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Proposal of Viridibacillus gen. nov. and reclassification of Bacillus arvi, Bacillus arenosi and Bacillus neidei as Viridibacillus arvi gen. nov., comb. nov., Viridibacillus arenosi comb. nov. and Viridibacillus neidei comb. nov.

Richard A. Albert^1,, Julieta Archambault^1,, Melissa Lempa¹, Beth Hurst¹, Christine Richardson^1,, Stephanie Gruenloh^1,2, Metin Duran³, Hanna Lucia Worliczek⁴, Birgit E. Huber⁴, Ramon Rosselló-Mora⁵, Peter Schumann⁶ and Hans-Jürgen Busse⁴

¹ Semco BioScience, 630 East Keefe, Milwaukee, WI 53212, USA
² Medical College, University of Wisconsin, Milwaukee, WI, USA
³ Department of Civil and Environmental Engineering, Villanova University, Villanova, PA, USA
⁴ Institut für Bakteriologie, Mykrologie und Hygiene, Veterinärmedizinische Universität Wien, Veterinärplatz 1, A-1210 Vienna, Austria
⁵ Grup de Microbiologia Marina, Institut Mediterrani d'Estudis Avançats, E-07190 Esporles, Mallorca, Spain
⁶ DSMZ – Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, D-38124 Braunschweig, Germany

Correspondence
Richard A. Albert
richard.albert{at}marquette.edu

A polyphasic study was undertaken to clarify the taxonomic position of endospore-forming strains 433-D9, 433-E17 and 121-X1. BOX-PCR-generated fingerprints indicated that they may be members of a single species. 16S rRNA gene sequence similarity demonstrated that a representative of this group, 433-D9, is affiliated closely with Bacillus arvi DSM 16317^T (100 %), Bacillus arenosi DSM 16319^T (99.8 %) and Bacillus neidei NRRL BD-87^T (97.1 %). Sequence similarities revealed Bacillus pycnus NRRL NRS-1691^T and several Kurthia species as the next nearest relatives. DNA–DNA hybridization results showed that strain 433-D9 is a member of B. arvi. Detection of L-Lys–D-Asp-based peptidoglycan in strain 433-D9, B. arvi DSM 16317^T and B. arenosi DSM 16319^T was in agreement with their close relationship, but differentiated these strains from B. neidei NRRL BD-87^T and B. pycnus NRRL NRS-1691^T, for which L-Lys–D-Glu was reported. A similar quinone system was detected in strains 433-D9, 433-E17, 121-X1, B. arvi DSM 16317^T, B. arenosi DSM 16319^T and B. neidei NRRL BD-87^T. This system, unusual for bacilli, consisted of the major compound menaquinone MK-8 (69–80 %) and moderate amounts of MK-7 (19–30 %). This observation was in contrast to the predominance of MK-7 of the closest relative B. pycnus NRRL NRS-1691^T, as also reported for representatives of the closely related non-endospore-forming genus Kurthia. Strains 433-D9, B. arvi DSM 16317^T and B. arenosi DSM 16319^T exhibited homogeneous and discriminative polar lipid profiles and fatty acid profiles consisting of major acids i-C_{15 : 0} and ai-C_{15 : 0} and moderate amounts of i-C_{17 : 1}10c and i-C_{17 : 1} I/ai-C_{17 : 1} B that discriminated them from closely related strains such as B. neidei NRRL BD-87^T. On the basis of clear-cut discriminative chemotaxonomic markers, we propose strains 433-D9, 433-E17 and 121-X1, B. arvi DSM 16317^T, B. arenosi DSM 16319^T and B. neidei NRRL BD-87^T to be reclassified within a separate genus. For this new taxon, we propose the name Viridibacillus gen. nov., and we propose the reclassification of Bacillus arvi, Bacillus arenosi and Bacillus neidei as Viridibacillus arvi gen. nov., comb. nov. (the type species of Viridibacillus, with the type strain DSM 16317^T =LMG 22165^T), Viridibacillus arenosi comb. nov. (type strain DSM 16319^T =LMG 22166^T) and Viridibacillus neidei comb. nov. (type strain NRRL BD-87^T =DSM 15031^T =JCM 11077^T).

Present address: Genesis Technologies International, 696 Winer Industrial Way, Lawrenceville, GA, USA.

The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain 433-D9 is AY266991.

BOX-PCR profiles, phase-contrast micrographs of cells of strain 433-D9 and photographs of growth of strain 433-D9 are available as supplementary material with the online version of this paper.

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