Methanospirillum, a New Genus of Methanogenic Bacteria, and Characterization of Methanospirillum hungatii sp.nov.

Abstract

A new genus of methanogenic bacteria is described. The colonies produced by these bacteria are yellow, circular, and convex with lobate margins; an optical pattern of regular, light and dark striations throughout the colonies is a most unique and distinguishing characteristic. These striations are two cell lengths apart. Cells are gram negative and occur in filaments up t o 100 pm in length. Tufts of polar flagella and a striated cell surface are revealed in electron micrographs; cell ends are blunt, not rounded. The deoxyribonucleic acid base composition of the type species is 45 mol% guanine plus cytosine. Formate or hydrogen and carbon dioxide serves as a substrate for methane formation and growth; acetate, pyruvate, methanol, ethanol, and benzoate do not. The name MethanospiriZZum is proposed for this new genus of spiral-shaped methanogenic bacteria. The type species, Methanospirilhm hungatii sp. nov., is named in honor of R. E. Hungate. The type strain of M. hungatii is JF1 (ATCC 27890). In 1966, P. H. Smith reported the isolation of a new spiral-shaped me thanogenic bacterium from sewage sludge (8). In the present paper we define the characteristics of and propose a name for this new organism. A report on some of these findings has appeared previously (J. G. Ferry and R. S . Wolfe, Abstr. Annu. Meet. Amer. SOC. Microbiol. 1973, G121, p. 46). MATERIALS AND METHODSBacterial strains. Strain BM was received from Barry C. McBride (University of British Columbia, Vancouver), strain 3-PS was received from Paul H. Smith (University of Florida, Gainesville), and strain JF 1 was isolated in the present study. The inoculum from which each strain was isolated was sewage sludge i 8 , 9 ) .Media. Sterile growth media were prepared, and the organisms were cultivated under a strictly anaerobic (80% H,-20% CO,) atmosphere by a modification of the Hungate technique (4) as described by Bryant and Robinson (3). The maintenance medium contained the following constituents in distilled water a t the indicated final percentage compositions (wt/vol): K, HPO, , 0.023; KH, PO,, 0.023; (NH, ), SO,, 0.023; NaCl, 0.046; MgSO, -7H, 0, 0.009; CaCl, *2H, 0, 0.006; sodium formate, 0.2; yeast extract (Difco), 0.2; Trypticase (BBL), 0.2; resazurin, 0.0001 ; Na, CO, , 0.4; cysteine hydrochloride, 0.025; Na, S*9H2 0, 0.025. In addition, 1% (vol/vol) each of vitamin solution and trace mineral solution were added to the medium (10). Before sterilization, the medium was Present address: Department of Biochemistry, University of Georgia, Athens, Ga. 30601. adjusted to pH 7.2 with 1 M HCl. The final pH after sterilization and equilibration with a gas mixture of 80% H, -20% CO, was 7.0. Solid medium for roll-tube cultures and slants was prepared by including Noble agar (Difco) at a final concentration of 2% (wtlvol). Cultures were incubated at 37 C. The liquid benzoateenrichment medium contained the following constituents in distilled water at the indicated final concentration (wt/vol) in percent: sodium benzoate, 0.2; NH,Cl, 0.075; K, HPO, , 0.04; ...

Methanolinea tarda gen. nov., sp. nov., a methane-producing archaeon isolated from a methanogenic digester sludge

“…(Paynter & Hungate, 1968); 4, Methanoplanus limicola M3 T (Wildgruber et al, 1982); 5, Methanogenium cariaci JR1 T (Romesser et al, 1979;Maestrojuán et al, 1990;Boone et al, 1993); 6, Methanoculleus borgensis MS2 T (Ollivier et al, 1985;Maestrojuán et al, 1990); 7, Methanofollis tationis Chile 9 T (Zabel et al, 1984;Boone et al, 1993;Zellner et al, 1999); 8, Methanospirillum hungatei JF-1 T (Ferry et al, 1974 Optimum pH 7 (6.7-8.0)~5 (4.0-5.8) 6.1-6.9 (5.9-7.7) 6.5-7.5 6.8-7.3 (6.0-8.3) 6.7 (5.5-8.0) 7 (6.6-8. …”

Phylogenetic Relationship of Symbiotic Archaea in the Gut of the Higher Termite Nasutitermes takasagoensis Fed with Various Carbon Sources

“…The clones affiliated with the Methanomicrobiales were observed in all libraries as the major phylogenetic group. Most of the Methanomicrobiales-related clones obtained in this study were affiliated with the genus Methanospirillum, which is a methanogenic group using H2 as an electron donor 11) , implying that H 2 is the primary substrate for methanogenesis in the gut of N. takasagoensis.…”

Microbial dynamics in upflow anaerobic sludge blanket (UASB) bioreactor granules in response to short-term changes in substrate feed

“…OTUs IV-VII of the class Methanococci were more divergent and each was most closely related to different organisms on the database. OTU IV, most closely related to Methanospirillum hungatei (Ferry et al, 1974), was the most numerous clone of this type for most treatments, with the others varied by treatment. In general, Methanospirillum-related clones were fewer in number than those related to the Methanobacteria-related clones.…”