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Bacillus farraginis sp. nov., Bacillus fortis sp. nov. and Bacillus fordii sp. nov., isolated at dairy farms

Patsy Scheldeman¹, Marina Rodríguez-Díaz², Johan Goris^1,3,, Annelies Pil^1,3,, Elke De Clerck³, Lieve Herman¹, Paul De Vos³, Niall A. Logan² and Marc Heyndrickx¹

¹ Ministry of the Flemish Community, Centre for Agricultural Research, Department of Animal Product Quality, Brusselsesteenweg 370, 9090 Melle, Belgium
² Glasgow Caledonian University, School of Biological and Biomedical Sciences, Cowcaddens Road, Glasgow G4 0BA, UK
³ Ghent University, Faculty of Sciences, Laboratory of Microbiology (WE10), K. L. Ledeganckstraat 35, 9000 Gent, Belgium

Correspondence
Patsy Scheldeman
P.Scheldeman{at}clo.fgov.be

Forty-eight bacterial strains were isolated at dairy farms from raw milk, the milking apparatus, green fodder or feed concentrate after a heat treatment of 30 min at 100 °C. In this way, spore-forming bacteria with a very high intrinsic heat resistance were selected for. The aerobic spore-forming isolates were subjected to a polyphasic taxonomical study, including repetitive element sequence-based PCR typing, whole-cell protein profiling, 16S rDNA sequence analysis, DNA–DNA hybridizations, DNA base composition, fatty acid analysis, and morphological and biochemical characteristics. A comparison of the REP- and (GTG)₅-PCR and whole-cell protein SDS-PAGE profiles resulted in three clusters of similar strains. Analysis of the 16S rDNA sequences and DNA–DNA relatedness data showed that these clusters represented three novel species. The highest 16S rDNA similarity to a recognized species found for the three groups was around 94 % with Bacillus lentus and Bacillus sporothermodurans. Further phenotypic characterization supported the proposal of three novel species in the genus Bacillus, Bacillus farraginis, Bacillus fortis and Bacillus fordii. The respective type strains are R-6540^T (=LMG 22081^T=DSM 16013^T), R-6514^T (=LMG 22079^T=DSM 16012^T) and R-7190^T (=LMG 22080^T=DSM 16014^T); their G+C DNA base contents are 43·7, 44·3 and 41·9 mol%, respectively. Although in variable amounts, a predominance of the branched fatty acids iso-C_{15 : 0} and anteiso-C_{15 : 0} was observed in all three novel species.

Published online ahead of print on 9 February 2004 as DOI 10.1099/ijs.0.63095-0.

The GenBank accession numbers for the 16S rRNA gene sequences of B. farraginis R-8039, R-6538, R-6540^T and R-6915, B. fortis R-6514^T and B. fordii R-7190^T are AY443034–AY443039, respectively.

Details of DNA–DNA hybridization results and photomicrographs of sporangia and vegetative cells of the type strains of B. farraginis, B. fortis and B. fordii are available as supplementary material in IJSEM Online.

Present address: Center for Microbial Ecology, Michigan State University, 540 Plant & Soil Science Building, East Lansing, MI 48824-1325, USA.

Present address: Department of Bacteriology and Immunology, Veterinary and Agrochemical Research Centre, VAR-CODA-CERVA, Groeselenberg 99, 1180 Ukkel, Belgium.

This article has been cited by other articles:

				P. Scheldeman, A. Pil, L. Herman, P. De Vos, and M. Heyndrickx Incidence and Diversity of Potentially Highly Heat-Resistant Spores Isolated at Dairy Farms Appl. Envir. Microbiol., March 1, 2005; 71(3): 1480 - 1494. [Abstract] [Full Text] [PDF]