1887

Abstract

Free-living bacteria with spherical cells 0.5–2.5 µm in diameter were isolated from freshwater sediment. 16S rRNA gene sequence analysis placed the new isolates within the phylum (‘spirochaetes’). The isolates never displayed a helical morphology or motility. Growth occurred in the presence of 100 mg ampicillin l in complex and defined mineral salts medium amended with vitamins, yeast extract and monosaccharides, disaccharides or soluble starch as fermentable substrates. Two distinct isolates, designated Buddy and Grapes, exhibited doubling times of 21±2 and 15±1 h in glucose-amended medium and grew at 15–37 and 15–30 °C. Optimum growth was observed between 25 and 30 °C and pH 6.5–7.5, with no growth below pH 5 or above pH 10. Hexose and pentose fermentation yielded ethanol, acetate and formate as major end products. Growth was strictly fermentative and anaerobic, but the isolates tolerated brief oxygen exposure. Nitrate, sulfate, thiosulfate and carbon dioxide were not used as electron acceptors, but soluble Fe(III) was reduced to Fe(II) in glucose-amended medium. The DNA G+C base contents of isolates Buddy and Grapes were 45.5–46.4 and 47.0–49.2 mol%, respectively. Phospholipid fatty acid (PLFA) profiles contained large proportions of C and C straight-chain saturated fatty acids; Cω7 and Cω9 dominated the mono-unsaturated PLFAs in isolate Grapes, whereas isolate Buddy also possessed Cω5, Cω7 and Cω9 fatty acids. Branched monoenoic acids accounted for up to 12.4 and 30 % of the total PLFA in isolates Grapes and Buddy, respectively. Based on their unique morphological features and the phylogenetic distance from their closest relatives, we propose the new genus, gen. nov., to accommodate the new isolates within the novel species sp. nov. (type strain Buddy  = DSM 22777  = ATCC BAA-1886) and sp. nov. (type strain Grapes  = DSM 22778  = ATCC BAA-1885). is the type species of the genus.

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2012-01-01
2019-08-25
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References

  1. Canale-Parola E. . ( 1981; ). Proposal that Spirochaeta stenostrepta Zuelzer replace Spirochaeta plicatilis Ehrenberg as the type species of the genus Spirochaeta Ehrenberg. Request for an Opinion. . Int J Syst Bacteriol 31:, 105–106. [CrossRef]
    [Google Scholar]
  2. Canale-Parola E. . ( 1984; ). The spirochetes. . In Bergey’s Manual of Systematic Bacteriology, vol. 1, pp. 38–70. Edited by Krieg N. R. , Holt J. G. . . Baltimore:: Williams & Wilkins;.
    [Google Scholar]
  3. Chun J. , Lee J.-H. , Jung Y. , Kim M. , Kim S. , Kim B. K. , Lim Y.-W. . ( 2007; ). EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. . Int J Syst Evol Microbiol 57:, 2259–2261. [CrossRef] [PubMed]
    [Google Scholar]
  4. Chung J. , Rittmann B. E. . ( 2007; ). Bio-reductive dechlorination of 1,1,1-trichloroethane and chloroform using a hydrogen-based membrane biofilm reactor. . Biotechnol Bioeng 97:, 52–60. [CrossRef] [PubMed]
    [Google Scholar]
  5. Cole J. R. , Chai B. , Farris R. J. , Wang Q. , Kulam S. A. , McGarrell D. M. , Garrity G. M. , Tiedje J. M. . ( 2005; ). The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis. . Nucleic Acids Res 33:, D294–D296. [CrossRef] [PubMed]
    [Google Scholar]
  6. Cole J. R. , Konstantinidis K. , Farris R. J. , Tiedje J. M. . ( 2010; ). Microbial diversity and phylogeny: extending from rRNAs to genomes. . In Environmental Molecular Microbiology, pp. 1–20. Edited by Liu W.-T. , Jansson J. K. . . Wymondham, UK:: Caister Academic Press;.
