Integration of ITS-2 rDNA nemabiome metabarcoding with Fecal Egg Count Reduction Testing (FECRT) reveals ivermectin resistance in multiple gastrointestinal nematode species, including hypobiotic Ostertagia ostertagi, in western Canadian beef cattle.

A large-scale Fecal Egg Count Reduction Test (FECRT) was integrated with ITS-2 rDNA nemabiome metabarcoding to investigate anthelmintic resistance in gastrointestinal nematode (GIN) parasites in western Canadian beef cattle. The study was designed to detect anthelmintic resistance with the low fecal egg counts that typically occur in cattle in northern temperate regions. Two hundred and thirty-four auction market-derived, fall-weaned steer calves coming off pasture were randomized into three groups in feedlot pens: an untreated control group, an injectable ivermectin treatment group, and an injectable ivermectin/oral fenbendazole combination treatment group. Each group was divided into six replicate pens with 13 calves per pen. Individual fecal samples were taken pre-treatment, day 14 post-treatment, and at monthly intervals for six months for strongyle egg counting and metabarcoding. Ivermectin treatment resulted in an 82.4% mean strongyle-type fecal egg count reduction (95% CI 67.8-90.4) at 14 days post-treatment, while the combination treatment was 100% effective, confirming the existence of ivermectin-resistant GIN. Nemabiome metabarcoding of third-stage larvae from coprocultures revealed an increase in the relative abundance of Cooperia oncophora, Cooperia punctata, and Haemonchus placei at 14 days post-ivermectin treatment indicating ivermectin resistance in adult worms. In contrast, Ostertagia ostertagi third-stage larvae were almost completely absent from day 14 coprocultures, indicating that adult worms of this species were not ivermectin resistant. However, there was a recrudescence of O. ostertagi third stage larvae in coprocultures at three to six months post-ivermectin treatment, which indicated ivermectin resistance in hypobiotic larvae. The calves were recruited from the auction market and, therefore, derived from multiple sources in western Canada, suggesting that ivermectin-resistant parasites, including hypobiotic O. ostertagi larvae, are likely widespread in western Canadian beef herds. This work demonstrates the value of integrating ITS-2 rDNA metabarcoding with the FECRT to enhance anthelmintic resistance detection and provide GIN species- and stage-specific information.

Effects of naturally acquired gastrointestinal nematode infection on bovine viral diarrhea virus vaccine-directed antibody response in western Canadian feedlot cattle.

Objective: To determine how gastrointestinal nematode (GIN) infection, reflected by fecal egg counts and Ostertagia ostertagi serum antibody titers, is associated with the antibody response to bovine viral diarrhea virus type 1 (BVDV-1) vaccine antigen in fall-weaned feedlot cattle from western Canada. Animals: Cross-sectional study with 240 steer calves derived from an auction market. Procedure: At feedlot arrival, calves were given a commercial vaccine containing modified live BVDV-1. Serum neutralization antibody titers against BVDV-1 antigens were determined in individual blood samples collected pre-vaccination and 21 d after vaccination. A modified Wisconsin sugar floatation method was used to obtain individual calf GIN egg counts in fecal samples on arrival. Antibody titers against O. ostertagi were determined using an enzyme-linked immunosorbent assay in on-arrival blood samples. Results: Fecal egg counts and O. ostertagi titers were not associated with vaccine antibody-fold changes. Similarly, fecal egg counts and O. ostertagi titers were not associated with vaccine-induced seroconversion. Conclusions: The relatively low GIN burdens, reflected by the overall low fecal egg counts in these fall-weaned feedlot calves, did not have measurable adverse effects on the humoral immune response to BVDV-1 vaccine antigens. Clinical relevance: An adequate response to vaccination is important for cattle welfare and productivity. Conditions that negatively affect this response may vary regionally, such as GIN infection. Understanding this is essential. Although subclinical intestinal parasitism did not noticeably affect the antibody response in these steers, higher GIN burdens and actual immune protection from clinical disease remain to be investigated.

