Frequency of isolation and phenotypic antimicrobial resistance of fecal Salmonella enterica recovered from dairy cattle in Canada.

Salmonellosis is one of the leading causes of gastrointestinal infections in humans. In Canada, it is estimated that approximately 87,500 cases of salmonellosis occur every year in humans, resulting in 17 deaths. In the United States, it is estimated that 26,500 hospitalizations and 420 deaths occur every year. In dairy cattle, infections caused by nontyphoidal Salmonella enterica can cause mild to severe disease, including enteritis, pneumonia, and septicemia. Our study objectives were to determine the proportion of fecal samples positive for Salmonella in dairy cattle in Canada and determine the resistance pattern of these isolates. We used data collected through the Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR). Pooled fecal samples from preweaning calves, postweaning heifers, lactating cows, and manure storage were cultured for Salmonella, and the isolates were identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibilities were determined using the minimum inhibitory concentration test, and resistance interpretation was made according to the Clinical and Laboratory Standards Institute. A 2-level, multivariable logistic regression model was built to determine the probability of recovering Salmonella from a sample, accounting for province, year, and sample source. The proportion of farms with at least one positive sample were 12% (17/140), 19% (28/144), and 17% (24/144) for the sampling years 2019, 2020, and 2021, respectively. Out of the 113 Salmonella isolates, 23 different serovars were identified. The occurrence of Salmonella appeared to be clustered by farms and provinces. The most common serovars identified were Infantis (14%) and Typhimurium (14%). Overall, 21% (24/113) of the Salmonella isolates were resistant to at least one antimicrobial. Resistance to tetracycline was commonly observed (17%); however, very limited resistance to category I antimicrobials (categorization according to Health Canada that includes third-generation cephalosporins, fluoroquinolones, polymyxins, and carbapenems) was observed, with one isolate resistant to amoxicillin and clavulanic acid. The proportion of Salmonella isolates resistant to 2 and 3 antimicrobial classes was 3.5% and 8.8%, respectively. Our study provided valuable information on the proportion of fecal samples positive for Salmonella, the serovars identified, and the associated resistance patterns across CaDNetASR herds, at regional and national levels.

Temporal patterns of bovine leukemia virus infection in dairy herds in Atlantic Canada.

Objective: The primary objective was to determine the youngest age group where bovine leukemia virus (BLV)-infected dairy animals were identified. The secondary objective was to investigate associations between age-specific management practices and BLV infection status of different age groups of dairy calves and heifers. Procedure: For enrolled herds, BLV status was determined using blood samples from pre-weaned calves, weaned calves, and breeding-age heifers; and bulk tank milk from the adult herd. A questionnaire investigating age-specific management factors was administered for each herd. Ordinal logistic regression was performed to identify management factors associated with the youngest age range in which BLV was identified. Results: Fifty-three dairy herds from the 4 provinces in Atlantic Canada were enrolled. Bovine leukemia virus was most commonly earliest identified in pre-weaned heifers (18 herds, 32.1%) and the adult herd (18 herds, 32.1%). Ordinal logistic regression revealed that BLV was first identified in older age groups more often than in younger age groups when herds regrouped weaned heifers at least once, when fly control was used for breeding-age heifers, when herds practiced foot trimming on breeding-age heifers, and when bred heifers were brought in. Conclusion: Producers can use results to identify the youngest age group(s) in which BLV is identified and to tailor management strategies to prevent new infections.

Tendances temporelles de l'infection par le virus de la leucémie bovine dans les troupeaux laitiers des provinces atlantiques canadiennes. Objectif: L'objectif principal était de déterminer le groupe d'âge le plus jeune dans lequel les animaux laitiers infectés par le virus de la leucémie bovine (BLV) ont été identifiés. L'objectif secondaire était d'étudier les associations entre les pratiques de gestion spécifiques à l'âge et le statut d'infection par le BLV de différents groupes d'âge de veaux et de génisses laitiers. Procédure: Pour les troupeaux inscrits, le statut BLV a été déterminé à l'aide d'échantillons de sang provenant de veaux présevrés, de veaux sevrés et de génisses en âge de se reproduire; et de lait de réservoir en vrac du troupeau adulte. Un questionnaire portant sur les facteurs de gestion spécifiques à l'âge a été administré pour chaque troupeau. Une régression logistique ordinale a été réalisée pour identifier les facteurs de gestion associés à la tranche d'âge la plus jeune dans laquelle le BLV a été identifié. Résultats: Cinquante-trois troupeaux laitiers des quatre provinces atlantiques canadiennes ont été inscrits. Le virus de la leucémie bovine a été le plus souvent identifié le plus tôt chez les génisses pré-sevrées (18 troupeaux, 32,1 %) et dans le troupeau adulte (18 troupeaux, 32,1 %). La régression logistique ordinale a révélé que le BLV a été identifié pour la première fois plus souvent dans les groupes d'âge plus âgés que dans les groupes d'âge plus jeunes lorsque les troupeaux regroupaient au moins une fois les génisses sevrées, lorsque le contrôle des mouches était utilisé pour les génisses en âge de se reproduire, lorsque les troupeaux pratiquaient le parage des pattes des génisses en âge de se reproduire., et quand les taures saillies étaient intégrées au troupeau. Conclusion: Les producteurs peuvent utiliser les résultats pour identifier le(s) groupe(s) d'âge le plus jeune dans lequel le BLV est identifié et pour adapter les stratégies de gestion afin de prévenir de nouvelles infections.(Traduit par Dr Serge Messier).

