Benzimidazole F167Y polymorphism in the canine hookworm, Ancylostoma caninum: Widespread geographic, seasonal, age, and breed distribution in United States and Canada dogs.

Surveillance data for Ancylostoma spp. and the A. caninum benzimidazole treatment resistance associated F167Y polymorphism using molecular diagnostics was obtained in a large population of dogs from the United States and Canada. Real-time PCR (qPCR) for Ancylostoma spp. and allele-specific qPCR detecting a single nucleotide polymorphism (SNP) F167Y was used in 262,872 canine stool samples collected between March and December of 2022. Ancylostoma spp. was found at an overall prevalence of 2.5% (6538/262,872), with the highest prevalence in the Southern US, 4.4% (4490/103,095), and the lowest prevalence in Canada 0.6% (101/15,829). The A. caninum F167Y polymorphism was found with the highest prevalence (13.4%, n = 46/343) in the Western US and the lowest in Canada at 4.1% (4/97). The F167Y polymorphism was detected every month over the 10-month collection period. Seasonal distribution showed a peak in June for both Ancylostoma spp. (3.08%, 547/17,775) and A. caninum F167Y (12.25%, 67/547). However, the A. caninum F167Y polymorphism prevalence was highest in September (13.9%, 119/856). Age analysis indicates a higher prevalence of both hookworm infections and occurrence of resistant isolates in puppies. The breeds with the highest F167Y polymorphism prevalence in Ancylostoma spp. detected samples were poodles (28.9%), followed by Bernese Mountain dogs (25%), Cocker spaniels (23.1%), and greyhounds (22.4%). Our data set describes widespread geographic distribution of the A. caninum benzimidazole resistance associated F167Y polymorphism in the United States and Canada, with no clear seasonality compared to the Ancylostoma spp. prevalence patterns. The F167 polymorphism was present in all geographic areas with detected hookworms, including Canada. Our study highlights that the F167Y polymorphism is represented in many dog breeds, including greyhounds.

Comparative study of a broad qPCR panel and centrifugal flotation for detection of gastrointestinal parasites in fecal samples from dogs and cats in the United States.

BACKGROUND: For decades, zinc sulfate centrifugal fecal flotation microscopy (ZCF) has been the mainstay technique for gastrointestinal (GI) parasite screening at veterinary clinics and laboratories. Elsewhere, PCR has replaced microscopy because of generally increased sensitivity and detection capabilities; however, until recently it has been unavailable commercially. Therefore, the primary aim of this study was to compare the performance of real-time PCR (qPCR) and ZCF for fecal parasite screening. Secondary aims included further characterization of markers for hookworm treatment resistance and Giardia spp. assemblages with zoonotic potential and qPCR optimization. METHODS: A convenience sampling of 931 canine/feline fecal samples submitted to a veterinary reference laboratory for routine ZCF from the Northeast US (11/2022) was subsequently evaluated by a broad qPCR panel following retention release. Detection frequency and agreement (kappa statistics) were evaluated between ZCF and qPCR for seven GI parasites [hookworm/(Ancylostoma spp.), roundworm/(Toxocara spp.), whipworm/(Trichuris spp.), Giardia duodenalis, Cystoisospora spp., Toxoplasma gondii, and Tritrichomonas blagburni] and detections per sample. Total detection frequencies were compared using a paired t-test; positive sample and co-infection frequencies were compared using Pearson's chi-squared test (p ≤ 0.05 significant) and qPCR frequency for hookworm benzimidazole (BZ) resistance (F167Y) and zoonotic Giardia spp. assemblage markers calculated. Confirmatory testing, characterization, and qPCR optimization were carried out with Sanger sequencing. RESULTS: qPCR detected a significantly higher overall parasite frequency (n = 679) compared to ZCF (n = 437) [p = < 0.0001, t = 14.38, degrees-of-freedom (df) = 930] and 2.6 × the co-infections [qPCR (n = 172) vs. ZCF (n = 66)], which was also significant (p = < 0.0001, X2 = 279.49; df = 1). While overall agreement of parasite detection was substantial [kappa = 0.74; (0.69-0.78], ZCF-undetected parasites reduced agreement for individual and co-infected samples. qPCR detected markers for Ancylostoma caninum BZ resistance (n = 5, 16.1%) and Giardia with zoonotic potential (n = 22, 9.1%) as well as two parasites undetected by ZCF (T. gondii/T. blagburni). Sanger sequencing detected novel roundworm species, and qPCR optimization provided detection beyond ZCF. CONCLUSIONS: These results demonstrate the statistically significant detection frequency advantage offered by qPCR compared to routine ZCF for both single and co-infections. While overall agreement was excellent, this rapid, commercially available qPCR panel offers benefits beyond ZCF with detection of markers for Giardia assemblages with zoonotic potential and hookworm (A. caninum) BZ resistance.

Emergence of canine hookworm treatment resistance: Novel detection of Ancylostoma caninum anthelmintic resistance markers by fecal PCR in 11 dogs from Canada.