Effets d'une infection par des nématodes gastro-intestinaux d'origine naturelle sur la réponse en anticorps dirigés par le vaccin contre le virus de la diarrhée virale bovine chez les bovins des parcs d'engraissement de l'Ouest canadien. Objectif: Déterminer comment l'infection par les nématodes gastro-intestinaux (GIN), reflétée par le nombre d'oeufs fécaux et les titres d'anticorps sériques d'Ostertagia ostertagi, est associée à la réponse en anticorps à l'antigène du vaccin contre le virus de la diarrhée virale bovine de type 1 (BVDV-1) chez les bovins en parc d'engraissement sevrés à l'automne de l'Ouest canadien. Animaux: Étude transversale auprès de 240 veaux bouvillons issus d'un marché aux enchères. Procédure: À leur arrivée au parc d'engraissement, les veaux ont reçu un vaccin commercial contenant du BVDV-1 vivant modifié. Les titres d'anticorps sériques neutralisants contre les antigènes BVDV-1 ont été déterminés dans des échantillons de sang individuels prélevés avant la vaccination et 21 jours après la vaccination. Une méthode de Wisconsin modifiée de flottation au sucre a été utilisée pour obtenir le nombre d'oeufs GIN de chaque veau dans les échantillons fécaux à l'arrivée. Les titres d'anticorps dirigés contre O. ostertagi ont été déterminés à l'aide d'un dosage immuno-enzymatique dans des échantillons de sang à l'arrivée. Résultats: Le nombre d'oeufs fécaux et les titres d'O. ostertagi n'étaient pas associés aux modifications du titre d'anticorps vaccinaux. De même, le nombre d'oeufs fécaux et les titres d'O. ostertagi n'étaient pas associés à la séroconversion induite par le vaccin. Conclusion: Les charges relativement faibles de GIN, reflétées par le faible nombre global d'oeufs fécaux chez ces veaux d'engraissement sevrés à l'automne, n'ont pas eu d'effets indésirables mesurables sur la réponse immunitaire humorale aux antigènes du vaccin BVDV-1. Pertinence clinique: Une réponse adéquate à la vaccination est importante pour le bien-être et la productivité des bovins. Les conditions qui affectent négativement cette réponse peuvent varier selon les régions, telles que l'infection par les GIN. Comprendre cela est essentiel. Bien que le parasitisme intestinal subclinique n'ait pas sensiblement affecté la réponse en anticorps chez ces bouvillons, des charges de GIN plus élevées et une protection immunitaire réelle contre la maladie clinique restent à étudier.(Traduit par Dr Serge Messier).

Morphometric, metabolic, and inflammatory markers across a cohort of client-owned horses and ponies on the insulin dysregulation spectrum.

In human metabolic syndrome and type II diabetes, methylglyoxal (MG), D-lactate, and several cytokines have been recognized as biomarkers of important metabolic and inflammatory processes. Equine metabolic syndrome (EMS) shares many similarities with these human counterparts. The objectives of this cross-sectional study were to compare body condition score (BCS), cresty neck score (CNS), resting insulin, MG, D-lactate, L-lactate, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) between horses with and without insulin dysregulation, as classified via combined glucose and insulin test (CGIT). 32 client-owned horses were included. History and morphometric data such as BCS and CNS were recorded. Subjects with abnormalities on physical examination or CBC, elevated ACTH or incomplete information were excluded. Baseline serum or plasma concentrations of biomarkers were tested via commercial ELISA or colorimetric assays. Characteristics of insulin dysregulated and insulin sensitive horses were compared by univariate analysis and forward logistic regression. 12 (38%) of the 32 horses were classified as insulin dysregulated. No significant difference between the 2 groups was found for age, BCS, baseline glucose, triglycerides, MG, D-lactate, L-lactate, TNF-α, IL-6, and MCP-1. Baseline insulin was significantly associated with insulin dysregulation in univariate analysis (P = 0.02), but not in the final model. Horses with CNS ≥ 3 had 11.3 times higher odds of having insulin dysregulation (OR 11.3, 95% C.I. 2.04 - 63.08, P = 0.006). In this population, horses with mild-moderate signs of EMS presented similar metabolic and inflammatory profiles to non-insulin dysregulated controls.

Pharmacokinetics and pharmacodynamics of doxycycline in a Streptococcusequi subsp. zooepidemicus infection model in horses.

