Additional diagnoses of Echinococcus multilocularis in eastern chipmunks (Tamias striatus) from southern Ontario - results from ongoing surveillance for E. multilocularis in intermediate hosts in Ontario, Canada.

Echinococcus multilocularis, a cestode with zoonotic potential, is now known to have a high prevalence in wild canid definitive hosts of southern Ontario. The distribution of E. multilocularis across this region in red foxes (Vulpes vulpes) and coyotes (Canis latrans) is widespread yet heterogenous. In contrast, confirmed diagnoses of E. multilocularis in wild free-ranging intermediate hosts within Ontario are currently limited to a single eastern chipmunk (Tamias striatus). These findings prompted ongoing surveillance efforts in intermediate host species, primarily rodents. Our report describes the results of passive surveillance through wildlife carcass submissions to the Canadian Wildlife Health Cooperative (CWHC) and targeted active sampling of small mammal species from 2018 to 2023; a second and third eastern chipmunk were found to be infected with E. multilocularis. However, these were the only occurrences from surveillance efforts which collectively totaled 510 rodents and other small mammals. Continued surveillance for E. multilocularis in intermediate hosts is of high importance in light of the recent emergence of this parasite in Ontario.

Local outbreak of spontaneous pneumothorax secondary to paragonimosis in southwestern Ontario dogs.

Three dogs were diagnosed with spontaneous pneumothorax and referred to the Ontario Veterinary College Health Sciences Centre for management. The 3 dogs were diagnosed with secondary spontaneous pneumothorax due to paragonimosis. The diagnosis was made by visualization of adult trematodes during surgical exploration with histopathological confirmation in 1 dog, and detection of trematode eggs via fecal sedimentation in the other 2 dogs. Two of the dogs developed unusual additional lesions, including hemoabdomen, muscle abscess, and abdominal adhesions. These were suspected to be secondary to aberrant fluke larval migration. All 3 dogs lived within a relatively small geographical area of Ontario and were hospitalized between December 2021 and March 2022. Each dog survived to discharge with surgical or medical management of the pneumothorax and treatment with a prolonged course of fenbendazole. Key clinical message: Paragonimosis should be considered as a differential diagnosis for canine spontaneous pneumothorax in areas where Paragonimus kellicotti is, or may be, endemic, or in dogs that have travelled to endemic areas - particularly if the patient has a history of cough or potential exposure to freshwater crayfish. Routine anthelmintic treatment does not prevent infection and standard fecal floatation methods may not detect the eggs. Therefore, diagnostic testing should include a fecal sedimentation test and thoracic radiographs to screen for P. kellicotti.

Éclosion locale de pneumothorax spontané secondaire à la paragonimose chez des chiens du sud-ouest de l'Ontario. Trois chiens ont reçu un diagnostic de pneumothorax spontané et ont été référés au Ontario Veterinary College Health Sciences Centre pour prise en charge. Les 3 chiens ont été diagnostiqués avec un pneumothorax spontané secondaire dû à la paragonimose. Le diagnostic a été posé par la visualisation des trématodes adultes lors de l'exploration chirurgicale avec confirmation histopathologique chez 1 chien, et la détection des oeufs de trématodes par sédimentation fécale chez les 2 autres chiens. Deux des chiens ont développé des lésions supplémentaires inhabituelles, notamment un hémoabdomen, un abcès musculaire et des adhérences abdominales. Ces lésions étaient soupçonnées d'être secondaires à une migration larvaire aberrante de la douve. Les 3 chiens vivaient dans une zone géographique relativement restreinte de l'Ontario et ont été hospitalisés entre décembre 2021 et mars 2022. Chaque chien a survécu jusqu'à sa sortie avec une prise en charge chirurgicale ou médicale du pneumothorax et un traitement avec un traitement prolongé au fenbendazole.Message clinique clé :La paragonimose doit être considérée comme un diagnostic différentiel du pneumothorax spontané canin dans les zones où Paragonimus kellicotti est, ou peut être, endémique, ou chez les chiens qui ont voyagé dans des zones endémiques ­ en particulier si le patient a des antécédents de toux ou une exposition potentielle aux écrevisses d'eau douce. Le traitement vermifuge de routine n'empêche pas l'infection et les méthodes de flottaison fécale standard peuvent ne pas détecter les oeufs. Par conséquent, les tests de diagnostic doivent inclure un test de sédimentation fécale et des radiographies thoraciques pour dépister P. kellicotti.(Traduit par Dr Serge Messier).

