ORIGINAL ARTICLE Noncontiguous finished genome sequences and description of Bacillus massiliglaciei, Bacillus mediterraneensis, Bacillus massilinigeriensis, Bacillus phocaeensis and Bacillus tuaregi, five new species identified by culturomics F. Cadoret 1 , M. T. Alou 1 , P. Afouda 1 , I. S. Traore 1 , L. Bréchard 1 , C. Michelle 1 , F. Di Pinto 1 , C. Andrieu 1 , J. Delerce 1 , A. Levasseur 1 , P.-E. Fournier 1 and D. Raoult 1,2 1) Aix-Marseille Université, URMITE, UM63, CNRS7278, IRD198, INSERM 1095, Institut Hospitalo-Universitaire Méditerranée-Infection, Faculté de médecine, Marseille, France and 2) Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia Abstract Microbial culturomics, which investigates microbial diversity by combining diversified culture conditions, matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry and 16S rDNA identification, allowed to identify five new species within the Bacillus genus. Bacillus massiliglaciei strain Marseille-P2600 T , Bacillus mediterraneensis strain Marseille-P2384 T , Bacillus massilinigeriensis strain Marseille- P2366 T , Bacillus tuaregi strain Marseille-P2489 T and Bacillus phocaeensis strain SIT16 T are each the type strain of the corresponding bacterial species. These strains, the genomes of which are described here, are facultative anaerobic Gram-positive bacilli. Here, we describe the main characteristics of each bacterium and present their complete genome sequence and annotation. © 2017 Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases. Keywords: Bacillus massiliglaciei, Bacillus massilinigeriensis, Bacillus mediterraneensis, Bacillus phocaeensis, Bacillus tuaregi, culturomics, emerging bacteria, human microbiota, taxonogenomics Original Submission: 1 December 2016; Revised Submission: 6 April 2017; Accepted: 13 April 2017 Article published online: 20 April 2017 Corresponding author: D. Raoult, Aix-Marseille Université, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), CNRS 7278, IRD 198, INSERM 1095, UM63, Institut Hospitalo-Universitaire Méditerranée-Infection, Faculté de médecine, 27 Boulevard Jean Moulin, 13385, Marseille cedex 5, France. E-mail: didier.raoult@gmail.com Introduction The genus Bacillus (Cohn, 1872), classified among the Firmicutes, was created in 1872 [1]. Bacillus species are strictly aerobic or anaerobic-tolerant rod-shaped bacteria that are able to form endospores [2 – 4]. Bacilli colonize a wide range of environ- ments (soil, water) and human organisms. Several Bacillus spe- cies present a biotechnologic interest because of their metabolism, and some of them, as Bacillus thuringiensis, are known to be pathogenic for human beings [3]. Since the creation of the Bacillus genus, 221 new species with validly published names standing in nomenclature have been identified (Fig. 1). The development of PCR techniques at the end of the 1980s and now faster genome sequencing allow the number of Bacillus species identified to significantly increase; it also permits some strains to be reclassified as only one species because their description was based only on phenotypic observation. In this study, we used a new approach including genome sequencing, matrix-assisted desorption ionization – time of flight mass spectrometry (MALDI-TOF MS) spectrum and main pheno- typic characteristics [5 – 9] to describe five new Bacillus species. Bacillus massiliglaciei strain Marseille-P2600 T , Bacillus medi- terraneensis strain Marseille-P2384 T , Bacillus massilinigeriensis strain Marseille-P2366 T , Bacillus tuaregi strain Marseille-P2489 T and Bacillus phocaeensis strain SIT16 T are the type strains of the corresponding species. There are all Gram-positive bacilli and facultatively anaerobic. They were respectively isolated from a Siberian permafrost sample (B. massiliglaciei), the stool sample of a healthy Senegalese boy (B. mediterraneensis), the stool sample of a healthy Nigerien girl (B. tuaregi and B. massilinigeriensis) and New Microbe and New Infect 2017; 19: 45– 59 © 2017 Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) http://dx.doi.org/10.1016/j.nmni.2017.04.005