Genus Firmicutes/“Clostridia”/Clostridiales/“Peptostreptococcaceae”/ Tepidibacter Slobodkin, Tourova, Kostrikina, Chernyh, Bonch-Osmolovskaya, Jeanthon and Jones 2003, 1133VP .......................................................................................................................................................................................... Alexander Slobodkin, Russian Academy of Sciences, Winogradsky Institute of Microbiology, 7/2, Prospekt 60-letiya Oktyabrya, Moscow 117312, Russia Te.pi.di.bac’ter. L. adj. tepidus warm; N.L. bacter masc. equivalent of Gr. neut. dim. n. bakterion rod; N.L. masc. n. Tepidibacter a warm rod. Straight to slightly curved rods 0.7–0.9 μm in diameter and 3.5–6.0 μm in length. Gram-positive type cell wall. Cells occur singly, in pairs or in short chains and exhibit tumbling motility due to peritrichous lagellation. Forms round or ovoid refractile endospores in terminally or subterminally swollen sporangia. Anaerobic. Moderately thermophilic, the temperature range for growth is 30–60∘ C. Neutrophilic. Optimal growth at marine salinity. Grows organotrophically on a number of proteinaceous substrates and carbohydrates. Elemental sulfur may be reduced, but sulfur reduction does not stimulate growth. DNA G+C content (mol%): 24–29 (Tm ). Type species: Tepidibacter thalassicus Slobodkin, Tourova, Kostrikina, Chernyh, Bonch-Osmolovskaya, Jeanthon and Jones 2003, 1133VP . .................................................................................. Straight to slightly curved rods 0.7–0.9 μm in diameter and 3.5–6.0 μm in length. Gram-positive type cell wall. Cells occur singly, in pairs or in short chains and exhibit tumbling motility due to peritrichous lagellation. Forms round or ovoid refractile endospores in terminally or subterminally swollen sporangia. Anaerobic. Moderately thermophilic, the temperature range for growth is 30–60∘ C. Neutrophilic. Optimal growth at marine salinity. Grows organotrophically on a number of proteinaceous substrates and carbohydrates. Elemental sulfur may be reduced, but sulfur reduction does not stimulate growth. DNA G+C content (mol%): 24–29 (Tm ). Type species: Tepidibacter thalassicus Slobodkin, Tourova, Kostrikina, Chernyh, Bonch-Osmolovskaya, Jeanthon and Jones 2003, 1133VP . Number of validated species: 2 Further descriptive information 16S rRNA gene sequence analysis places the genus Tepidibacter within cluster XI of the clostridia (nomenclature of Collins et al., 1994). Members of the genus Tepidibacter are currently represented by two species, Tepidibacter thalassicus and Tepidibacter formicigenes, which share 95% of 16S rRNA gene sequence similarity. Both species of the genus Tepidibacter form terminal or subterminal endospores. In the late-exponential phase of growth, up to 30% of the cells contain spores. The best growth of Tepidibacter can be obtained on complex proteinaceous substrates such as tryptone, casein, and peptone. Tepidibacter thalassicus is able to perform the Stickland reaction with alanine and proline. Carbohydrates, with an exception of starch, slightly stimulate growth of Tepidibacter thalassicus. Growth of Tepidibacter formicigenes on sugars is more eficient. Both species produce ...................................................................................................................................................................................................... Bergey’s Manual of Systematics of Archaea and Bacteria, Online © 2015 Bergey’s Manual Trust. This article is © 2009 Bergey’s Manual Trust. DOI: 10.1002/9781118960608.gbm00669. Published by John Wiley & Sons, Inc., in association with Bergey’s Manual Trust. 2 acetate and ethanol from glucose, and either H2 /CO2 , (Tepidibacter thalassicus) or formate (Tepidibacter formicigenes). External electron acceptors do not enhance growth, however, elemental sulfur can be reduced to hydrogen sulide. Both members of the genus Tepidibacter were isolated from deep-sea hydrothermal vents. Tepidibacter thalassicus was isolated from the outer wall of a ‘black smoker’ covered with the polychetous annelid Alvinella species (13∘ N hydrothermal ield on the East-Paciic Rise, depth 2650 m); Tepidibacter formicigenes was isolated from hydrothermal luid (the Menez-Gwen hydrothermal site, Mid-Atlantic Ridge, depth 800–1000 m). Location of these vents, one of which is in Paciic and other in Atlantic Ocean, suggests wide geographical distribution of Tepidibacter in marine hydrothermal