The objectives of this study were to investigate the pharmacokinetics (PK), pharmacodynamics (PD), and the efficacy of oral administration of doxycycline (DXC) in horses with Streptococcus zooepidemicus tissue infections. Tissue chambers (TC) were implanted subcutaneously in the cervical region of 7 horses and inoculated with a single S. zooepidemicus isolate with a minimum inhibitory concentration (MIC) of 0.25 µg/ml, determined by agar dilution. Doxycycline hyclate (10 mg/kg, orally, q 12 h, for 5 days) mixed with poloxamer gel was started following inoculation. The TC fluid was sampled prior to and following inoculation for cytology analysis, quantitative culture, and DXC determination. Plasma DXC concentrations were measured over 48 h following the last dose of DXC administered. The mean plasma peak concentration (Cmax ) of DXC was 0.32 µg/ml, and concentrations above the MIC were only reached in 3 TC samples. In plasma, mean T > MIC was 2.4 h, mean Cmax /MIC was 1.30, and mean AUClast /MIC was 11.63 h. These PK/PD indices did not reach the suggested targets for DXC treatments of infections, and the TC abscessed in all horses. This is the first study to evaluate the recommended dose of DXC in horse in an infection model.

Septic fibrinous pericarditis in 4 horses in Saskatchewan following an outbreak of forest tent caterpillars in 2017.

In the summer of 2017, 4 horses were diagnosed with septic fibrinous pericarditis at the Western College of Veterinary Medicine, Saskatoon. This case series occurred after a significant outbreak of forest tent caterpillars (Malacosoma disstria) in the province during that spring. Three horses were immediately euthanized, and treatment was attempted in 1 mare. This is the first case series of pericarditis possibly associated with the ingestion of forest tent caterpillars to be reported in western Canada. Although cause-effect is not proven, it is prudent to prevent the ingestion of caterpillars by horses. Key clinical message: Septic fibrinous pericarditis, a rare condition in horses, has previously been linked to outbreaks of eastern tent caterpillars. A similar link might exist in this case series.

Péricardites fibrineuses septiques chez quatre chevaux saskatchewanais consécutive à une infestation de chenilles de livrée des forêts en 2017. Au cours de l'été 2017, quatre chevaux ont été diagnostiqués avec une péricardite fibrineuse septique au Western College of Veterinary Medicine de Saskatoon. Ces cas ont été présentés après une sévère infestation printanière de chenilles de livrée de forêts (Malacosoma disstria) dans la province de la Saskatchewan, reportée au printemps. Trois chevaux ont été immédiatement euthanasiés et une jument a été hospitalisée pour traitement. Ceci est le premier rapport décrivant la possible association entre des cas de péricardite chez des chevaux et l'ingestion de chenilles de livrée de forêts dans l'Ouest canadien. Même si le lien de cause à effet n'est pas prouvé, il est prudent d'éviter l'ingestion de ces chenilles processionnaires par les chevaux.Message clinique clé :La péricardite fibrineuse septique, une condition rare chez les chevaux, a précédemment été liée à des flambées de livrées des forêts. Un lien similaire pourrait exister dans la présente série de cas.(Traduit par les auteurs).

Gastrointestinal nematode prevalence and fecal egg counts in beef cattle from western Canada.

Fecal samples were collected from cows (n = 1458), calves (n = 1188), and replacement heifers (n = 921) between 2012 and 2014 from 199 herds and generalized estimating equations were used to predict mean fecal egg counts and prevalence of egg-positive samples. Replacement heifers had the highest prevalence of Trichostrongylid-type eggs at 83% [95% confidence interval (CI): 78% to 87%], and cows had the lowest at 75% (95% C: 70% to 81%). Nematodirus spp. was most frequently present in calves [predicted prevalence: 34% (95% CI: 28% to 40%)]. Mean fecal egg counts were highest in calves with 5.9 (95% CI: 3.9 to 7.8) Trichostrongylid-type eggs per gram (EPG) of feces and 1.0 (95% CI: 0.7 to 1.4) Nematodirus spp. EPG. Although mean egg counts were low to moderate, the high prevalence highlights the need to further investigate the epidemiology of gastrointestinal nematodes in western Canada. This is particularly relevant considering management changes, increasing herd sizes, climate change, and threatening anthelmintic resistance.

