culturelist.php
PCC(PasteurCultureCollectionofCyanobacteria)
http://www.pasteur.fr/ip/easysite/go/03b00001200g/collectionofcyanobacteriapcc/
ALGOBANK (la collection de cultures de microalgues de l'Universit de Caen BasseNormandie)
http://www.unicaen.fr/ufr/ibfa/algobank
CCBA(CultureCollectionofBalticAlgaeattheUniversityofGdansk)
http://ocean.ug.edu.pl/~ccba/ien.php?id=ccba
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa)
http://www.sccap.dk/search/
RCC(RoscoffCultureCollection)
http://www.sbroscoff.fr/Phyto/RCC
BCCM(BelgianCoordinatedCollectionsofMicroorganisms)DiatomCollectionattheGhentUniversity
F&MCultureCollection,Italy
http://www.femonline.it/
CSIRO(CollectionofLivingMicroalgae)
http://www.marine.csiro.au/algaedb/default.htm
CCCM(CanadianCenterfortheCultureofMicroorganisms)including:
NEPCC(NorthEastPacificCultureCollection)
FWAC(FreshwaterAlgalCultureCollection)
http://botany.ubc.ca/cccm
CPCC(CanadianPhycologicalCultureCentre)(formerlyknownasUTCC)
http://www.phycol.ca/cultures
ATCC(AmericanTypeCultureCollection)
http://www.lgcstandardsatcc.org/
SERImicroalgaeculturecollection
http://www.tpub.com/content/altfuels05/3814/38140091.htm
FACHB(FreshwaterAlgaeCultureCollection)
http://www.ctcccas.ac.cn/typecc/danshui/database.html
JapanNITE(holdingformerMBICstrains)
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?lang=en
JapanNIES(NationalInstituteforEnvironmentalStudies)
http://mcc.nies.go.jp
Theavailabilityofeachmicroalgaandcyanobacteriumconsideredinthedocumentisreportedinthe
following section. No availability was found for Parietochloris incisa. Note that some strains are shared
betweencollections.NotealsothatmanystrainsareownedbyResearchInstitutesandUniversitiesandare
notdepositedinCultureCollections,sothattheiravailabilityisnotknown.
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Arthrospirasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCMP (ProvasoliGuillard National Center for Culture of
MarinePhytoplankton)
UTEX (The Culture Collection of Algae at the University of
TexasAustin)
http://www.pasteur.fr/ip/easysite/go/03b000012
00g/collectionofcyanobacteriapcc/
CPCC (Canadian Phycological Culture Centre) (formerly
knownasUTCC)
CCAP 1406/1 (A. brevis); CCAP 1475/8 (A.

fusiformis);CCAP1475/9(A.maxima)
SAG31.96(A.massartii)
CCMP1295(A.platensis)
UTEXLB2720;LB2721(A.fusiformis)
CCALA 023; 025 (A. fusiformis); CCALA 027 to 030

(A.maxima)
PCC7345;7939to7940;8005to8006;9108;9223
K0966(Arthrospirasp.)
CS328(A.maxima)
CPCC29408(A.platensis)
ATCC29408(A.platensis)
FACHB314;350;439;790to794(A.platensis)
Phormidiumsp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 1462/6; 1462/10 (P. autumnale); CCAP

1446/8 (P. foveolarum); CCAP 1462/3 (P.
minnesotense); CCAP 1462/8; 1462/11; 1462/13
(Phormidiumsp.)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 14596 (P. animale); SAG 35.90; 78.79 (P.
autumnale); SAG 60.90 (P. ectocarpi); SAG 14621
(P. foveolarum); SAG 79.79 (P. inundatum); SAG
26.99 (P. molle); SAG 80.79 (P. persicinum); SAG
75.79 (P. tergestinum); SAG 81.79 (P. uncinatum);
SAG 2012; 14.92; 14631e; 212.80; 37.90; 47.90;
61.90;82.79;9.92(Phormidiumsp.)
CCMP (ProvasoliGuillard National Center for Culture of CCMP2591(P.breve);CCMP2607(P.janthiforum);
CCMP 2544 (P. keutzingianum); CCMP 2574 (P.
okenii); CCMP 638 (P. persicinum); CCMP 1230 to
1231 (P. tenue); CCMP 2068; CCMP 2118; 2137;
2162;2521;2523;2554(Phormidiumsp.)
UTEX (The Culture Collection of Algae at the University of UTEX B 1580 (P. autumnale); UTEX B 427 (P.
TexasAustin)
foveolarum); UTEX LB 2426 (P. fragile); UTEX LB
2517 (P. inundatum); UTEX LB SP38 (P.
keutzingianum); UTEX LB 2425 (P. persicinum);
UTEXB1540;BEE19;BEE29;BEE38;BSP43;LB
2584toLB2586(Phormidiumsp.)
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CCALA 139; 140; 761 (P. animale); CCALA 143;