Factors associated with Giardia infection in dogs in southern Ontario, Canada.

Information concerning risk factors associated with Giardia infection in dogs in southern Ontario, Canada, is currently lacking. This study therefore aimed to identify risk factors for Giardia infection in dogs that visit off-leash dog parks in southern Ontario. From May-November 2018, fecal samples were collected from 466 dogs in 12 off-leash dog parks in the Niagara and Hamilton regions of Ontario. A survey that asked questions pertaining to travel history (i.e., area of residence, locations and regions visited in the previous 6 months), basic medical history (i.e., spay/neuter status, veterinary visits, use of deworming medication), consumption of a raw diet, and the physical (i.e., age, sex, breed) and behavioral characteristics (i.e., off-leash activities, hunting activities) of each dog sampled was administered to the respective owner. All fecal samples were examined with the Giardia plate ELISA (IDEXX Laboratories) for parasite antigen. Multivariable logistic regression analyses were conducted on the survey data to investigate putative risk factors for Giardia infection. Overall, 11.8% (95% CI: 9.2-15.1%) of samples tested positive for Giardia antigen. Results from the multivariable logistic regression analyses identified an interaction between dog age and spay/neuter status that was significantly associated with Giardia infection. The odds of infection were greater in intact as compared to neutered adult dogs (OR: 3.6, 95% CI: 1.7-7.9, p = 0.001), and in neutered juvenile dogs as compared to neutered adults (OR: 5.2, 95% CI: 2.2-12.2, p < 0.001). The results provide veterinarians with evidence-based information for identifying dogs at greatest risk of Giardia infection in southern Ontario.

Novel molecular diagnostic (PCR) diagnosis and outcome of intestinal Echinococcus multilocularis in a dog from western Canada.

OBJECTIVE: To describe the novel PCR diagnosis and outcome of intestinal Echinococcus multilocularis in a dog. ANIMAL: A 13-month-old female intact dog with naturally occurring intestinal E multilocularis. CLINICAL PRESENTATION, PROGRESSION, AND PROCEDURES: The 13-month-old dog initially presented with a reduced appetite and weight loss and then developed hematochezia. The clinical history included a lack of endoparasite preventive care (fecal testing, deworming), exposure to coyotes, fox, sheep, and rodents and the dog had intermittently been fed a raw food diet. Physical examination revealed a thin dog, with a 2/9 body condition score, that was otherwise clinically unremarkable. A fecal sample was submitted for screening for gastrointestinal parasites as part of an infectious disease assessment. The fecal PCR test reported detection of E multilocularis. This result was sequenced as the European haplotype E3/E4. Centrifugal flotation (same sample) did not detect taeniid eggs. TREATMENT AND OUTCOME: The dog was treated with metronidazole, maropitant, and milbemycin oxime/praziquantel. Clinical improvement was noted within 48 hours. No DNA of E multilocularis was detected in a fecal sample collected approximately 10 days after treatment. The dog's owner was advised to provide monthly deworming (praziquantel) for all dogs on the property and to contact their human health-care provider due to potential zoonotic exposure risk. CLINICAL RELEVANCE: Increasing detection of E multilocularis is occurring in dogs in Canada and the US. Alveolar echinococcosis can cause severe disease in dogs and humans. Fecal PCR detection and surveillance may alert practitioners to canine intestinal cases and allow dogs to serve as sentinels for human exposure risk.

Identifying hepatic genes regulating the ovine response to gastrointestinal nematodes using RNA-Sequencing.

