Downloaded from www.microbiologyresearch.org by IP: 54.191.40.80 On: Fri, 25 Aug 2017 01:15:30 Bacillus cytotoxicus sp. nov. is a novel thermotolerant species of the Bacillus cereus Group occasionally associated with food poisoning Marie-He ´ le ` ne Guinebretie ` re, 1,2 Sandrine Auger, 3,4 Nathalie Galleron, 3,4 Matthias Contzen, 5 Benoit De Sarrau, 1,2 Marie-Laure De Buyser, 6 Gilles Lamberet, 3,4 Annette Fagerlund, 7,8 Per Einar Granum, 7 Didier Lereclus, 3,4 Paul De Vos, 9 Christophe Nguyen-The 1,2 and Alexei Sorokin 3,4 Correspondence Marie-He ´ le ` ne Guinebretie `re marie-helene.guinebretiere@ avignon.inra.fr 1 INRA, UMR408 Se ´ curite ´ et Qualite ´ des produits d’Origine Ve ´ ge ´ tale, F-84000 Avignon, France 2 Universite ´ d’Avignon, UMR408 Se ´ curite ´ et Qualite ´ des produits d’Origine Ve ´ ge ´ tale, F-84000 Avignon, France 3 INRA, UMR1319 MICALIS, F-78352 Jouy-en-Josas, France 4 AgroParisTech, UMR1319 MICALIS, F-78352 Jouy-en-Josas, France 5 Chemisches und Veterina ¨ runtersuchungsamt Stuttgart, Schaflandstrasse 3/2, 70736 Fellbach, Germany 6 ANSE, LERQUAP, Unite ´ Caracte ´ risation et Epide ´ miologie Bacte ´ rienne, F-94706 Maisons-Alfort cedex, France 7 Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, PO Box 8146, N-0033 Oslo, Norway 8 Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, PO Box 5003, N-1432 Aas, Norway 9 Laboratory for Microbiology, Department Biochemistry and Microbiology, Ghent University, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium An aerobic endospore-forming bacillus (NVH 391-98 T ) was isolated during a severe food poisoning outbreak in France in 1998, and four other similar strains have since been isolated, also mostly from food poisoning cases. Based on 16S rRNA gene sequence similarity, these strains were shown to belong to the Bacillus cereus Group (over 97 % similarity with the current Group species) and phylogenetic distance from other validly described species of the genus Bacillus was less than 95 %. Based on 16S rRNA gene sequence similarity and MLST data, these novel strains were shown to form a robust and well-separated cluster in the B. cereus Group, and constituted the most distant cluster from species of this Group. Major fatty acids (iso-C 15 : 0 , C 16 : 0 , iso-C 17 : 0 , anteiso-C 15 : 0 , iso-C 16 : 0 , iso-C 13 : 0 ) supported the affiliation of these strains to the genus Bacillus, and more specifically to the B. cereus Group. NVH 391-98 T taxon was more specifically characterized by an abundance of iso-C 15 : 0 and low amounts of iso-C 13 : 0 compared with other members of the B. cereus Group. Genome similarity together with DNA–DNA hybridization values and physiological and biochemical tests made it possible to genotypically and phenotypically differentiate NVH 391-98 T taxon from the six current B. cereus Group species. NVH 391-98 T therefore represents a novel species, for which the name Bacillus cytotoxicus sp. nov. is proposed, with the type strain NVH 391-98 T (5DSM 22905 T 5CIP 110041 T ) . Abbreviations: DDH, DNA–DNA hybridization; MLST, multilocus sequence typing. The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene of strains are described in Tables 1 and S1. The sequences of the MLST genes can be found at http://mlstoslo.uio.no/index.html. A supplementary figure and three supplementary tables are available with the online version of this paper. International Journal of Systematic and Evolutionary Microbiology (2013), 63, 31–40 DOI 10.1099/ijs.0.030627-0 030627 G 2013 IUMS Printed in Great Britain 31