1887

Abstract

Two Gram-positive, rod-shaped, endospore-forming bacteria (strains CR-119 and CR-95) were isolated from brackish sediments in the mouth of the river Vélez in Málaga, southern Spain, and subjected to a polyphasic taxonomic study. Phenotypic tests showed that these strains were related to other species at a similarity level of less than 87·6 %. Both strains are halotolerant, aerobic, chemoheterotrophic, motile with peritrichous flagella and biosurfactant producers. Their endospores are oval, subterminal and non-deforming structures. The predominant menaquinone in both strains is MK-7. The fatty-acid profiles of both strains contain large quantities of branched and saturated fatty acids. The major fatty acids (%) are 15 : 0 anteiso (32·4), 15 : 0 iso (16·8), 17 : 0 iso (13·4), 16 : 0 (11·5) and 17 : 0 anteiso (10·2) in strain CR-119 and 15 : 0 anteiso (37·5), 17 : 0 iso (16·0) and 17 : 0 anteiso (15·8) in strain CR-95. The G+C contents of strains CR-119 and CR-95 are 41·0 and 42·5 mol%, respectively. RAPD analysis confirmed the low degree of similarity between the two strains and also amongst other species. 16S rRNA gene analysis of strain CR-119 showed the highest sequence similarity to be 97·4 %, with and subsp. . In the case of strain CR-95, the maximum similarity value was 99·5 %, with . DNA–DNA hybridization of strains CR-119 and CR-95 with the above species produced values lower than 46·9 %. Therefore, on the basis of phenotypic characteristics, phylogenetic data and genomic distinctiveness, we conclude that these strains merit classification as novel species, for which we propose the names sp. nov. (type strain CR-119=CECT 5688=LMG 22476) and sp. nov. (type strain CR-95=CECT 5687=LMG 22477).

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2005-05-01
2019-10-21
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References