Gastrointestinal nematode (GIN) infections are considered the most important disease of grazing sheep and due to increasing anthelmintic resistance, chemical control alone is inadequate. Resistance to Gastrointestinal nematode infection is a heritable trait, and through natural selection many sheep breeds have higher resistance. Studying the transcriptome from GIN-exposed and GIN-unexposed sheep using RNA-Sequencing technology can provide measurements of transcript levels associated with the host response to Gastrointestinal nematode infection, and these transcripts may harbor genetic markers that can be used in selective breeding programs to enhance disease resistance. The objective of this study was to compare liver transcriptomes of sheep naturally exposed to Gastrointestinal nematode s, with either high or low parasite burdens, to GIN-unexposed control sheep in order to identify key regulator genes and biological processes associated with Gastrointestinal nematode infection. Differential gene expression analysis revealed no significant differentially expressed genes (DEG) between sheep with a high or low parasite burden (p-value ≤0.01; False Discovery Rate (FDR) ≤ 0.05; and Fold-Change (FC) of > ±2). However, when compared to the control group, low parasite burden sheep showed 146 differentially expressed genes (64 upregulated and 82 downregulated in the low parasite burden group relative to the control), and high parasite burden sheep showed 159 differentially expressed genes (57 upregulated and 102 downregulated in the low parasite burden group relative to the control) (p-value ≤0.01; FDR ≤0.05; and FC of > ±2). Among these two lists of significant differentially expressed genes, 86 differentially expressed genes (34 upregulated, 52 downregulated in the parasited group relative to the control) were found in common between the two parasite burden groups compared to the control (GIN-unexposed sheep). Functional analysis of these significant 86 differentially expressed genes found upregulated genes involved in immune response and downregulated genes involved in lipid metabolism. Results of this study offer insight into the liver transcriptome during natural Gastrointestinal nematode exposure that helps provide a better understanding of the key regulator genes involved in Gastrointestinal nematode infection in sheep.

Vector-borne and other pathogens of potential relevance disseminated by relocated cats.

Large populations of unowned cats constitute an animal welfare, ecological, societal and public health issue worldwide. Their relocation and homing are currently carried out in many parts of the world with the intention of relieving suffering and social problems, while contributing to ethical and humane population control in these cat populations. An understanding of an individual cat's lifestyle and disease status by veterinary team professionals and those working with cat charities can help to prevent severe cat stress and the spread of feline pathogens, especially vector-borne pathogens, which can be overlooked in cats. In this article, we discuss the issue of relocation and homing of unowned cats from a global perspective. We also review zoonotic and non-zoonotic infectious agents of cats and give a list of practical recommendations for veterinary team professionals dealing with homing cats. Finally, we present a consensus statement consolidated at the 15th Symposium of the Companion Vector-Borne Diseases (CVBD) World Forum in 2020, ultimately to help veterinary team professionals understand the problem and the role they have in helping to prevent and manage vector-borne and other pathogens in relocated cats.

Prevalence of intestinal parasites in dogs in southern Ontario, Canada, based on fecal samples tested using sucrose double centrifugation and Fecal Dx® tests.

In southern Ontario, Canada, there is a lack of information concerning the prevalence of intestinal parasites in dogs. As such, this study aimed to characterize the prevalence of intestinal parasites in dogs visiting off-leash parks in the region using sucrose double centrifugation and Fecal Dx® tests. Additionally, data obtained via the sucrose double centrifugation method were used to evaluate the performance of the Fecal Dx® tests. Fecal samples were collected from 466 dogs aged ≥6 months from May to November 2018 (mean age = 3.7 years). Overall, eleven intestinal parasites were identified using sucrose double centrifugation. Roundworm eggs (Toxocara canis and Baylisascaris procyonis), hookworm eggs (Ancylostoma caninum and Uncinaria stenocephala), and whipworm eggs (Trichuris vulpis) were identified in 1.07% (95% confidence interval [CI] 0.38-2.56%), 5.79% (95% CI 3.85-8.31%), and 5.15% (95% CI 3.33-7.57) of samples, respectively. Using the Fecal Dx® tests, 1.07% (95% CI 0.38-2.56%), 4.29% (95% CI 2.64-6.55%), and 2.15% (95% CI 1.03-3.91) of the samples tested positive for roundworm, hookworm, and whipworm antigen, respectively. To assess the level of agreement between the Fecal Dx® tests and sucrose double centrifugation, three methods were used. Cohen's kappa indicated a fair-to-moderate level of agreement between Fecal Dx® tests and sucrose double centrifugation. In contrast, the prevalence-adjusted bias-adjusted kappa and Gwet's first-order agreement coefficient indicated almost perfect agreement between these tests, ranging from 0.87 to 0.99 among the parasites examined. This study provides valuable information on the prevalence of intestinal parasites in mature dogs in southern Ontario that will help guide parasite control recommendations for dogs in this region.