145; 697; 757; 816; 860 to 861 (P. autumnale);
CCALA 882 (P. favosum); CCALA 759; 815 (P.
irriguum);CCALA147(P.nigrum);CCALA144;149
(P.setchellianum);CCALA152;154(P.subfuscum);
CCALA 849 (P. uncinatum); CCALA 726; 771; 845;
850;858;881;923(Phormidiumsp.)
CCBA (Culture Collection of Baltic Algae at the University of BA0013 (P. amphibium); BA0141 (Phormidium
Gdansk)
sp.)
http://ocean.ug.edu.pl/~ccba/ien.php?id=ccba
CPCC (Canadian Phycological Culture Centre) (formerly 16strainsof12differentspeciesofPhormidium
knownasUTCC)
ATCC 700613 (P. corium); ATCC 29409 to 29410;
39161(Phormidiumsp.)
FACHB 239 (P. foveolarum); FACHB 238 (P.
luridum);FACHB723(P.mucicola)
NBRC 102691; 102696; 102724; 102728; 102751;
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l 102835(Phormidiumsp.)
ang=en
NIES2119 to 2122 (P. ambiguum); NIES 2123 (P.
angustissimus); NIES 32 to 34; 503 to 505 (P.
foveolarum);NIES2124(P.henningsii);NIES506to
507(P.jenkelianum);NIES2125(P.luridum);NIES
509; 2126 (P. molle); NIES 510 (P. mucicola); NIES
305(P.ramosum);NIES2128(Phormidiumsp.)
Anabaenasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP1403/7(A.ambigua);CCAP1403/2;1403/2B;
1403/30 (A. cylindrica); CCAP 1403/13B; 1403/3D
to 1403/3G; 1446/1C (A. flosaquae); CCAP
1446/1A (A. inaequalis); CCAP 1403/10 (A.
oscillaroides); CCAP 1403/19; 1403/27 (A.
solitaria); CCAP 1403/24 (A. spiroides); CCAP
1403/4B; 1403/12 (A. variabilis); CCAP 1403/4A;
1403/13A;1403/16;1403/21to1403/23;1403/29;
1453/30(Anabaenasp.)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 14037 (A. ambigua); SAG 14031 (A.
catenula);SAG14032(A.cylindrica);SAG30.87(A.
flosaquae); SAG 140310 (A. inaequalis); SAG
25.79(A.lutea);SAG26.79(A.torulosa);SAG1403
4b (A. variabilis);; SAG 27.79 (A. viguieri); SAG
12.82;25.82;28.79;29.79(Anabaenasp.)
CCMP (ProvasoliGuillard National Center for Culture of CCMP2066(Anabaenasp.)
UTEX (The Culture Collection of Algae at the University of 20strainsof12differentspeciesofAnabaena
TexasAustin)
CCALA 001 (A. augstumnalis); CCALA 755 (A.
catenula);CCALA758(A.doliolum);CCALA008(A.
gracile); CCALA 009 (A. klebhanii); CCALA 805 (A.
laxa); CCALA 002 (A. oscillaroides); CCALA 003 (A.
torulosa);CCALA010(A.rivularis);CCALA760;006
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ALGOBANK (la collection de cultures de microalgues de
l'Universit
de
Caen
BasseNormandie)
CCBA (Culture Collection of Baltic Algae at the University of
Gdansk)http://ocean.ug.edu.pl/~ccba/ien.php?id=ccba
to007(Anabaenasp.)
PCC 6309; 7108; 9217; 9109; 9208; 7122; 73105;
7938
AC163(A.circinalis)
BA0097(Anabaenasp.)
K1333toK1338(A.circinalis);K0544;K0599(A.
lemmermannii)
CS337/01 to CS337/02; CS530/05; CS533/02 to
CS533/03; CS534/02; CS534/05 to CS534/06;
CS536/01; CS537/01 to CS537/03; CS537/05;
CS537/11; CS537/13; CS538/01; CS538/05 to
CS538/06; CS539; CS539/08 to CS539/09; CS
541/06; CS545/17; CS547 (A. circinalis); CS53;
CS172 (A. cylindrica); CS548 to CS549 (A. flos
aquae); CS546 (A. spiroides); CS550; CS542/02;
CS542/04(Anabaenasp.)
FWAC1710(Anabaenasp.)
FWAC(FreshwaterAlgalCultureCollection)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC 64; 67 (A. flosaquae); CPCC 437 (A.
knownasUTCC)
sphaerica); CPCC 68 (A. spiroides); CPCC 45; 105;
342 (A. variabilis); CPCC 387; 426 to 427; 543 to
544;631(Anabaenasp.)
ATCC55755(A.affinis);ATCC29414(A.cylindrica);
ATCC 43530 (A. doliolum); ATCC 22664 (A. flos
aquae); ATCC 29413; 33801 (A. variabilis); ATCC
27898to27899;29211(Anabaenasp.)
46strainsof16differentspeciesofAnabaena
183 strains of 23 different species, the most
representedofwhichare:A.crassa(49strains);A.
reniformis (19 strains); A. circinalis (15 strains); A.
planctonica(15strains)
Synechococcussp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 1479/7 (S. bacillaris); CCAP 1499/3 (S.

capitatus); CCAP 1479/1A to 1479/1B (S.
elongatus); CCAP 1405/1 (S. leopoliensis); CCAP
1479/8 (S. linearis); CCAP 1479/5; 1479/9 to
1479/14(Synechococcussp.)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 88.79 (S. cedrorum); SAG 15.90; 89.79 (S.
elongatus); SAG 14021 (S. leopoliensis); SAG 3.81
(S.rubescens);SAG2156(Synechococcussp.)
CCMP (ProvasoliGuillard National Center for Culture of CCMP1261(S.bacillus);CCMP1333(S.bacillaris);
CCMP 1379; 1628 to 1631 (S. elongatus); CCMP
1284(S.nigra);CCMP833;835to842;1183;1282;
1334;1632;1768to1769;2161;2163;2165;2370;
2515; 2606; 2669; 2713; 2943 to 2946; 3074 to
3075;3195(Synechococcussp.)
UTEX (The Culture Collection of Algae at the University of UTEX 1191 (S. cedrorum); UTEX LB 563 (S.
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knownasUTCC)
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l
ang=en
elongatus); UTEX 2434; B 625 (S. leopoliensis);

UTEX LB 2380; LB 2389 to LB 2390; LB 2537; LB
2625(Synechococcussp.)
CCALA 187 (S. bigranulatus); CCALA 188 (S.
nidulans)
PCC543;712(Synechococcussp.)
PCC6301;6311to6312;6603;6706to6709;6715
to6717;6908;7002to7003;7117;73109;7335to
7336; 7502; 7901 to 7902; 7917 to 7918; 7920;
7922 to 7923; 7942; 7942/1; 7943; 7951; 8806 to
8807,8908;8943;8978;8991;9001to9004
K0408(Synechococcussp.)
79strainsofSynechococcussp.
CS450 (Synechococcus like); CS94; 197; 197/02;
NEPCC550(S.lividus)
CPCC 79 (S. cedrorum); CPCC 97 (S. elongatus);
CPCC 102 (S. leopoliensis); CPCC 246; 434; 477 to
480;660to661(Synechococcussp.)
ATCC 27144 to 27148; 27167 to 27168; 27177;
27180; 27192; 27194; 27264; 27344; 39138;
29140;29203;29403to29404(Synechococcussp.)
FACHB 283 (S. cedrorum); FACHB 347 (S.
elongatus);FACHB805(Synechococcussp.)
NBRC 102692; 102694; 102726; 102731; 102734;
102774; 102777; 102789; 102838 to 102839;
102864 to 102865; 102908 to 102909; 102952 to
Synechocystissp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 1480/5 (S. limnetica); CCAP 1480/1 (S.

minima);CCAP1480/4(Synechocystissp.)
SAG90.79(S.aquatilis);SAG258.80(S.minuscula);
SAG 91.79 (S. pevalekii); SAG 37.92; 45.90; 51.79;
92.79(Synechocystissp.)
CCMP (ProvasoliGuillard National Center for Culture of CCMP843(Synechocystissp.)
UTEX (The Culture Collection of Algae at the University of UTEX LB 2587 (S. nigrescens); UTEX 2470
TexasAustin)
(Synechocystissp.)
CCALA189to190(S.aquatilis);CCALA191;810(S.
fuscopigmentosa);CCALA192(S.cf.salina)
PCC6308;6701to6702;6711;6714;6803to6806;
6808; 6902; 6905 to 6906; 7008; 7201; 7338 to
7339; 7509; 7511; 7719; 7809; 7903; 7919; 7921;
8010;8805;8906;8912;8915;8931to8932;8938;
8962;8974;8981;8990;9027;9115;9218to9220;
9227
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CS95(Synechocystissp.)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC346;352;534(Synechocystissp.)
knownasUTCC)
ATCC27150;27170to27171;27175;27178;27184
to 27187; 27266; 29108 to 29110; 29152; 29235;
35679to35680;43105(Synechocystissp.)
Ostreococcussp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCMP (ProvasoliGuillard National Center for Culture of CCMP2407;2972(Ostreococcussp.)

