Bacillus cereus is ubiquitous and commonly found in a wide range of environments,including food. In this study, we analysed 114 foodborne B. cereus strains isolated mainly from starchy and dairy products in order to investigate their phenotypic diversity (API system), antimicrobial resistance and toxigenic profiles (hblA, nheA, hlyII,cereolysin O, cytK2, cytK1 and EM1 genes). All isolates were confirmed as B. cereus using 16S-23S ribosomal DNA intergenic transcribed spacers (ITS) signature. They were shown as Gram-positive, catalase and caseinase positive, haemolytic (97%), and positive for lecithin hydrolysis and motility (97 and 87 %, respectively). PCR detection of the B. cereus specific toxin genes revealed the occurrence rates of 100 % for cereolysin O, 98 % for nheA, 74 % for cytk2, 52 % for hblA, 28 % for hlyII, and the absence of cytK1. Only two strains (1.75%), isolated from intestine of boar and pheasant, carried the emetic toxin genetic determinants (ces). Antimicrobial susceptibility of isolates was tested towards 15 different antimicrobial agents. We detected susceptibility of all strains to most antibiotics, intermediate resistance to clindamycin and resistance to β lactam antibiotics with 83% of the resistant isolates producing β lactamase enzyme. This large phenotypic diversity combined with the toxigenic traits and antibiotic resistance; emphasize the high potential risk of food poisoning of B. cereus isolates. Beside, a clear correlation between the metabolic features and the origin of isolation was shown. Most of starchy isolates were able to hydrolyse starch while dairy strains were not able to produce amylases. Our overall results point out that the metabolic flexibility and toxigenic potential represent the main drivers for B. cereus ubiquity and adaptation in a given ecological niche
ABRIOUEL HIKMATE, BEN OMAR NABIL, LÓPEZ ROSARIO LUCAS, CAÑAMERO MAGDALENA MARTÍNEZ, ORTEGA ELENA, GÁLVEZ ANTONIO, Differentiation and Characterization by Molecular Techniques of Bacillus cereus Group Isolates from Poto Poto and Dégué, Two Traditional Cereal-Based Fermented Foods of Burkina Faso and Republic of Congo, 10.4315/0362-028x-70.5.1165
AFNOR (1996) NF ISO 7932. Microbiologie. Directives générales pour le dénombrement de Bacillus cereus Méthode par comptage des colonies à 30°C In: Analyse Microbiologique. Tome 1. Méthodes Horizontales. Contrôle de la Qualité des Produits Alimentaires. pp. 209–217. Paris La Défense, Association Française de Normalisation
Agaisse Herve, Gominet Myriam, Okstad Ole Andreas, Kolsto Anne-Brit, Lereclus Didier, PlcR is a pleiotropic regulator of extracellular virulence factor gene expression in Bacillus thuringiensis, 10.1046/j.1365-2958.1999.01419.x
Al-Khatib Mariam Saleh, Khyami-Horani Hala, Badran Eman, Shehabi Asem A., Incidence and characterization of diarrheal enterotoxins of fecal Bacillus cereus isolates associated with diarrhea, 10.1016/j.diagmicrobio.2007.06.014
Altayar M., Sutherland A.D., Bacillus cereus is common in the environment but emetic toxin producing isolates are rare, 10.1111/j.1365-2672.2005.02764.x
Andreeva Zhanna I., Nesterenko Vladimir F., Fomkina Maria G., Ternovsky Vadim I., Suzina Natalia E., Bakulina Anastasia Yu, Solonin Alexander S., Sineva Elena V., The properties of Bacillus cereus hemolysin II pores depend on environmental conditions, 10.1016/j.bbamem.2006.11.004
Andrews J. M., Susceptibility testing of Bacillus species, 10.1093/jac/dkf063
Ankolekar Chandrakant, Rahmati Talat, Labbé Ronald G., Detection of toxigenic Bacillus cereus and Bacillus thuringiensis spores in U.S. rice, 10.1016/j.ijfoodmicro.2008.10.006
