Comparison of DNA Fingerprinting Methods for Use in Investigation of Type E Botulism Outbreaks in the Canadian Arctic

Leclair, Daniel; Pagotto, Franco; Farber, Jeffrey M.; Cadieux, Brigitte; Austin, John W.

doi:10.1128/jcm.44.5.1635-1644.2006

Cited by 55 publications

(27 citation statements)

References 49 publications

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“…Isolation of C. botulinum type E from positive cultures was conducted on selective Clostridium botulinum Isolation (CBI) agar plates [16] using a colony blot immunoassay [17]. Up to three confirmed colonies were purified and kept for PFGE analysis using SmaI and XhoI [18]. …”

Section: Methodsmentioning

confidence: 99%

Tracking sources ofClostridium botulinumtype E contamination in seal meat

Leclair

Farber

Pagotto

et al. 2017

International Journal of Circumpolar Health

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Botulism in Nunavik, Quebec is associated with the consumption of aged marine mammal meat and fat. The objective was to identify meat handling practices presenting a risk of contamination of seal meat with C. botulinum. Potential sources of contamination were assessed through interviews with igunaq producers from five communities of Nunavik. These sources were verified by detection and isolation of C. botulinum from igunaq prepared in the field from seal carcasses. Interviews indicated practices presenting a risk for contamination included: placing meat or fat on coastal rocks, using seawater for rinsing, and ageing meat in inverted seal skin pouches. Although the presence of C. botulinum type E spores was detected in only two of 32 (6.3%) meat or fat samples collected during the butchering process, two of four igunaq preparations from these samples contained type E botulinum toxin. Analysis of C. botulinum type E isolates recovered from these preparations indicated that shoreline soil may be a source of contamination. Seal meat and fat may be contaminated with C. botulinum type E during the butchering process. Measures can be adopted to reduce the risks of contamination in the field and possibly decrease the incidence of type E botulism in Nunavik.

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Section: Methodsmentioning

confidence: 99%

Tracking sources ofClostridium botulinumtype E contamination in seal meat

Leclair

Farber

Pagotto

et al. 2017

International Journal of Circumpolar Health

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“…These methods include PFGE of strains from the Baltic Sea and the arctic region of Canada, amplified fragment length polymorphism (AFLP) analysis, and RAPD analysis (9,10,12,15,17). The molecular characterizations illustrate the diversity of serotype E bacterial strains in comparison to other C. botulinum groups.…”

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Analysis of Clostridium botulinum Serotype E Strains by Using Multilocus Sequence Typing, Amplified Fragment Length Polymorphism, Variable-Number Tandem-Repeat Analysis, and Botulinum Neurotoxin Gene Sequencing

MacDonald

Helma

Shou

et al. 2011

Appl Environ Microbiol

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A total of 41 Clostridium botulinum serotype E strains from different geographic regions, including Canada, Denmark, Finland, France, Greenland, Japan, and the United States, were compared by multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) analysis, variable-number tandem-repeat (VNTR) analysis, and botulinum neurotoxin (bont) E gene sequencing. The strains, representing environmental, food-borne, and infant botulism samples collected from 1932 to 2007, were analyzed to compare serotype E strains from different geographic regions and types of botulism and to determine whether each of the strains contained the transposon-associated recombinase rarA, involved with bont/E insertion. MLST examination using 15 genes clustered the strains into several clades, with most members within a cluster sharing the same BoNT/E subtype (BoNT/E1, E2, E3, or E6). Sequencing of the bont/E gene identified two new variants (E7, E8) that showed regions of recombination with other E subtypes. The AFLP dendrogram clustered the 41 strains similarly to the MLST dendrogram. Strains that could not be differentiated by AFLP, MLST, or bont gene sequencing were further examined using three VNTR regions. Both intact and split rarA genes were amplified by PCR in each of the strains, and their identities were confirmed in 11 strains by amplicon sequencing. The findings suggest that (i) the C. botulinum serotype E strains result from the targeted insertion of the bont/E gene into genetically conserved bacteria and (ii) recombination events (not random mutations) within bont/E result in toxin variants or subtypes within strains.

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“…Although physiological traits, biochemical tests, and toxin serotyping are still used to characterize C. botulinum strains, this information does not possess the discrimination required for source attribution and epidemiological investigations. To perform such analysis, it is essential to investigate the strains at the genetic level using methods such as randomly amplified polymorphic DNA (RAPD) analysis, amplified rRNA gene restriction analysis, pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP), single-locus and multilocus sequence typing, multilocus variable-number tandem-repeat analysis (MLVA), or real-time PCR (4)(5)(6)(7)(8). Despite the large range of technical methods available, C. botulinum group III, responsible for animal botulism, has not been as intensively studied as strains responsible for human botulism.…”

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confidence: 99%