Prévalence des nématodes gastro-intestinaux et dénombrements des oeufs dans les fèces chez les bovins d'embouche dans l'ouest canadien. Des échantillons de fèces furent prélevés de vaches (n = 1458), veaux (n = 1188) et génisses de remplacement (n = 921) entre 2012 et 2014 dans 199 troupeaux et des équations d'estimation généralisée furent utilisées pour prédire les dénombrements moyens d'oeufs dans les fèces et la prévalence d'échantillons positifs pour la présence d'oeufs. Les génisses de remplacement avaient la prévalence la plus élevée d'oeufs de type Trichostrongylide avec 83 % [intervalle de confiance 95 % (IC) : 78 % à 87 %], et les vaches avaient la plus faible avec 75 % (95 % IC : 70 % à 81 %). Les Nematodirus spp. étaient présents le plus fréquemment chez les veaux [prévalence prédite : 34 % (95 % IC : 28 % à 40 %)]. Les dénombrements moyens d'oeufs dans les fèces étaient les plus élevés chez les veaux avec 5,9 (95 % IC : 3,9 à 7,8) oeufs de type Trichostrongylide par gramme (EPG) de fèces et 1,0 (95 % IC : 0,7 à 1,4) EPG de Nematodirus spp. Bien que les dénombrements moyens d'oeufs étaient faibles à modérés, la prévalence élevée met en évidence le besoin de continuer d'examiner l'épidémiologie des nématodes gastro-intestinaux dans l'ouest du Canada. Ceci est particulièrement approprié considérant les changements dans la gestion, l'augmentation de la taille des troupeaux, les changements climatiques et la menace de la résistance aux produits anthelmintiques.(Traduit par Dr Serge Messier).

Gastrointestinal nematode management in western Canadian cow-calf herds.

There is a paucity of information from western Canadian beef cow-calf producers about how they control gastrointestinal nematodes. The objectives of this study were to describe cow-calf producers' management practices related to control of gastrointestinal nematodes including pasture management and use of parasite control products. A questionnaire was distributed to 105 producers in May 2015. Responses from 97 producers revealed the almost uniform dependence on the use of a pour-on macrocyclic lactone parasite control product in the fall as part of a routine farm management program. Control of external parasites was the primary reason for treatment, while none of the producers chose to treat specifically to manage internal parasites. The predominant management practices identified through this study increase the risk of development of anthelmintic resistance. The results also highlight the need to raise awareness of the importance of an evidence-based gastrointestinal nematode control program in beef cow-calf herds.

Gestion des nématodes gastro-intestinaux dans les troupeaux vaches-veaux de l'ouest canadien. Il y a une rareté d'informations provenant des producteurs de vaches-veaux de l'ouest canadien sur la façon dont ils maitrisent les nématodes gastro-intestinaux. Les objectifs de la présente étude étaient de décrire les pratiques de gestion des producteurs de vaches-veaux relativement à la maitrise des nématodes gastro-intestinaux incluant la gestion des pâturages et l'utilisation des produits antiparasitaires. Un questionnaire fut distribué à 105 producteurs en mai 2015. Des réponses en provenance de 97 producteurs ont révélé la dépendance presque généralisée sur l'utilisation d'un produit antiparasitaire à verser sur l'animal à base de lactone macrocyclique à l'automne comme faisant partie d'un programme de routine de gestion à la ferme. La maitrise des parasites externes était la principale raison pour le traitement, alors qu'aucun des producteurs n'avait choisi de traiter spécifiquement pour gérer les parasites internes. Les pratiques de gestion prédominantes identifiées dans la présente étude augmentent le risque de développement de résistance aux anthelmintiques. Les résultats mettent également en lumière le besoin d'augmenter la sensibilisation de l'importance d'une approche factuelle à un programme de gestion des nématodes dans les troupeaux vaches-veaux.(Traduit par Dr Serge Messier).

Investigation of a Yersinia enterocolitica outbreak in a commercial alpaca farm in Saskatchewan.

An outbreak of enterocolitis and sudden deaths occurred in an alpaca herd in Saskatchewan (mortality rate: 18.6%). Yersinia enterocolitica was isolated from the intestine, liver, and spleen of the index case, the kidney of a second case, and the feces of affected animals. The initial source of infection could not be identified. Treatment with oxytetracycline was followed by resolution of diarrhea and arrest of fatalities. The end of the outbreak was determined by absence of new cases and negative fecal cultures. This is the first report of a presumed herd outbreak of yersiniosis in camelids in North America.