Relationship between quantitative real-time PCR cycle threshold and culture for detection of Streptococcus equi subspecies equi.

Objective: To compare PCR and culture results for the detection of Streptococcus equi subspecies equi (S. equi). Animals: Respiratory tract samples (N = 158) from horses being tested for S. equi. Procedure: Bacterial culture was carried out on samples from which S. equi was detected by quantitative real-time PCR. Results: S. equi was isolated from 12 (7.6%) samples: 4/9 (44%) samples when the PCR cycle threshold (CT) was ≤ 30, 7/30 (23%) when the CT was 30.1 to 35, and 1/119 (0.8%) when the CT was 35.1 to 40. The highest CT sample from a sample that yielded a positive culture was 36.9. The optimal Youden's J value was at a CT of 34.2, the same value as determined by number needed to misdiagnose when the cost of a false negative is deemed to be either 5 or 10 × that of a false positive. Conclusions: Viable S. equi was only detected in a minority of quantitative PCR (qPCR) positive samples. A qPCR CT of 34.2 was a reasonable breakpoint for likelihood of the presence of culturable S. equi. Clinical relevance: Evaluation of CT values may be useful as a proxy to indicate the likelihood of cultivable S. equi being present and could be useful as part of risk assessments.

Relation entre le seuil du cycle de PCR quantitatif en temps réel et la culture pour la détection de Streptococcus equi sous-espèce equi. Objectif: Comparer les résultats de PCR et de culture pour la détection de Streptococcus equi sous-espèce equi (S. equi). Animaux: Échantillons des voies respiratoires (N = 158) de chevaux testés pour S. equi. Procédure: La culture bactérienne a été réalisée sur des échantillons à partir desquels S. equi a été détecté par PCR quantitatif en temps réel. Résultats: S. equi a été isolé à partir de 12 échantillons (7,6 %) : 4/9 (44 %) échantillons lorsque le seuil du cycle de PCR (CT) était ≤ 30, 7/30 (23 %) lorsque le CT était de 30,1 à 35 et 1/119 (0,8 %) lorsque le CT était de 35,1 à 40. L'échantillon CT le plus élevé d'un échantillon ayant donné une culture positive était de 36,9. La valeur J optimale de Youden était à un CT de 34,2, la même valeur que celle déterminée par le nombre nécessaire pour un mauvais diagnostic lorsque le coût d'un faux négatif est estimé à 5 ou 10 × celui d'un faux positif. Conclusion: Du S. equi viable n'a été détecté que dans une minorité d'échantillons positifs pour le PCR quantitatif (qPCR). Un CT qPCR de 34,2 était un seuil raisonnable pour la probabilité de la présence de S. equi cultivable. Pertinence clinique: L'évaluation des valeurs CT peut être utile comme approximation pour indiquer la probabilité de présence de S. equi cultivable et pourrait être utile dans le cadre d'une évaluation des risques.(Traduit par Dr Serge Messier).

Comparison of chlorhexidine and alcohol-based antisepsis on the paralumbar fossa in cattle.

OBJECTIVE: To determine skin reaction, post-treatment reduction (immediate effect), and 1 hour post-treatment reduction (sustained effect) of aerobic bacterial colony forming units (CFU) following three antiseptic protocols in cattle. STUDY DESIGN: Prospective, randomized experimental study. ANIMALS: Eighteen cows. METHODS: Three sites in each paralumbar fossa were clipped and randomly assigned to one of three treatment groups: 5 minute 4% chlorhexidine gluconate scrub (CHG); 90 second 80% ethanol scrub (ET); 90 second 70% isopropyl alcohol scrub (IPA). All sites were monitored at all sampling time points and at 24 hours following treatment for adverse skin reaction. Samples were collected pre-, immediately post-, and 1 hour post-treatment and plated in duplicate. Bacterial counts were shifted to eliminate zeroes, log10 transformed, and averaged. ANOVA was used to compare differences in mean reduction in log10 CFU/ml between groups. RESULTS: Reduction in log10CFU/ml was more pronounced immediately after application of IPA (p = .001) and ET (p = .001) than CHG. This reduction was better sustained after preparation with CHG than ET (p = .005) but not IPA. Immediate and sustained reductions in bacterial loads did not differ after application of IPA or ET. No adverse skin reactions were noted. CONCLUSIONS: Skin preparation with alcohol-based antiseptics was well tolerated and improved immediate bacterial reduction compared to CHG. This reduction was better sustained 1 hour after application of CHG than ET, but no difference was detected between CHG and IPA. CLINICAL RELEVANCE: Lack of adverse skin reaction and performance provide evidence to support skin preparation with alcohol-based antiseptics in cattle.