Apetroaie Camelia, Andersson Maria A., Spröer Cathrin, Tsitko Irina, Shaheen Ranad, Jääskeläinen Elina L., Wijnands Luc M., Heikkilä Ritva, Salkinoja-Salonen Mirja S., Cereulide-producing strains of Bacillus cereus show diversity, 10.1007/s00203-005-0032-1
Auger S., Galleron N., Bidnenko E., Ehrlich S. D., Lapidus A., Sorokin A., The Genetically Remote Pathogenic Strain NVH391-98 of the Bacillus cereus Group Is Representative of a Cluster of Thermophilic Strains, 10.1128/aem.02242-07
Banerjee M, Nair GB, Ramamurthy T (2011) Phenotypic and genetic characterization of Bacillus cereus isolated from the acute diarrhoeal patients. Indian J Med Res 133:88–95
Bartoszewicz M., Hansen B.M., Swiecicka I., The members of the Bacillus cereus group are commonly present contaminants of fresh and heat-treated milk, 10.1016/j.fm.2008.02.001
Carlin Frédéric, Brillard Julien, Broussolle Véronique, Clavel Thierry, Duport Catherine, Jobin Michel, Guinebretière Marie-Hélène, Auger Sandrine, Sorokine Alexei, Nguyen-Thé Christophe, Adaptation of Bacillus cereus, an ubiquitous worldwide-distributed foodborne pathogen, to a changing environment, 10.1016/j.foodres.2009.10.024
Chaves Jeane Quintanilha, Pires Eder Soares, Vivoni Adriana Marcos, Genetic diversity, antimicrobial resistance and toxigenic profiles of Bacillus cereus isolated from food in Brazil over three decades, 10.1016/j.ijfoodmicro.2011.02.029
Cherif A., Borin S., Rizzi A., Ouzari H., Boudabous A., Daffonchio D., Bacillus anthracis Diverges from Related Clades of the Bacillus cereus Group in 16S-23S Ribosomal DNA Intergenic Transcribed Spacers Containing tRNA Genes, 10.1128/aem.69.1.33-40.2003
Choma C., Guinebretiere M.H., Carlin F., Schmitt P., Velge P., Granum P.E., Nguyen-The C., Prevalence, characterization and growth of Bacillus cereus in commercial cooked chilled foods containing vegetables, 10.1046/j.1365-2672.2000.00998.x
Chon Jung-Whan, Kim Jong-Hyun, Lee Sun-Jin, Hyeon Ji-Yeon, Seo Kun-Ho, Toxin profile, antibiotic resistance, and phenotypic and molecular characterization of Bacillus cereus in Sunsik, 10.1016/j.fm.2012.06.003
Clinical and Laboratory Standards Institute (2010) Performance standards for antimicrobial susceptibility testing. Twentieth informational supplement. Document M100S20, CLSI. Wayne, PA
Drew WL, Barry AL, O’Toole R, Sherris JC (1972) Reliability of the Kirby-Bauer disc diffusion method for detecting methicillin-resistant strains of Staphylococcus aureus. Appl Microbiol 24:240–247
Ehling-Schulz Monika, Fricker Martina, Scherer Siegfried, Bacillus cereus, the causative agent of an emetic type of food-borne illness, 10.1002/mnfr.200400055
Ehling-Schulz M., Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains, 10.1099/mic.0.27607-0
Ettoumi Besma, Raddadi Noura, Borin Sara, Daffonchio Daniele, Boudabous Abdellatif, Cherif Ameur, Diversity and phylogeny of culturable spore-forming Bacilli isolated from marine sediments, 10.1002/jobm.200800306
Fagerlund A., Genetic and functional analysis of the cytK family of genes in Bacillus cereus, 10.1099/mic.0.26975-0
Fagerlund Annette, Lindbäck Toril, Granum Per Einar, Bacillus cereus cytotoxins Hbl, Nhe and CytK are secreted via the Sec translocation pathway, 10.1186/1471-2180-10-304
J. Finlay W.J., Logan N.A., Sutherland A.D., Bacillus cereus emetic toxin production in cooked rice, 10.1006/fmic.2002.0505
Godič Torkar K., Seme K., Antimicrobial susceptibility, β-lactamase and enterotoxin production in Bacillus cereus isolates from clinical and food samples, 10.1007/s12223-009-0037-2
Gohar Michel, Økstad Ole Andreas, Gilois Nathalie, Sanchis Vincent, Kolst⊘ Anne-Brit, Lereclus Didier, Two-dimensional electrophoresis analysis of the extracellular proteome ofBacillus cereusreveals the importance of the PlcR regulon, 10.1002/1615-9861(200206)2:6<784::aid-prot784>3.0.co;2-r
