Surveillance of Babesia odocoilei in wild and farmed cervid populations of Saskatchewan, Canada.

Objective: To investigate whether Babesia odocoilei could be detected from farmed and wild cervid diagnostic submissions prior to its first reported occurrence in Saskatchewan. Procedure: Polymerase chain reaction (PCR) for B. odocoilei was used to survey 85 fresh-frozen samples and 112 formalin-fixed, paraffin-embedded samples from Saskatchewan cervids submitted for necropsy between 2000 and 2014. Results: The PCR was positive for B. odocoilei in 1/84 white-tailed deer, 1/41 moose, 0/37 mule deer, and 1/35 elk. The positive elk was from a farmed herd, but the remaining 2 positive samples were from wild cervids. The positive moose sample was the earliest confirmed infection, dating back to 2008. Therefore, 1.5% of the study population tested positive over the 14-year period. Conclusion: There were low numbers of cervids infected with B. odocoilei in the study population. Clinical relevance: Babesiosis should be included as a differential diagnosis for disease in susceptible cervids when clinical signs are compatible; however, a lack of suggestive clinical signs or necropsy findings does not preclude infection. Thus, monitoring prevalence of the disease within Saskatchewan (and Canada) will likely require targeted surveillance.

Objectif: Déterminer si Babesia odocoilei pouvait être détectée dans les soumissions de diagnostic de cervidés d'élevage et sauvages avant sa première occurrence signalée en Saskatchewan. Procédure: La réaction d'amplification en chaîne par la polymérase (PCR) pour B. odocoilei a été utilisée pour étudier 85 échantillons fraîchement congelés et 112 échantillons fixés au formol et inclus en paraffine de cervidés de la Saskatchewan soumis à l'autopsie entre 2000 et 2014. Résultats: La PCR était positive pour B. odocoilei chez 1/84 cerf de Virginie, 1/41 orignal, 0/37 cerf mulet et 1/35 wapiti. Le wapiti positif provenait d'un troupeau d'élevage, mais les deux autres échantillons positifs provenaient de cervidés sauvages. L'échantillon d'orignal positif était la première infection confirmée, remontant à 2008. Par conséquent, 1,5 % de la population étudiée a été testée positive au cours de la période de 14 ans. Conclusion: Il y avait un faible nombre de cervidés infectés par B. odocoilei dans la population étudiée. Pertinence clinique: La babésiose devrait être incluse comme diagnostic différentiel de maladie chez les cervidés sensibles lorsque les signes cliniques sont compatibles; cependant, l'absence de signes cliniques évocateurs ou de résultats d'autopsie n'exclut pas l'infection. Ainsi, la surveillance de la prévalence de la maladie en Saskatchewan (et au Canada) nécessitera probablement une surveillance ciblée.(Traduit par Dr Serge Messier).

Species-specific PCR assay for the detection of Babesia odocoilei.

We developed a PCR assay for the detection of Babesia odocoilei based on the 18S rRNA gene. Multiple specimens of B. odocoilei were examined, and the assay consistently produced a small specific PCR product of 306 bp. The PCR assay was also challenged with DNA from 13 other Babesia species and 2 Theileria species, originating from 10 different host species; however, nonspecific DNA amplification and multiple banding patterns were observed, and the amplicon banding patterns varied between different isolates of the same species. Sensitivity was determined to be 6.4 pg of DNA, and an estimated 0.0001% parasitism. This assay can be utilized for species-specific differential detection of B. odocoilei.

Use of slide scrape lysates for polymerase chain reaction confirmation of disseminated Mycobacterium avium infection in a cat.

Disseminated mycobacteriosis in a 3-year-old domestic medium-haired cat was diagnosed on lymph node cytology. Slide scrape lysates from the cytology submission were used to confirm Mycobacterium avium by polymerase chain reaction (PCR) and sequencing and proved a simple technique that could be a valuable tool in veterinary diagnostics and research.

Utilisation d'un lysat de grattage de lame pour la confirmation par amplification en chaîne par la polymérase d'une infection disséminée à Mycobacterium avium chez un chat. Une mycobactériose disséminée chez un chat domestique à poil moyen âgé de 3 ans a été diagnostiquée à l'aide d'une cytologie des ganglions lymphatiques. La soumission d'un lysat de grattage d'une lame provenant de la soumission de cytologie a été utilisé pour confirmer Mycobacterium avium par amplification en chaîne par la (PCR) et séquençage et elle s'est avérée une technique simple qui pourrait être un outil utile dans les diagnostics et la recherche vétérinaires.(Traduit par Isabelle Vallières).

Use of the polymerase chain reaction assay for the detection of Babesia odocoilei 18S ribosomal RNA in formalin-fixed tissues.

The effect of fixation and storage conditions on the performance of polymerase chain reaction (PCR) assays for Babesia odocoilei were examined using 3 different primer sets targeting the eukaryotic 18S ribosomal RNA gene, with variably sized products of 1,723 base pairs (bp), 483 bp, and 306 bp. All primer sets performed well on fresh-frozen tissue, and storage for 1 year at -20°C did not affect PCR performance. Formalin fixation markedly affected the amplicon length that could be amplified. However, DNA was successfully amplified after storage in formalin for 2 months using the primer set with a 483-bp product, and up to 6 months using the primer set with a 306-bp product. The latter primer set successfully differentiated B. odocoilei and Babesia microti DNA; however, further evaluation is required to confirm its specificity. Treatment of tissues with formic acid, at concentrations typically used to denature prions, degraded the DNA and made it unsuitable for PCR testing.

Babesia odocoilei infection in a Saskatchewan elk (Cervus elaphus canadensis) herd.

An 8-year-old female elk (Cervus elaphus canadensis) cow, presented for chronic severe weight loss and unthriftiness, was diagnosed with Babesia odocoilei infection based on blood smear evaluation, polymerase chain reaction (PCR), and DNA sequence analysis. Subsequently, velvet antler from a male that died acutely on the same farm was also PCR positive for Babesia spp. Both animals originated from a game ranch of Saskatchewan-bred and -raised animals with no known history of tick exposure, but with a history of numerous sudden deaths of unknown etiology. The presence of B. odocoilei in Canada might be a result of a recent introduction that could have deleterious effects on local wild ungulates or may represent discovery of a previously unrecognized endemic disease in local wildlife.

Equine laryngeal rhinosporidiosis in western Canada.

A 12-year-old female Argentinean Warmblood mare was evaluated because of respiratory noise. The horse resided in Calgary, Alberta, Canada, but had been imported from Argentina 28 months prior to presentation. Endoscopy of the upper respiratory tract revealed a single polypoid mass on the left arytenoid. The mass was surgically excised and was diagnosed histologically as rhinosporidiosis. Polymerase chain reaction and DNA sequencing were used to confirm the etiological agent. Four weeks postoperatively, endoscopy was repeated, revealing recurrence of the original lesion with multiple additional polypoid masses on the larynx and in the oropharynx. Resolution of the disease had not been attained at the time of publication. The current report outlines a case of rhinosporidiosis in an unusual anatomical and geographic location. The infection most likely originated in Argentina, with a prolonged subclinical phase. Due to increased travel of human beings and animals, there is potential for the introduction of exotic diseases into nonendemic areas.