Champika Fernando

Sentinel Surveillance for Zoonotic Parasites in Companion Animals in Indigenous Communities of Saskatchewan

Indigenous communities may have increased risk of exposure to zoonotic parasites, including Echinococcus granulosus, Toxocara canis, Toxoplasma gondii, Diphyllobothrium spp., and Giardia duodenalis, for which dogs may serve as sentinels for or sources of human infection. Canid fecal samples were collected from dogs and the environment in five indigenous communities across Saskatchewan and Alberta (N = 58, 62, 43, 66, and 25). Parasites in individual fecal samples were quantified using fecal flotation and a commercial immunofluorescent antibody test for Giardia and Cryptosporidium. Overall, the prevalence of canine intestinal parasitic infection was 20-71%, which is 5-16 times higher in indigenous communities than a nearby urban center in Saskatchewan. The overall prevalences of T. canis, Diphyllobothrium, and taeniid eggs in dog feces were, respectively, 11.8%, 4.9%, and 1.2% in our study compared with 0-0.2% in urban dogs. Giardia cysts present in 21% of samples were identified as zoonotic genotype Assemblage A.

Fecal shedding of Brachyspira spp. on a farrow-to-finish swine farm with a clinical history of “Brachyspira hampsonii”-associated colitis

BackgroundBrachyspira associated diarrhea is a re-emerging concern for Canadian swine producers. To identify critical control points for reducing the impact of Brachyspira on production, improved diagnostic tools and a better understanding of the on-farm epidemiology of these pathogens are required. A cross-sectional study was conducted for the detection of Brachyspira on a commercial, two-site, farrow-to-finish pork production unit in Saskatchewan, Canada with a clinical history of mucohaemorrhagic colitis associated with “B. hampsonii”.ResultsRectal swabs from pigs at all production stages were collected over 13 weeks (n = 866). Two swabs were collected per pig for culture and Gram stain, and for PCR. Ninety-one culture positive samples were detected, with the highest prevalence of Brachyspira shedding in grower pigs (21%). No Brachyspira were detected in pre-weaned piglets. PCR and Gram stain of rectal swabs detected fewer positive samples than culture. The most prevalent species detected was B. murdochii; other species detected included B. pilosicoli, B. innocens, and “Brachyspira hampsonii”. Phylogenetic analysis revealed that several of the isolates, including some strongly beta-haemolytic isolates, might represent novel taxa.ConclusionsOur results indicate that apparently healthy pigs can be colonized with diverse Brachyspira species, including some potential pathogens, and that frequency of shedding peaks in the grower stage. Difference in the detection rates of Brachyspira amongst culture, Gram stain or PCR on rectal swabs have implications for choice of detection methods and surveillance approaches that may be most effective in Brachyspira control strategies.

In vitro attenuation of a virulent swine isolate of Brachyspira hampsonii

Brachyspira hampsonii causes dysentery-like disease in infected pigs. Serial passage of a virulent swine isolate (P13) one-hundred times in laboratory culture medium was conducted to produce an attenuated strain, and to identify genomic determinants of virulence through comparison of genome sequences of the original and passaged strains. The resulting strain, P113, did not differ from P13 in terms of diagnostic biochemical characteristics but had an enhanced growth rate in culture, indicating laboratory adaptation. Whole genome sequencing of P113 revealed several single-nucleotide changes including a T to C transition that results in an R to G amino acid change in a putative mannose-1-phosphate guanylytransferase that is implicated in production of lipo-oligosaccharide. P113 was partially attenuated in a mouse model of infection, indicated by significantly fewer observations of abnormal feces in mice infected with P113 relative to P13. No differences were detected in bacterial shedding in feces, demonstrating that the ability of the organism to colonize mice was not affected. Passage through a mouse did not further alter the virulence of P113. Results of this study provide insight into genomic determinants of virulence in B. hampsonii and a live attenuated vaccine candidate.