Investigation of  Clostridium botulinum  group III's mobilome content

Woudstra, Cédric; Maréchal, Caroline Le; Souillard, Rozenn; Anniballi, Fabrizio; Auricchio, Bruna; Bano, Luca; Bayon-Auboyer, Marie Hélène; Koene, Miriam; Mermoud, Isabelle; Brito, Roseane B.; Lobato, Francisco Carlos Faria; Silva, Rodrigo; Dorner, Martin B.; Fach, Patrick

doi:10.1016/j.anaerobe.2017.12.009

Cited by 6 publications

(9 citation statements)

References 22 publications

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“…Samples positive for all primers had also been reported in some other avian outbreaks [38]. It appears that type D is uncommon in avian botulism outbreaks and has not been reported in other available studies [16,21,22,38]. No type C BoNTs were detected either in the samples analyzed during our study or in previously published studies that distinguished type C/D from type C BoNTs [14,16].…”

Section: Discussionsupporting

confidence: 74%

“…Type D/C (7.84%), type D (3.27%), and samples positive for several or all primers (C, D, C/D, and D/C) (8.50%) were detected far more scarcely. Type D/C had been reported in studies conducted in Italy, where it had been found either in the intestinal contents of a stork [16] or from coot, duck, or heron samples [38]. All the type D/C-positive samples in our study were detected in turkeys and not in wild birds.…”

Section: Discussionsupporting

confidence: 68%

“…All the type D/C-positive samples in our study were detected in turkeys and not in wild birds. Samples positive for all primers had also been reported in some other avian outbreaks [38]. It appears that type D is uncommon in avian botulism outbreaks and has not been reported in other available studies [16,21,22,38].…”

Section: Discussionmentioning

confidence: 63%

“…The authors therefore compiled an overview of the avian botulism outbreaks diagnosed since 2013 and the enrichment broths were selected on this basis so as to cover, insofar as possible, the avian species, geographical areas and BoNT types involved. As in most European countries [14,16,35,38], BoNT type C/D (80.39%) was the most frequently detected in avian botulism outbreaks. Type D/C (7.84%), type D (3.27%), and samples positive for several or all primers (C, D, C/D, and D/C) (8.50%) were detected far more scarcely.…”

Section: Discussionmentioning

confidence: 86%

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Development of An Innovative and Quick Method for the Isolation of Clostridium botulinum Strains Involved in Avian Botulism Outbreaks

Gratiet

Poëzévara

Rouxel

et al. 2020

Toxins

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Avian botulism is a serious neuroparalytic disease mainly caused by a type C/D botulinum neurotoxin produced by Clostridium botulinum group III, one of the entwined bacterial species from the Clostridium novyi sensu lato genospecies. Its isolation is very challenging due to the absence of selective media and the instability of the phage carrying the gene encoding for the neurotoxin. The present study describes the development of an original method for isolating C. botulinum group III strains. Briefly, this method consists of streaking the InstaGene matrix extraction pellet on Egg Yolk Agar plates and then collecting the colonies with lipase and lecithinase activities. Using this approach, it was possible to isolate 21 C. novyi sensu lato strains from 22 enrichment broths of avian livers, including 14 toxic strains. This method was successfully used to re-isolate type C, D, C/D, and D/C strains from liver samples spiked with five spores per gram. This method is cheap, user-friendly, and reliable. It can be used to quickly isolate toxic strains involved in avian botulism with a 64% success rate and C. novyi sensu lato with a 95% rate. This opens up new perspectives for C. botulinum genomic research, which will shed light on the epidemiology of avian botulism.

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Section: Discussionsupporting

confidence: 74%

Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 63%

Section: Discussionmentioning

confidence: 86%

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Development of An Innovative and Quick Method for the Isolation of Clostridium botulinum Strains Involved in Avian Botulism Outbreaks

Gratiet

Poëzévara

Rouxel

et al. 2020

Toxins

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“…Although these MGEs belong to different groups of mobile elements (p1_Cst and p1_BKT015925 are pseudolysogenic BoNT phages; Skarin et al, 2011 , p4_BKT015925 and p6_Cst have the characteristics of prophage-plasmid hybrids; Pfeifer et al, 2021 , CWou-2020a is a circular prophage and p2_BKT015925 is a plasmid; Skarin et al, 2011 ), they appeared to be genuinely recognized by the CRISPR systems. In a few cases, CRISPR protection was incomplete (e.g., against p4_BKT015925), because the strain had spacers as well as the immunized MGE as part of their genome ( Woudstra et al, 2017 ). This may suggest that these MGEs persist even in the presence of spacers matching with 100% homology, using an unidentified strategy (screening for anti-CRISPR using AcRanker was negative, data not shown; Eitzinger et al, 2020 ).…”

Section: Resultsmentioning

confidence: 99%

Exploration of the Diversity of Clustered Regularly Interspaced Short Palindromic Repeats-Cas Systems in Clostridium novyi sensu lato

et al. 2021

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Classified as the genospecies Clostridium novyi sensu lato and distributed into four lineages (I–IV), Clostridium botulinum (group III), Clostridium novyi, and Clostridium haemolyticum are clostridial pathogens that cause animal diseases. Clostridium novyi sensu lato contains a large mobilome consisting of plasmids and circular bacteriophages. Here, we explored clustered regularly interspaced short palindromic repeats (CRISPR) arrays and their associated proteins (Cas) to shed light on the link between evolution of CRISPR-Cas systems and the plasmid and phage composition in a study of 58 Clostridium novyi sensu lato genomes. In 55 of these genomes, types I-B (complete or partial), I-D, II-C, III-B, III-D, or V-U CRISPR-Cas systems were detected in chromosomes as well as in mobile genetic elements (MGEs). Type I-B predominated (67.2%) and was the only CRISPR type detected in the Ia, III, and IV genomic lineages. Putative type V-U CRISPR Cas14a genes were detected in two different cases: next to partial type-IB CRISPR loci on the phage encoding the botulinum neurotoxin (BoNT) in lineage Ia and in 12 lineage II genomes, as part of a putative integrative element related to a phage-inducible chromosomal island (PICI). In the putative PICI, Cas14a was associated with CRISPR arrays and restriction modification (RM) systems as part of an accessory locus. This is the first time a PICI containing such locus has been detected in C. botulinum. Mobilome composition and dynamics were also investigated based on the contents of the CRISPR arrays and the study of spacers. A large proportion of identified protospacers (20.2%) originated from Clostridium novyi sensu lato (p1_Cst, p4_BKT015925, p6_Cst, CWou-2020a, p1_BKT015925, and p2_BKT015925), confirming active exchanges within this genospecies and the key importance of specific MGEs in Clostridium novyi sensu lato.

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