MORBIDITY AND MORTALITY IN ONTARIO RODENTS AND LAGOMORPHS: A 30-YEAR RETROSPECTIVE REVIEW.

Passive surveillance is an important component of wildlife health surveillance that allows for the identification of emerging pathogens as well as population-level threats. We investigated the most common causes of morbidity and mortality in rodents and lagomorphs submitted to the Canadian Wildlife Health Cooperative (CWHC) in Ontario and the Ontario Veterinary College (OVC) over a 30-yr period. A total of 836 cases representing 13 species of rodents and three species of lagomorph were submitted to the CWHC and the OVC wildlife pathology service. Infectious or inflammatory diagnoses were most common in our data set, followed by trauma and unknown diagnoses. The most frequently identified primary diagnosis was encephalitis with histological lesions consistent with neural larva migrans including the presence of inflammation and malacia of brain tissue and, in some cases, characteristic nematode larvae. Other infectious diagnoses were squirrel fibroma virus and Toxoplasma gondii infections. Knowledge of common pathogens observed in various species of rodents and lagomorphs can aid in triage and treatment decisions at veterinary clinics and wildlife rehabilitation centers, and guide sample collection and test requisition at post-mortem examination.

Investigation of the occurrence of Angiostrongylus vasorum in coyotes in southern Ontario, Canada.

In North America, the only endemic focus for Angiostrongylus vasorum (French heartworm) was historically thought to occur in the southeastern part of the island of Newfoundland. However, reports of A. vasorum infection in wild canids in West Virginia, USA, and Nova Scotia, Canada, suggest the introduction of the parasite to mainland North America. We screened for A. vasorum in coyotes from across southern Ontario. Additionally, we evaluated the performance of ELISAs for detection of circulating A. vasorum antigen (Ag-ELISA) and antibodies against A. vasorum (Ab-ELISA) designed for use in sera or blood of foxes for use with coyotes in this region. Autopsies were performed on 397 coyotes, and lung tissue extract prepared from each carcass was tested via both ELISAs. The sensitivity and specificity for both tests were estimated in the absence of a gold standard using a 2-test single population Bayesian model; sensitivity and specificity priors were based on the performance of the assays in foxes in Switzerland. Eight coyotes tested positive for A. vasorum antigen; no animal was antibody positive. The estimated sensitivity and specificity of the Ag-ELISA were 90.8% (95% credible interval [CrI]: 83.8-95.6%) and 95.5% (95% CrI: 93.4-97.2%), respectively. For the Ab-ELISA, the estimated sensitivity and specificity were 41.9% (95% CrI: 32.1-51.9%) and 98.0% (95% CrI: 96.3-99.0%), respectively. Based on these findings and negative postmortem data for the same animals, there is insufficient evidence to suggest the presence of A. vasorum in southern Ontario coyotes.

Evaluation of the prevalence of Echinococcus multilocularis in dogs that visit off-leash dog parks in southern Ontario, Canada.

Prior to 2012, Echinococcus multilocularis was not known to occur in any host in Ontario, Canada. However, since that year, five cases of alveolar echinococcosis have been diagnosed in dogs that resided at the western end of Lake Ontario. In addition, E. multilocularis has been shown to be a common infection in wild canids (i.e. coyotes and foxes) across southern Ontario with a high-risk infection cluster in the area surrounding the western shores of Lake Ontario and northern shores of Lake Erie. In regions endemic for E. multilocularis, dog ownership is considered a risk factor for human alveolar echinococcosis. A study was therefore carried out to determine the prevalence of E. multilocularis intestinal infections in dogs within the high-risk infection cluster. From May to November 2018, faecal samples were collected from 477 dogs aged ≥6 months that visited 12 off-leash dog parks in the Halton, Hamilton and Niagara public health units. Faecal samples were analysed via a magnetic capture probe DNA extraction and real-time PCR method for E. multilocularis DNA. Overall, 0% (97.5% CI: 0%-0.80%) of samples tested positive. This result informs preventive recommendations for E. multilocularis infections in dogs in this region.

