Echinococcus multilocularis infection in a red fox (Vulpes vulpes) on Prince Edward Island, Canada.

Objective: Molecular identification of small cestodes, morphologically consistent with Echinococcus multilocularis, recovered at necropsy from the gastrointestinal tract contents of a red fox, was accomplished by PCR using published species-specific n ad1 primers and methods. Animal: Red fox (Vulpes vulpes). Procedure: Small cestodes recovered from intestinal contents of a red fox trapped on Prince Edward Island in December 2020 (frozen at -20°C before being processed for parasite recovery in June 2021) were morphologically identified. Species identity confirmation and haplotyping of the cestodes were done via PCR and DNA sequencing of the n ad1, nad2, and cob genes. Results: Small cestodes morphologically consistent with E. multilocularis were detected in the gastrointestinal tract contents of a red fox trapped near Montague, PEI. The species identity was confirmed via PCR. Haplotyping revealed that they were of the European E1 haplotype. Conclusion: In Canada, E. multilocularis has been reported as far east as Québec, with most reports being in central and western provinces and territories. This is the first report of E. multilocularis infection in a canid host east of Ontario, Canada and illustrates the need for regular wildlife disease surveillance to enhance our understanding of emerging pathogens of veterinary and medical importance. Clinical Relevance: Echinococcus multilocularis is a highly pathogenic zoonotic cestode from the family Taeniidae that can cause alveolar echinococcosis (AE) when rodents, dogs, horses, pigs, non-human primates, or humans ingest its eggs. Alveolar echinococcosis is challenging to treat, and survival rates for untreated individuals are low.

Objectif: L'identification moléculaire de petits cestodes, morphologiquement compatibles avec Echinococcus multilocularis, récupérés à l'autopsie du contenu du tractus gastro-intestinal d'un renard roux, a été réalisée par PCR à l'aide d'amorces nad1 spécifiques à l'espèce et de méthodes publiées. Animal: Renard roux (Vulpes vulpes). Procédure: De petits cestodes récupérés du contenu intestinal d'un renard roux piégé à l'Île-du-Prince-Édouard en décembre 2020 (congelés à −20 °C avant d'être traités pour la récupération des parasites en juin 2021) ont été morphologiquement identifiés. La confirmation de l'identité des espèces et l'haplotypage des cestodes ont été effectués par PCR et séquençage de l'ADN des gènes nad1, nad2 et cob. Résultats: De petits cestodes morphologiquement compatibles avec E. multilocularis ont été détectés dans le contenu du tractus gastro-intestinal d'un renard roux piégé près de Montague, Î.-P.-É. L'identité de l'espèce a été confirmée par PCR. L'haplotypage a révélé qu'ils étaient de l'haplotype européen E1. Conclusion: Au Canada, E. multilocularis a été signalé aussi loin à l'est que le Québec, la plupart des signalements ayant été rapportés dans les provinces et territoires du centre et de l'ouest. Il s'agit du premier rapport d'infection à E. multilocularis chez un canidé hôte à l'est de l'Ontario, au Canada, et illustre la nécessité d'une surveillance régulière des maladies de la faune pour améliorer notre compréhension des agents pathogènes émergents d'importance vétérinaire et médicale. Pertinence clinique: Echinococcus multilocularis est un cestode zoonotique hautement pathogène de la famille des Taeniidae qui peut provoquer une échinococcose alvéolaire (EA) lorsque des rongeurs, des chiens, des chevaux, des porcs, des primates non humains ou des humains ingèrent ses oeufs. L'échinococcose alvéolaire est difficile à traiter et les taux de survie des personnes non traitées sont faibles.(Traduit par Dr Serge Messier).

First report of Angiostrongylus vasorum (French heartworm) in red foxes (Vulpes vulpes) on Prince Edward Island.

Objective: To identify first-stage nematode larvae (L1) recovered from a red fox scat sample and adult female worms recovered from 2 red fox lungs at necropsy, using published molecular methods to confirm a morphological diagnosis of Angiostrongylus vasorum (French heartworm). Animal: Red fox (Vulpes vulpes). Procedure: Nematode larvae recovered from a Baermann examination survey of wild canid scats (n = 101) conducted from January 2017 to August 2020, were identified by size and morphology and subjected to PCR and DNA sequencing of the small subunit (SSU) rRNA gene, the large subunit (LSU) rRNA gene, or the second internal transcribed spacer (ITS2). In addition, these techniques were applied to adult female worms recovered from the heart/lungs of 2 red foxes (obtained from PEI trappers and stored frozen at -20°C since December of 2018 and 2020). Results: Size and morphology of L1 recovered by Baermann examination from a wild canid scat sample (presumed to be red fox) collected near Montague, PEI and adult female worms recovered at necropsy from 2 red fox carcasses were identified as A. vasorum. Molecular analysis confirmed the larvae and adult worms were A. vasorum. Conclusion: These findings indicated that A. vasorum has become endemic in the red fox population on PEI. Clinical relevance: Angiostrongylus vasorum infection is potentially fatal in dogs. Veterinarians and regional diagnostic laboratories in the Maritime provinces should consider the possibility of A. vasorum infection in dogs with clinical signs of cardiopulmonary and/or central nervous system disease or bleeding disorders.