    [Google Scholar]
  7. Dröge S. , Fröhlich J. , Radek R. , König H. . ( 2006; ). Spirochaeta coccoides sp. nov., a novel coccoid spirochete from the hindgut of the termite Neotermes castaneus . . Appl Environ Microbiol 72:, 392–397. [CrossRef] [PubMed]
    [Google Scholar]
  8. Drummond A. J. , Ashton B. , Cheung M. , Heled J. , Kearse M. , Moir R. , Stones-Havas S. , Thierer T. , Wilson A. . ( 2009; ). Geneious version 4.7. . http://www.geneious.com/
  9. Fernandez A. S. , Hashsham S. A. , Dollhopf S. L. , Raskin L. , Glagoleva O. , Dazzo F. B. , Hickey R. F. , Criddle C. S. , Tiedje J. M. . ( 2000; ). Flexible community structure correlates with stable community function in methanogenic bioreactor communities perturbed by glucose. . Appl Environ Microbiol 66:, 4058–4067. [CrossRef] [PubMed]
    [Google Scholar]
  10. Franzmann P. D. , Dobson S. J. . ( 1992; ). Cell wall-less, free-living spirochetes in Antarctica. . FEMS Microbiol Lett 76:, 289–292. [CrossRef] [PubMed]
    [Google Scholar]
  11. Franzmann P. D. , Rohde M. . ( 1992; ). Characteristics of a novel, anaerobic, mycoplasma-like bacterium from Ace Lake, Antarctica. . Antarct Sci 4:, 155–162. [CrossRef]
    [Google Scholar]
  12. Grabowski A. , Nercessian O. , Fayolle F. , Blanchet D. , Jeanthon C. . ( 2005; ). Microbial diversity in production waters of a low-temperature biodegraded oil reservoir. . FEMS Microbiol Ecol 54:, 427–443. [CrossRef] [PubMed]
    [Google Scholar]
  13. Gu A. Z. , Hedlund B. P. , Staley J. T. , Strand S. E. , Stensel H. D. . ( 2004; ). Analysis and comparison of the microbial community structures of two enrichment cultures capable of reductively dechlorinating TCE and cis-DCE. . Environ Microbiol 6:, 45–54. [CrossRef] [PubMed]
    [Google Scholar]
  14. Guindon S. , Gascuel O. . ( 2003; ). A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. . Syst Biol 52:, 696–704.[CrossRef]
    [Google Scholar]
  15. He J. , Sung Y. , Krajmalnik-Brown R. , Ritalahti K. M. , Löffler F. E. . ( 2005; ). Isolation and characterization of Dehalococcoides sp. strain FL2, a trichloroethene (TCE)- and 1,2-dichloroethene-respiring anaerobe. . Environ Microbiol 7:, 1442–1450. [CrossRef] [PubMed]
    [Google Scholar]
  16. Joseph R. , Holt S. C. , Canale-Parola E. . ( 1973; ). Peptidoglycan of free-living anaerobic spirochetes. . J Bacteriol 115:, 426–435.[PubMed]
    [Google Scholar]
  17. Kondo E. , Ueta N. . ( 1972; ). Composition of fatty acids and carbohydrates in Leptospira . . J Bacteriol 110:, 459–467.[PubMed]
    [Google Scholar]
  18. Li C. , Wolgemuth C. W. , Marko M. , Morgan D. G. , Charon N. W. . ( 2008; ). Genetic analysis of spirochete flagellin proteins and their involvement in motility, filament assembly, and flagellar morphology. . J Bacteriol 190:, 5607–5615. [CrossRef] [PubMed]
    [Google Scholar]
  19. Livermore B. P. , Johnson R. C. . ( 1974; ). Lipids of the Spirochaetales: comparison of the lipids of several members of the genera Spirochaeta, Treponema, and Leptospira . . J Bacteriol 120:, 1268–1273.[PubMed]
    [Google Scholar]
  20. Löffler F. E. , Sanford R. A. , Ritalahti K. M. . ( 2005; ). Enrichment, cultivation, and detection of reductively dechlorinating bacteria. . Methods Enzymol 397:, 77–111. [CrossRef] [PubMed]
    [Google Scholar]
  21. MacKenzie S. L. . ( 1987; ). Gas chromatographic analysis of amino acids as the N-heptafluorobutyryl isobutyl esters. . J Assoc Off Anal Chem 70:, 151–160.[PubMed]
    [Google Scholar]