Granum Per Einar, Bacillus cereus, Food Microbiology: Fundamentals and Frontiers, Third Edition ISBN:9781555814076 p.445-455, 10.1128/9781555815912.ch20
Guinebretiere M.-H., Broussolle V., Nguyen-The C., Enterotoxigenic Profiles of Food-Poisoning and Food-Borne Bacillus cereus Strains, 10.1128/jcm.40.8.3053-3056.2002
Guinebretière Marie-Hélène, Thompson Fabiano L., Sorokin Alexei, Normand Philippe, Dawyndt Peter, Ehling-Schulz Monika, Svensson Birgitta, Sanchis Vincent, Nguyen-The Christophe, Heyndrickx Marc, De Vos Paul, Ecological diversification in the Bacillus cereus Group, 10.1111/j.1462-2920.2007.01495.x
Guinebretiere M.-H., Velge P., Couvert O., Carlin F., Debuyser M.-L., Nguyen-The C., Ability of Bacillus cereus Group Strains To Cause Food Poisoning Varies According to Phylogenetic Affiliation (Groups I to VII) Rather than Species Affiliation, 10.1128/jcm.00921-10
Guinebretiere M.-H., Auger S., Galleron N., Contzen M., De Sarrau B., De Buyser M.-L., Lamberet G., Fagerlund A., Granum P. E., Lereclus D., De Vos P., Nguyen-The C., Sorokin A., Bacillus cytotoxicus sp. nov. is a novel thermotolerant species of the Bacillus cereus Group occasionally associated with food poisoning, 10.1099/ijs.0.030627-0
Hansen B. M., Hendriksen N. B., Detection of Enterotoxic Bacillus cereus and Bacillus thuringiensis Strains by PCR Analysis, 10.1128/aem.67.1.185-189.2001
Helgason E., Tourasse N. J., Meisal R., Caugant D. A., Kolsto A.-B., Multilocus Sequence Typing Scheme for Bacteria of the Bacillus cereus Group, 10.1128/aem.70.1.191-201.2004
Hoton F. M., The cereulide genetic determinants of emetic Bacillus cereus are plasmid-borne, 10.1099/mic.0.28069-0
Hoton Florence M., Fornelos Nadine, N'Guessan Elise, Hu Xiaomin, Swiecicka Izabela, Dierick Katelijne, Jääskeläinen Elina, Salkinoja-Salonen Mirja, Mahillon Jacques, Family portrait ofBacillus cereusandBacillus weihenstephanensiscereulide-producing strains, 10.1111/j.1758-2229.2009.00028.x
Jensen MA, Webster JA, Straus N (1993) Rapid identification of bacteria on the basis of polymerase chain reaction—amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol 59:945–952
Jensen Lars Bogø, Baloda Suraj, Boye Mette, Aarestrup Frank M., Antimicrobial resistance among Pseudomonas spp. and the Bacillus cereus group isolated from Danish agricultural soil, 10.1016/s0160-4120(01)00045-9
Kalman S, Kiehne LK, Libs LJ, Yamamota T (1993) Cloning of novel CrylC-Type gene from a strain of Bacillus thuringiensis subsp. Galleriae. Appl Environ Microbiol 59:1131–1137
Kim Jung-Beom, Kim Jai-Moung, Kim Cheon-Hyeon, Seo Kyu Seok, Park Yong-Bae, Choi Na-Jung, Oh Deog-Hwan, Emetic toxin producing Bacillus cereus Korean isolates contain genes encoding diarrheal-related enterotoxins, 10.1016/j.ijfoodmicro.2010.08.021
Kim Jung-Beom, Park Jong-Suk, Kim Moon-Seok, Hong Seong-Cheol, Park Joong-Hyun, Oh Deog-Hwan, Genetic diversity of emetic toxin producing Bacillus cereus Korean strains, 10.1016/j.ijfoodmicro.2011.07.014
Lapidus Alla, Goltsman Eugene, Auger Sandrine, Galleron Nathalie, Ségurens Béatrice, Dossat Carole, Land Miriam L., Broussolle Veronique, Brillard Julien, Guinebretiere Marie-Helene, Sanchis Vincent, Nguen-the Christophe, Lereclus Didier, Richardson Paul, Wincker Patrick, Weissenbach Jean, Ehrlich S. Dusko, Sorokin Alexei, Extending the Bacillus cereus group genomics to putative food-borne pathogens of different toxicity, 10.1016/j.cbi.2007.03.003
Lund Terje, De Buyser Marie-Laure, Granum Per Einar, A new cytotoxin from Bacillus cereus that may cause necrotic enteritis, 10.1046/j.1365-2958.2000.02147.x
