Adoption and decision factors regarding selective treatment of clinical mastitis on Canadian dairy farms.

As clinical mastitis (CM) treatments are responsible for a large portion of antimicrobial use on dairy farms, many selective CM treatment protocols have been developed and evaluated against a blanket treatment approach of CM cases. Selective treatment protocols use outcomes of diagnostic tests to exclude CM cases from antimicrobial treatment when they are unlikely to benefit. To tailor interventions to increase uptake of selective treatment strategies, a comprehension of current on-farm treatment practices and factors affecting treatment decisions is vital. Two questionnaires were conducted among 142 farms across 5 provinces participating in the Canadian Dairy Network for Antimicrobial Stewardship and Resistance in this cross-sectional study. Self-reported adoption of selective CM treatments by dairy farmers was 64%, with median of 82% of cows treated in those herds using selective treatment. Using logistic regression models, the odds to implement a selective CM treatment protocol increased with a decreasing average cow somatic cell count. No other associations were identified between use of a selective CM treatment protocol and farm characteristics (herd size, CM incidence, province, milking system, and housing system). Three subsets of farmers making cow-level CM treatment decisions were identified using a cluster analysis approach: those who based decisions almost exclusively on severity of clinical signs, those who used various udder health indicators, and farmers who also incorporated more general cow information such as production, age, and genetics. When somatic cell count was considered, the median threshold used for treating was >300,000 cells/mL at the last Dairy Herd Improvement test. Various thresholds were present among those considering CM case history. Veterinary laboratories were most frequently used for bacteriological testing. Test results were used to start, change, and stop treatments. Regardless of protocol, reasons for antimicrobial treatment withheld included cow being on a cull list, having a chronic intramammary infection, or being at end of lactation (i.e., close to dry off). If clinical signs persisted after treatment, farmers indicated that they would ask veterinarians for advice, stop treatment, or continue with the same or different antibiotics. Results of this study can be used to design interventions targeting judicious mastitis-related antimicrobial use, and aid discussions between veterinarians and dairy producers regarding CM-related antimicrobial use.

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.

Occurrence of extended-spectrum ß-lactamase and AmpC-producing Escherichia coli in retail meat products from the Maritime Provinces, Canada.

This study was conducted to determine the occurrence of antimicrobial resistance to the extended-spectrum cephalosporins (ESC) in Escherichia coli isolates. The isolates were collected from retail meat products collected in the Maritime Provinces of Canada. Our analyses involved the use of both selective and traditional culture methods; we also conducted genotype analyses using multiplex polymerase chain reactions. ESC-resistant (ESC-R) E. coli were detected in 33 of 559 samples (5.9%) using the traditional culture method, compared with 151 of 557 samples (27.1%) using the selective culture method. We recovered more isolates of ESC-R E. coli from poultry compared with beef and pork (P < 0.001). Multidrug resistance, extended-spectrum ß-lactamase (ESBL), and AmpC phenotypes were more common in chicken-derived isolates than other retail meat products (P < 0.001). From the 98 isolates examined, 76 isolates (77.6%) were positive for either ESBL and AmpC ß-lactamases or both. Among the 76 isolates, blaCMY-2 (78.9%), blaCTXM (46.1%), blaTEM (21.1%), and blaSHV (1.3%) genes were detected. Among the blaCTXM-producing isolates, blaCTXM-1, blaCTXM-2, and blaCTXM-9 phylogenetic groups were detected. ß-lactamase genes were more commonly detected in chicken-derived isolates compared with other meat types (P < 0.01). This study demonstrates the occurrence of ESBL- and AmpC-resistance genes in retail meat products in the Maritime Provinces of Canada. We found that selective culture significantly improved the recovery of ESC-R E. coli isolates from retail meat samples.

Characterization of the microbiota and resistome of bulk tank milk samples from Prince Edward Island dairy farms.

Bulk tank milk (BTM) is regularly used for surveillance on dairy farms for disease conditions such as mastitis and Johne's disease. In this study, we used 16S rRNA sequencing and bait-capture enrichment to characterize the microbiota and resistome of BTM, and investigate potential differences between the cream or pellet fractions. A total of 12 BTM samples were taken from 12 Prince Edward Island dairy farms, in Atlantic Canada, in duplicates. The DNA was analyzed by high-throughput sequencing of the 16S rRNA gene and a suite of antimicrobial resistance (AMR) genes. Target-capture enrichment of AMR genes was conducted before shotgun sequencing. The bioinformatics pipelines QIIME 2 and AMR++ were used for microbiota and resistome analysis, respectively. Differences between microbiotae were evaluated qualitatively with nonmetric multidimensional scaling and quantitatively with permutational ANOVA of UniFrac distances. A total of 47 phyla were present across the BTM samples. Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria were the most abundant phyla. At the genus level, Corynebacterium, Acinetobacter, Lactobacillus, and Turicibacter were the most abundant. There was no significant difference in the Faith's phylogenetic diversity between the cream and pellet fraction. Faith's phylogenetic diversity differed marginally by stall type. There were 10,217 hits across 80 unique AMR genes, with tetracycline resistance being the most common class.