  22. Magot M. , Fardeau M. L. , Arnauld O. , Lanau C. , Ollivier B. , Thomas P. , Patel B. K. . ( 1997; ). Spirochaeta smaragdinae sp. nov., a new mesophilic strictly anaerobic spirochete from an oil field. . FEMS Microbiol Lett 155:, 185–191. [CrossRef] [PubMed]
    [Google Scholar]
  23. Mesbah M. , Premachandran U. , Whitman W. B. . ( 1989; ). Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. . Int J Syst Bacteriol 39:, 159–167. [CrossRef]
    [Google Scholar]
  24. Paster B. J. , Canale-Parola E. . ( 1980; ). Involvement of periplasmic fibrils in motility of spirochetes. . J Bacteriol 141:, 359–364.[PubMed]
    [Google Scholar]
  25. Paster B. J. , Dewhirst F. E. . ( 2000; ). Phylogenetic foundation of spirochetes. . J Mol Microbiol Biotechnol 2:, 341–344.[PubMed]
    [Google Scholar]
  26. Rademaker J. L. , Hoste B. , Louws F. J. , Kersters K. , Swings J. , Vauterin L. , Vauterin P. , de Bruijn F. J. . ( 2000; ). Comparison of AFLP and rep-PCR genomic fingerprinting with DNA–DNA homology studies: Xanthomonas as a model system. . Int J Syst Evol Microbiol 50:, 665–677. [CrossRef] [PubMed]
    [Google Scholar]
  27. Rhuland L. E. , Work E. , Denman R. F. , Hoare D. S. . ( 1955; ). The behavior of the isomers of α,ϵ-diaminopimelic acid on paper chromatograms. . J Am Chem Soc 77:, 4844–4846. [CrossRef]
    [Google Scholar]
  28. Ritalahti K. M. , Löffler F. E. . ( 2002; ). Ecology and characterization of novel, free-living, non-spiral spirochetes. In Abstracts of the 102nd General Meeting of the American Society for Microbiology, 19–23 May 2002, Salt Lake City, UT, USA. . Washington, DC:: American Society for Microbiology;.
    [Google Scholar]
  29. Ritalahti K. M. , Löffler F. E. . ( 2004; )., Characterization of novel free-living pleomorphic spirochetes [FLiPS]. In Abstracts of the 10th International Symposium on Microbial Ecology, 22–27 August 2004, Cancun, Mexico. Wageningen: International Society for Microbial Ecology. .
    [Google Scholar]
  30. Satomi M. , Kimura B. , Takahashi G. , Fuji T. . ( 1997; ). Microbial diversity in Kusaya gravy. . Fish Sci 63:, 1019–1023.
    [Google Scholar]
  31. Stackebrandt E. , Frederiksen W. , Garrity G. M. , Grimont P. A. D. , Kämpfer P. , Maiden M. C. J. , Nesme X. , Rosselló-Mora R. , Swings J. . & other authors ( 2002; ). Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology. . Int J Syst Evol Microbiol 52:, 1043–1047. [CrossRef] [PubMed]
    [Google Scholar]
  32. Sung Y. , Ritalahti K. M. , Sanford R. A. , Urbance J. W. , Flynn S. J. , Tiedje J. M. , Löffler F. E. . ( 2003; ). Characterization of two tetrachloroethene-reducing, acetate-oxidizing anaerobic bacteria and their description as Desulfuromonas michiganensis sp. nov.. Appl Environ Microbiol 69:, 2964–2974. [CrossRef] [PubMed]
    [Google Scholar]
  33. Wayne L. G. . ( 1982; ). Actions of the Judicial Commission of the International Committee on Systematic Bacteriology on requests for opinions published between July 1979 and April 1981. . Int J Syst Bacteriol 32:, 464–465.[CrossRef]
    [Google Scholar]
  34. White D. C. , Geyer R. , Peacock A. D. , Hedrick D. B. , Koenigsberg S. S. , Sung Y. , He J. , Löffler F. E. . ( 2005; ). Phospholipid furan fatty acids and ubiquinone-8: lipid biomarkers that may protect Dehalococcoides strains from free radicals. . Appl Environ Microbiol 71:, 8426–8433. [CrossRef] [PubMed]
    [Google Scholar]
  35. Wolin E. A. , Wolin M. J. , Wolfe R. S. . ( 1963; ). Formation of methane by bacterial extracts. . J Biol Chem 238:, 2882–2886.[PubMed]
    [Google Scholar]
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