environments where they probably function as decomposers of organic matter produced by deepsea biota. Table 1 shows differential characteristics for Tepidibacter from other members of Peptostreptococcaceae and related genera. Enrichment and isolation procedures Members of the genus Tepidibacter have been isolated from deep-sea hydrothermal vents (Slobodkin et al., 2003; Urios et al., 2004). There are no reports that members of this genus occur in other environments. Employment of the marine anaerobic media rich in proteinaceous substrates and incubation in the temperature range of 45–55∘ C may favor the enrichment of Tepidibacter species. Tepidibacter thalassicus rapidly hydrolyzes casein (Hammerstein grade) that results in visual disappearance of the casein locks and may help in the detection of growth. Tepidibacter forms colonies in 1.5% (w/v) agar. Maintenance procedures Members of the genus Tepidibacter may be maintained on the medium of Slobodkin et al. (2003) with peptone or casein as a substrate or on the glucose/yeast extract/peptone medium of Urios et al. (2004). Good and reproducible growth can also be obtained in liquid medium lacking sulide as a reducing agent and prepared anaerobically (Slobodkin et al., 2003). Freeze-drying of the cultures results in good recovery. Liquid cultures may be stored at +4∘ C for 10–12 months without loss of viability. Bergey’s Manual of Systematics of Archaea and Bacteria List of species of the genus Tepidibacter Tepidibacter thalassicus Slobodkin, Tourova, Kostrikina, Chernyh, Bonch-Osmolovskaya, Jeanthon and Jones 2003, 1133VP ................................................................................... tha.las’si.cus. Gr. fem. n. thalassa the sea; N.L. masc. adj. thalassicus of the sea. Cells are straight to slightly curved rods, 0.7–0.9 μm in diameter and 3.5–6.0 μm in length, which form round, refractile endospores in terminally swollen sporangia. Cells occur singly or in short chains and exhibit tumbling motility due to peritrichous lagellation. The temperature range for growth is 33–60∘ C, with an optimum at 50∘ C. The pH range for growth is 4.8–8.5, with an optimum at 6.5–6.8. Growth occurs at NaCl concentrations in the range 1.5–6% (w/v). Anaerobic. Substrates utilized include casein, peptone, albumin, yeast extract, beef extract, alanine plus praline, and starch. Glucose, maltose, pyruvate, valine, and arginine slightly stimulate growth in the presence of yeast extract. Fructose, sucrose, xylose, cellobiose, L-arabinose, glycerol, sorbitol, acetate, butyrate, lactate, formate, methanol, fumarate, glycine, alanine, proline, alanine plus glycine, betaine, olive oil, xylan, carboxymethylcellulose, ilter paper, chitin, keratin, and H2 /CO2 are not utilized. The products of glucose fermentation are ethanol, acetate, and molecular hydrogen. Reduces elemental sulfur to hydrogen sulide. Does not use nitrate, fumarate, sulfate, sulite, thiosulfate, amorphous Fe(III) oxide, Fe(III) citrate, or oxygen as electron acceptors. DNA G+C content (mol%): 24 (Tm ). Type strain: SC 562, DSM 15285, UNIQEM 215. GenBank accession number (16S rRNA gene): AY158079. Tepidibacter formicigenes Urios, Cueff, Pignet and Barbier 2004, 442VP ................................................................................... for.mi.ci’ge.nes. N.L. adj. formicicum from L. n. formica ant; Gr. v. gennaio produce; N.L. adj. formicigenes producing formic acid. Rod-shaped cells, 0.8 μm in diameter and 4.0 μm in length; motile by means of peritrichous lagella. Cells stain Gram-stain-positive. Forms ovoid refractile subterminal endospore. Growth occurs between 35 and 55∘ C (optimum, 45∘ C), between pH 5.0 and 8.0 (optimum, 6.0) and at 2–6% (w/v) sea salts (optimum, 3%). Anaerobic, able to ferment ...................................................................................................................................................................................................... This article is © 2009 Bergey’s Manual Trust. Published by John Wiley & Sons, Inc., in association with Bergey’s Manual Trust. Morphology Oxygen requirement Peptidoglycan (position 3, bridge) Sugar fermented Major metabolic product from PYG Peptostreptococcus Filifactor Tepidibacter Anaerococcus Finegoldia Fusibacter Gallicola Helcococcus Micromonas Peptoniphilus Sedimentibacter Sporanaerobacter Tissierella Characteristic GenBank accession number (16S rRNA)b DNA G+C content (mol%) Growth temperature (optimum) (°C) Habitats