Effect of housing improvement and other factors on the growth of heifer calves on Kenyan smallholder dairy farms.

This trial evaluated average daily gain (ADG) effects of heifer calves (< 1 year old) from affordable housing improvements to the roof and flooring on 150 randomly allocated smallholder dairy farms. During the 16-month data collection period, bimonthly farm visits were used to measure weight and other animal- and farm-level factors on the 187 study calves. Multivariable linear regression was used to model ln ADG and ADG during pre-weaning and post-weaning periods, respectively. Median pre-weaning and post-weaning ADGs were 0.307 (interquartile range (IQR): 0.227-0.398) and 0.487 (IQR: 0.354-0.675) kg/d, respectively. In the final pre-weaning model (p<0.050), factors positively associated with ln ADG were calf age at first acaricide application, and total number of calf pens, while factors negatively associated with ln ADG included calf mortality risk over the last 5 years and calf age at first ad lib access to water. In an interaction term, for calves from parity 3+ dams, ADG was lower when milk was fed twice/day than thrice/day, with no difference in calves of lower parity dams. In the final post-weaning model, housing improvements increased ADG by 5.6%. Other factors positively associated with post-weaning ADG were feeding of calf pellets, wheat bran, maize bran, and hay. Calf age at first introduction of concentrate and calf mortality risk over the last 5 years were negatively associated with ADG. In an interaction term, ADG was high when there were faecal coccidia oocysts and when calves had visual or physical contact with their dams, but low when faecal coccidia cysts were present, and these dam-calf connections were absent. In a second interaction term, ADG increased with more calf pens for female principal farmers, while remaining low for male principal farmers. In conclusion, while controlling for other factors of ADG, making affordable calf housing improvements enhanced ADG, particularly during the post-weaning period.

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.

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.

Salmonella enterica and extended-spectrum cephalosporin-resistant Escherichia coli recovered from Holstein dairy calves from 8 farms in New Brunswick, Canada.

This study was carried out to determine the frequency of fecal carriage, antimicrobial susceptibility, and resistance genes in Salmonella enterica and Escherichia coli with reduced susceptibility to extended-spectrum cephalosporins (ESC) isolated from 488 dairy calves from 8 farms in New Brunswick, Canada. Both S. enterica and E. coli with reduced susceptibility to ESC were isolated using selective culture. Minimum inhibitory concentrations to a panel of antimicrobial drugs were determined for randomly selected E. coli isolates and all of the Salmonella isolates. Multiplex PCR were conducted on the selected ESC-resistant E. coli to assess the ß-lactamase resistance genes (blaCTX-M, blaCMY-2, blaSHV, and blaTEM) and plasmid-mediated qnrB and qnrS resistant genes. Information on ceftiofur use and other farm management practices were collected by the use of a questionnaire to determine the risk factors for the fecal recovery of E. coli with reduced susceptibility to ESC. Salmonella enterica frequency in calves' fecal samples was 3.3%, and all were pansusceptible. Salmonella isolates belonged to 3 serovars namely Salmonella Senftenberg, Salmonella Typhimurium, and Salmonella Derby. The frequency of fecal carriage of E. coli with reduced susceptibility to ESC in calves was 81.2%. Of the selected isolates (n = 100), all were multi-drug resistant, whereas 88% were ESC resistant based on minimum inhibitory concentration testing. From the selected ESC-resistant E. coli isolates, blaTEM was detected in 84.1%, blaCMY-2 was detected in 52.2%, blaCTXM groups were detected in 30.7%, and blaSHV was detected in 1.1% of isolates. Plasmid-mediated quinolone resistance genes were identified in 7 of 9 isolates resistant to quinolones. Five isolates were positive for qnrB, whereas 2 isolates were positive for both qnrB and qnrS. Whereas neonatal calves [odds ratio (OR) = 2.42, 95% confidence interval (CI): 1.87-3.12], regular ceftiofur use on the farm (OR = 3.83, 95% CI: 2.29-6.39), feeding of unpasteurized nonsalable milk (OR = 1.6, 95% CI: 1.18-2.18), and use of florfenicol (OR = 2.02, 95% CI: 1.43-2.86) were statistically associated with fecal recovery of E. coli with reduced susceptibility to ESC, use of ceftiofur for the treatment of respiratory diseases (OR = 0.57, 95% CI: 0.41-0.79) was statistically associated with decreased recovery of E. coli with reduced susceptibility to ESC. This study has provided information on the resistance genes associated with the occurrence of ESC and fluoroquinolone resistance in dairy calves within this region.