Objectif: Identifier les larves de nématodes de premier stade (L1) récupérées à partir d'un échantillon d'excréments de renard roux et les vers femelles adultes récupérés à partir de deux poumons de renard roux à l'autopsie, en utilisant des méthodes moléculaires publiées pour confirmer un diagnostic morphologique d'Angiostrongylus vasorum (ver du coeur français). Animal: Renard roux (Vulpis vulpis). Procédure: Les larves de nématodes récupérées lors d'une enquête sur des excréments de canidés sauvages (n = 101) par examen Baermann menée de janvier 2017 à août 2020, ont été identifiées par taille et morphologie et soumises à la PCR et au séquençage de DNA de la petite sous-unité (SSU) du gène de rRNA, de la grande sous-unité (LSU) du gène de rRNA ou du deuxième espaceur interne transcrit (ITS2). De plus, ces techniques ont été appliquées à des vers femelles adultes récupérés du coeur/poumons de deux renards roux (obtenus auprès de trappeurs de l'Î.-P.-É. et conservés congelés à −20 °C depuis décembre 2018 et 2020). Résultats: La taille et la morphologie de L1 récupérées par examen Baermann à partir d'un échantillon d'excréments de canidés sauvages (présumé être du renard roux) prélevé près de Montague, Î.-P.-É. et des vers adultes femelles récupérés des carcasses lors de la nécropsie de deux renards roux ont été identifiés comme étant A. vasorum. L'analyse moléculaire a confirmé que les larves et les vers adultes étaient A. vasorum. Conclusion: Ces résultats indiquent qu'A. vasorum est devenu endémique dans la population de renards roux de l'Î.-P.-É. Pertinence clinique: L'infection à A. vasorum est potentiellement mortelle chez le chien. Les vétérinaires et les laboratoires de diagnostic régionaux des provinces maritimes devraient envisager la possibilité d'une infection à A. vasorum chez les chiens présentant des signes cliniques de maladie cardio-pulmonaire et/ou du système nerveux central ou de troubles de la coagulation.(Traduit par Dr Serge Messier).

An in vitro larval motility assay characterizes anthelmintic efficacy against Crenosoma vulpis, Angiostrongylus vasorum, and Aelurostrongylus abstrusus.

OBJECTIVE: This study determined the in vitro efficacy of 6 common anthelmintics (eprinomectin, ivermectin, milbemycin oxime, moxidectin, selamectin, and fenbendazole) on motility (viability) of infectious third-stage larvae (L3) of Crenosoma vulpis, Angiostrongylus vasorum, and Aelurostrongylus abstrusus, which are important causes of canine and feline cardiopulmonary disease. SAMPLES: First-stage larvae (L1) from C vulpis, An vasorum, and Ae abstrusus. PROCEDURES: Naïve Limax maximus slugs were fed 1,000 to 2,000 L1 and held at 16 °C for at least 4 weeks to produce live L3. Approximately 50 to 100 L3/well were subsequently incubated in culture media alone or media containing 6 separate test anthelmintics at 4 concentrations, to bracket expected in vivo drug plasma levels in anthelmintic-treated dogs and cats. Drug effects on L3 motility (viability) were analyzed by multilevel logistic models, generating dose-response relationships. Experiments were completed 1-9/2019. RESULTS: Drug concentration estimates corresponding to a 50% larval mortality rate identified that C vulpis was the most sensitive species to the anthelmintics tested. Ae abstrusus was most susceptible to moxidectin and selamectin, while An vasorum was insusceptible to all anthelmintics tested, except for selamectin at high drug concentrations. CLINICAL RELEVANCE: The in vitro anthelmintic response to antiparasitic agents may guide and improve disease therapy and prevention. Considering the observed lack of efficacy against L3, monthly anthelmintic treatment for protection against An vasorum infection in dogs would primarily rely on the anthelmintic's adulticidal activity. Maximal preventive control for An vasorum would, therefore, require at least 1 treatment administered a minimum of 1 week after the end of the transmission season.