Spatio-temporal clustering of Baylisascaris procyonis, a zoonotic parasite, in raccoons across different landscapes in southern Ontario.

Baylisascaris procyonis, the raccoon roundworm, is a parasite found throughout North America and parts of Europe. More than 150 species of mammals and birds including humans can develop neurological disease following infection with the larval stage of this parasite. To investigate whether B. procyonis infections in raccoons cluster in space, time, or space-time, we used data from 1353 Ontario raccoons submitted to the Canadian Wildlife Health Cooperative between 2013 and 2016. We identified a significant spatial cluster of increased infection prevalence in southern Ontario centered over a major metropolitan area, as well as a significant cluster of decreased infection prevalence in a primarily agricultural region in southwestern Ontario. Furthermore, we identified statistically significant temporal clusters in the fall in annual scans of data from 2014, 2015 and 2016. Examination of both Bernoulli and space-time permutation models for space-time analysis suggested that the purely spatial and temporal clusters were not explained by relatively short and spatially discrete events in space-time. The identified annual temporal clusters are consistent with previous research on the seasonality of B. procyonis infection in raccoons. Recognition of the spatial infection clusters will help identify potential geographic and anthropogenic factors associated with the occurrence of B. procyonis infection in raccoons. Given the zoonotic potential of this parasite, identification of a cluster of high infection prevalence in a major metropolitan area has implications for public education and risk management strategies.

Correlation of subclinical gastrointestinal nematode parasitism with growth and reproductive performance in ewe lambs in Ontario.

Infection with gastrointestinal nematode parasites (GINs) is an important cause of productivity loss on sheep farms in Ontario and worldwide. However, efforts to quantify the effect of GIN infection on growth have demonstrated mixed results. Furthermore, there has been limited investigation of their effect on reproductive performance. This study evaluated the effect of subclinical GIN parasitism on growth and reproductive performance of ewe lambs under Ontario grazing conditions. Rideau cross ewe lambs (n = 140) born in spring 2016 on a farm in central Ontario were followed for two years from before weaning through to November 2017, including their first lambing and lactation. These animals grazed from May to November of each year and were sampled every 6-8 weeks during both grazing seasons and once at mid-gestation in March 2017. At each sampling the ewe lambs were weighed, body condition scores assigned, fecal egg counts (FECs) performed, and pasture samples collected to assess number of infective GIN larvae. Study animals with a FEC of 500 eggs per gram or higher were selectively treated with anthelmintics to prevent morbidity and mortality. Fecal samples were cultured to determine infecting GIN species, and climate data were obtained from a weather station 26 km away from the farm. Precipitation levels and numbers of infective larvae on pasture were low during the first grazing season but were more typical of Ontario conditions in the second grazing season. The three most common GIN species were Haemonchus contortus, Teladorsagia circumcincta, and Trichostrongylus spp. General linear mixed models were generated for weight change over time, litter size at lambing, and weaning weights of offspring. Despite moderate peak GIN burdens in both grazing seasons, FEC was not significantly associated with weight change or litter size, apart from periparturient egg rise in study ewe lambs with larger litters (p = 0.05). Significant positive quadratic and negative linear associations were identified between late lactation FECs and offspring weaning weights; the association between FECs and weaning weights changed from negative to positive at a FEC of 361 eggs per gram. These results indicate that when GIN burdens are moderate as evidenced by fecal egg counts and infection is subclinical, there appears to be low to no impact on growth and reproductive performance in ewe lambs in the first 18 months of life. This suggests that when GIN parasitism is regularly monitored and controlled using targeted selective treatment, animal performance is minimally affected.

Correlation of salivary antibody to carbohydrate larval antigen (CarLA) with health and gastrointestinal nematode parasitism in sheep under Ontario grazing conditions.