OBJECTIVE: To describe dogs with detected Ancylostoma caninum anthelmintic treatment resistance markers in Canada. ANIMALS: 11 client-owned dogs with fecal quantitative PCR (qPCR) assay detected A caninum with benzimidazole (BZ) resistance genotypic markers. METHODS: Signalment, presenting concern, duration of clinical signs, fecal testing, treatment, and outcomes were obtained. Where available, follow-up data were collected via telephone or email with the primary veterinarian. RESULTS: Ancylostoma spp was detected from 184/32,205 dog fecal samples by reference laboratory qPCR surveillance, between May 15, 2022, and April 26, 2023. 11 of these 184 samples had A caninum with genetic BZ F167Y resistance marker detection. 4 dogs had not traveled outside Canada, 6 had been imported from the US, and the travel history was unclear in 1 dog. 7 of the dogs had gastro-intestinal signs (diarrhea or soft stool) on initial presentation. Clinical improvement was reported in 6 of these dogs (resolution of diarrhea and soft stool), with 1 dog lost to follow-up. All 11 dogs received anthelmintic treatment (varied drugs and duration). CLINICAL RELEVANCE: Identification of genetic markers of BZ resistance raises concerns about the potential animal and human impacts of resistant hookworms. 4 dogs lacked an origin from or travel history to the US, indicating true emergence and/or novel spread within Canada, not just importation from an area where resistance has been reported. Fecal surveillance was performed with a qPCR test incorporating treatment (BZ) resistance markers. There is a need to raise clinician awareness around treatment-resistant hookworm in dogs and the capability of fecal surveillance for genotypic and phenotypic resistance.

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.

Impact of a Spectrum of Care Elective Course on Third-Year Veterinary Students' Self-Reported Knowledge, Attitudes, and Competencies.

Veterinary services' rising cost is an increasing barrier to pet care. Spectrum of care (SpOC) refers to evidence-based veterinary medicine options along the socioeconomic spectrum. To meet growing pet owner financial constraints and pet care needs, training to equip veterinarians with competencies to provide SpOC as Day One graduates is argued to be added as part of the veterinary curriculum. Objectives of our prospective pre- and post-survey study were to (a) determine baseline self-reported knowledge, attitudes, and competencies (KACs) surrounding SpOC in third-year DVM students; (b) develop and assess impact of a SpOC course on student self-reported SpOC KACs; and (c) obtain student feedback on the course and future SpOC training. Enrolled students (n = 35) completed the pre-survey (n = 35) and post-survey (n = 33). Results indicated that students were aware of the need for SpOC training within the veterinary curriculum, and positive changes occurred in self-reported KAC from pre- to post-survey. Students tended (p = .08) to predict better outcomes in SpOC cost-barrier scenarios from pre- (34%) to post-survey (76%), such as reduced perceived likelihood of euthanasia (63%-39%) and unsuccessful outcomes (40%-27%). Most students (31/33, 94%) predicted the course would benefit them in clinical practice and had preferred future training preferences (online modules [70%], seminars [60%], webinars [58%]). Data indicate benefits in student self-reported KACs following the SpOC course, warranting formal course inclusion, with tracking of students into clinical practice to document objective KAC impacts and perhaps similar course rollout to other institutions.

Investigation of spatio-temporal clusters of positive leptospirosis polymerase chain reaction test results in dogs in the United States, 2009 to 2016.

BACKGROUND: Leptospirosis is a zoonotic disease of concern and an investigation of recent spatio-temporal trends of leptospirosis in dogs in the United States is needed. Leptospira PCR testing has become increasingly used in veterinary clinical medicine and these data might provide information on recent trends of disease occurrence. OBJECTIVES: To identify and describe clusters of PCR-positive Leptospira test results in dogs in the United States. ANIMALS: Leptospira real-time PCR test results from dogs (n = 40 118) in the United States from IDEXX Laboratories, Inc., between 2009 and 2016 were included in the analysis. METHODS: In this retrospective study, spatio-temporal clusters for a real-time PCR-positive test were identified using the space-time permutation scan statistic and the centroid of the zip code reported for each test. A maximum spatial window of 20% of the population at risk, and a maximum temporal window of 6 months were used. RESULTS: Seven statistically significant space-time clusters of Leptospira real-time PCR-positive test results were identified across the United States: 1 each located within the states of Arizona (2016), California (2014-2015), Florida (2010), South Carolina (2015), and 1 each located within the south-central region (2015), midwest region (2014), and northeast region (2011). Clusters ranged from 3 to 108 dogs and were identified during all years under study, except 2009, 2012, and 2013. CONCLUSIONS AND CLINICAL IMPORTANCE: The spatial and temporal components of leptospirosis in dogs in this study are similar to those in previous work. However, clusters were identified in new areas, demonstrating the complex epidemiology of this disease.

A cross-sectional study of environmental, dog, and human-related risk factors for positive canine leptospirosis PCR test results in the United States, 2009 to 2016.