RCC 116; 614; 745; 1108 to 1110; 1115; 1117 to
1118;1558to1559;1625(O.taurii);RCC343;410;
422;426to427;429;453to454;467to468;675;
690;747;754to756;788to789;796to797;802;
809 to 810; 1106 to 1107; 1121 to 11123; 1521;
1565to1566;1616;1645;1662(Ostreococcussp.)
NBRC102845(Ostreococcussp.)
ang=en
Tetraselmissp.
COLLECTION+WEBSITE
STRAINNUMBERS
73 strains of which 27 Tetraselmis sp.; other

species: T. apiculata; T. chuii; T. convolutae; T.
gracilis; T. inconspicua; T. levis; T. maculata; T.
marina; T. striata; T. subcordiformis; T. suecica; T.
tetrabrachia;T.tetrathele;T.verrucosa
SAG 1.96; 86 (T. chuii); SAG 50.87 (T. contracta);
SAG26.82(T.cordiformis);SAG202.80(T.marina);
SAG 41.85 (T. striata); SAG 1611a (T.
subcordiformis); SAG 1612b to 1612c (T.
tetrathele); SAG 1613; 3.98; 35.93 (Tetraselmis
sp.)
119 strains of which 110 Tetraselmis sp.; other
species:T.astigmatica;T.convolutae;T.marina;T.
rubens;T.suecica;T.wertsteinii;T.striata
UTEXB2562(T.apiculata);UTEXLB232(T.chuii);
UTEXB2563(T.gracilis);UTEXB2565;BSP22(T.
striata);UTEXLB2286(T.suecica);UTEXLB557(T.
tetrathele);UTEX2767;LB2767(Tetraselmissp.)
PCC 372; 372A; 581 (T. convolutae); PCC 429 (T.
impellucida); PCC 308;308A; 570 (T. marina); PCC
443 (T. striata); PCC 305 (T. suecica); PCC 272 (T.
tetrathele);PCC456(T.verrucosa);PCC315;511A
to511B;512to513;514B(Tetraselmissp.)
AC 726 (T. chuii); AC 258 (T. marina); AC 725 (T.
striata);AC254(T.suecica);AC261(T.tetrathele);
AC255;257;264(Tetraselmissp.)
K0011(T.contracta);K0937(T.levis);K0298(T.

TexasAustin)

l'Universit
de
Caen
BasseNormandie)
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CoordinationAction
FP7ENERGY20091
aff. maculata); K0377 (T. marina); K0297 (T.

suecica);K0243;K0950(T.verrucosa);K0296;K
0380;K0524;K0930;K0935(Tetraselmissp.)
RCC 128 to 129 (T. chuii); RCC 1563 to 1564 (T.
convolutae); RCC 132 to 133 (T. rubens); RCC 130
to131(T.striata);RCC119to127;233;235;348;
500;1947;1975to1976(Tetraselmissp.)
CS691/01 (T. antarctica); CS26 (T. chuii); CS56;
CS187 (T. suecica); CS87; CS91; CS317; CS352
(Tetraselmissp.)
NEPCC485;502(T.apiculata);NEPCC484;501(T.
chuii);NEPCC486(T.convolutae);NEPCC489;551
(T. gracilis); NEPCC 503 (T. inconspicua); NEPCC
488 (T. levis); NEPCC 487; 497 (T. striata); NEPCC
494 (T. suecica); NEPCC 483; 500 (T. tetrathele);
NEPCC496(T.verrucosa);NEPCC46;86;365;498
(Tetraselmissp.)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC196(Tetraselmissp.)
knownasUTCC)
NBRC 102994; 102997 to 102998; 103003
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l (Tetraselmissp.)
ang=en
NIES 18; 533 (T. cordiformis); NIES 1430 (T. levis);
NIES 1019 (T. striata); NIES 1836 (T. verrucosa);
NIES1421;1431to1434(Tetraselmissp.)
Botryococcussp.
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
CCAP807/1to807/2(B.braunii)
SAG30.81;8071(B.braunii)
CCMP2742(Botryococcussp.)
UTEX572;2441;LB572(B.braunii);UTEX2629(B.
sudeticus)
CCALA220;777to778;835(B.braunii);CCALA779
(B.protuberans);CCALA933(Botryococcussp.)
K1489(B.braunii)
FACHB357;762to766;768;774;777;1049;1079;
11041108(B.braunii)
NIES836;2199(B.braunii)
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 11/32A; 11/32B; 11/32C; 11/32CW15+;

11/32D;11/45
Other: 125 strains of 35 different species of
Page243of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
Chlamydomonas
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 1131; 1132a; 1132aM; 1132b; 1132c;
18.79; 23.90; 32.89; 33.89; 37.89; 38.89; 53.72;
54.72;7.73;73.72;77.81;81.72;83.81
Chlamydomonas
CCMP (ProvasoliGuillard National Center for Culture of 29strainsofChlamydomonassp.andC.euryale(1
strain)andC.parkeae(2strains)
UTEX (The Culture Collection of Algae at the University of UTEX89to90;2243to2244;2247;2337;B2246;
TexasAustin)
LB2607;LB2608
Chlamydomonas
CCALA928
Chlamydomonas
3strains(C.reginae;C.Concordia;Chlamydomonas
sp.)
ALGOBANK (la collection de cultures de microalgues de AC726;609
l'Universit
de
Caen
BasseNormandie) Other:Chlamydomonassp.(1strain);C.pertusa(1
strain)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K1016toK1017
Other:Chlamydomonassp.(1strain);C.debaryana
(1strain);C.geitlerii(1strain)
3 strains (C. concordia; C. reginae and
Chlamydomonassp.)
CS51
2strainsofChlamydomonassp.
FWAC/FreshwaterAlgalCultureCollection)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC11to12;84;243to244;532to533
knownasUTCC)
Chlamydomonas
FACHB265;355to356;359;479;1121
Chlamydomonas
9strains,ofwhich8ofChlamydomonassp.and1
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l strainofC.parkeae
ang=en
NIES2235to2239;2463to2464
Chlamydomonas
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP34/1D;34/1F;34/6;34/7;34/12to34/14
Other: 5 strains (H. buetschlii; H. capensis; H.
droebakensis)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG192.80;341ato341f;341h;341kto341n;
44.96;49.94
Other: 9 strains (H. buetschlii; H. capensis; H.
droebakensis;H.zimbabwensis)
UTEX (The Culture Collection of Algae at the University of UTEX2505
Page244of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
TexasAustin)
l'Universit
de
Caen
BasseNormandie)
knownasUTCC)
Other:3strains(H.droebakensis,H.capensis)
CCALA357to358;793;840
Other:Haematococcussp.(1strain)
AC136;143;146;587;588
K0084
Haematococcussp.(1strain)
FWAC7039;7072
Haematococcuslacustris(1strain)
FACHB712;797;847;871to878;954to955;1164
Haematococcuslacustris(4strains)
Dunaliellasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 19/35 (D. acidophila); CCAP 19/4 (D.