Leveraging mucosal immunity is a promising method for controlling gastrointestinal nematode (GIN) parasitism in sheep. Salivary antibody to carbohydrate larval antigen (sCarLA), a heritable measure of immunity to third-stage GIN larvae (L3), has been successfully applied to genetic improvement programs in New Zealand. However, sCarLA levels wane in the absence of ongoing GIN exposure. New Zealand's temperate climate permits year-round exposure to L3, but cold winters in boreal regions such as Ontario, Canada interrupt exposure for five months or more. This study investigated associations between sCarLA levels, GIN parasitism, and indicators of overall health in sheep grazing under Ontario conditions. A commercial flock of 140 Rideau cross ewe lambs were followed from approximately 30 days of age in May 2016 until November 2017, including lambing and lactation in the spring of 2017. Every 6-8 weeks during the grazing season and at mid-gestation in March 2017, fecal egg counts were performed, blood collected to assess serum albumin, globulin, and hematocrit, and pasture samples obtained to confirm exposure to infective larvae. Measurements of sCarLA level were performed at the beginning, middle, and end of each grazing season, and at mid-gestation. Spearman's rank correlation coefficients were calculated to compare sCarLA levels over time, and general linear mixed models created to evaluate associations between sCarLA levels, GIN fecal egg count, hematocrit, serum albumin, and serum globulin. Levels of sCarLA followed a similar seasonal pattern to GIN fecal egg counts with a 6-8 week delay; much higher sCarLA levels were observed in the second grazing season. The proportion of the flock with detectable sCarLA (≥ 0.3 units/mL) was 68.3 % by the end of the first grazing season, declined over winter to 43.9 % at lambing, and approached 100 % after 3 months of grazing in the second grazing season. Correlations between sCarLA levels over time were consistently positive, of weak to moderate strength, and significant (p < 0.05). At all time points, sCarLA level was significantly (p < 0.001) and negatively associated with fecal egg counts. The flock displayed minimal variability in hematocrit, serum albumin, and serum globulin; none of which were significantly associated with sCarLA levels. These results suggest that sCarLA can be maintained over winter and is a useful measure of immunity to GINs in sheep under Ontario grazing conditions.

Evaluation of the SNAP® 4Dx® plus test for the detection of Dirofilaria immitis antigen and characterization of exposure to tick-borne pathogens in wild canids in southern Ontario.

Population-level surveys for Dirofilaria immitis in wild canids typically rely on identification of the parasite at necropsy. More recently, some studies have employed the use of the SNAP® 4Dx® Plus test. However, since the assay was designed for use with domestic dogs it needs to be validated for use with wild canids for accurate interpretation of results. We therefore evaluated the performance of the SNAP® 4Dx® Plus test for detection of D. immitis in wild canids in southern Ontario. Overall, 199 wild canid carcasses were collected from across the region and assessed for the presence of D. immitis parasites at necropsy; ten were infected. Lung tissue extract (LE) and thoracic fluid filter paper extract (TFE) prepared from each wild canid were tested via the SNAP® 4Dx® Plus test, which simultaneously tests for the presence of D. immitis antigen and antibodies to Borrelia burgdorferi, Anaplasma spp., and Ehrlichia spp. The prevalence adjusted bias adjusted kappa (PABAK) and Gwet's first-order-agreement coefficient (AC1) were used to assess the level of agreement between sample pairs. The PABAK and AC1 between LF and TFE applied to the SNAP® 4Dx® Plus test and the necropsy-confirmed D. immitis status indicated a very high level of agreement for all sample pairs. Compared to necropsy-confirmed D. immitis status, the estimated sensitivity and specificity of the SNAP® 4Dx® Plus test for D. immitis antigen in LE was 80 % (95 % CI 44.4-97.5%) and 98.9 % (95 % CI 96.2-99.9%), respectively. For the TFE, the sensitivity was 70 % (95 % CI 34.8-93.3%) and the specificity was 97.9 % (95 % CI 94.7-99.4%). With respect to the tick-borne pathogen components, 1.5 % (3/199; 95 % CI 0.3-4.5%) of wild canids tested positive for B. burgdorferi antibody; 1.0 % (2/199; 95 % CI 0-3.8%) of LE samples were positive and 0.5 % (1/199; 95 % CI 0-3.1%) of TFE samples were positive. No samples tested positive for antibody to Anaplasma spp. (95 % CI 0-2.3%) or Ehrlichia spp. (95 % CI 0-2.3%). Collectively, the results suggest that the SNAP® 4Dx® Plus test may be a suitable test for use with LE and TFE for the detection of D. immitis antigen in wild canids from southern Ontario.