Short communication: Whole-genome sequence analysis of 4 fecal blaCMY-2-producing Escherichia coli isolates from Holstein dairy calves.

This study was carried out to determine the antimicrobial resistance (AMR) genes and mobile genetic elements of 4 fecal blaCMY-2-producing Escherichia coli isolated from Holstein dairy calves on the same farm using whole-genome sequencing. Genomic analysis revealed that 3 of the 4 isolates shared similar genetic features, including sequence type (ST), serotype, plasmid characteristics, insertion ST, and virulence genes. In addition to genes encoding for complex multidrug resistance efflux systems, all 4 isolates were carriers of genes conferring resistance to ß-lactams (blaCMY-2, blaTEM-1B), tetracyclines (tetA, tetB, tetD), aminoglycosides [aadA1, aph(3")-lb, aph(6)-ld], sulfonamides (sul2), and trimethoprim (dfrA1). We also detected 4 incompatibility plasmid groups: Inc.F, Inc.N, Inc.I, and Inc.Q. A novel ST showing a new purA and mdh allelic combination was found. The 4 isolates were likely enterotoxigenic pathotypes of E. coli, based on serotype and presence of the plasmid Inc.FII(pCoo). This study provides information for comparative genomic analysis of AMR genes and mobile genetic elements. This analysis could give some explanation to the multidrug resistance characteristics of bacteria colonizing the intestinal tract of dairy calves in the first few weeks of life.

Compliance with the Code of Practice for the Care and Handling of Equines on 50 horse farms in Prince Edward Island.

The objective of this study was to determine the proportion of horse farms on Prince Edward Island, Canada that comply with the requirements of the Code of Practice for the Care and Handling of Equines (Code). An investigator performed on-farm assessments while administering a questionnaire to owners of 50 horse farms. The percentage of farms in compliance with specific requirements in the Code ranged from 20% to 100% per requirement. The largest areas of non-compliance regarding facilities and housing were the lack of the ability to segregate sick or injured animals and the lack of an emergency action plan. It was determined that 72% of farms were in compliance with body condition scores and 54% reported to have taken corrective action when required. Farm owners who were aware of the Code were more likely to have good quality air in their barns as well as an emergency action plan in place.

L'objectif de la présente étude était de déterminer la proportion de fermes équines sur l'Île-du-Prince-Édouard, Canada qui se conforme aux exigences du Code de pratiques pour les soins et la manipulation des chevaux (Code). Un enquêteur effectua des évaluations à la ferme tout en procédant à un questionnaire auprès des propriétaires de 50 fermes équines. Le pourcentage de fermes en conformité avec des exigences spécifiques du Code variait de 20 % à 100 % selon l'exigence. Les secteurs les plus importants de nonconformité en regard des facilités et de logement étaient l'absence de la capacité à isoler les animaux malades ou blessés et l'absence d'un plan d'action d'urgence. Il fut déterminé que 72 % des fermes étaient en conformité pour les pointages d'état de chair et 54 % rapportèrent d'avoir pris les actions correctives lorsque requises. Les propriétaires de ferme qui étaient au courant du Code étaient plus susceptibles d'avoir un air de bonne qualité dans leurs fermes ainsi qu'un plan d'action d'urgence en place.(Traduit par Dr Serge Messier).

Determination of antimicrobial resistance to extended-spectrum cephalosporin, quinolones, and vancomycin in selected human enteric pathogens from Prince Edward Island, Canada.