  1. Arima, K., Kakinuma, A. & Tamura, G. ( 1968; ). Surfactin, a crystalline peptide lipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochem Biophys Res Commun 31, 488–494.[CrossRef]
    [Google Scholar]
  2. Ash, C., Priest, F. G. & Collins, M. D. ( 1993; ). Molecular identification of rRNA group 3 bacilli (Ash, Farrow, Wallbanks and Collins) using a PCR probe test. Proposal for the creation of a new genus Paenibacillus. Antonie van Leeuwenhoek 64, 253–260.
    [Google Scholar]
  3. Banat, I. M., Makkar, R. S. & Cameotra, S. S. ( 2000; ). Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol 53, 495–508.[CrossRef]
    [Google Scholar]
  4. Barrow, G. I. & Feltham, R. K. A. ( 1993; ). Cowan and Steel's Manual for the Identification of Medical Bacteria, 3rd edn. Cambridge: Cambridge University Press.
  5. Bauer, A. W., Kirby, W. M. M., Sherris, J. C. & Turck, M. ( 1966; ). Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45, 493–496.
    [Google Scholar]
  6. Bouchotroch, S., Quesada, E., Del Moral, A., Llamas, I. & Béjar, V. ( 2001; ). Halomonas maura sp. nov., a novel, moderately halophilic, exopolysaccharide-producing bacterium. Int J Syst Evol Microbiol 51, 1625–1632.[CrossRef]
    [Google Scholar]
  7. Brosius, J., Palmer, M. L., Kennedy, P. J. & Noller, H. F. ( 1978; ). Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A 75, 4801–4805.[CrossRef]
    [Google Scholar]
  8. Claus, D. & Berkeley, R. C. W. ( 1986; ). Genus Bacillus Cohn 1872174AL. In Bergey's Manual of Systematic Bacteriology, vol. 2, pp. 1105–1139. Edited by P. H. A. Sneath, N. S. Mair, M. E. Sharpe & J. G. Holt. Baltimore: Williams & Wilkins.
  9. Euzéby, J. P. ( 2004; ). List of Bacterial Names with Standing in Nomenclature. http://www.bacterio.cict.fr/
  10. Ferragut, C. & Leclerc, H. ( 1976; ). Etude comparative des methodes de determination du Tm de l'ADN bacterien. Ann Microbiol 127, 223–235 (in French).
    [Google Scholar]
  11. Fortina, M. G., Pukall, R., Schumann, P., Mora, D., Parini, C., Manachini, P. L. & Stackebrandt, E. ( 2001; ). Ureibacillus gen. nov., a new genus to accommodate Bacillus thermosphaericus (Andersson et al. 1995), emendation of Ureibacillus thermosphaericus and description of Ureibacillus terrenus sp. nov. Int J Syst Evol Microbiol 51, 447–455.
    [Google Scholar]
  12. Heyndrickx, M., Lebbe, L., Kersters, K., De Vos, P., Forsyth, G. & Logan, N. A. ( 1998; ). Virgibacillus: a new genus to accommodate Bacillus pantothenticus (Proom and Knight 1950). Emended description of Virgibacillus pantothenticus. Int J Syst Bacteriol 48, 99–106.[CrossRef]
    [Google Scholar]
  13. Jain, D. K., Collins-Thompson, D. L., Lee, H. & Trevors, J. T. ( 1991; ). A drop collapsing test for screening surfactant-producing microorganisms. J Microbiol Methods 13, 271–279.[CrossRef]
    [Google Scholar]
  14. Kumar, S., Tamura, K. & Nei, M. ( 2004; ). mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef]
    [Google Scholar]
  15. Lind, E. & Ursing, J. ( 1986; ). Clinical strains of Enterobacter agglomerans (synonyms, Erwinia herbicola, Erwinia milletiae) identified by DNA-DNA hybridization. Acta Pathol Microbiol Immunol Scand Sect B 94, 205–213.
    [Google Scholar]
  16. Logan, N. A. & Berkeley, R. C. ( 1984; ). Identification of Bacillus strains using the API system. J Gen Microbiol 130, 1871–1882.
    [Google Scholar]
  17. Marmur, J. ( 1961; ). A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3, 208–212.[CrossRef]
    [Google Scholar]
  18. Marmur, J. & Doty, P. ( 1962; ). Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol 5, 109–118.[CrossRef]
    [Google Scholar]
  19. Moraine, R. A. & Rogovin, P. ( 1966; ). Kinetics of polysaccharide B-1459 fermentation. Biotechnol Bioeng 8, 511–524.[CrossRef]
    [Google Scholar]
  20. Nakamura, L. K. ( 1989; ). Taxonomic relationship of black-pigmented Bacillus subtilis strains and a proposal for Bacillus atrophaeus sp. nov. Int J Syst Bacteriol 39, 295–300.[CrossRef]
    [Google Scholar]
  21. Naruse, N., Tenmyo, O., Kobaru, S., Kamei, H., Miyaki, T., Konishi, M. & Oki, T. ( 1990; ). Pumilacidin, a complex of new antiviral antibiotics. Production, isolation, chemical properties, structure and biological activity. J Antibiot 43, 267–280.[CrossRef]
    [Google Scholar]
  22. Nazina, T. N., Tourova, T. P., Poltaraus, A. B. & 8 other authors ( 2001; ). Taxonomic study of aerobic thermophilic bacilli: descriptions of Geobacillus subterraneus gen. nov., sp. nov. and Geobacillus uzenensis sp. nov. from petroleum reservoirs and transfer of Bacillus stearothermophilus, Bacillus thermocatenulatus, Bacillus thermoleovorans, Bacillus kaustophilus, Bacillus thermodenitrificans to Geobacillus as the new combinations G. stearothermophilus, G. thermocatenulatus, G. thermoleovorans, G. kaustophilus and G. thermodenitrificans. Int J Syst Evol Microbiol 51, 433–446.
    [Google Scholar]