Prevalence of Baylisascaris procyonis in raccoon latrines in southern Ontario, Canada.

Raccoon latrines represent sites of potential infection by the zoonotic parasite Baylisascaris procyonis for wildlife and humans. Our objective was to determine the prevalence of B. procyonis at raccoon latrine sites in southern Ontario. Thirty raccoon latrines were sampled between June - July 2018; multiple scats were collected and homogenized to form a representative sample of each latrine. To determine the presence or absence of B. procyonis eggs in each sample, we used the Cornell-Wisconsin centrifugal floatation technique. Twenty-three percent (7/30) of homogenized samples tested positive for B. procyonis. Eggs per gram of feces ranged from 1 to 388 (median = 1.28, IQR = 0.32-232.5). Baylisascaris procyonis positive latrines were found in conservation areas heavily used by people, which may represent a possible source of exposure for humans in these areas.

Factors associated with Echinococcus multilocularis infection in coyotes in southern Ontario.

Echinococcus multilocularis was recently reported in wild canids across southern Ontario, a newly recognized endemic area in Canada. In such areas, a comprehensive understanding of factors associated with infection in definitive hosts (wild canids) is critical for mitigating risk of transmission to humans. However, little is known about the transmission dynamics of the parasite in definitive hosts for this region. A study was therefore carried out to investigate the association of host-level (sex, body condition), environmental (southern Ontario region, land cover), temporal (season, hunting season, calendar year) and extraneous factors (submitter type) with E. multilocularis infection in coyotes in southern Ontario. Between November 2015 and March 2017, 416 coyotes were collected from across the region as part of a study that investigated the prevalence and distribution of the parasite in wild canids; approximately 24% of coyotes were positive for E. multilocularis. Associations between infection and factors of interest were assessed via a mixed-effects logistic regression model with a random intercept for submitter to account for clustering. Coyotes with poor body condition were at greater odds of E. multilocularis infection than those in good condition (odds ratio [OR] 2.14; 95% CI: 1.08-4.26; p = .030). A negative association was observed between infection in coyotes and the proportion of natural land in a coyote's estimated home range (OR: 0.67; 95% CI: 0.52-0.85; p = .001). Coyotes from the western region of southern Ontario had lower odds of infection compared to coyotes from the central region (OR: 0.26; 95% CI: 0.12-0.55; p < .001). These results can be used to help guide future public health prevention strategies for human alveolar echinococcosis.

Comparison of ITS-2 rDNA nemabiome sequencing with morphological identification to quantify gastrointestinal nematode community species composition in small ruminant feces.

Mixed gastrointestinal nematode (GIN) infections are a common and significant cause of financial loss for small ruminant producers. Morphologic examination of third-stage larvae (L3) can be used to identify species composition in feces but has limitations due to the requirement for specialized expertise and the extensive time (8-15 d depending on method used) and labour involved. Moreover, differential development and survival of larvae during coproculture to the third stage often occurs. Deep amplicon sequencing of the ITS-2 rDNA locus of first-stage larvae (L1) allows for higher throughput with reduced specialist labour and reduces the risk of misidentification. Harvesting of L1 soon after hatching is also faster and further reduces labour as well as biases that can occur due to differential larval development and survival. This study compares the results of morphologic examination of L3 with those of ITS-2 rDNA deep amplicon sequencing of L1 from a set of pooled fecal samples. The proportions of eggs that were successfully recovered as larvae following culture to L3 and L1 were also compared. Larval recovery rate was significantly lower from L3 cultures than from L1 cultures (p < 0.001); eggs were 238.7 times less likely to develop to L3 than to L1 (95 % confidence interval for odds ratio 80.0-712.0). Significantly lower proportions of Teladorsagia circumcincta (odds ratio = 3.1, p = 0.008) and higher proportions of Trichostrongylus spp. (p = 0.009) were identified using morphologic examination of L3 compared with deep amplicon sequencing of L1 on the same samples. This is consistent with previous reports of differential survival of these species in L3 cultures. These results indicate that deep amplicon sequencing of L1 may reduce bias introduced by differential GIN survival to L3 in small ruminants.