The aim of this study was to determine the frequency of fecal carriage of vancomycin-resistant Enterococcus spp. and Escherichia coli with reduced susceptibilities to extended-spectrum cephalosporins (ESCs) and quinolones in humans on Prince Edward Island, Canada. Convenience fecal samples from individuals on Prince Edward Island were screened phenotypically using selective culture and genotypically using multiplex polymerase chain reactions to detect E. coli and Enterococcus spp. resistant to critically important antimicrobials. Twenty-six (5.3%) of 489 individuals had E. coli with reduced susceptibility to ESCs. Twenty-five (96.2%) of the 26 isolates harbored blaTEM, 18 (69.2%) harbored blaCMY-2, 16 (61.5%) harbored blaCTX-M groups, 2 (7.7%) harbored blaSHV genes. None of the ESC-resistant E. coli was positive for carbapenem resistance. Twenty-one (8.3%) of 253 individuals had E. coli isolates with reduced quinolone susceptibility. All 21 isolates were positive for at least 1 qnr gene, with 3 (14.3%) isolates positive for qnrB, 5 (23.8%) positive for qnrS, and 13 (61.9%) positive for both qnrB and qnrS genes. All the enterococci isolates were vancomycin-susceptible. Higher susceptibility to the critically important antimicrobials was found in this study. This study can serve as a baseline for future antimicrobial resistance surveillance within this region.

Antimicrobial resistance in bacteria isolated from cats and dogs from the Atlantic Provinces, Canada from 1994-2013.

Antimicrobial susceptibility patterns and trends in bacteria isolated from cats and dogs were determined from diagnostic laboratory data from the Atlantic Veterinary College Diagnostic Services Bacteriology Laboratory over a 20-year period. Clinical samples were most commonly from the urinary tract and the ear. Staphylococcus spp. and Escherichia coli were the bacteria that were most frequently isolated. Increases in percentage resistant were seen with E. coli to cephalexin (57% to 61%), Pasteurella spp. to erythromycin (35% to 53%), and Pseudomonas aeruginosa (31% to 39%), and Streptococcus spp. (46% to 53%) to enrofloxacin. The frequency of resistance did not change significantly over the study period; however, increased enrofloxacin resistance was identified for canine isolates of Staphylococcus spp., Streptococcus spp., Enterococcus spp., E. coli, P. aeruginosa, and Proteus spp. Multidrug resistance was observed in 12% and 9% of the isolates from dogs and cats, respectively. Data from this study could be used to guide empirical antimicrobial selection in companion animal veterinary practices in Atlantic Canada.

Résistance antimicrobienne des bactéries isolées chez des chats et des chiens des provinces de l'Atlantique, Canada, de 1994 à 2013. Les tendances de susceptibilité antimicrobienne des bactéries isolées chez des chats et des chiens ont été déterminées d'après les données des laboratoires de diagnostic du Diagnostic Services Bacteriology Laboratory de l'Atlantic Veterinary College pendant une période de 20 ans. Les échantillons cliniques provenaient le plus communément des voies urinaires et de l'oreille. Staphylococcus spp. et Escherichia coli étaient les bactéries les plus fréquemment isolées. Une hausse du pourcentage de la résistance a été observée pour E. coli à la céphalexine (de 57 % à 61 %), pour Pasteurella spp. à l'érythromycine (de 35 % à 53 %) ainsi que pour Pseudomonas aeruginosa (de 31 % à 39 %) et Streptococcus spp. (de 46 % à 53 %) à l'enrofloxacine. La fréquence de la résistance n'a pas changé significativement pendant la période de l'étude. Cependant, une résistance accrue à l'enrofloxacine a été identifiée pour les isolats canins de Staphylococcus spp., de Streptococcus spp., d'Enterococcus spp., d'E. coli, de P. aeruginosa, et de Proteus spp. La multirésistance aux médicaments a été observée dans 12 % et 9 % des isolats des chiens et des chats, respectivement. Les données de cette étude pourraient être utilisées pour guider le choix empirique des antimicrobiens dans les pratiques vétérinaires pour animaux de compagnie des provinces de l'Atlantique.(Traduit par Isabelle Vallières).

Association of unmeasured strong ions with outcome of hospitalized beef and dairy diarrheic calves.

Increased systemic concentrations of L-lactate and unmeasured strong ions (USI) are associated with an increased risk of mortality in human neonates and adults suffering from various diseases. This exploratory study aimed to investigate if values of certain acid-base parameters, especially L-lactate and USI, on admission to hospital are associated with mortality in diarrheic calves. Fifty-five calves < 28 days old admitted to 2 teaching hospitals for diagnosis and treatment of diarrhea were included. Admission demographic, physical examination, blood gas and biochemistry analysis, and outcome data were recorded. Admission acid-base values associated with outcome were assessed using multivariable regression modeling. Calves with elevated plasma L-lactate (OR: 1.30, 95% CI: 1.08 to 1.55; P = 0.005) and USI (OR: 1.40, 95% CI: 1.12 to 1.74; P = 0.003) at admission were more likely to die or to be euthanized. This study revealed that elevated concentrations of L-lactate and USI at admission were positively associated with mortality.