bioculata); CCAP 19/5 (D. minuta); CCAP 19/1;
19/9 to 19/10; 19/26 (D. parva); CCAP 19/2 (D.
piercei); CCAP 19/7A to 19/7C (D. polymorpha);
CCAP 11/34 (D. primolecta); CCAP19/8 (D.
quartolecta); CCAP 19/3; 19/18; 19/20; 19/25;
19/30 to19/31 (D. salina);CCAP 19/24;19/27(D.
tertiolecta); CCAP 19/12; 19/14 to 19/15; 19/19;
19/21to19/23;19/32to19/34(Dunaliellasp.)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) 26strainsof12differentspeciesofDunaliella
CCMP (ProvasoliGuillard National Center for Culture of CCMP 362 (D. parva); CCMP 364; 1302; 1320 (D.
tertiolecta);CCMP220;366to367;1641;1883to
1886;1892;1922to1925;2545;2575;2577;2589
to2590;2605(Dunaliellasp.)
UTEX (The Culture Collection of Algae at the University of UTEXLB2538(D.bardawil);LB199(D.bioculata);
TexasAustin)
UTEXLB1983(D.parva);UTEXLB2192(D.piercei);
UTEX LB 1000 (D. primolecta); UTEX LB 1644; LB
200(D.salina);UTEXLB999(D.tertiolecta);UTEX
BSP20(Dunaliellasp.)
CCALA 337 (D. bioculata); CCALA 339 to 340; 821
(D.salina)
PCC 430 (D. minuta); PCC 81 (D. primolecta); PCC
83(D.tertiolecta)
ALGOBANK (la collection de cultures de microalgues de AC144(D.salina);AC148(D.tertiolecta)
l'Universit
de
Caen
BasseNormandie)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K0591(D.tertiolecta)
RCC6(D.tertiolecta);RCC5(Dunaliellasp.)
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AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
knownasUTCC)
CS265; CS744/01 (D. salina); CS14; CS175 (D.

tertiolecta); CS353; CS631 to CS635 (Dunaliella
sp.)
NEPCC1(D.tertiolecta)
CPCC 307 (D. acidophila); CPCC 197 (D. salina);
CPCC420(D.tertiolecta);CPCC457(Dunaliellasp.)
FACHB 949 (D. bioculata); FACHB 815 (D. parva);
FACHB818(D.primolecta);FACHB435(D.salina);
FACHB 821 (D. tertiolecta); FACHB 748 (Dunaliella
sp.)
NIES 2253 (D. bioculata); NIES 2254 (D. parva);
NIES 2255 (D. piercei); NIES 2256 (D. primolecta);
NIES2257(D.salina);NIES2258(D.tertiolecta)
Chlorococcumsp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 213/5 (C. echinozygotum); CCAP 213/11 (C.

humicola); CCAP 213/6; 237/1 (C. hypnosporum);
CCAP213/2A;213/2B(C.infusionum);CCAP213/9
(C. macrostigmatum); CCAP 213/7 (C. minutum);
CCAP 213/10 (C. submarinum); CCAP 213/12 (C.
tetrasporum); CCAP 213/8 (C. vacuolatum); CCAP
11/52(Chlorococcumsp.)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) 28strainsof16differentspeciesofChlorococcum
UTEX (The Culture Collection of Algae at the University of 39strainsof36differentspeciesofChlorococcum
TexasAustin)
CCALA 281 (C. echinozygotum); CCALA 283 to 284
(C. ellipsoideum); CCALA 285; 772 (C.
hypnosporum); CCALA 286 to 287 (C. infusionum);
CCALA 288 (C. lobatum); CCALA 289 (C.
macrostigmatum);CCALA290(C.minutum);CCALA
291 to 293 (C. scabellum); CCALA 294 (C.
vacuolatum)
ALGOBANK (la collection de cultures de microalgues de AC727(Chlorococcumsp.)
l'Universit
de
Caen
BasseNormandie)
CCBA (Culture Collection of Baltic Algae at the University of BA0019(Chlorococcumsp.)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K1301toK1303(Chlorococcumsp.)
NEPCC478;499;505(Chlorococcumsp.)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC 178 (C. hypnosporum); CPCC 315; 513
knownasUTCC)
(Chlorococcumsp.)
FACHB 21 to 23 (C. humicola); FACHB 1225 (C.
infusionum);FACHB957(Chlorococcumsp.)
NBRC 102705 to 102708 (C. dorsiventrale); NBRC
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l 102761 (C. littorale); NBRC 102999 (Chlorococcum
ang=en
sp.)
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AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
NIES 2249 (C. echinozygotum); NIES 2250 (C.

elkhartiense);NIES2251(C.humicola)
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
10strainsof10differentspeciesofNeochloris
2strains(N.conjuncta,N.vigensis)
UTEX1185
Neochloris
5strainsof5differentspeciesofNeochloris
Scenedesmussp.
COLLECTION+WEBSITE
STRAINNUMBERS
74 strains of29 different species ofScenedesmus,

the most represented of which is S. obliquus (19
strains)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) 29strainsof11differentspeciesofScenedesmus
CCMP (ProvasoliGuillard National Center for Culture of CCMP1625;2399(Scenedesmussp.);CCMP2254;
2257(Scenedesmuslike)
UTEX (The Culture Collection of Algae at the University of 29strainsof17differentspeciesofScenedesmus
TexasAustin)
20strainsof11differentspeciesofScenedesmus
ALGOBANK (la collection de cultures de microalgues de AC147(S.armatus);AC139(S.subspicatus)
l'Universit
de
Caen
BasseNormandie)
CCBA (Culture Collection of Baltic Algae at the University of BA0007 (S. acuminatus); BA0008; BA0147 (S.
acutus); BA0006 (S. armatus); BA0024
(Scenedesmussp.)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K1010 (S. armatus); K1149; K1305 (S.
quadricauda)
FWAC276(S.quadricauda)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC6,8;10;282;285;353(S.acutus);CPCC153
knownasUTCC)
(S. denticulatus); CPCC 5 157 (S. obliquus); CPCC
158;163(S.quadricauda);CPCC20;286;297;316
to319;367;423;531(Scenedesmussp.)
FACHB1221;1234to1235(S.acuminatus);FACHB
76 to 77 (S. bijuga); FACHB 496; 959; 963; 1248;
1266(S.dimorphus);FACHB12to14;39;276;416
to 417 (S. obliquus); FACHB 43 to 45; 506 to 508;
1117; 1297 (S. quadricauda); FACHB 1268 (S.
spinosus); FACHB 489 to 490; 492; 933; 1229
(Scenedesmussp.)
NIES92(S.acuminatus);NIES94to95;120;2269
to 2270 (S. acutus); NIES 2271 to 2272 (S.
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AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
basiliensis); NIES 2273 (S. bijuga); NIES 2274 (S.

