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• We report discovery and phylogeography of a cryptic species of Trichinella, proposed as T. chanchalensis n. sp (T13). • Specimens of this cryptic species were initially identified as T. nativa based on multiplex PCR. • Exploration of Alaskan and Siberian isolates may further resolve the phylogeographically complex history of Trichinella. Understanding parasite diversity and distribution is essential in managing the potential impact of parasitic diseases in animals and people. Imperfect diagnostic methods, however, may conceal cryptic species. Here, we report the discovery and phylogeography of a previously unrecognized species of Trichinella in wolverine (Gulo gulo) from northwestern Canada that was indistinguishable from T. nativa using the standard multiplex PCR assay based on the expansion segment 5 (ESV) of ribosomal DNA. The novel genotype, designated as T13, was discovered when sequencing the mitochondrial genome. Phylogenetic analyses of the mitochondrial genome and of 15 concatenated single copy orthologs of nuclear DNA indicated a common ancestor for the encapsulated clade is shared by a subclade containing Trichinella spiralis and Trichinella nelsoni, and a subclade containing T13 and remaining taxa: T12 + (T2 + T6) + [(T5 + T9) + (T3 + T8)]. Of 95 individual hosts from 12 species of mammalian carnivores from northwestern Canada from which larvae were identified as T. nativa on multiplex PCR, only wolverines were infected with T13 (14 of 42 individuals). These infections were single or mixed with T. nativa and/or T6. Visual examination and motility testing confirmed that T13 is encapsulated and likely freeze-tolerant. We developed a new Polymerase Chain Reaction-Restriction Fragment Length Polymorphism which unequivocally distinguishes between T13 and T. nativa. We propose Trichinella chanchalensis n. sp. for T13, based on significant genetic divergence from other species of Trichinella and broad-based sampling of the Trichinella genome. Exploration of Alaskan and Siberian isolates may contribute to further resolution of a phylogeographically complex history for species of Trichinella across Beringia, including Trichinella chanchalensis n. sp. (T13). [ABSTRACT FROM AUTHOR]

Coyotes (Canis latrans) rapidly expanded across North America during the 20th century and in 1987 colonized insular Newfoundland, Canada. Their arrival brought the potential for new predator-prey interactions and the potential for transmission of parasites to naïve populations. Trichinella spp. and Echinococcus spp. are zoonotic parasites not previously reported from the island of Newfoundland, Canada. Muscle samples (diaphragm and tongue) from 153 coyotes and feces from 35/153 coyotes were collected. Larvae of Trichinella spp. were recovered by muscle digestion from 6/153 coyotes (3.9%) and identified using multiplex PCR and Sanger sequencing as T. nativa. Fecal samples were screened for DNA of Echinococcus spp. using qPCR, and intestines from positive animals were examined for adult cestodes. No fecal samples were positive for DNA of E. multilocularis, and 2/35 (5.7%) samples were positive for E. canadensis, of which one was successfully genotyped as the G10 cervid strain. Echinococcus canadensis has not previously been reported on the island of Newfoundland, historically the only region of Canada where Echinococcus spp. was not known to occur. No species of Trichinella have previously been reported on the island. Both parasites are zoonotic, and hunters, trappers, dog owners, and the general public should be aware of these new risks for public health., (© Wildlife Disease Association 2024.)

Prolonged eosinophilia is characteristic of trichinellosis. To determine the optimal eosinophil threshold for reflex Trichinella testing, we examined all 43 cases in Nunavik, Quebec, Canada, during 2009-2019. Using receiver operating characteristic analysis, we determined that eosinophil counts >0.8 × 10 9 cells/L should prompt consideration of trichinellosis and testing to rapidly identify potential outbreaks.

Abstract: Residents of Arctic communities are at increased risk of contracting infectious diseases transmitted by wildlife. Data collected from the International Polar Year Inuit Health Survey were used to determine the seroprevalence of 4 parasitic zoonoses in three Inuit jurisdictions of the Canadian Arctic and to assess risk factors of infection. To date, this is the most comprehensive survey of its kind. Immunoenzymatic methods were used for the detection of antibodies against Toxocara canis, Echinococcus granulosus, Trichinella sp., and Toxoplasma gondii. We determined the weighted prevalence of parasitic infections in 36 Inuit communities across the Inuvialuit settlement region, Nunavut, and Nunatsiavut. Our results indicate infrequent exposure to Toxocara and Echinococcus (1.7 and 6.3%, respectively). Exposure to T. gondii (27.2%) and Trichinella (18.6%) was more prevalent and was generally higher in Nunavut compared to other northern regions. Overall, seropositivity was related to age, education, and consumption of marine mammals and seafood. [Copyright &y& Elsevier]