Association des ions forts non mesurés avec les résultats des veaux de boucherie et laitiers diarrhéiques hospitalisés. Des concentrations systémiques accrues de L-lactate et des ions forts non mesurés (IFN) sont associées à un risque accru de mortalité chez les nouveau-nés humains et les adultes souffrant de diverses affections. Cette étude exploratoire visait à vérifier si les valeurs de certains paramètres acides, particulièrement L-lactate et IFN, à l'admission à l'hôpital sont associées à la mortalité chez les veaux diarrhéiques. Cinquante-cinq veaux âgés de < 28 jours admis à deux hôpitaux d'enseignement pour le diagnostic et le traitement ont été inclus. Les données démographiques, l'examen physique, les valeurs des gaz sanguins et de l'analyse biochimique du sang ainsi que l'issu des animaux ont été consignés. Les valeurs de l'équilibre acide-base à l'admission associées à l'issu des animaux ont été évaluées en utilisant une modélisation de régression multi-variable. Il était plus probable que les veaux avec du L-lactate plasmatique élevé (RC : 1,30, IC 95 %, 1,08 à 1,55; P = 0,005) et IFN (RC : 1,40, IC de 95 %, 1,12 à 1,74; P = 0,003), à l'admission meurent ou soient euthanasiés. Cette étude a révélé que des concentrations élevées de L-lactate et d'IFN à l'admission étaient positivement associées à la mortalité.(Traduit par Isabelle Vallières).

A rapid field test for the measurement of bovine serum immunoglobulin G using attenuated total reflectance infrared spectroscopy.

BACKGROUND: Following the recent development of a new approach to quantitative analysis of IgG concentrations in bovine serum using transmission infrared spectroscopy, the potential to measure IgG levels using technology and a device better designed for field use was investigated. A method using attenuated total reflectance infrared (ATR) spectroscopy in combination with partial least squares (PLS) regression was developed to measure bovine serum IgG concentrations. ATR spectroscopy has a distinct ease-of-use advantage that may open the door to routine point-of-care testing. Serum samples were collected from calves and adult cows, tested by a reference RID method, and ATR spectra acquired. The spectra were linked to the RID-IgG concentrations and then randomly split into two sets: calibration and prediction. The calibration set was used to build a calibration model, while the prediction set was used to assess the predictive performance and accuracy of the final model. The procedure was repeated for various spectral data preprocessing approaches. RESULTS: For the prediction set, the Pearson's and concordance correlation coefficients between the IgG measured by RID and predicted by ATR spectroscopy were both 0.93. The Bland Altman plot revealed no obvious systematic bias between the two methods. ATR spectroscopy showed a sensitivity for detection of failure of transfer of passive immunity (FTPI) of 88 %, specificity of 100 % and accuracy of 94 % (with IgG <1000 mg/dL as the FTPI cut-off value). CONCLUSION: ATR spectroscopy in combination with multivariate data analysis shows potential as an alternative approach for rapid quantification of IgG concentrations in bovine serum and the diagnosis of FTPI in calves.

The association of detection method, season, and lactation stage on identification of fecal shedding in Mycobacterium avium ssp. paratuberculosis infectious dairy cows.

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative organism of Johne's disease. Although fecal culture is considered the standard diagnostic test, the long incubation times, costs, and intermittent shedding of MAP hinder efficient screening programs based on culture results. The primary objectives of this study were to determine the detection ability of solid culture, broth culture, and real-time PCR (qPCR) for MAP in fecal samples and to assess how shedding patterns of MAP in feces vary with lactation stage and season. This knowledge could improve the use of these diagnostic assays in Johne's management programs. For this study, 51 MAP-infectious cows from 7 Atlantic Canadian dairy farms had fecal samples collected monthly over a 12-mo period. Samples were analyzed for MAP bacterial load via solid culture, broth culture, and qPCR. For all fecal samples, 46% [95% confidence interval (CI): 40 to 51%] were positive by solid culture, 55% (95% CI: 50 to 60%) by broth culture, and 78% (95% CI: 73 to 82%) by qPCR. Sensitivity of qPCR was numerically higher in the dry and postpartum lactation periods, and qPCR detection in summer and fall was 85% of that in winter and spring. Furthermore, culture-determined moderate or light shedding categories generally corresponded to qPCR cycle threshold values <35, but heavy shedding categories corresponded to qPCR values <29. Direct fecal qPCR is a MAP detection method that is quick and less costly than culture techniques, and it avoids the use of decontamination steps that could decrease numbers of bacteria in a sample below the detection limit. This study indicates that, for known MAP-positive cows, fecal qPCR had high sensitivity of MAP detection, thereby supporting the use of direct fecal qPCR as part of a Johne's herd control program.

Long-read sequencing reveals the shell microbiome of apparently healthy American lobsters Homarus americanus from Atlantic Canada.