Massive hepatic trematodosis in 5 juvenile bald eagles.

Hepatic trematodosis by opisthorchiid flukes has been reported sporadically in North American fish-eating raptors. Bald eagles (Haliaeetus leucocephalus) infected by these flukes often have various degrees of granulomatous cholangitis, pericholangitis, necrosis of adjacent hepatocytes, and subsequent hepatic fibrosis. Species identification has been complicated by the inability to dissect intact specimens from liver tissue. Between 2007 and 2018, 5 juvenile bald eagles with massive hepatic trematodosis were identified at autopsy. Histologically, flukes were non-spinous. Parasitologic identification revealed ventral suckers (80-93 µm diameter), and uteri containing golden, operculated eggs (~25.0 × 12.0 µm). An unfixed frozen liver sample of one eagle was analyzed by PCR and DNA sequencing targeting the large subunit rRNA, ITS region, and cox1 genes of the parasite. The fluke DNA sequences shared 99.6%, 98.4%, and 87.0% similarity, respectively, with Erschoviorchis anuiensis, a newly described opisthorchiid species infecting the liver and pancreas of fish-eating birds in Europe and Asia. Infection by E. anuiensis is highly pathogenic in several piscivorous bird species. The clinical significance of trematodosis in our 5 cases is uncertain because all birds had comorbidities.

Clinical crenosomosis in a black bear (Ursus americanus).

An orphaned black bear (Ursus americanus) cub, estimated to be 9 months-of-age was presented to a wildlife rehabilitation facility in December of 2016. The cub was afebrile, under-weight (6.8 kg) and had a cough condition. Centrifugal sugar fecal flotation examination failed to detect any gastrointestinal helminth or protozoan parasites, but revealed the presence of first-stage nematode larvae (L1). Large numbers of L1 (>8000 L1/g) identified as Crenosoma sp. based on morphology were recovered using the Baermann technique. Three species (Crenosoma petrowi, Crenosoma potos, Crenosoma vulpis) have been reported from black bears. Based on larval length measurements (range = 253-277 µm; mean = 263 µm; n = 8), the L1 were tentatively identified as C. petrowi. Further molecular characterization using Polymerase Chain Reaction (PCR) and DNA sequencing of the small subunit (SSU) RNA gene and two regions of the large subunit (LSU) rRNA gene did not match any submissions in GenBank, but were most similar to Crenosoma mephiditis. There is a paucity of molecular data for members of the genus Crenosoma, with only information for Crenosoma vulpis (red fox), C. mephitidis (skunks), Crenosoma striatum (hedgehog) and Crenosoma sp. (red panda) in GenBank. Molecular analysis eliminates C. vulpis as a possibility in this case but due to the lack of submissions in GenBank, the identification of the L1 as C. petrowi based on length measurements could not be confirmed. Receiving in total, three separate courses of treatment with fenbendazole (50 mg/kg, oral, once a day for 3 days), fecal larval shedding ceased and clinical signs resolved. The black bear cub was released into the wild in June 2017. This is the first report of clinical chronic respiratory disease due to Crenosoma sp. infection in a black bear.

Endoparasites in dogs and cats diagnosed at the Veterinary Teaching Hospital (VTH) of the University of Prince Edward Island between 2000 and 2017. A large-scale retrospective study.

Although many studies on the frequency of endoparasites in dogs and cats in Canada have been reported, seasonal and/or annual patterns are often not evaluated. The frequency and risk factors of endoparasite infections from fecal samples of cats and dogs submitted to the Veterinary Teaching Hospital of the Atlantic Veterinary College, University of Prince Edward Island-Canada were determined, using univariable and multivariable logistic regression analyses. Investigated predictors of endoparasitism available in the 2000-2017 database included sex, age, geographic origin and seasonality. A total of 15,016 dogs and 2,391 cats were evaluated for endoparasite status using specific diagnostic tests: direct smear, Baermann, and/or 33 % zinc sulfate solution in a standardized centrifugal flotation method. Overall, twelve and eight parasite genera were detected in dogs and cats, respectively. The overall proportional infection was 14.6 %, and the cat population showed a higher frequency of positivity to parasites compared to the dog population (P < 0.001). The most frequent genera recovered in the whole population (dogs and cats), were Giardia duodenalis (5.2 %), Cystoisospora spp. (3.3 %) and Toxocara spp. (3.2 %). Endoparasitism levels were diagnosed more in feces submitted from young, female intact dogs from PEI compared to the baselines of mature, sterilized male dogs from other provinces, respectively, and diagnoses occurred more often in autumn months than in winter months. There was no significant diagnostic trend across the years for the individual parasites models. The frequency of detected potentially zoonotic parasites in this study highlights the veterinary public health and One Health context of parasitic infections in pets. Although the presented results are not from a random sample and therefore frequency results should be interpreted with caution, the model relationship results may still be relevant. In addition, results are of value to estimate parasite impact and to assist researchers, veterinarians and pet-owners with suitable information to control parasites.