Enquête à la suite d'une épidémie due à Yersinia enterocolitica dans une ferme commerciale d'alpagas en Saskatchewan. Une épidémie d'entérocolite et de mortalités soudaines est survenue dans un troupeau d'alpaga en Saskatchewan (taux de mortalité : 18,6 %). Yersinia enterocolitica fut isolé à partir de l'intestin, du foie et de la rate du cas index, du rein d'un second cas, et des fèces des animaux affectés. La source initiale d'infection ne put être identifiée. Un traitement avec de l'oxytétracycline fut suivi d'une résolution de la diarrhée et un arrêt des mortalités. La fin de l'épidémie fut déterminée par l'absence de nouveaux cas et des cultures de fèces négatives. Ceci constitue le premier rapport d'une épidémie présumée de yersiniose chez des camélidés en Amérique du Nord.(Traduit par Dr Serge Messier).

The One Health Leadership Experience at the University of Saskatchewan, Canada.

Competency in One Health (OH) leadership was emphasized in the Roadmap for Veterinary Medical Education in the 21st Century in 2011. Since then, several educational interventions have been aimed at increasing awareness and capacity for inter-professional collaboration. At the University of Saskatchewan, Canada, a 3-day event, the One Health Leadership Experience (OHLE), was initiated in 2012 and continues to the present. The event targets students entering their first year of a health professional program and consists of presentations by invited OH guest speakers, networking sessions, small-group case discussions of OH scenarios, and leadership development through panel discussions and interactive small-group dialogues. Post-conference surveys, a 5-year follow-up survey, and two focus groups were conducted to evaluate the impact of participation in the OHLE. After the event, the proportion of students who said they clearly understood OH and its goals was substantially higher than before: 86% versus 14% in 2012, 91% versus 23% in 2013, and 69% versus 24% in 2014. In the 5-year follow-up survey, most respondents (90%) indicated that attending the OHLE increased their interaction with other students from health sciences colleges or schools on campus. Also, most (81%) believed that OH should be formally taught in their program and 80% anticipated implementing, or had already implemented, OH practices after graduation. The OHLE increased participants' awareness of the importance of interdisciplinary approaches and is a successful educational model that can be adapted to health professional curricula at other institutions.

Observations of mortality in farmed bison in the Canadian prairies: 2103 - 2016.

The present study is a continuation of a previous mortality study on Saskatchewan bison farms with special emphasis on Malignant Catarrhal Fever. The updated objective of the study was to estimate the most common causes of mortality in farmed bison herds in Western Canada. Results were compared to the previous Saskatchewan study to assess the similarities and differences in the etiology associated with farmed bison deaths across the Prairie Provinces of Canada. The most common cause of death was respiratory disease associated with Mycoplasma bovis, although this was restricted to Alberta and Saskatchewan farm locations. This was in contrast to the previous Saskatchewan based study which did not identify any deaths involving this pathogen. An updated overall assessment of the risks of Malignant Catarrhal Fever in farmed bison at various proximities to sheep operations further confirmed the low risk of occurrence on farms within a 1 km boundary fence distance.

Seroprevalence of Cache Valley virus and related viruses in sheep and other livestock from Saskatchewan, Canada.

Cache Valley virus, an orthobunyavirus, is an important cause of ovine neonatal malformations. Information on the seroprevalence of this virus in Saskatchewan livestock populations is lacking. The objectives of this study were to determine the seroprevalence of Cache Valley virus and closely related viruses in sheep, cattle, goats, horses, and mule deer in Saskatchewan by performing a plaque-reduction neutralization test using Cache Valley virus. In total, sera from 130 sheep from 50 flocks were tested. Seroprevalence in sheep was 64.6% (84/130) and 94.0% (47/50) of flocks had 1 or more seropositive sheep. Antibodies to Cache Valley virus or closely related viruses were also detected in serum samples collected from cattle, goats, horses, and mule deer with seroprevalences of 20.0% (5/25), 33.3% (8/24), 69.0% (40/58), and 50.8% (33/65), respectively. These results suggest widespread exposure to Cache Valley virus or closely related viruses in domestic animals and mule deer in Saskatchewan.