D14150 X73452 AY158079 D14139 D14149 AF050099 AB038361 X69837 AF542231 D14138 L11305 AF358114 X80833 34–36 34 24–29 30–35 32–34 43 32–34 29–29.5 28–30 30–34 34 32.2 28–32 30–39 (37) (37) 50 30–39 (37) 30–39 (37) 20–40 (37) 30–39 (37) 30–39 (37) 30–39 (37) 30–39 (37) (37) 25–50 (40) 20–39 (37) Bergey’s Manual of Systematics of Archaea and Bacteria Freshwater Anaerobic Human intestine, Human Human Human Oil-producHuman Chicken HydrotherCat skin Human Human sediment sludge vagina, abscess intestine, oral cavity, abscess, ing well intestine, intestine, abscess, mal vent intestine, intestine, vagina, human otorrhea human vagina, vagina, vagina, human ginabscess abscess abscess abscess abscess abscess gival sulcus Large Gram-stain- Gram-stain- Small Gram- Gram-stain- Gram-stain- Gram-stain- Short rods, pairs, SpindleGramGram-stainGram-stain- Gram-stainpositive stain-positive positive, positive Gram-stain- shaped rod, positive positive positive positive stainoften stained positive motile, rod cocci (pairs, cocci (pairs, cocci (pairs, cocci (pairs, curved rods, rod, motile, positive cocci (pairs, negative, rods (single, cocci (pairs, negative chains) motile, mass) mass) mass) tetrads) cocci (pairs, in pairs ilamentous, chains) sporeformsporeformmass) or short spore variing ing chains) able Obligate Obligate Facultative Obligate Obligate Anaerobic Obligate Obligate Obligate Obligate Obligate Obligate Obligate anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe anaerobe Lys, D -Asp Orn, D -Asp ND Lys, D -Glu ND Lys, D -Asp Orn, D -Glu No data No data m-DAP, Orn, D -Glu Lys, Gly No data Orn, D -Asp w Butyric acid, caproic acid − ND + ND w Butyric acid − Acetic acid w No data − D Acetic acid, Acetic acid, butyric acid lactic acid − Acetic acid − Butyric acid, acetic acid − No data + No data − Acetic acid a Symbols: +, >85% positive; d, different strains give different reactions (16–84% positive); −, 0–15% positive; D, different among species; w, weak acid produced; ND, not determined. b GenBank accession number given for type species of the genus. 3 ...................................................................................................................................................................................................... This article is © 2009 Bergey’s Manual Trust. Published by John Wiley & Sons, Inc., in association with Bergey’s Manual Trust. TABLE 1. Characteristics differentiating members of the family Peptostreptococcus fam. nov. and related generaa 4 Bergey’s Manual of Systematics of Archaea and Bacteria mainly complex proteinaceous substrates and carbohydrates. Grows on glucose/yeast extract/peptone medium. Tryptone, glucose, sucrose, fructose, maltose, and pyruvate support growth in the presence of yeast extract. Poor growth on ethanol, mannose, and peptone. Cellobiose, xylose, starch, cellulose, dextran, xylan, succinate, lactate, trehalose, lactose, arabinose, galactose, ribose, rhamnose, mannitol, sorbitol, glycerol, urea, and olive oil are not utilized. The products of glucose fermentation (in decreasing order) are formate, acetate, and ethanol. Elemental sulfur, polysulides, thiosulfate, sulite, sulfate, nitrite, nitrate, and FeCl3 do not enhance growth. DNA G+C content (mol%): 29 (Tm ). Type strain: DV1184, CIP 107893, DSM 15518. GenBank accession number (16S rRNA gene): AY245527. J.A.E. Farrow. 1994. The phylogeny of the genus Clostridium: proposal of ive new genera and eleven new species combinations. Int. J. Syst. Bacteriol 44: 812–826. Slobodkin, A.I., T.P. Tourova, N.A. Kostrikina, N.A. Chernyh, E.A. Bonch-Osmolovskaya, C. Jeanthon and B.E. Jones. 2003. Tepidibacter thalassicus gen. nov., sp. nov., a novel moderately thermophilic, anaerobic, fermentative bacterium from a deep-sea hydrothermal vent. Int. J. Syst. Evol. Microbiol. 53: 1131–1134. Urios, L., V. Cueff, P. Pignet and G. Barbier. 2004. Tepidibacter formicigenes sp. nov., a novel spore-forming bacterium isolated from a Mid-Atlantic Ridge hydrothermal vent. Int. J. Syst. Evol. Microbiol. 54: 439–443. References Collins, M.D., P.A. Lawson, A. Willems, J.J. Cordoba, J. Fernández-Garayzábal, P. Garcia, J. Cai, H. Hippe and ...................................................................................................................................................................................................... This article is © 2009 Bergey’s Manual Trust. Published by John Wiley & Sons, Inc., in association with Bergey’s Manual Trust.