Antimicrobial resistance in bacteria isolated from horses from the Atlantic Provinces, Canada (1994 to 2013).

This study determined the antimicrobial susceptibility patterns and trends for selected bacteria isolated from horses using diagnostic data from the Atlantic Veterinary College Diagnostic Services Bacteriology Laboratory, Charlottetown, Prince Edward Island over a 20-year period. Streptococcus equi subsp. zooepidemicus and Escherichia coli were the most commonly isolated bacteria over the study period. Clinical samples were most frequently submitted from respiratory and reproductive systems. Most bacterial isolates were susceptible to ceftiofur. Resistance was most common in Gram-negative enteric bacteria, while streptococci were frequently susceptible to most of the antimicrobials tested including penicillin and trimethoprim-sulfamethoxazole. The antimicrobial resistance trends over the study period were relatively stable. Multi-drug resistance was observed in 9% of the bacterial isolates. Information provided in this study could be used to help guide rational, empirical antimicrobial treatment selection in equine practices in Atlantic Canada.

Résistance aux antimicrobiens dans les bactéries isolées des chevaux dans les provinces de l'Atlantique, au Canada, de 1994 à 2013. Cette étude a déterminé les modèles et les tendances de susceptibilité antimicrobienne pour certaines bactéries isolées des chevaux en s'appuyant sur des données diagnostiques du Laboratoire de bactériologie des services diagnostiques de l'Atlantic Veterinary College à Charlottetown, à l'Île-du-Prince-Édouard, pendant une période de 20 ans. Streptococcus equi ssp. zooepidemicus et Escherichia coli ont été les bactéries les plus communément isolées pendant la période à l'étude. Des échantillons cliniques des systèmes respiratoires et reproducteurs étaient les plus fréquemment soumis. La plupart des isolats bactériens étaient susceptibles au ceftiofur. La résistance était la plus courante chez les bactéries entériques à Gram négatif, tandis que les streptocoques étaient fréquemment susceptibles à la plupart des antimicrobiens testés incluant la pénicilline et le triméthoprime-sulfaméthoxazole. Les tendances de résistance aux antimicrobiens pendant la période à l'étude étaient relativement stables. Une multirésistance aux médicaments a été observée dans 9 % des isolats bactériens. Les renseignements fournis dans cette étude pourraient servir à guider un choix rationnel et empirique pour les traitements antimicrobiens dans les pratiques équines des provinces de l'Atlantique.(Traduit par Isabelle Vallières).

Antimicrobial resistance in mastitis, respiratory and enteric bacteria isolated from ruminant animals from the Atlantic Provinces of Canada from 1994-2013.

Diagnostic laboratory antimicrobial susceptibility data for bacteria isolated from clinical samples of cattle, sheep, and goats from 1994 to 2013 were evaluated retrospectively. Among bacteria from bovine mastitis, Staphylococcus aureus and Streptococcus uberis were the most commonly isolated organisms. Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni were commonly isolated from the respiratory tract, while Escherichia coli isolates were frequently recovered from the intestinal tract. Isolates from mastitis were generally highly susceptible to the antimicrobials tested, except neomycin and oxytetracycline. Respiratory tract isolates were highly susceptible to trimethoprim-sulfamethoxazole, penicillin, florfenicol, and ceftiofur, while enteric bacteria were frequently susceptible to ceftiofur. Antimicrobial resistance trends over the study period were generally stable for small ruminant and cattle isolates. Multi-drug resistance was more common among respiratory isolates from small ruminants compared to those from cattle but more common in enteric bacteria from cattle compared to those from small ruminants. This information may guide clinicians when they are choosing empirical therapies for the treatment of ruminant animals in Atlantic Canada.