  23. Owen, R. J. & Hill, L. R. ( 1979; ). The estimation of base compositions, base pairing and genome size of bacterial deoxyribonucleic acids. In Identification Methods for Microbiologists, pp. 277–296, 2nd edn. Edited by F. A. Skinner & D. W. Lovelock. London: Academic Press.
  24. Owen, R. J. & Pitcher, D. ( 1985; ). Current methods for estimating DNA composition and levels of DNA-DNA hybridization. In Chemical Methods in Bacterial Systematics, pp. 67–93. Edited by M. Goodfellow & D. E. Minnikin. London: Academic Press.
  25. Pinchuk, I. V., Bressollier, P., Sorokulova, I. B., Verneuil, B. & Urdaci, M. C. ( 2002; ). Amicoumacin antibiotic production and genetic diversity of Bacillus subtilis strains isolated from different habitats. Res Microbiol 153, 269–276.[CrossRef]
    [Google Scholar]
  26. Rhodes, M. E. ( 1958; ). The cytology of Pseudomonas spp. as revealed by a silver-plating staining method. J Gen Microbiol 18, 639–648.[CrossRef]
    [Google Scholar]
  27. Roberts, M. S., Nakamura, L. K. & Cohan, F. M. ( 1994; ). Bacillus mojavensis sp. nov., distinguishable from Bacillus subtilis by sexual isolation, divergence in DNA sequence, and differences in fatty acid composition. Int J Syst Bacteriol 44, 256–264.[CrossRef]
    [Google Scholar]
  28. Roberts, M. S., Nakamura, L. K. & Cohan, F. M. ( 1996; ). Bacillus vallismortis sp. nov., a close relative of Bacillus subtilis isolated from soil in Death Valley, California. Int J Syst Bacteriol 46, 470–475.[CrossRef]
    [Google Scholar]
  29. Rodríguez-Valera, F., Ruiz-Berraquero, F. & Ramos-Cormenzana, A. ( 1981; ). Characteristics of the heterotrophic bacterial populations in hypersaline environments of different salt concentrations. Microb Ecol 7, 235–243.[CrossRef]
    [Google Scholar]
  30. Ruiz-García, C., Béjar, V., Martínez-Checa, F., Llamas, I. & Quesada, E. ( 2005; ). Bacillus velezensis sp. nov., a surfactant-producing bacterium isolated from the river Vélez in Málaga, southern Spain. Int J Syst Evol Microbiol 55, 191–195.[CrossRef]
    [Google Scholar]
  31. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B. & Erlich, H. A. ( 1988; ). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487–491.[CrossRef]
    [Google Scholar]
  32. Shida, O., Takagi, H., Kadowaki, K. & Komagata, K. ( 1996; ). Proposal for two new genera, Brevibacillus gen. nov. and Aneurinibacillus gen. nov. Int J Syst Bacteriol 46, 939–946.[CrossRef]
    [Google Scholar]
  33. Sneath, P. H. A. & Sokal, R. R. ( 1973; ). Numerical Taxonomy. The Principles and Practice of Numerical Classification. San Francisco: Freeman, Williams & Wilkins.
  34. Sokal, R. R. & Michener, C. D. ( 1958; ). A statistical method for evaluating systematic relationships. Univ Kansas Sci Bull 38, 1409–1438.
    [Google Scholar]
  35. Stackebrandt, E., Fredricksen, W., Garrity, G. M. & 10 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]
    [Google Scholar]
  36. Thompson, J. D., Gibson, T. J., Plewniak, K., Jeanmougin, F. & Higgins, D. G. ( 1997; ). The clustal_x Windows interface: flexible strategies for multiple sequence alignments aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.[CrossRef]
    [Google Scholar]
  37. Waino, M., Tindall, B. J., Schumann, P. & Ingvorsen, K. ( 1999; ). Gracilibacillus gen. nov., with description of Gracilibacillus halotolerans gen. nov., sp. nov.; transfer of Bacillus dipsosauri to Gracilibacillus dipsosauri comb. nov., and Bacillus salexigens to the genus Salibacillus gen. nov. as Salibacillus salexigens comb. nov. Int J Syst Bacteriol 49, 821–831.[CrossRef]
    [Google Scholar]
  38. Wisotzkey, J. D., Jurtshuk, P., Jr, Fox, G. E., Deinhard, G. & Poralla, K. ( 1992; ). Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov. Int J Syst Bacteriol 42, 263–269.[CrossRef]
    [Google Scholar]
  39. Yakimov, M. M., Timmis, K. N., Wray, V. & Fredrickson, H. L. ( 1995; ). Characterization of a new lipopeptide surfactant produced by thermotolerant and halotolerant subsurface Bacillus licheniformis BAS50. Appl Environ Microbiol 61, 1706–1713.
    [Google Scholar]
  40. Yoon, J.-H., Weiss, N., Lee, K.-C., Lee, I.-S., Kang, K. H. & Park, Y.-H. ( 2001; ). Jeotgalibacillus alimentarius gen. nov., sp. nov., a novel bacterium isolated from jeotgal with l-lysine in the cell wall, and reclassification of Bacillus marinus Rüger 1983 as Marinibacillus marinus gen. nov., comb. nov. Int J Syst Evol Microbiol 51, 2087–2093.[CrossRef]
    [Google Scholar]
  41. Ziemke, F., Höfle, M. G., Lalucat, J. & Rossello-Mora, R. ( 1998; ). Reclassification of Shewanella putrefaciens Owen's group II as Shewanella baltica sp. nov. Int J Syst Bacteriol 48, 179–186.[CrossRef]
    [Google Scholar]
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DNA-DNA hybridization between strains CR-95 and CR-119 and other related species. [PDF](73 KB)

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