Séroprevalence du virus de la Vallée Cache ou de virus connexes chez les moutons et d'autres animaux de cheptel en Saskatchewan, Canada. Le virus de la Vallée Cache, un orthobunyavirus, est une cause importante de malformations néonatales ovines. Il manque des renseignements sur la séroprévalence de ce virus dans les populations des cheptels de la Saskatchewan. Les objectifs de cette étude consistaient à déterminer la séroprévalence du virus de la Vallée Cache et des virus étroitement apparentés chez les moutons, les bovins, les chèvres, les chevaux et les cerfs mulets en Saskatchewan en réalisant un test de séro-neutralisation par réduction des plages en utilisant le virus de la Vallée Cache. Au total, le sérum provenant de 130 moutons dans 50 troupeaux a été testé. Chez les moutons, la séroprévalence était de 64,6 % (84/130) et 94,0 % (47/50) des troupeaux avaient un mouton ou plusieurs moutons séropositifs. Les anticorps pour le virus de la Vallée Cache ou les virus étroitement apparentés ont aussi été détectés dans les échantillons de sérum prélevés auprès des bovins, des chèvres, des chevaux et des cerfs mulets avec une séroprévalence de 20,0 % (5/25), de 33,3 % (8/24), de 69,0 % (40/58) et de 50,8 % (33/65), respectivement. Ces résultats suggèrent une vaste exposition au virus de la Vallée Cache ou à des virus étroitement apparentés chez les animaux domestiques et les cerfs mulets en Saskatchewan.(Traduit par Isabelle Vallières).

Comparison of five diagnostic tests for Giardia duodenalis in fecal samples from young dogs.

Five diagnostic tests were compared for the diagnosis of Giardia duodenalis in fecal samples of young dogs. Fecal samples were collected from 136 healthy dogs <1year old and examined using immunofluorescence antibody microscopy (IFA) after sucrose gradient centrifugation, zinc sulfate centrifugal flotation technique (ZSCT), SNAP®Giardia test, and ProSpecT®Giardia EZ Microplate assay. In addition, polymerase chain reaction (PCR) of the 16S rRNA gene was performed. Kappa (κ) statistic was calculated to assess diagnostic agreement between the IFA and each test. Using the IFA as the gold standard, the relative sensitivity and specificity of each test were determined. Subsequently, a Bayesian approach was used to estimate the sensitivity and specificity of each test in comparison to the IFA results. Giardia duodenalis was detected in 41% of the samples examined by IFA. The ZSCT resulted in 37% of positive samples, with a relative sensitivity and specificity of 86 and 98%, respectively. The SNAP®Giardia test was positive in 40% of the samples, with a relative sensitivity and specificity of 91 and 96%, respectively. The ProSpecT® test was positive in 51% of the samples, with a relative sensitivity and specificity of 100 and 83%, respectively. The relative sensitivity and specificity for PCR were 58 and 56%, respectively, with 55% of samples being PCR-positive. While the sensitivity and specificity estimates of each test in comparison to the IFA changed when using a Bayesian approach, the conclusions remained the same. While the ProSpecT® test was the most sensitive test in this study, it is not designed for dogs and more costly than the other tests. The SNAP®Giardia test performed similar to the ZSCT but may be more favorable because it is fast and easy to perform. Performance of the PCR was poor and the benefit of PCR may be in determining genotypes for evaluating zoonotic transfer between dogs and humans.

Zoonotic potential of Giardia duodenalis and Cryptosporidium spp. and prevalence of intestinal parasites in young dogs from different populations on Prince Edward Island, Canada.

The prevalence of Giardia duodenalis, Cryptosporidium spp. and other intestinal parasites was determined in dogs <1 year old from Prince Edward Island, Canada. Fecal samples were collected from the local animal shelter (n=62), private veterinary clinics (n=78) and a pet store (n=69). Intestinal parasites isolated included G. duodenalis, Cryptosporidium spp., Toxocara canis, Isospora spp. and Uncinaria stenocephala. To estimate the zoonotic risk associated with these infections, genotypes of G. duodenalis and Cryptosporidium spp. were determined using 16S rRNA and Hsp70 gene sequencing, respectively. Dogs from the pet store had the highest prevalence of intestinal parasites (78%, 95% CI: 68-88%), followed by the private veterinary clinics (49%, 95% CI: 37-60%), and the local animal shelter (34%, 95% CI: 22-46%). The majority G. duodenalis belonged to host-adapted assemblages D (47%, 95% CI: 31-64%) and C (26%, 95% CI: 13-43%), respectively. Zoonotic assemblages A and B were isolated alone or in mixed infections from 16% (95% CI: 6-31%) of G. duodenalis-positive dogs. All Cryptosporidium spp. were the host-adapted C. canis. While host-adapted, non-zoonotic G. duodenalis genotypes were more common, the presence of G. duodenalis assemblages A and B, T. canis, and U. stenocephala suggests that these dogs may present a zoonotic risk. The zoonotic risk from Cryptosporidium-infected dogs was minimal.