chlorelloides); NIES 2275 (S. coelastroides); NIES
2276 (S. costulatus); NIES 93; 119 (S. dimorphus);
NIES2279to2280(S.obliquus)
Desmodesmussp.
COLLECTION+WEBSITE
STRAINNUMBERS
298 strains of 43 different species of

Desmodesmus,themostrepresentedofwhichare
D. armatus (47 strains) and D. communis (34
strains); D. opoliensis (17 strains); D. intermedius
(15strains)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) 21strainsof14differentspeciesofDesmodesmus
CCMP (ProvasoliGuillard National Center for Culture of CCMP 2256 (D. ultrasquamatus); CCMP 2450 (D.
hystrix); CCMP 2489 (D. pikollei); CCMP 2491
(Desmodesmussp.)
UTEX (The Culture Collection of Algae at the University of UTEX 2892 (D. brasiliensis); UTEX B 2894 (D.
TexasAustin)
serratus)
16strainsof14differentspeciesofDesmodesmus
CS899; CS905 (D. asymmetricus lineage); CS904
(D.opoliensislineage)
NIES685(D.abundans);NIES97(D.serratus);NIES
797to802(D.subspicatus);NIES96;2277to2278
(Desmodesmussp.)
Chlorellasp.
COLLECTION+WEBSITE
STRAINNUMBERS
110strainsofwhich31Chlorellasp.andtheothers
of 17 different species of Chlorella, the most
represented of which are: C. vulgaris (21 strains);
C.saccharophila(12strains);C.protothecoides(10
strains);C.emersonii(9strains)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) 78 strains of 13 different species of Chlorella, the
most represented of which are: C. vulgaris (15
strains); C. luteoviridis (12 strains); C.
protothecoides (10 strains); C. saccharophila (9
strains);C.sorokiniana(7strains)
CCMP (ProvasoliGuillard National Center for Culture of 37 strains of which 34 of Chlorella sp. and C.
capsulata(3strains)andC.autotrophica(1strain)
UTEX (The Culture Collection of Algae at the University of 71strainsofwhich10Chlorellasp.andtheothers
TexasAustin)
representedofwhichare:C.vulgaris(9strains);C.
sorokiniana (9 strains); C. protothecoides (8
strains); C. kessleri (7 strains); C. luteoviridis (6
strains)
CCALA714;733;741;884to885;914to915;884
to 885 (C. homosphaera); CCALA 250 to 255 (C.
kessleri); CCALA 723; 745; 886; 916 (C.
minutissima);CCALA257;917(C.mirabilis);CCALA
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AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
258 (C. saccharophila); CCALA 259 to 560; 918 (C.

sorokiniana); CCALA 256; 261 to 269; 788; 896 to
898;902;924(C.vulgaris);CCALA715to717;936
(Chlorellasp.)
PCC 476 (C. saccharophila); PCC 309 (C. salina);
PCC85(C.stigmatophora)
ALGOBANK (la collection de cultures de microalgues de AC 586; 628 to 629 (C. saccharophila); AC 149 to
l'Universit
de
Caen
BasseNormandie) 150;659(C.vulgaris)
CCBA (Culture Collection of Baltic Algae at the University of BA0018(C.fusca);BA0012(C.minutissima);BA
0002;BA0043;BA0046;BA0080(C.vulgaris);BA
0020toBA0023;BA0025;BA0059;BA0079;BA
0103(Chlorellasp.)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K1006(C.vulgaris)
RCC 661 (C. stigmatophora); RCC 288; 347; 396;
533;537(Chlorellasp.)
CS41 to CS42 (C. vulgaris); CS122; CS903
(Chlorella sp.); CS195; CS247 to CS248; CS436
(Chlorellalike)
FWAC 7066 (C. pyrenoidosa); NEPCC 893; 896
(Chlorellasp.)
CPCC (Canadian Phycological Culture Centre) (formerly CPCC 86 (C. emersonii); CPCC 89; 394 (C. fusca);
knownasUTCC)
CPCC 266 (C. kessleri); CPCC 87 (C. luteoviridis);
CPCC 88 (C. miniata); CPCC 89 to 90; 138 (C.
pyrenoidosa); CPCC 91; 272 (C. saccharophila);
CPCC 138 (C. sorokiniana); CPCC 90; 92; 111; 139
to 147 (C. vulgaris); CPCC 23; 247; 493; 522; 560;
603(Chlorellasp.)
52strainsofwhich14Chlorellasp.andtheothers
represented of which are: C. pyrenoidosa (15
strains);C.vulgaris(13strains)
NBRC 102700 to 102704; 102721; 102740; 10895
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet?l to 102896 (C. saccharophila); NBRC 102737;
ang=en
102741; 102768; 102831; 102892 to 102894;
102897;102902;102949;102951(Chlorellasp.)
NIES 2150(C. ellipsoidea); NIES 640; 2352 (C.
saccharophila);NIES2167to2169(C.sorokiniana);
NIES 227; 641 to 642; 686; 1269; 2170; 2172 to
2173(Chlorellasp.)
Protothecasp.
COLLECTION+WEBSITE
STRAINNUMBERS
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 2064 (P. blaschkeae); SAG 26311

(P.wickerhamii); SAG 2021; 2063; 2631; 2633 to
2634; 2636 to 2638; 26310; 26312; 43.80;
44.80;45.80(P.zopfii)
UTEX (The Culture Collection of Algae at the University of UTEX329(P.kruegeri);UTEX288,14341437,1441
TexasAustin)
(P. moriformis); UTEX 289, 327 (P. portoricensis);
UTEX 14424 to1443 (P. stagnora); UTEX 1440,
1533(P.wickerhamii);UTEX1438(P.zopfii)
NBRC 6994 (P. ciferrii); NBRC 32449 (P.
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AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
ang=en
eriobotryae); NBRC 6995 (P. moriformis); NBRC

