title: Molecular gene profiling of Clostridium botulinum group III and its detection in naturally contaminated samples originating from various European countries.
abstract: We report the development of real-time PCR assays for genotyping Clostridium botulinum group III targeting the newly defined C. novyi sensu lato group; the nontoxic nonhemagglutinin (NTNH)-encoding gene ntnh; the botulinum neurotoxin (BoNT)-encoding genes bont/C, bont/C/D, bont/D, and bont/D/C; and the flagellin (fliC) gene. The genetic diversity of fliC among C. botulinum group III strains resulted in the definition of five major subgroups named fliC-I to fliC-V. Investigation of fliC subtypes in 560 samples, with various European origins, showed that fliC-I was predominant and found exclusively in samples contaminated by C. botulinum type C/D, fliC-II was rarely detected, no sample was recorded as fliC-III or fliC-V, and only C. botulinum type D/C samples tested positive for fliC-IV. The lack of genetic diversity of the flagellin gene of C. botulinum type C/D would support a clonal spread of type C/D strains in different geographical areas. fliC-I to fliC-III are genetically related (87% to 92% sequence identity), whereas fliC-IV from C. botulinum type D/C is more genetically distant from the other fliC types (with only 50% sequence identity). These findings suggest fliC-I to fliC-III have evolved in a common environment and support a different genetic evolution for fliC-IV. A combination of the C. novyi sensu lato, ntnh, bont, and fliC PCR assays developed in this study allowed better characterization of C. botulinum group III and showed the group to be less genetically diverse than C. botulinum groups I and II, supporting a slow genetic evolution of the strains belonging to C. botulinum group III.
authors: Anses (French Agency for Food, Environmental and Occupational Health and Safety), Food Safety Laboratory, Maisons-Alfort, France.
authors: Cedric
authors: C
authors: Woudstra
authors: Anses, Ploufragan-Plouzané Laboratory, Hygiene and Quality of Poultry and Pig Products Unit, Ploufragan, France Université Européenne de Bretagne, Rennes, France.
authors: Caroline
authors: C
authors: Le Maréchal
authors: Anses, Ploufragan-Plouzané Laboratory, Hygiene and Quality of Poultry and Pig Products Unit, Ploufragan, France Université Européenne de Bretagne, Rennes, France.
authors: Rozenn
authors: R
authors: Souillard
authors: LABOCEA, Ploufragan, France.
authors: Marie-Hélène
authors: MH
authors: Bayon-Auboyer
authors: Istituto Superiore di Sanità (ISS) Department of Veterinary Public Health and Food Safety, National Reference Centre for Botulism, Rome, Italy.
authors: Fabrizio
authors: F
authors: Anniballi
authors: Istituto Superiore di Sanità (ISS) Department of Veterinary Public Health and Food Safety, National Reference Centre for Botulism, Rome, Italy.
authors: Bruna
authors: B
authors: Auricchio
authors: Istituto Superiore di Sanità (ISS) Department of Veterinary Public Health and Food Safety, National Reference Centre for Botulism, Rome, Italy.
authors: Dario
authors: D
authors: De Medici
authors: Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Laboratorio di Treviso, Treviso, Italy.
authors: Luca
authors: L
authors: Bano
authors: Central Veterinary Institute (CVI) of Wageningen, UR, Lelystad, The Netherlands.
authors: Miriam
authors: M
authors: Koene
authors: LNC, Veterinary Diagnostic Laboratory, Paita, New Caledonia.
authors: Marie-Hélène
authors: MH
authors: Sansonetti
authors: LNC, Veterinary Diagnostic Laboratory, Paita, New Caledonia.
authors: Denise
authors: D
authors: Desoutter
authors: Robert Koch-Institut (RKI) Centre for Biological Threats and Special Pathogens, Biological Toxins, Consultant Laboratory for Clostridium botulinum, Berlin, Germany.
authors: Eva-Maria
authors: EM
authors: Hansbauer
authors: Robert Koch-Institut (RKI) Centre for Biological Threats and Special Pathogens, Biological Toxins, Consultant Laboratory for Clostridium botulinum, Berlin, Germany.
authors: Martin B
authors: MB
authors: Dorner
authors: Robert Koch-Institut (RKI) Centre for Biological Threats and Special Pathogens, Biological Toxins, Consultant Laboratory for Clostridium botulinum, Berlin, Germany.
authors: Brigitte G
authors: BG
authors: Dorner
authors: Anses (French Agency for Food, Environmental and Occupational Health and Safety), Food Safety Laboratory, Maisons-Alfort, France patrick.fach@anses.fr.
authors: Patrick
authors: P
authors: Fach
mesh_terms: D000818:Animals; D000829:Animals, Domestic; D003014:Clostridium botulinum / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D019143:Evolution, Molecular; D005798:Genes, Bacterial; D014644:Genetic Variation; D005838:Genotype; D008969:Molecular Sequence Data; D058889:Molecular Typing / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial
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