Granulocytic anaplasmosis in a horse from Nova Scotia caused by infection with Anaplasma phagocytophilum.

A 16-year-old Paint stallion was presented with intermittent fever, inappetance, lethargy, icterus, distal limb edema, and submandibular lymphadenopathy. The horse was native to Nova Scotia and had never left that province. Morulae were detected in granulocytes. Ananaplasma phagocytophilum infection was confirmed by serology and polymerase chain reaction (PCR). The horse responded to treatment with oxytetracycline.

Concurrent nephrosplenic entrapment and acquired inguinal herniation of the jejunum in a Standardbred stallion.

A stallion presenting for surgical correction of an acquired inguinal hernia was also diagnosed with a nephrosplenic entrapment (NSE) intraoperatively. Surgical intervention resulted in a successful outcome. To date, these conditions have not been reported to occur simultaneously.

Prevalence and genotypes of Giardia duodenalis in dairy and beef cattle in farms around Charlottetown, Prince Edward Island, Canada.

Prevalence of Giardia duodenalis in dairy and beef cattle on farms around Charlottetown, Prince Edward Island (Canada) was determined by analyzing feces using direct immunofluorescence antibody microscopy. Genotypes were determined by 16S-rRNA sequencing. Fecal samples (n = 892) were collected from adult cattle in dairy tie-stall, dairy free-stall, and beef herds (10 herds each), and from calves (n = 183) from 11 dairy farms. Prevalence rates were 38% and 51% in cows and calves, respectively. Giardia duodenalis was present in all dairy herds, in 9/10 beef herds and in calves from 10/11 herds examined. Prevalence rates were 40% and 41% for cows in tie- and free-stall herds, respectively, and 27% for beef cows. Zoonotic Assemblage A was found in 12.2% of calves concomitantly infected with Assemblage E. All successfully sequenced samples (114/128) from cows corresponded to Assemblage E. Giardia duodenalis is highly prevalent in cattle herds in Prince Edward Island and Assemblage A in calves is a potential public health concern.

Prevalence and molecular characterization of Cryptosporidium spp. in dairy calves from 11 farms in Prince Edward Island, Canada.

Cryptosporidium spp. are common intestinal protozoan parasites that infect a wide range of hosts, including humans and livestock, worldwide. The objective of this study was to determine the prevalence of Cryptosporidium spp. in dairy calves in Prince Edward Island, Canada, and the potential for transmission of this parasite between dairy calves and humans. Fecal samples were collected from 183 dairy calves from 11 farms in Prince Edward Island. The prevalence of Cryptosporidium spp. infections in these animals was determined by examining for the presence of oocysts in the fecal samples, using immunofluorescence microscopy. Molecular characterization was done using a nested-PCR protocol to amplify fragments of the Cryptosporidium heat-shock protein 70 gene, followed by DNA sequencing. Ten calves (6.2%), representing 4 out of 11 farms tested, were positive for Cryptosporidium spp. DNA sequence analysis on five PCR positive samples demonstrated that Cryptosporidium parvum was the only species present in the calves tested, suggesting that there is a potential risk of zoonotic transmission between dairy calves and humans in this region.

Comparison of flow cytometry and immunofluorescence microscopy for the detection of Giardia duodenalis in bovine fecal samples.