6996 (P. trispora); NBRC 6997 (P.wickerhamii);
NBRC6998,7532to7536(P.zopfii)
COLLECTION+WEBSITE
STRAINNUMBERS
P.marinum(1strain);P.purpureum(4strains);P.
sordidum(2strains)
P.aerugineum(4strains);P.purpureum(8strains);
P.sordidum(2strains)
CCMP675;1328
Other: P. aerugineum (2 strains); P. grisea (1
strain); P. purpureum (1 strain); P. sordidum (1
strain);Porphyridiumsp.(2strains)
UTEX (The Culture Collection of Algae at the University of UTEX161
TexasAustin)
Other: P. aerugineum (3 strains); P. purpureum (1
strain);P.sordidum(1strain);Porphyridiumsp.(1
strain)
CCALA415
Other: P. aerugineum (2 strains); P. purpureum (1
strain);P.sordidum(1strain)
P.purpureum(1strain)
ALGOBANK (la collection de cultures de microalgues de P.aerugineum(3strains);P.purpureum(3strains);
l'Universit
de
Caen
BasseNormandie) Porphyridiumsp.(9strains)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) P.aerugineum(1strain);P.purpureum(2strains)
P.purpureum(1strain)
P.purpureum(1strain);Porphyridiumsp.(1strain)
CPCC (Canadian Phycological Culture Centre) (formerly P.purpureum(1strain)
knownasUTCC)
P. aerugineum (1 strain); P. purpureum (1 strain);
Porphyridiumsp.(1strain)
P.sordidum(1strain);Porphyridiumsp.(5strains)
ang=en
P.aerugineum(4strains);P.purpureum(3strains);
Porphyridiumsp.(5strains)
Amphorasp.
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
CCAP1001/3
CCAP1001/1to1001/2(A.coffeaeformis)
CCMP129;1389;1523;2107;2522;2531;2910;
CCMP126to128;1405(A.coffeaeformis)
UTEX2038;2080;B2036;B2040;BSP1;LBFD57;
LBFD71;LBFD75;LBFD9(A.coffeaeformis)
Page250of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
l'Universit
de
Caen
BasseNormandie)
http://www.tpub.com/content/altfuels05/3814/38140091.ht
m
knownasUTCC)
ang=en
PCC545;547(A.coffeaeformis)
AC711;
AC713(A.coffeaeformis)
BA0016(A.coffeaeformis)
K1250
CS10;CS307;CS366;CS518toCS521
NEPCC762to763(A.coffeaeformis)
AMPHO*strainsofA.coffeaeformisand
A.delicatissima
CPCC58(A.coffeaeformis)
FACHB363(A.coffeaeformis)
NBRC102745
Amphiprorahyalina
COLLECTION+WEBSITE
STRAINNUMBERS
http://www.tpub.com/content/altfuels05/3814/38140091.ht
m
CCAP1003/4(A.paludosavar.hyalina)
SAG15.83(A.paludosa)
CCMP467(Amphiprorasp.)
NEPCC266
ENTOM1
Chaetocerosmuelleri
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
CCAP1010/3
CCMP194;1316;1318
UTEXLBFD74
PCC586
Page251of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
F&MM43
CS176
CPCC (Canadian Phycological Culture Centre) (formerly CPCC485
knownasUTCC)
CHAET*strains
Cyclotellacryptica
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP1070/2;1070/6
CCMP (ProvasoliGuillard National Center for Culture of CCMP331to333
CYCLO*strains
Cylindrothecasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 1017/8 to 1017/11 (C. closterium); CCAP

1017/2(C.fusiformis);CCAP1017/7(Cylindrotheca
sp.)
CCMP (ProvasoliGuillard National Center for Culture of CCMP 339 to 340; 1554; 1725; 1855; 2086 (C.
closterium);CCMP343to344(C.fusiformis);CCMP
1989(Cylindrothecasp.)
UTEX (The Culture Collection of Algae at the University of UTEXB2085toB2087(C.fusiformis);UTEXLBFD30
TexasAustin)
(C.gracilis)
ALGOBANK (la collection de cultures de microalgues de AC170(C.closterium)
lUniversit
de
Caen
BasseNormandie)
SCCAP(ScandinavianCultureCollectionofAlgae&Protozoa) K1342;K0520(C.closterium)
RCC782;1937;1950(C.closterium)
BCCM(BelgianCoordinatedCollectionsofMicroorganisms) SeveralstrainsofCylindrothecasp.,C.fusiformisen
DiatomCollectionattheGhentUniversity
C.closterium
CS13(C.fusiformis)
NEPCC417;425(C.fusiformis)
NIES1045 (C. closterium); NIES1046 (C.
fusiformis);NIES1047(Cylindrothecasp.)
NBRC103017
ang=en
Naviculasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP1050/8(N.hanseni);CCAP1050/9(N.
Page252of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
pelliculosa);CCAP1050/10(N.incerta);
CCAP1050/12(N.radiosa)
SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG10503(N.pelliculosa);SAG40.96(N.
salinicola)
15strainsofwhich5ofNaviculasp.
58strainsofwhich1ofNaviculasp.
TexasAustin)
NAVIC*strains
Naviculasaprophila
COLLECTION+WEBSITE
STRAINNUMBERS
NAVIC*strains
Nitzschiadissipata
COLLECTION+WEBSITE
STRAINNUMBERS

TexasAustin)
UTEXLBFD253
BA0040
http://www.tpub.com/content/altfuels05/3814/38140091.ht NITZS13
m
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
l'UniversitdeCaenBasseNormandie)
CCAP 1052/1A; 1052/1B; 1052/6; 1055/1 to

1055/8
SAG10901a;10901b;10906
CCMP630to633;1327;2928;2557to2561
UTEXB2089;646;L642;640
PCC100;670
AC590to591;171
AA0004;AA0079;AA0142
K1280
RCC69;641
F&MM40
Page253of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
BCCM(BelgianCoordinatedCollectionsofMicroorganisms)
knownasUTCC)
1055/1;1052/1A
CS29
NEPCC31;640;738;552;860
CPCC162
PHAEO1,2
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
l'Universit
de
Caen
BasseNormandie)
BCCM(BelgianCoordinatedCollectionsofMicroorganisms)
CCAP1085/12
SAG10201b
CCMP1007;1011to1015;1335
UTEXLBFD2
PCC693
AC589
AA0078
K1282
severalstrains
CS20;CS173
NEPCC58;58b;709
Odontellaaurita
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP1054/1
CCMP145;595;1108;1796;816(O.cf.aurita)
K1251;K1252
CS19
NEPCC553
Page254of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
Skeletonemasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 1077/1B, 1077/1C, 1077/3 to 1077/5 (S.

