Molucular gene profiling of Clostridium botulinum group III and their detection in naturally contaminated samples originating from various European countries

C. Woudstra, C. LeMarechal, R. Souillard, M.H. Bäyon-Auboyer, F. Anniballi, B. Auricchio, D. De Medici, L. Bano, M.G.J. Koene, M.H. Sansonetti, E.M. Hansbauer, D. Desoutter, M.B. Dorner, P. Fach, B.G. Dorner

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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.
Original languageEnglish
Pages (from-to)2495-2505
JournalApplied and Environmental Microbiology
Volume81
DOIs
Publication statusPublished - 2015

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Clostridium botulinum
Clostridium botulinum D
Clostridium botulinum C
gene
Dilatation and Curettage
flagellin
Clostridium botulinum type D
Clostridium botulinum type C
Genes
genes
Flagellin
assay
Molecular Evolution
Clostridium novyi
sampling
botulinum toxin
genetic variation
targeting
assays
genotyping

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Woudstra, C. ; LeMarechal, C. ; Souillard, R. ; Bäyon-Auboyer, M.H. ; Anniballi, F. ; Auricchio, B. ; De Medici, D. ; Bano, L. ; Koene, M.G.J. ; Sansonetti, M.H. ; Hansbauer, E.M. ; Desoutter, D. ; Dorner, M.B. ; Fach, P. ; Dorner, B.G. / Molucular gene profiling of Clostridium botulinum group III and their detection in naturally contaminated samples originating from various European countries. In: Applied and Environmental Microbiology. 2015 ; Vol. 81. pp. 2495-2505.
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title = "Molucular gene profiling of Clostridium botulinum group III and their 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.",
author = "C. Woudstra and C. LeMarechal and R. Souillard and M.H. B{\"a}yon-Auboyer and F. Anniballi and B. Auricchio and {De Medici}, D. and L. Bano and M.G.J. Koene and M.H. Sansonetti and E.M. Hansbauer and D. Desoutter and M.B. Dorner and P. Fach and B.G. Dorner",
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journal = "Applied and Environmental Microbiology",
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Woudstra, C, LeMarechal, C, Souillard, R, Bäyon-Auboyer, MH, Anniballi, F, Auricchio, B, De Medici, D, Bano, L, Koene, MGJ, Sansonetti, MH, Hansbauer, EM, Desoutter, D, Dorner, MB, Fach, P & Dorner, BG 2015, 'Molucular gene profiling of Clostridium botulinum group III and their detection in naturally contaminated samples originating from various European countries', Applied and Environmental Microbiology, vol. 81, pp. 2495-2505. https://doi.org/10.1128/AEM.03915-14

Molucular gene profiling of Clostridium botulinum group III and their detection in naturally contaminated samples originating from various European countries. / Woudstra, C.; LeMarechal, C.; Souillard, R.; Bäyon-Auboyer, M.H.; Anniballi, F.; Auricchio, B.; De Medici, D.; Bano, L.; Koene, M.G.J.; Sansonetti, M.H.; Hansbauer, E.M.; Desoutter, D.; Dorner, M.B.; Fach, P.; Dorner, B.G.

In: Applied and Environmental Microbiology, Vol. 81, 2015, p. 2495-2505.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Molucular gene profiling of Clostridium botulinum group III and their detection in naturally contaminated samples originating from various European countries

AU - Woudstra, C.

AU - LeMarechal, C.

AU - Souillard, R.

AU - Bäyon-Auboyer, M.H.

AU - Anniballi, F.

AU - Auricchio, B.

AU - De Medici, D.

AU - Bano, L.

AU - Koene, M.G.J.

AU - Sansonetti, M.H.

AU - Hansbauer, E.M.

AU - Desoutter, D.

AU - Dorner, M.B.

AU - Fach, P.

AU - Dorner, B.G.

PY - 2015

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N2 - 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.

AB - 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.

U2 - 10.1128/AEM.03915-14

DO - 10.1128/AEM.03915-14

M3 - Article

VL - 81

SP - 2495

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JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

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