The performance of flow cytometry (FC) was compared with immunofluorescence microscopy (IM) for detection of Giardia duodenalis in bovine feces. Samples from 36 adult dairy cows and 208 dairy calves were collected. Flow cytometry test characteristics were calculated using continuous, ordinal, and dichotomized results. Spearman correlation coefficients comparing the results of the 2 tests were 0.47 and 0.68 for cows and calves, respectively. Using IM as indicative of presence or absence of G. duodenalis cysts in each sample, likelihood ratios of FC results with 0, 1, and > or = 2 gated events indicated that samples with 1 gated event were likely to be positive in the cows but not in the calves. Immunofluorescence microscopy detected G. duodenalis in 69.7% and 48.1% of cows and calves, respectively. When dichotomizing the FC results at a cut-off point of 1 or 2 gated events, 46.3% and 19.9% of the cow and 51.9% and 35.1% of the calf samples, respectively, were classified as G. duodenalis-positive. Relative to IM, the sensitivity in the cows was 0.59 and 0.28, respectively, and 0.76 and 0.64, respectively, in the calves. At a cut-off point of 1, 65.7% and 73.1% of the cow and calf samples, respectively, were correctly classified in FC, and at a cut-off point of 2, 49.3% and 78.4% were correctly classified in the cows and calves, respectively. Flow cytometry was less sensitive than IM. Possible reasons and research needed to improve FC for G. duodenalis detection are discussed.

Efficacy of vaccination in preventing giardiasis in calves.

The objective of this study was to evaluate the efficacy of a vaccine in the prevention of Giardia duodenalis infection in calves. Six 2-week old calves were vaccinated subcutaneously with a sonicated G. duodenalis trophozoite vaccine. Six 2-week old control calves received a subcutaneous injection of sterile phosphate-buffered-saline mixed with adjuvant. Injections were repeated after 28 days. Eleven days after the second injection, calves were challenged orally with 1x10(5) purified G. duodenalis cysts from a naturally infected calf. Throughout the study, fecal samples were collected at regular intervals and examined for the presence of G. duodenalis cysts. Blood samples were collected weekly until G. duodenalis challenge and bi-weekly following challenge. Calves were euthanized 14 days after challenge and G. duodenalis trophozoites within the small intestines were enumerated. Serum antibody titers were significantly higher in vaccinated compared to non-vaccinated calves. Vaccinated calves tended to excrete more G. duodenalis cysts in their feces than non-vaccinated calves. The number of trophozoites in the small intestine was not different between vaccinated and non-vaccinated calves. Changes consistent of moderate enteritis were found in the intestines of one vaccinated and one non-vaccinated calf. Despite a serological immune response following vaccination, this vaccine was not efficacious in preventing giardiasis or reducing cyst shedding in calves.

Giardia duodenalis and Cryptosporidium spp. in a veterinary college bovine teaching herd.

In a preliminary study, we commonly identified Giardia duodenalis in adult dairy cattle from a veterinary college teaching herd. Therefore, the present study was carried out in order to better understand the potential of adult cattle to act as a source for G. duodenalis infections for students and staff at the veterinary college. Fecal samples were collected bi-weekly from this herd of adult cattle (n=30) over an 8-month period to determine the prevalence of G. duodenalis and Cryptosporidium spp. within the herd. Nested PCR followed by DNA sequencing was then performed on a subset of positive samples in order to better understand the zoonotic potential of these infections. Every cow was sampled between 11 and 18 times, depending on the date the animal joined the teaching herd. In total, 507 fecal samples were collected from 30 different cows and examined for cysts and oocysts using epifluorescence microscopy. G. duodenalis prevalence during the course of the study ranged from 37% (11/30) to 64% (18/28), with a mean of 49%. Cumulative G. duodenalis prevalence was 73% (22/30). Zoonotic G. duodenalis assemblage A genotype was identified in 43% (6/14) of the G. duodenalis-positive samples on which PCR and genetic sequencing were successfully performed. G. duodenalis assemblage E was identified in 57% (8/14) of these samples. Cryptosporidium spp. oocysts were not detected in the feces of any cows during the study period. The presence of the zoonotic G. duodenalis assemblage A in 43% of the sequenced samples indicates that there is a potential risk of infection for students and staff at this research and teaching facility, although the roles of cows as sources of giardiasis in humans remain uncertain. Furthermore, due to the large amount of feces they produce, adult cattle may serve as important sources for G. duodenalis infections in young cattle, or other animals in the facility, despite relatively low numbers of cysts excreted per gram of feces. In contrast, the results of this study indicate that this herd posed a negligible risk of transmitting Cryptosporidium parvum infections to humans.