3. Introduction
• The actinomycetes are very diverse and contain a variety
of subdivisions, as well as yet-unclassified isolates,
mainly because some genera are very difficult to classify
because of a highly niche-dependent phenotype.
• For example, Nocardia contains several phenotypes first
believed to be distinct species before their differences
were shown to be entirely dependent on their growth
conditions.
• Actinomycetales are generally gram-positive and
anaerobic and have mycelium in a filamentous and
branching growth pattern. Some actinobacteria can form
rod- or coccoid-shaped forms, while others can form
spores on aerial hyphae.
4. Introduction…..
• Actinomycetales bacteria can be infected
by bacteriophages, which are called actinophages.
• Actinomycetales can range from harmless bacteria to
pathogens with resistance to antibiotics.
• Important genera:
Actinomyces
Nocardia
Actinomadura
Streptomyces
thermophilic
5. Habitat
• Predominantly soil bacteria Good at degrading
recalcitrant compounds such as chitin &
cellulose. Often active at higher pH (contrast
to fungi who may dominate at lower pH)
• Give soil the “earthy” smell
• Almost all species are commensals of the oral
covity and tonsils.
• Some of the form the normal flora of mouth
and female genital tract.
6. Morphology
• Diverse group
• Gram positive
• Non-motile
• Non-sporing
• Non-capsulated
• Straight, curved or pleomorphic
• Arranged in chains or branching filaments
• Related to Mycobacteria and Corynebacteria
• Mycelium -- tangled mass of hyphae, found in
nature
7. • Unicellular like bacteria but produce mycelium
which is non septate and more slender
• Like bacteria they don’t like distinct cell wall
• Don’t contain chitin and cellulose
• They produce hyphae and conidia / sporangia
like fungi.
• Certain actinomycetes whose hyphae undergo
segmentation resemble bacteria, both
morphologically and physiologically.
• Note: Morphology resembles to fungi, cellular
organization typical of bacteria
10. Difference between fungi and
Actinomycetes
Actinomycetes are non-motile filamentous
gram positive bacteria belonging to the
genus of the Actinobacteria class of
bacteria.
Fungi are a group of microorganism which
includes single cell and complex
multicellular organisms such as yeast,
mushrooms, moulds, etc.
Actinomycetes are prokaryotic
organisms.
Fungi are eukaryotic organisms.
Actinomycetes contain peptidoglycan in
their cell walls.
Fungi contain chitin in their cell wall
Actinomycetes filaments are smaller. Fungi filaments are bigger
GC content in actinomycetes DNA is less
than fungi.
Fungi have more GC bases in DNA.
11. Slimarlities between Actinomycetes and fungi
1. Actinomycetes and fungi are filamentous.
2. Both produce spores.
3. Both types are good decomposers.
4. Both groups include antibiotic producing species.
12.
13. Similarities Between Actinomycetes and Bacteria
Actinomycetes and bacteria are prokaryotes.
They do not have a membrane-bound nucleus and organelles.
Both have a cell wall made up of murine.
They are microorganisms that form colonies on the solid media.
Both form endospores.
They occur in the environment and as a part of the normal microbiota.
Both can be pathogens.
14. Culture characteristic
• Anaerobic or microaerophilic bacteria and
grows well in presence of 5-10% co2.
• Optimum temperature 35-37 degree Celsius
• Grow on brain heart infusion agar/broth and
thioglycollate agar containing o.12%-0.2%
rabbit blood.
• Incudation time: 3-4 days mostely but for few
speciesit extended from 1 to 2 week
• Colony can be rough, pigmented with chalky
appearance.
18. Pathogenic
• Actinomycetes normally reside in human mouth, throat, gastrointestinal
tract, and urogenital tract without producing disease.
• Since the organisms cannot invade a human or animal body, they must
be introduced by a deep puncture wound or trauma such as dental
extraction or jaw trauma, aspiration of dental debris, surgery (removal of
the appendix), or prolonged use of intrauterine devices.
• Actinomycetes require dead or devitalized tissue to facilitate their
invasion and proliferation into deeper tissues. Establishment of human
infection by Actinomycetes always requires the presence of
companion bacteria.
• These companion bacteria help in initiation of infection by producing a
toxin or an enzyme or by inhibiting host immunity.
19. • Once the infection by Actinomycetes is established, the immune
system of the infected human host stimulates an intense
inflammation. Bacteria from the infected site may disseminate to
distant organs of the body.