Martínez-Blanch J. F., Sánchez G., Garay E., Aznar R., Evaluation of phenotypic and PCR-based approaches for routine analysis of Bacillus cereus group foodborne isolates, 10.1007/s10482-010-9545-y
Ouoba Labia Irene I., Thorsen Line, Varnam Alan H., Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments, 10.1016/j.ijfoodmicro.2008.03.026
PARK YONG-BAE, KIM JUNG-BEOM, SHIN SANG-WOON, KIM JONG-CHAN, CHO SEUNG-HAK, LEE BOK-KWON, AHN JUHEE, KIM JAE-MYUNG, OH DEOG-HWAN, Prevalence, Genetic Diversity, and Antibiotic Susceptibility of Bacillus cereus Strains Isolated from Rice and Cereals Collected in Korea, 10.4315/0362-028x-72.3.612
Pirttijärvi Tuija S.M., Andersson Maria A., Scoging Anne C., Salkinoja-Salonen Mirja S., Evaluation of Methods for Recognising Strains of the Bacillus cereus Group with Food Poisoning Potential Among Industrial and Environmental Contaminants, 10.1016/s0723-2020(99)80036-8
Pruss BM, Dietrich R, Nibler B, Martlbauer E, Scherer S (1999) The hemolytic enterotoxin HBL is broadly distributed among species of the Bacillus cereus group. Appl Environ Microbiol 65:5436–5442
Roy Arindam, Moktan Bijoy, Sarkar Prabir K., Characteristics of Bacillus cereus isolates from legume-based Indian fermented foods, 10.1016/j.foodcont.2006.12.006
Setlow B, Setlow P, Heat inactivation of Bacillus subtilis spores lacking small, acid-soluble spore proteins is accompanied by generation of abasic sites in spore DNA., 10.1128/jb.176.7.2111-2113.1994
Shadrin AM, Shapyrina EV, Siunov AV, Severinov KV, Solonin AS (2007) Bacillus cereus pore-forming toxins hemolysin II and cytotoxin K: polymorphism and distribution of genes among representatives of the cereus group. Mikrobiologiia 76:462–470
Slamti L., A cell-cell signaling peptide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group, 10.1093/emboj/cdf450
Svensson Birgitta, Monthán Amanda, Shaheen Ranad, Andersson Maria A., Salkinoja-Salonen Mirja, Christiansson Anders, Occurrence of emetic toxin producing Bacillus cereus in the dairy production chain, 10.1016/j.idairyj.2005.07.002
Swiecicka Izabela, Van der Auwera Géraldine A., Mahillon Jacques, Hemolytic and Nonhemolytic Enterotoxin Genes are Broadly Distributed among Bacillus thuringiensis Isolated from Wild Mammals, 10.1007/s00248-006-9122-0
Valero M., Hernández-Herrero L.A., Fernández P.S., Salmerón M.C., Characterization of Bacillus cereus isolates from fresh vegetables and refrigerated minimally processed foods by biochemical and physiological tests, 10.1006/fmic.2002.0507
van der Voort Menno, Kuipers Oscar P, Buist Girbe, de Vos Willem M, Abee Tjakko, Assessment of CcpA-mediated catabolite control of gene expression in Bacillus cereus ATCC 14579, 10.1186/1471-2180-8-62
Vilain S., Luo Y., Hildreth M. B., Brozel V. S., Analysis of the Life Cycle of the Soil Saprophyte Bacillus cereus in Liquid Soil Extract and in Soil, 10.1128/aem.03076-05
Wunschel David, Fox Karen F., Black Gavin E., Fox Alvin, Discrimination Among the B. Cereus Group, in Comparison to B. Subtilis, by Structural Carbohydrate Profiles and Ribosomal RNA Spacer Region PCR, 10.1016/s0723-2020(11)80085-8
YANG I-CHEN, SHIH DANIEL YANG-CHIH, HUANG TSUI-PING, HUANG YUN-PU, WANG JAN-YI, PAN TZU-MING, Establishment of a Novel Multiplex PCR Assay and Detection of Toxigenic Strains of the Species in the Bacillus cereus Group, 10.4315/0362-028x-68.10.2123
Bibliographic reference
Chaabouni, Ines ; Barkallah, Insaf ; Belaouis, Abir ; Hamdi, Chadlya ; Jouini, Ahlem ; et. al. Metabolic capacities and toxigenic potential as key drivers of Bacillus cereus ubiquity and adaptation. In: Annals of Microbiology, Vol. 65, no. 2, p. 975-983 (2014)