The shell microbial community of lobsters-a key factor in the development of epizootic shell disease (ESD)-is still insufficiently researched in Atlantic Canada and many knowledge gaps remain. This study aimed to establish a baseline description and analysis of the shell microbiome of apparently healthy lobsters from four locations in the region. More than 180 lobster shell swab samples were collected from New Brunswick, Nova Scotia and Prince Edward Island (PEI). PacBio long-read 16S rDNA sequencing and bioinformatic analyses in QIIME2 identified the shell-associated bacteria. The shell microbiome of healthy lobsters consisted mainly of the bacterial classes Gammaproteobacteria, Saprospiria, Verrucomicrobiae, Alphaproteobacteria, Flavobacteriia, Acidimicrobiia and Planctomycetia. The microbial composition differed regionally and seasonally, with some classes showing decreased or increased relative abundances in the PEI samples as well as in the winter and spring samples in Nova Scotia. The core shell microbiome included potentially pathogenic as well as beneficial bacterial taxa, of which some were present only in certain regions. Bacterial taxa that have previously been associated with ESD were present on healthy lobsters in Atlantic Canada, but their frequency differed by location, sampling time, and moult stage. This study indicated that geographical and seasonal factors influenced the shell microbiome of apparently healthy lobsters more than host factors such as sex, size, and moult stage. Our results provide valuable reference microbial data from lobsters in a disease-free state.

Antimicrobial use and its association with the isolation of and antimicrobial resistance in Campylobacter spp. recovered from fecal samples from Canadian dairy herds: A cross-sectional study.

Campylobacteriosis is one of the most common zoonotic diseases in North America. As opposed to humans, animal infections caused by Campylobacter spp. are often asymptomatic. In this study, data collected through the Canadian Dairy Network for Antimicrobial Stewardship surveillance system were used to determine the proportion of Campylobacter spp. and antimicrobial resistant isolates recovered from dairy cattle herds. Additionally, the association of antimicrobial use (AMU) with fecal carriage and antimicrobial resistance (AMR) of Campylobacter spp. were investigated. Pooled fecal samples from 5 animals from each production phase (pre-weaned calves, post-weaned heifers, lactating cows), and a manure storage sample were collected from 140 dairy herds across Canada. Samples were cultured using selective media, and Campylobacter isolates were speciated using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibilities were determined using the minimum inhibitory concentration test, and interpretation was made according to the Clinical and Laboratory Standards Institute. Two multilevel logistic regression models were used to investigate the association between the AMU with the isolation and antimicrobial resistance in Campylobacter spp. Of 560 samples, 63.8% were positive for Campylobacter spp., and 96% of the participating farms had at least one sample source (i.e., calves, heifers, lactating cows, or manure storage) positive for Campylobacter spp. Overall, 54.3% of the Campylobacter spp. isolates were resistant to at least one antimicrobial. Resistance to tetracycline was observed in 49.7% of the Campylobacter spp. isolates, followed by ciprofloxacin (19.9%) and nalidixic acid (19.3%). The proportion of multi-drug resistant (≥3 antimicrobial classes) Campylobacter spp. isolates was low (0.3%); however, 15.6% were resistant to two different classes of antimicrobials. Samples collected from lactating cows, heifers, and manure storage were more likely to be positive for Campylobacter spp. compared to calves. Total AMU was associated with a decreased probability of recovering Campylobacter spp. In addition, AMR to either tetracycline or ciprofloxacin had an interaction with antimicrobial use. The probability of resistance to tetracycline increased for each unit increase in the total AMU (Defined Course Dose/100 animal-years), while the probability of resistance to ciprofloxacin decreased. Campylobacter coli isolates were more likely to be resistant to ciprofloxacin and tetracycline when compared to C. jejuni. Our study demonstrated that Campylobacter spp. is widespread among Canadian dairy farms, and a higher proportion of resistance to tetracycline was identified. The total AMU was associated with increased resistance to tetracycline in Campylobacter spp. isolates; however, for ciprofloxacin the AMU was associated with decreased resistance.

Intramammary and systemic use of antimicrobials and their association with resistance in generic Escherichia coli recovered from fecal samples from Canadian dairy herds: A cross-sectional study.