costatum);CCAP1077/7(S.pesudocostatum);CCAP
1077/8(S.subsalsum)
SAG (Sammlung von Algenkulturen der Universitt SAG19.99(S.costatum),SAG8.94(S.subsalsum)
Gttingen)
CCMP (ProvasoliGuillard National Center for Culture of CCMP 794 (S. ardens); CCMP 2092 (S. costatum);
CCMP 779, 782, 785, 789, 1715, 2479 to 2481 (S.
dohrnii);CCMP775to776,780,2508to2509,2801
(S. grethae); CCMP 1685 (S. grevillei); CCMP 784,
1281, 1283, 2506 (S. japonicum); CCMP 781, 791,
1009,1016,1223to1226,1332,2497,2501to2505,
2507(S.marinoi);CCMP786to787,790,792to793,
795, 2799 (S. menzelii); CCMP 2472 to 2478 (S.
pseudocostatum);CCMP778,788,2070,2157,2798,
2800,2802to2803(S.tropicum)
UTEX (The Culture Collection of Algae at the University of UTEXLB2308(S.costatum)
TexasAustin)
PCC612(S.dohrnii);PCC106,582(S.marinoi);PCC
627(Skeletonemasp.)
ALGOBANK (la collection de cultures de microalgues de AC174,714(S.marinoi);AC715(Skeletonemasp.)
l'Universit
de
Caen
BasseNormandie)
CCBA(CultureCollectionofBalticAlgaeattheUniversityof BA0011,BA0098(S.costatum)
SCCAP (Scandinavian Culture Collection of Algae & K0669(S.costatum)
Protozoa)
CS347toCS348(S.ardens);CS408(S.dohrnii);CS
864 (S. japonicum); CS181 (S. marinoi); CS76, CS
167,CS252(S.pseudocostatum)
NEPCC 755; 782 (S. costatum); NEPCC 910 (S.
marinoi);NEPCC904(Skeletonemasp.)
FACHB1114to1115(S.costatum)
Monodussp.
COLLECTION+WEBSITE
STRAINNUMBERS
TexasAustin)
l'Universit
de
Caen
BasseNormandie)
CCAP848/1(M.subterraneus)
SAG8.83(M.unipapilla)
UTEX151(M.subterraneus);BSNO46,76,8384,93
95, 9899, 104, 110, 112, 114, 127, 131, 139, 142,
145,SNO26,29,34,44(Monodussp.)
CCALA825to826;828(M.guttula);CCALA830(M.
subterraneus);CCALA827(Monodussp.)
AC733(M.subterraneus)
Page255of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
CPCC(CanadianPhycologicalCultureCentre)(formerlyknown CPCC116(M.subterraneus)
asUTCC)
FACHB819(M.subterraneus)
Nannochloropsissp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 849/5 (N. gaditana); CCAP 849/10 (N.

oceanica); CCAP 849/1, 849/7 (N. oculata); CCAP
849/3,849/4,849/6(N.salina);CCAP211/78,849/8,
849/9(Nannochloropsissp.)
SAG (Sammlung von Algenkulturen der Universitt SAG 2.99 (N. gaditana); SAG 18.99 (N. limnetica);
Gttingen)
SAG38.85(N.oculata);SAG40.85(N.salina)
CCMP (ProvasoliGuillard National Center for Culture of CCMP 526 to 527, 532, 536, 1775 (N. gaditana);
CCMP 529, 534 to 535, 1662 (N. granulata); CCMP
505,2260,2267,2271to2272,2392(N.limnetica);
CCMP525,2195(N.oculata);CCMP369,537to538,
1776 to 1778 (N. salina); CCMP 531, 821, 2001
(Nannochloropsis
sp.);
CCMP
1997
(cf.
Nannochloropsis)
UTEX (The Culture Collection of Algae at the University of LB2164(N.oculata)
TexasAustin)
PCC663(N.oculata);PCC561(N.salina);PCC588to
590(Nannochloropsissp.)
ALGOBANK (la collection de cultures de microalgues de AC223(N.gaditana);AC227to228(N.oculata);AC
lUniversit
de
Caen
BasseNormandie) 224(N.salina);AC225to226,593(Nannochloropsis
sp.)
CCBA(CultureCollectionofBalticAlgaeattheUniversityof AA0083 (N. gaditana); AA0105 (Nannochloropsis
sp.)
SCCAP (Scandinavian Culture Collection of Algae & K1281(N.oculata)
Protozoa)
CS246, CS702 (N. oceanica); CS179, CS189, CS
192, CS216 (N. oculata); CS190 to CS191 (N.
salina)
NEPCC631(N.oculata)
NBRC 102713 (N. granulata); NBRC 102738 (N.
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet oceanica);NBRC102719(N.salina)
?lang=en
Isochrysissp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP 927/1 (I. galbana.); CCAP 927/14 (Isochrysis

sp.)
SAG 13.92 (I. galbana); SAG 9272, 9273
(Isochrysissp.)
CCMP 462, 1323, 1611 (I. galbana.); CCMP 463,
1324(Isochrysissp.);CCMP355,1244,1257,1406,
2164(cf.Isochrysis)
Page256of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091

TexasAustin)
l'Universit
de
Caen
BasseNormandie)
knownasUTCC)
ang=en
LB 987 (I. galbana); LB 1292 (Isochrysis sp.); LB

2307(I.aff.galbana)
PCC8, 553 (I. aff. galbana); PCC240, 506AC, 507,
562(Isochrysissp.);PCC352,401B(cf.Isochrysis)
AC102(I.aff.galbana);AC34,101(I.galbana);AC
18(I.litoralis);AC66,620(Isochrysissp.)
AA0067,AA0097(I.galbana)
K1355(I.galbana);K0633(Isochrysissp.)
CS254(cf.Isochrysis);CS177(Isochrysissp.clone
TISO)
NEPCC633(I.galbana);NEPCC601(Isochrysissp.)
CPCC690(I.galbana)
NBRC 102813 (I. galbana); NBRC 102814, 102815,

102941(Isochrysissp.)
Pavlovasp.
COLLECTION+WEBSITE
STRAINNUMBERS
CCAP940/1C(P.gyrans);CCAP931/1,931/6,931/7
(P.lutheri);CCAP940/2(P.pinguis);CCAP940/3(P.
salina)
SAG (Sammlung von Algenkulturen der Universitt SAG926.1(P.lutheri)
Gttingen)
CCMP (ProvasoliGuillard National Center for Culture of CCMP 607 to 608, 1279 (P. gyrans); CCMP 1325 (P.
lutheri); CCMP 609 (P. pinguis); CCMP 611 to 613,
616to617,619to620,1209,1228to1229,1233to
1234, 1255 to 1256, 1390, 1394, 1416 to 1417
(Pavlova sp.); CCMP 317, 459, 583, 614, 618, 1222,
1265(cf.Pavlova)
UTEX (The Culture Collection of Algae at the University of LB992(P.gyrans);LB1293(P.lutheri)
TexasAustin)
PCC 552 (P. granifera); PCC 93 (P. gyrans); PCC 75,
554 (P. lutheri); PCC 471 (P.pinguis); PCC 154, 465,
486(P.salina);PCC515(P.virescens);PCC463,482,
484to485,487,600to601(Pavlovasp.)
ALGOBANK (la collection de cultures de microalgues de AC 89, 539 (P. granifera); AC 88 (P. noctivaga); AC
l'Universit
de
Caen
BasseNormandie) 19,696(P.pinguis);AC16(P.virescens);AC33,35,
37, 245 to 247, 250 to 252, 538, 699701 (Pavlova
sp.)
CCBA(CultureCollectionofBalticAlgaeattheUniversityof AA0026,AA0104(P.lutheri)
SCCAP (Scandinavian Culture Collection of Algae & K1308 to K1310 (P. gyrans); K0013, K0465
Protozoa)
(Pavlovasp.)
CS213(P.gyrans);CS23,CS182(P.lutheri);CS286,
Page257of258
AQUAFUELFP72413012
CoordinationAction
FP7ENERGY20091
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServlet
?lang=en
CS375 (P. pinguis); CS49 (P. salina); CS50, CS63