DEMOGRAPHIC AND ENVIRONMENTAL FACTORS ASSOCIATED WITH BAYLISASCARIS PROCYONIS INFECTION OF RACCOONS (PROCYON LOTOR) IN ONTARIO, CANADA.

The raccoon (Procyon lotor) roundworm, Baylisascaris procyonis, is an emerging wildlife zoonosis of public health significance in North America. Although the adult stage typically causes no disease in raccoons, the larval stage can cause significant disease in a variety of species, including humans. Raccoons often use human environments, which may increase the risk of B. procyonis exposure in people, particularly in urban settings. Because of this, our objectives were to identify host and environmental risk factors associated with the prevalence and intensity of B. procyonis infection in raccoons in Ontario, Canada. Between 2013 and 2016, 1,539 raccoons were collected and examined for the presence of B. procyonis. Thereafter, we analyzed our data for the influence of age, sex, fat stores, human population size, land use classification, season, and year of collection on the prevalence and intensity of infection. With multilevel logistic regression models, we identified significant associations between prevalence and host age, prevalence and amount of fat stores, and prevalence and season of collection; a significant two-way interaction was also identified between host sex and land use classification. Additionally, by using multilevel negative binomial regression models, we identified significant associations between the intensity of parasite infection and season of collection, as well as three significant two-way interactions: host sex and land use classification, host age and land use classification, and host sex and amount of fat stores. These findings help provide a more complete understanding of B. procyonis ecology in raccoons, including identifying associations between different environments and B. procyonis, which may assist in the development of future risk management strategies.

Prevalence and distribution of Dirofilaria immitis infection in wild canids in southern Ontario.

Wild canids represent a potential reservoir host for Dirofilaria immitis infection in dogs in Ontario. Since wild canids are not protected by chemoprophylaxis, understanding the epidemiology of D. immmitis in these populations may help elucidate the background risk of infection for dogs. The aim of this study was to determine the prevalence and distribution of D. immitis infection in wild canids across southern Ontario. From February 2016 to March 2017, 290 wild canid carcasses (273 coyotes and 17 foxes) were collected from across the region and assessed for the presence of D. immitis at the time of necropsy. Overall, D. immitis infection was identified in 4.8% (95% CI 2.8-8.0%) of these wild canid carcasses. Among coyotes, 5.1% (95% CI 3.0-8.5%) were positive; no evidence of D. immitis was found in the 17 foxes. Dirofilaria immitis infections in wild canids were detected in two regions of southern Ontario: 12 of the 14 D. immitis infections were detected in the south-western region and two were detected in the eastern region. Our findings provide preliminary insights into the prevalence and geographical distribution of D. immitis in coyotes and foxes in southern Ontario.

Epidemiology of canine heartworm (Dirofilaria immitis) infection in domestic dogs in Ontario, Canada: Geographic distribution, risk factors and effects of climate.

Dirofilaria immitis is the causal agent of heartworm, a mosquito-borne parasite that primarily infects domestic and wild canids. The infection is endemic in parts of Canada, and Ontario has been identified as the province where the majority of heartworm infections occur. Test results for blood samples submitted by veterinary clinics for the years 2007-2016 were used to conduct a spatial risk analysis of heartworm among domestic dogs in Ontario. The geographic extent of the apparent heartworm prevalence was examined through smoothed choropleth maps for all 49 census division regions. Furthermore, the regions were assessed for local clusters in apparent prevalence using the flexible spatial scan statistic. Three clusters were found and located in western, southern and eastern Ontario, respectively. A spatial Poisson regression model for heartworm prevalence among pet dog populations in southern Ontario census divisions was fit to determine the association between human population size, heartworm development units (HDUs), climate moisture index (CMI), precipitation and directions, east or north, with heartworm infection. The model identified the spatial distribution of HDUs and CMI as positively associated with heartworm infection and therefore important predictors of the infection. In contrast, human population size, increasing northern latitude and drier areas were negatively associated with heartworm infection. The east direction and precipitation were not significant.