Two novel toxin variants revealed by whole-genome sequencing of 175 Clostridium botulinum type E strains.

We sequenced 175 Clostridium botulinum type E strains isolated from food, clinical, and environmental sources from northern Canada and analyzed their botulinum neurotoxin (bont) coding sequences (CDSs). In addition to bont/E1 and bont/E3 variant types, neurotoxin sequence analysis identified two novel BoNT type E variants termed E10 and E11. Strains producing type E10 were found along the eastern coastlines of Hudson Bay and the shores of Ungava Bay, while strains producing type E11 were only found in the Koksoak River region of Nunavik. Strains producing BoNT/E3 were widespread throughout northern Canada, with the exception of the coast of eastern Hudson Bay.

Proposal of Bisgaardia hudsonensis gen. nov., sp. nov. and an additional genomospecies, isolated from seals, as new members of the family Pasteurellaceae.

Phenotypic and phylogenetic studies were performed on eight Gram-negative-staining, rod-shaped bacteria isolated from seals. Biochemical and physiological studies showed identical profiles for all of the isolates and indicated that they were related to the family Pasteurellaceae. 16S rRNA gene sequencing demonstrated that the organism represented a distinct cluster with two sublines within the family Pasteurellaceae with <96% sequence similarity to any recognized species. Multilocus sequence analysis (MLSA) including rpoB, infB and recN genes further confirmed these findings with the eight isolates forming a genus-like cluster with two branches. Genome relatedness as deduced from recN gene sequences suggested that the isolates represented a new genus with two species. On the basis of the results of the phylogenetic analysis and phenotypic criteria, it is proposed that these bacteria from seals are classified as Bisgaardia hudsonensis gen. nov., sp. nov. (the type species) and Bisgaardia genomospecies 1. The G+C content of the DNA was 39.5 mol%. The type strain of Bisgaardia hudsonensis gen. nov., sp. nov. is M327/99/2(T) (=CCUG 43067(T)=NCTC 13475(T)=98-D-690B(T)) and the reference strain of Bisgaardia genomospecies 1 is M1765/96/5 (=CCUG 59551=NCTC 13474).

Isolation and genetic characterization of Toxoplasma gondii from raccoons (Procyon lotor), cats (Felis domesticus), striped skunk (Mephitis mephitis), black bear (Ursus americanus), and cougar (Puma concolor) from Canada.

Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats (Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus) from Kuujjuaq, northern Quebec, Canada. Genotyping of these T. gondii isolates using polymorphisms at 10 nuclear markers including SAGI, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico revealed 4 genotypes. None of the isolates was clonal archetypal Types I, II, and III found in the United States. These results are in contrast with the Type II genotype that is widespread in domestic animals and humans throughout the United States and Europe. This is the first genotyping of T. gondii isolates from this part of North America.

Lead concentrations in ruffed grouse, rock ptarmigan, and willow ptarmigan in Québec.

Between 1996 and 1998, ruffed grouse, spruce grouse, willow ptarmigan, and rock ptarmigan harvested by hunters in Québec were examined for lead contamination. On examination of the gizzards of these birds, lead shot was found only in ruffed grouse (1.2%). The probability of ingestion of lead shot by grouse and ptarmigans is low. Analyses of the lead concentrations in the wing bones of grouse and ptarmigans and in the muscle tissue of ptarmigans were conducted. Although differences were observed between individuals based on age and sex, the mean concentrations measured were in the range of those that occur naturally at background levels (<6 microg/g dw). Lead concentrations in muscle tissue were low, often at the limit of detection. However, the few high concentrations detected were probably related to a lead pellet or bullet fragment. Based on an analysis of the health risk associated with consumption of ptarmigan muscle, we conclude that the use of lead ammunition for hunting gallinaceous birds may pose an unnecessary risk of lead poisoning because of the possible ingestion of lead shot, bullets, fragments or embedded shot.

Adenocarcinoma of the small intestine in a ringed seal from Hudson Bay.

Nineteen wild ringed seals (Phoca hispida) were killed in winter 1999 to assess the health status of seals harvested in eastern Hudson Bay (Quebec, Canada). One of these seals, an 11-yr-old male, had a poorly differentiated adenocarcinoma that severely constricted the lumen of the distal small intestine. The tumor was characterized by proliferation of polygonal epithelial cells that formed closely packed acini and cords. This appears to be the first reported case of adenocarcinoma of the small intestine in Pinnipedia.

Evaluation of the fluorescence polarization assay and comparison to other serological assays for detection of brucellosis in cervids.

The complement fixation test (CFT), competitive enzyme immunoassay (CELISA), indirect enzyme immunoassay (IELISA) and fluorescence polarization assay (FPA) were evaluated for the detection of antibodies to Brucella abortus and Brucella suis biotype 4 in caribou (Rangifer tarandus caribou), elk (Cervus elapus), red deer (Cervus elapus), and reindeer (Rangifer tarandus tarandus). When combining the data the FPA and the CELISA were determined to be the most suitable tests for serodiagnosis of Cervidae. The overall actual sensitivity of the CFT and the IELISA was 100%. The overall actual sensitivity for the CELISA and FPA was 99%. The overall relative specificity of the CFT (including treatment of anti-complementary data as positive or negative for analysis), the CELISA, the IELISA and the FPA were 65%, 93%, 99%, 99%, and 99%, respectively. The specificities of the buffered plate agglutination test (BPAT), the CFT, the CELISA, the FPA and the IELISA for 55 elk vaccinated with B. abortus strain 19 and tested 4 mo post vaccination were 14%, 31%, 51%, 84%, and 2%, respectively. The FPA is the diagnostic test of choice because it has sensitivity and specificity values comparable to the CELISA; it has the capability to distinguish vaccinal antibody and antibody resulting from exposure to cross-reacting organisms such as Yersinia enterocolitica 0:9 from antibody to Brucella spp. in most cases; it is technically simple to do; it is adaptable to field use and it is relatively inexpensive.

Movement preparation in adults with and without Down syndrome.

This study was designed to determine whether adults with mental retardation, with and without Down syndrome, display flexibility in the preparation of limb movements. A four-alternative target-aiming task involving movements with either the right or left hand to near or far targets was employed. Prior information about the movement to be made was manipulated by precuing specific features. Although reaction times and movement times were longer for subjects with than for those without mental retardation, the pattern of results was similar. Specifically, both retarded groups were able to use advance information to reduce movement preparation time. Moreover, information about movement distance was useful to subjects whether or not hand was precued. This finding indicates that the individuals with mental retardation were not limited by a fixed-order movement preparation process.