Antimicrobial resistance (AMR) in animals, including dairy cattle, is a significant concern for animal and public health worldwide. In this study, we used data collected through the Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR) to: (1) describe the proportions of AMR in fecal E. coli, and (2) investigate the relationship between antimicrobial use (AMU) (intramammary and systemic routes, while accounting for confounding by other variables) and AMR/multidrug resistance (MDR - resistance to ≥ 3 antimicrobial classes) in fecal E. coli from Canadian dairy farms. We hypothesized that an increase of the AMU was associated with an increase in AMR in E. coli isolates. A total of 140 dairy farms across five provinces in Canada were included in the study. Fecal samples from pre-weaned calves, post-weaned heifers, lactating cows, and farm manure storage were cultured, and E. coli isolates were identified using MALDI-TOF MS. The minimum inhibitory concentrations (MIC) to 14 antimicrobials were evaluated using a microbroth dilution methodology. AMU was quantified in Defined Course Dose (DCD - the dose for a standardized complete treatment course on a standard size animal) and converted to a rate indicator - DCD/100 animal-years. Of 1134 fecal samples collected, the proportion of samples positive for E. coli in 2019 and 2020 was 97.1% (544/560) and 94.4% (542/574), respectively. Overall, 24.5% (266/1086) of the E. coli isolates were resistant to at least one antimicrobial. Resistance towards tetracycline was commonly observed (20.7%), whereas resistance to third-generation cephalosporins, fluoroquinolones, and carbapenems was found in 2.2%, 1.4%, and 0.1% of E. coli isolates, respectively. E. coli isolates resistant to two or ≥ 3 antimicrobial classes (MDR) was 2.7% and 15%, respectively. Two multilevel models were built to explore risk factors associated with AMR with AMU being the main exposure. Systemic AMU was associated with increased E. coli resistance. For an increase in systemic AMU equivalent to its IQR, the odds of resistance to any antimicrobial in the model increased by 18%. Fecal samples from calves had higher odds of being resistant to any antimicrobial when compared to other production ages and farm manure storage. The samples collected in 2020 were less likely to be resistant when compared to samples collected in 2019. Compared to previous studies in dairy cattle in North America, AMR in E. coli was lower.

Knowledge, attitudes and influencers of cat owners in North America around antimicrobials and antimicrobial stewardship.

OBJECTIVES: The primary aims of this study were to determine preferences of North American cat owners when they are prescribed an antimicrobial for their cat with regard to cost, method of administration and the importance of antibiotics for treating infections in people, and to establish baseline knowledge, attitudes and influencers of cat owners on antimicrobial resistance and stewardship. METHODS: An online questionnaire was used for data collection from two cat-owner groups: US cat owners and Canadian cat owners. Participants were queried on antimicrobial resistance and stewardship, and their preferences for their own cat when prescribed an antimicrobial, with respect to cost, method of drug administration and the importance of a drug for treating infections in people. Responses were evaluated through conjoint analysis and Likert-type questions. Data were analyzed using descriptive and analytic statistics. RESULTS: A total of 630 complete responses were included in the final analysis. Cost (37%) and method of administration (38%) were of similar participant preference when assessed using conjoint analysis. The importance of a drug for treating infections in people was lower priority (21%). The majority of cat owners preferred an antimicrobial that was 'very important' in treating human infections. A low proportion (21%) of participants responded that antimicrobial use in pets posed a risk to humans. Participants with a university education were more likely to respond that antimicrobial use in pets was a concern for people (31%; P <0.001). CONCLUSIONS AND RELEVANCE: Cat owners prioritize antimicrobial cost and method of administration equally. Few cat owners recognized the human antimicrobial resistance risks associated with antimicrobial use in pets.

Development of a predictive model for bovine leukemia virus proviral load.

BACKGROUND: There is currently no commercially available method in Canada to identify bovine leukemia virus (BLV)-positive cows with high proviral load (PVL). OBJECTIVES: First, develop a model to predict PVL using common, commercially available, cost-effective diagnostic tests. Second, investigate the relationship between lymphocyte count and PVL in BLV-positive cows. ANIMALS: A total of 339 BLV-positive and 62 BLV-seronegative cows on 15 dairy farms. METHODS: Cross-sectional study. Blood and milk samples were collected from all lactating BLV-positive cows on each farm and 5 to 10 BLV-seronegative cows depending on herd size. Blood and milk samples were tested for anti-BLV antibodies using enzyme-linked immunosorbent assay (ELISA). Complete blood counts were performed on blood samples, and standard components analyses were obtained for milk samples. Proviral load was determined by quantitative polymerase chain reaction for each cow. RESULTS: The inverse of lymphocyte count, the square of the inverse of lymphocyte count, and milk ELISA percent positivity were positively associated with increasing PVL in BLV-positive cows. For BLV-positive cows, lymphocyte count >5.2 × 109 /L predicted a high PVL (BLV:Bovine DNA of >1 in blood) with a sensitivity of 92.4% and a specificity of 79.8%. For BLV-positive cows, white blood cell count >10.8 × 109 /L predicted a high PVL, with a sensitivity of 85.5% and a specificity of 83.6%. CONCLUSIONS AND CLINICAL IMPORTANCE: Based on these results, producers can implement commonly available diagnostic tests to identify cows with high probability of having high PVL, which may help in designing effective disease control strategies for BLV-positive herds.

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.

Comparison of chlorhexidine and alcohol-based antisepsis of the distal limbs of horses.