BACKGROUND: Canine leptospirosis is a reemerging zoonotic disease concern in North America, and a better understanding of its epidemiology is needed. Wide-scale use and subsequent analyses of polymerase chain reaction (PCR) testing may provide additional insight into leptospirosis. This study aimed to describe temporal trends, to descriptively map, and to identify environmental, dog, and human-level factors associated with positive canine leptospirosis PCR test results in the United States. RESULTS: Data obtained from IDEXX Laboratories, Inc. on 40,118 canine leptospirosis PCR tests run in the United States between 2009 and 2016 were evaluated. Climate and socioeconomic (e.g. urban influence, income) data were obtained from public databases. Choropleth maps were created to identify high test-positive proportion areas and a cross-sectional analysis was completed with generalized (univariable, followed by multivariable) mixed logistic regression models accounting for county within state to identify significant predictors for a positive test. Overall test-positive proportion was 5.4% across the United States, with the regional point estimate highest in the southwest (8.1%). In the final multivariable model, the odds of a positive test were greater for male dogs (Odds Ratio [OR] = 1.28) and dogs 0-4 years of age (ORs ranged from 0.35-0.71 for the other age groups). The odds of a positive test were greater for dogs living in areas with wet environmental conditions (OR = 1.24). Season and temperature, as well as the interaction between them, were significant predictors of a positive test. Dogs had a greater probability of testing positive during cool temperatures (< 4 °C) compared to the other temperature categories in the fall season. CONCLUSIONS: These findings based on PCR testing allow for an improved understanding of factors influencing a positive canine leptospirosis PCR test and will assist targeted education and prevention efforts.

A diet with a struvite relative supersaturation less than 1 is effective in dissolving struvite stones in vivo.

Magnesium ammonium phosphate (struvite) is one of the most common minerals found in feline uroliths. Previous studies have shown the efficacy of acidifying calculolytic diets (inducing urine pH < 6.5), in dissolving struvite stones in cats. Recent work in our laboratory found that wet and dry test diets induce a struvite urinary relative supersaturation (RSS) < 1 and that the urine of healthy cats fed the dry test diet dissolved feline struvite stones in vitro. The objective of the present study was to demonstrate the efficacy of those test diets on naturally occurring struvite urocystoliths in cats. A total of twenty-one cats were used, of which seventeen completed the study. Of the seventeen cats, eight were fed the wet test diet and nine the dry test diet. Uroliths dissolved in a median of 18 (10-55) d. In the remaining four cats, uroliths failed to dissolve and were removed surgically. Quantitative analysis showed that these uroliths contained either calcium oxalate or calcium phosphate. The present study demonstrates that diets that induce a struvite RSS < 1 result in struvite stone dissolution in vivo.

Eosinophilic cystitis in a female German wire-haired pointer.

A 7-month-old, intact female, German wire-haired pointer presented with a 3-week history of stranguria, pollakiuria, and dysuria that was nonresponsive to antibiotics. Two prior episodes of dysuria-stranguria appeared to respond to antibiotic therapy. Bladder wall biopsies revealed eosinophilic cystitis and the dog responded well to medical management.

Suspect osteogenesis imperfecta in a male kitten.

A 4.5-month-old, male domestic shorthair was presented with bilateral femoral fractures after falling from a low height. Radiographs revealed reduced radio-opacity and thin cortices of all long bones. A presumptive diagnosis of osteodystrophy, secondary to osteogenesis imperfecta, was made on postmortem examination.

Alterations in thyroid hormone concentrations in healthy sled dogs before and after athletic conditioning.

OBJECTIVE: To determine effects of athletic conditioning on thyroid hormone concentrations in a population of healthy sled dogs. ANIMALS: 19 healthy adult sled dogs. PROCEDURE: Serum concentrations of thyroxine (T4), triiodothyronine (T3), thyroid-stimulating hormone (TSH), free T4 (fT4), free T3 (fT3), and autoantibodies directed against T3, T4, and thyroglobulin were measured in sled dogs that were not in training (ie, nonracing season) and again after dogs had been training at maximum athletic potential for 4 months. RESULTS: Analysis revealed significant decreases in T4 and fT4 concentrations and a significant increase in TSH concentration for dogs in the peak training state, compared with concentrations for dogs in the untrained state. Serum concentrations of T4 and fT4 were less than established reference ranges during the peak training state for 11 of 19 and 8 of 19 dogs, respectively; fT4 concentration was greater than the established reference range in 9 of 19 dogs in the untrained state. CONCLUSIONS AND CLINICAL RELEVANCE: Decreased total T4 and fT4 concentrations and increased serum concentrations of TSH were consistently measured during the peak training state in healthy sled dogs, compared with concentrations determined during the untrained state. Although thyroid hormone concentrations remained within the established reference ranges in many of the dogs, values that were outside the reference range in some dogs could potentially lead to an incorrect assessment of thyroid status. Endurance training has a profound impact on the thyroid hormone concentrations of competitive sled dogs.