Biodiversity of Clostridium botulinum Type E Associated with a Large Outbreak of Botulism in Wildlife from Lake Erie and Lake Ontario

Abstract

The genetic relatedness of Clostridium botulinum type E isolates associated with an outbreak of wildlife botulism was studied using random amplification of polymorphic DNA (RAPD). Specimens were collected from November 2000 to December 2008 during a large outbreak of botulism affecting birds and fish living in and around Lake Erie and Lake Ontario. In our present study, a total of 355 wildlife samples were tested for the presence of botulinum toxin and/or organisms. Type E botulinum toxin was detected in 110 samples from birds, 12 samples from fish, and 2 samples from mammals. Sediment samples from Lake Erie were also examined for the presence of C. botulinum. Fifteen of 17 sediment samples were positive for the presence of C. botulinum type E. Eighty-one C. botulinum isolates were obtained from plants, animals, and sediments; of these isolates, 44 C. botulinum isolates produced type E toxin, as determined by mouse bioassay, while the remaining 37 isolates were not toxic for mice. All toxin-producing isolates were typed by RAPD; that analysis showed 12 different RAPD types and multiple subtypes. Our study thus demonstrates that multiple genetically distinct strains of C. botulinum were involved in the present outbreak of wildlife botulism. We found that C. botulinum type E is present in the sediments of Lake Erie and that a large range of bird and fish species is affected.

Clostridium botulinum Group II Isolate Phylogenomic Profiling Using Whole-Genome Sequence Data

“…Several nucleic acid-based typing methods, including pulsed-field gel electrophoresis (PFGE), random amplification of polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), variable number tandem repeat (VNTR), multiple-locus sequence typing (MLST), DNA microarrays, and sequence analysis of the bont gene and the flagellin gene variable region (flaVR), have all been used for genetic characterization of C. botulinum group II strains (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24).…”

In Vitro Detection and Quantification of Botulinum Neurotoxin Type E Activity in Avian Blood

“…BoNT/E outbreaks in the Great Lakes region typically occur between June and December and correlate with low mean annual water levels and increased surface water temperatures (25). It has been hypothesized that BoNT/E is mobilized through Great Lakes aquatic food webs consisting of exotic species (19,25,32,33), providing a potential link between sediment-inhabiting clostridia and the intoxication of fish-eating birds. Using the BoTest Matrix E assay to conduct enhanced epidemiological investigations of avian BoNT/E outbreaks, including the analysis of aquatic food web components, would aid in our understanding of toxin mobilization pathways and provide critical insights for the management and conservation of bird species in the Great Lakes region.…”

“…Intoxication by BoNT serotype C (BoNT/C) and BoNT/E is a significant contributor to avian mortality worldwide (14,19,21,29,45,46). BoNT/E mortality events were first documented among fish-eating birds of the North American Great Lakes in the early 1960s, and outbreaks reemerged during the fall of 1999, with highest mortality documented among common loons, mergansers, long-tailed ducks, and gulls (34).…”

An Atypical Outbreak of Food-Borne Botulism Due to Clostridium botulinum Types B and E from Ham

“…C. botulinum type E strains show a high genetic variation, as tested by pulsedfield gel electrophoresis and randomly amplified polymorphic DNA (RAPD) (14,15). A previous analysis by MLST indicates that C. botulinum type E strains are split into 4 clades, with most strains belonging to the same clade (13).…”