• The Actinomycetes are particularly common type of bacteria found on
moldy hay. Farmers may be routinely exposed to very high
concentrations of Actinomyces and may inhale as many as 750,000
spores per minute. Frequent exposure to Actinomyces is the cause of
Farmers Lung respiratory problems.
21. • Cervicofacial infection, which accounts for
more than half of reported cases; the jaw is
often involved. The disease is endogenous in
origin; dental caries is a predisposing factor,
and infection may follow tooth extractions or
other dental procedures.
22. Who Get Infected
• Men are affected more frequently than
women, and in some regions the disease is
more common in rural agricultural workers
than in town dwellers, probably owing to
lower standards of dental care in the former.
23. Thoracic actinomycosis
• Thoracic actinomycosis commences in the
lung, probably as a result of aspiration of
actinomyces from the mouth.
• Sinuses often appear on the chest wall, and
the ribs and spine may be eroded.
• Primary endobronchial actinomycosis is an
uncommon complication of an inhaled foreign
body.
24. Abdominal Actinomyctes
• Abdominal cases commence in the appendix
or, less frequently, in colonic diverticulae.
• Pelvic actinomycosis occurs occasionally in
women fitted with plastic intra-uterine
contraceptive devices.
25. Clinical significance
• Are part of the NF found in the cavities of humans and
other animals.
• All may cause actinomycosis or “lumpy jaw” which is a
cervicofacial infection that used to occur following
tooth extractions or dental surgery which provided
traumatized tissue for growth of the microorganism
which may also invade the bone. This is rare today
because of prophylactic antibiotic therapy.
• May cause thoracic or abdominal infections
• May cause meningitis, endocarditis, or genital
infections
26. • Every kind of infection is characterized by
draining sinuses, usually containing
characteristic granules which are colonies of
bacteria that look like dense rosettes of club-
shaped filaments in radial arrangement
27. Diagnosis
• Specimens should be obtained directly from
lesions by open biopsy, needle aspiration or,
sputum, sinus discharge, bronchial secretion
by fibreoptic bronchoscopy.
• Examination of sputum is of no value as it
frequently contains oral actinomycetes.
• Material from suspected cases is shaken with
sterile water in a tube.
• Sulphur granules settle to the bottom and may
be removed with a Pasteur pipette.
28. • Granules crushed between two glass slides are
stained by the Gram and Ziehl-Neelsen (modified
by using 1% sulphuric acid for decolorization)
methods, which reveal the Gram positive mycelia
and the zone of radiating acid-fast clubs.
• Sulphur granules and mycelia in tissue sections
are identifiable by use of fluoresce inconjugated
specific antisera.
• In-situ PCR has been used to detect A. israelii in
tissue biopsies.
29. Culture
• For culture, suitable media, such as blood agar or
brain-heart infusion agar, glucose broth and enriched
thioglycollate broth, are inoculated with washed and
crushed granules.
• Cultures are incubated aerobically and anaerobically
for up to 14 days.
• After several days on agar medium, A. israelii may
form so called spider colonies that resemble molar
teeth.
30. culturing
• Cultures are incubated aerobically and
anaerobically for up to 14 days.
• After several days on agar medium, A. israelii
may form socalled spider colonies that resemble
molar teeth.
• The identity may be confirmed by biochemical
tests, by staining with specific fluorescent
antisera or by gas chromatography of metabolic
products of carbohydrate fermentation.
31. Procedure: Various steps for the Isolation and characterization of actinomycetes were
performed which are mentioned below:
samples collection
Enrichment of samples using
thioglycollate broth
Sample were culture on BA,
BHIA, glucose broth
Isolate was characterized by Morphological,
Physiological, Biochemical and Molecular
method
32. Strains of actinomycetes isolated
Morphological
methods:
(a)Macroscopic
methods
•Cover slip
culture
(b)Microscopic
methods
•Slide culture
method
Physiological
methods:
• Range of
pH for
growth
• Optimum
temperature
for growth
•Salinity
Biochemical methods:
•Catalase production
•Urease production
•Hydrogen sulfide
production •Nitrate
reduction •Starch
hydrolysis •Gelatin
liquefaction •Methyl red
test •Vogues-proskauer
test •Indole production
•Citrate utilization
•Casein hydrolysis
Molecular
methods:
•RFLP using any
one of genomic
DNA
•RAPD
•PFGE
•ARDRA
•Use of genus
specific primers