BACKGROUND: An alcohol-based rub has been confirmed effective at reducing bacterial counts on equine skin. Skin sites with expected high bacterial burden have not been tested or has a comparison to a common protocol been performed. OBJECTIVES: To determine if ethanol-based antisepsis reduces bacterial counts on the equine distal limb comparable to a current chlorhexidine scrub method and determine the most effective application technique for the product. STUDY DESIGN: Randomised trial. METHODS: Forty-one horses were used in the study. By horse, each limb was randomly assigned to a treatment group: 5min scrub using 4% chlorhexidine gluconate to a clipped site (CHG); 90s scrub using 80% ethanol to a clipped site (ETC); 90s contact with 80% ethanol applied as a spray to a clipped site (ETS) and 90s scrub using 80% ethanol to an unclipped site (ETUC). Samples were collected pre- and post-treatment and plated in duplicate. Bacterial counts were log10 transformed and averaged between duplicates. A linear mixed model was used to compare mean log10 CFU/mL reduction between groups. A cost-benefit analysis was performed. RESULTS: There was no significant difference in mean log10 CFU/mL reduction between CHG and ETC in either fore- or hindlimbs. In forelimbs, there was no significant difference in mean log10 CFU/mL reduction between any groups. In hindlimbs, CHG had significantly greater mean log10 CFU/mL reduction than ETUC and ETS. No significant difference in cost-benefit was found between CHG and ETC. Significant differences were noted between CHG and both ETUC and ETS. MAIN LIMITATIONS: Researchers were not blinded to treatment group during sample collection. CONCLUSIONS: This study showed no significant difference in reduction in bacterial counts on the distal limb of horses between CHG and ethonol (ET) when applied as a scrub to a clipped site and there was no significant difference in cost-benefit between these treatments.

Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR): An On-Farm Surveillance System.

Canada has implemented on-farm antimicrobial resistance (AMR) surveillance systems for food-producing animals under the Canadian Integrated Program for Antimicrobial Resistance (CIPARS); however, dairy cattle have not been included in that program yet. The objective of this manuscript was to describe the development and implementation of the Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR). An Expert Panel (EP) of researchers was created to lead the development of the dairy surveillance system. The EP initiated a draft document outlining the essential elements of the surveillance framework. This document was then circulated to a Steering Committee (SC), which provided recommendations used by the EP to finalize the framework. CaDNetASR has the following components: (1) a herd-level antimicrobial use quantification system; (2) annually administered risk factor questionnaires; and (3) methods for herd-level detection of AMR in three sentinel enteric pathogens (generic Escherichia coli, Campylobacter spp., and Salmonella spp.) recovered from pooled fecal samples collected from calves, heifers, cows, and the manure pit. A total of 144 dairy farms were recruited in five Canadian provinces (British-Columbia, Alberta, Ontario, Québec, and Nova-Scotia), with the help of local herd veterinarians and regional field workers, and in September 2019, the surveillance system was launched. 97.1 and 94.4% of samples were positive for E. coli, 63.8, and 49.1% of samples were positive for Campylobacter spp., and 5.0 and 7.7% of samples were positive for Salmonella spp., in 2019 and 2020, respectively. E. coli was equally distributed among all sample types. However, it was more likely that Campylobacter spp. were recovered from heifer and cow samples. On the other hand, it was more common to isolate Salmonella spp. from the manure pit compared to samples from calves, heifers, or cows. CaDNetASR will continue sampling until 2022 after which time this system will be integrated into CIPARS. CaDNetASR will provide online access to farmers and veterinarians interested in visualizing benchmarking metrics regarding AMU practices and their relationship to AMR and animal health in dairy herds. This will provide an opportunity to enhance antimicrobial stewardship practices on dairy farms in Canada.

The selenium and vitamin E status of horses in Prince Edward Island.

Serum selenium (Se), vitamin E, and resting thyroid hormone concentrations were measured in 201 horses in Prince Edward Island (PEI). Selenium concentrations were either marginal (0.0053 to 0.1200 ppm) or deficient (< 0.0053 ppm) in 79% of horses based on current reference ranges for Se in serum. Aged and young adult pleasure horses had a higher prevalence of inadequate Se concentrations compared to racehorses and broodmares (82% and 97% versus 45% and 72%, respectively). Overall, 13% of horses had inadequate (< 200 µg/dL) serum vitamin E concentrations; most of these were young pleasure horses. No horses were hypothyroid and, contrary to findings in other species, there was a positive relationship between serum thyroxine and Se concentrations (P < 0.05). We conclude that Se deficiency is widespread in PEI horse populations, especially in pleasure horses, and vitamin E deficiency is more common in young pleasure horses. Micronutrient supplementation practices employed by PEI horse owners appear inadequate to ensure sufficiency.