(Pavlovasp.)
NEPCC 438 (P. gyrans); NEPCC 2, 5, 274, 634 (P.
lutheri)
NBRC 102809 to 102810 (P. gyrans); NBRC 102808
(P.lutheri);NBRC102807(P.pinguis);NBRC102727,
102742, 102764, 102773, 102776, 102787, 102848,
102856,102912to102914,103007(Pavlovasp.)
COLLECTION+WEBSITE
STRAINNUMBERS
ATCC 30021, 30334 to 30348, 30541 to 30542,

30555 to 30557, 30571 to 30572, 30772 to 30775,
30812,40750,50050to50060,50297to50300
FACHB272
Schizochytriumsp.
COLLECTION+WEBSITE
STRAINNUMBERS
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServl
et?lang=en
ATCC 28209 (S. aggregatum); MYA1381 (S.

limacinum);ATCC20888to20889(Schizochytriumsp.)
NBRC102615to102617
Thraustochytriumsp.
COLLECTION+WEBSITE
STRAINNUMBERS
ATCC 34304 (T. aureum); PRA210, 211, 307 (T.

caudivorum);PRA308(T.pachydermum);ATCC28210
(T. roseum); ATCC 24473 (T. striatum); ATCC 18907,
20891 to 20892, 26185, PRA295 to 296
(Thraustochytriumsp.)
Ulkeniasp.
COLLECTION+WEBSITE
STRAINNUMBERS
http://www.nbrc.nite.go.jp/NBRC2/NBRCDispSearchServl
et?lang=en
NBRC 104106 (U. amoeboidea); NRBC 104108 (U.

sarkariana);NRBC102975(U.minuta)
Page258of258

Algae and Aquatic Biomass For A Sustainable Production of 2nd Generation Biofuels

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Algae and Aquatic Biomass For A Sustainable Production of 2nd Generation Biofuels

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4.2.3 Sustainability of different cultivation systems............................................................................................ 35

8.3.2 Phaeodactylum tricornutum...................................................................................................................... 126

Ulva sp. ..................................................................................................................................................... 170

Cladophora sp. ......................................................................................................................................... 178

Codium fragile ......................................................................................................................................................................180

Ascophyllum nodosum .............................................................................................................................. 197

Himanthalia elongata ............................................................................................................................... 202

Halidrys siliquosa ..................................................................................................................................... 207

Laminaria sp. ........................................................................................................................................................................210

10 OTHER AQUATIC BIOMASS ........................................................................................................................ 221

Some species of green algae, particularly of genera Trebouxia and Pseudotrebouxia

Thraustochytrids are a commercial source of polyunsaturated fatty acids, in particular doicosahexaenoic

Modified: +boron + vitamins, N

is generated by the production of extracellular polymeric substances (EPS), primarily polysaccharides

Biotechnology of Thraustochytrium is similar to that of Schizochytrium, although no strain of this genus

Biofiltration; growth rate; Vandermeulen

Jensen & Haug, 1956; Indergaard & Minsaas,

Jensen & Haug, 1956; Indergaard & Minsaas,

CCAP 1406/1 (A. brevis); CCAP 1475/8 (A.

CCALA 023; 025 (A. fusiformis); CCALA 027 to 030

CCAP 1462/6; 1462/10 (P. autumnale); CCAP

CCALA 139; 140; 761 (P. animale); CCALA 143;

CCAP 1479/7 (S. bacillaris); CCAP 1499/3 (S.

elongatus); UTEX 2434; B 625 (S. leopoliensis);

CCAP 1480/5 (S. limnetica); CCAP 1480/1 (S.

CCMP (ProvasoliGuillard National Center for Culture of CCMP2407;2972(Ostreococcussp.)

73 strains of which 27 Tetraselmis sp.; other

CCMP (ProvasoliGuillard National Center for Culture of

ALGOBANK (la collection de cultures de microalgues de

aff. maculata); K0377 (T. marina); K0297 (T.

CCAP 11/32A; 11/32B; 11/32C; 11/32CW15+;

CCAP 19/35 (D. acidophila); CCAP 19/4 (D.

CS265; CS744/01 (D. salina); CS14; CS175 (D.

CCAP 213/5 (C. echinozygotum); CCAP 213/11 (C.

NIES 2249 (C. echinozygotum); NIES 2250 (C.

74 strains of29 different species ofScenedesmus,

basiliensis); NIES 2273 (S. bijuga); NIES 2274 (S.

298 strains of 43 different species of

258 (C. saccharophila); CCALA 259 to 560; 918 (C.

SAG(SammlungvonAlgenkulturenderUniversittGttingen) SAG 2064 (P. blaschkeae); SAG 26311

eriobotryae); NBRC 6995 (P. moriformis); NBRC

CCAP 1017/8 to 1017/11 (C. closterium); CCAP

UTEX (The Culture Collection of Algae at the University of

CCAP 1052/1A; 1052/1B; 1052/6; 1055/1 to

CCAP 1077/1B, 1077/1C, 1077/3 to 1077/5 (S.

CCAP 849/5 (N. gaditana); CCAP 849/10 (N.

CCAP 927/1 (I. galbana.); CCAP 927/14 (Isochrysis

UTEX (The Culture Collection of Algae at the University of

LB 987 (I. galbana); LB 1292 (Isochrysis sp.); LB

NBRC 102813 (I. galbana); NBRC 102814, 102815,

CS375 (P. pinguis); CS49 (P. salina); CS50, CS63

ATCC 30021, 30334 to 30348, 30541 to 30542,

ATCC 28209 (S. aggregatum); MYA1381 (S.

ATCC 34304 (T. aureum); PRA210, 211, 307 (T.

NBRC 104106 (U. amoeboidea); NRBC 104108 (U.

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