Indirect enzyme-linked immunosorbent assay (ELISA) utilizing excretory-secretory (E-S) antigens of Trichinella spiralis is currently the method of choice for testing pigs and wild boars for exposure to Trichinella spp. The E-S proteins are released by first-stage larvae (L1) of this parasitic nematode maintained in vitro. However, the production of these antigens is cumbersome and time-consuming. The process requires animals to be experimentally infected with the parasite as the source of L1. Antigen production using recombinant technology would be more time- and cost-effective. In this study, we produced a Serpin of T. spiralis as a recombinant protein secreted by the yeast Pichia pastoris. The diagnostic performance of indirect ELISA with purified Serpin antigen was compared to that of E-S ELISA. Both Serpin ELISA and E-S ELISA demonstrated 98 % diagnostic specificity in testing 1056 pigs from the Canadian Trichinella-free commercial herd. Twenty of 21 pigs with non-negative test results in E-S ELISA tested negative by the confirmatory Western blot (WB) assay. Therefore, the diagnostic specificity of combined E-S ELISA and WB was 99.9 %. Forty-five sera collected at or after six weeks from 34 pigs experimentally infected with various numbers of T. spiralis L1 produced positive results in both E-S and Serpin ELISA, resulting in 100 % diagnostic sensitivity. However, testing of sera serially collected from four pigs experimentally infected with various low doses of T. spiralis L1 demonstrated a delayed Serpin-specific antibody response compared to seroconversion detected by E-S ELISA in three animals. Moreover, Serpin ELISA demonstrated significantly lower sensitivity for detecting antibodies induced by experimental infections of pigs with T. britovi, T. nativa, Trichinella T6 and T. pseudospiralis, suggesting that it will not provide consistent detection of exposure to sylvatic Trichinella spp. The validation data support the application of Serpin ELISA in seroepidemiological surveys for detecting exposure to T. spiralis in swine., (Crown Copyright © 2022. Published by Elsevier B.V. All rights reserved.)

Abstract: Freedom from infection modeling, using scenario trees, has become an established methodology and is well described in the literature. However, standards for organizing and reporting the surveillance information incorporated into such models are less developed. Canada has been routinely testing for Trichinella spiralis in market hogs in federally inspected slaughter plants since the late 1990s. By way of presenting our work on T. spiralis in Canadian hogs, we propose that information in surveillance models be organized in distinct categories, each with specific parameters and values that are thoroughly described and justified. The proposed categories are: (1) definitions for the objectives, (2) initial time period, (3) inputs, (4) data, (5) model settings, (6) outputs, and (7) validation. Having a standardized manner of reporting such studies will facilitate their validation and expedite their evaluation by experts in the field and their use in trade negotiations. [Copyright &y& Elsevier]

Abstract: A survey of wild carnivores in Canada was conducted over a 10-year period to determine the prevalence and genotypes of Trichinella. Muscle samples collected from 1409 animals representing 15 hosts species were enzymatically digested to recover Trichinella larvae. Larvae were recovered from a total of 287 (20.4%) animals and PCR identified four genotypes of Trichinella. Trichinella nativa was found in 5 host species and was the most commonly found genotype. Trichinella T6 was present in 7 species of carnivores, and coyote and badger are new host records for this genotype. The recovery of T. pseudospiralis and T. murrelli from cougars is the first documentation of these species in Canada and in cougars. The cougar was also the only host species in which all four genotypes of Trichinella were identified. Black bears and walruses had the highest tissue levels of larvae in this study and are also the species most frequently associated with human trichinellosis in Canada. This work identifies additional host species and expanded geographic ranges for 4 genotypes of Trichinella in North America. Failure to demonstrate T. spiralis in wildlife and continued negative results from ongoing surveillance activities in swine provide additional evidence that T. spiralis is not present in Canada. [Copyright &y& Elsevier]

Abstract  Infection of Trichinella spp. is common among animals in the Canadian Arctic. We determined the prevalence of Trichinella spp. infection in wolverines (Gulo gulo) from Nunavut, Canada. Diaphragms from 41 wolverines were examined by artificial digestion. Trichinella spp. larvae were detected in 36 (87.8%) examined animals. Trichinella T6 was detected in 33 (91.7%), Trichinella nativa in only one (2.8%), and a mixed Trichinella T6 and T. nativa infections were detected in two (5.6%) wolverines. This is the first report of Trichinella spp. infection in wolverines from Nunavut and the first report of sympatric Trichinella T6 and T. nativa in any host. The high prevalence of Trichinella spp. infection in combination with the natural history of wolverines suggests that the mustelid may be a key species in the natural cycle of these parasites in Arctic and Subarctic areas. [ABSTRACT FROM AUTHOR]

The population of grey seals Halichoerus grypus in Canadian waters is currently used as a commercial source of meat for human consumption. As with domestic livestock, it is important to understand the occurrence in these seals of infectious agents that may be of public health significance and thus ensure appropriate measures are in place to avoid zoonotic transmission. This study examined the prevalence of antibodies against Brucella spp., Erysipelothrix rhusiopathiae, 6 serovars of Leptospira interrogans, and Toxoplasma gondii in 59 grey seals and determined by polymerase chain reaction (PCR) the presence of these potentially zoonotic agents in specific organs and tissues of seropositive animals. The presence of encysted Trichinella spp. larvae was also investigated by digestion of tongue, diaphragm and other muscle samples, but none were detected. Seroprevalence against Brucella spp. and E. rhusiopathiae was low (5 and 3%, respectively). All 59 seals tested had antibodies against L. interrogans, but no carrier of this bacterium was detected by PCR. Seroprevalence against T. gondii was 53%, and DNA of this protozoan was detected by PCR in 11/30 (37%) seropositive animals. Standard sanitary measures mandatory for commercialization of meat products for human consumption should greatly reduce the potential for exposure to these infectious agents. However, special consideration should be given to freezing seal meat for at least 3 d to ensure destruction of tissue cysts of T. gondii.

Repeated outbreaks of trichinellosis caused by the consumption of Trichinella-infected walrus (Odobenus rosmarus) meat, which have sometimes led to serious morbidity, have stimulated Inuit communities in Nunavik (northern Quebec), Canada, to develop an innovative trichinellosis prevention program. The program involves preconsumption testing of meat samples from harvested walrus at a regional laboratory and the rapid dissemination of the results of such testing to communities. Local health authorities in Inukjuak conducted an epidemiological investigation after testing identified Trichinella-positive walrus meat in September 1997. This report describes the events that occurred before, during, and after the trichinellosis outbreak and also documents how the prevention program contributed to successful resolution of the outbreak.