A new trichosomoidid from the skin of Sebastes spp. (Pisces) from British Columbia, Canada.

A new species of trichosomoidid nematode, Huffinanela canadensis n. sp., is described from the skin of rockfish (Sebastes spp.) from the Pacific Ocean off the coast of Vancouver Island (Clayoquot Sound region), British Columbia, Canada, on the basis of the morphology of the adult worms and their eggs in the host's tissue and the biological characters. The species is characterized mainly by the shape and structure of the fully developed eggs (absence of surface envelope, surface with transverse ridges), by their small size (48-63 x 24-27 microm), and by the site of infection (skin). Besides Huffmanela huffmani, this is the second Huffmanela species in which adult worms are known in addition to the eggs from the host's tissues (most Huffmanela spp. have been described only from their conspicuous eggs occurring in various tissues of fishes). Adults of H. canadensis differ morphologically from those of H. huffmani, mainly in the structure of the male caudal end and in the distinctly elevated anterior vulvar lip of the female.

Susceptibility of rainbow trout Oncorhynchus mykiss, Atlantic salmon Salmo salar and coho salmon Oncorhynchus kisutch to experimental infection with sea lice Lepeophtheirus salmonis.

Physiological, immunological and biochemical parameters of blood and mucus, as well as skin histology, were compared in 3 salmonid species (rainbow trout Oncorhynchus mykiss, Atlantic salmon Salmo salar and coho salmon O. kisutch) following experimental infection with sea lice Lepeophtheirus salmonis. The 3 salmonid species were cohabited in order to standardize initial infection conditions. Lice density was significantly reduced on coho salmon within 7 to 14 d, while lice persisted in higher numbers on rainbow trout and Atlantic salmon. Lice matured more slowly on coho salmon than on the other 2 species, and maturation was slightly slower on rainbow trout than on Atlantic salmon. Head kidney macrophages from infected Atlantic salmon had diminished respiratory burst and phagocytic capacity at 14 and 21 d post-infection (dpi), while infected rainbow trout macrophages had reduced respiratory burst and phagocytic capacities at 21 dpi, compared to controls. The slower development of lice, coupled with delayed suppression of immune parameters, suggests that rainbow trout are slightly more resistant to lice than Atlantic salmon. Infected rainbow trout and Atlantic salmon showed increases in mucus lysozyme activities at 1 dpi, which decreased over the rest of the study. Mucus lysozyme activities of infected rainbow trout, however, remained higher than controls over the entire period. Coho salmon lysozyme activities did not increase in infected fish until 21 dpi. Mucus alkaline phosphatase levels were also higher in infected Atlantic salmon compared to controls at 3 and 21 dpi. Low molecular weight (LMW) proteases increased in infected rainbow trout and Atlantic salmon between 14 and 21 dpi. Histological analysis of the outer epithelium revealed mucus cell hypertrophy in rainbow trout and Atlantic salmon following infection. Plasma cortisol, glucose, electrolyte and protein concentrations and hematocrit all remained within physiological limits for each species, with no differences occurring between infected and control fish. Our results demonstrate that significant differences in mucus biochemistry and numbers of L. salmonis occur between these species.

Pulmonary trematodosis (Pharyngostomoides sp.) in a juvenile raccoon (Procyon lotor).

A lethargic, wild, juvenile raccoon died and was subsequently presented for necropsy. The lungs had widely disseminated, finely granular, yellow-tan foci with moderate pulmonary, as well as, thoracic lymphadenopathy. Histopathology revealed numerous cross-sections of larval trematodes morphologically consistent with diplostomid mesocercariae and metacercariae as well as moderate alveolointerstitial pneumonia with a mixed population of eosinophils, macrophages, lymphocytes, and plasma cells. The presence of larval trematodes within lymphatics in multiple organs indicates a route of fluke migration. Live specimens of mesocercariae and metacercariae recovered from refrigerated lung tissue after 7 days were identified as Pharyngostomoides sp., an intestinal diplostomid parasite (trematode) of raccoons and other mammals that is similar in appearance and life cycle to Alaria spp.