Résistance antimicrobienne des bactéries respiratoires et entériques et de la mammite isolées des ruminants dans les provinces de l'Atlantique de 1994 à 2013. Les données de susceptibilité antimicrobienne des laboratoires diagnostiques, de 1994 à 2013, pour les bactéries isolées d'échantillons cliniques de bovins, de moutons et de chèvres, ont été évaluées rétrospectivement. Parmi les bactéries trouvées en lien avec la mammite bovine, Staphylococcus aureus et Streptococcus uberis ont été les organismes les plus communément isolés. Pasteurella multocida, Mannheimia haemolytica et Histophilus somni ont été communément isolés de l'appareil respiratoire, tandis que des isolats d'Escherichia coli étaient fréquemment récupérés du tractus intestinal. Les isolats de la mammite étaient généralement hautement susceptibles aux antimicrobiens testés, sauf pour la néomycine et l'oxytétracycline. Les isolats de l'appareil respiratoire étaient hautement susceptibles au triméthoprime-sulfaméthoxazole, à la pénicilline, au florfénicol et au ceftiofur, tandis que les bactéries entériques étaient fréquemment susceptibles au ceftiofur. Les tendances d'antibiorésistance pendant la période de l'étude étaient généralement stables pour les isolats des petits ruminants et des bovins. La multirésistance aux médicaments était plus commune pour les isolats respiratoires provenant des petits ruminants comparativement à ceux des bovins et plus commune dans les bactéries entériques des bovins par rapport à celles des petits ruminants. Ces données pourront guider les cliniciens qui choisissent des thérapies empiriques pour le traitement des ruminants dans les provinces de l'Atlantique.(Traduit par Isabelle Vallières).

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).

Performance and Cost-Efficiency of Single Hormonal Treatment Protocols in Tropical Anestrous Dairy Cows.

This retrospective study aimed to evaluate the performance of hormone treatment protocols, determine the factors associated with pregnancy success after hormone treatment, and compare the cost-efficiencies of two types of hormone treatment among cyclic and noncyclic anestrous dairy cows. The clinical records of 279 anestrous cows that received hormone treatment for artificial insemination (AI) from 64 herds in the western region of Thailand were obtained from Kasetsart University Veterinary Teaching Hospital from January to August 2017. The performance of the hormone treatment protocols, fixed-time AI (TAI) and estrus detection before AI (EAI), showed that the pregnancy risk for the TAI protocol was higher than that for the EAI protocol, but pregnancy per AI did not differ significantly between the two protocols in cyclic and noncyclic cows. Multivariate logistic regression analysis showed that cows receiving the TAI protocol were more likely to be pregnant compared to those treated with the EAI protocol. Cows with a 3.00 body condition score (BCS) < 3.75 after treatment and loose-housed cows were more likely to become pregnant. Treatment during winter showed higher pregnancy success than that in the summer and rainy seasons. The cost-efficiency analysis showed that the TAI protocol was the most cost-efficient option for noncyclic cows, whereas the EAI protocol was the most cost-efficient option for cyclic cows.

To seal or not to seal following an antimicrobial infusion at dry-off? A systematic review and multivariate meta-analysis of the incidence and prevalence of intramammary infections post-calving in dairy cows.

Teat sealants (TSs) consist of sterile formulations with no antibacterial activity. Alone or in combination with antimicrobial (AM) or non-AM treatments, TSs have been commonly used in dairy cows at dry-off to prevent intra-mammary infections (IMIs) during the dry period. This study aimed to identify and synthesise the available evidence on the efficacy of combining TSs with AM treatments on the incidence and prevalence of IMIs. A comprehensive search of three electronic databases, two relevant conference proceedings, and reference lists of reviews and eligible articles was conducted to retrieve and identify studies that could answer the following question: in dairy cows, how does the efficacy of an AM-TS combination administered at dry-off compare with an AM alone for preventing new IMI? In addition to the general IMIs, bacterial species-specific data were extracted and combined into nine distinct pathogen groups: coagulase-positive and negative staphylococci; S. dysgalactiae; non-dysgalactiae Streptococci; E. coli; non-E. coli Enterobacteriaceae; Corynebacterium spp.; yeast and other frequent mastitis pathogens. The structural relationship between each study's prevalence and incidence, as the new (incidence) and persistent (uncured) infections make up the prevalence, was utilised to approximate a variance-covariance matrix for the within-study correlation between their study-specific log odds ratios (ORs). A bivariate random-effects meta-analysis was employed, utilising the within- and between-study correlations to synthesise both outcomes simultaneously. The risk of bias was assessed using the Cochrane ROBINS-I tool, and the quality of the body of evidence was rated using the GRADE approach. A total of 17 trials (16 studies), providing either IMIs incidence (n = 4), prevalence (n = 3) or both (n = 10), were identified. Overall, quarters infused with AM-TS combinations showed lower odds of new IMIs post-calving (OR=0.70; 95% CI=0.57-0.86; Wald test P < 0.001) than those which received only AMs. Across the pathogen groups, varying levels of reduction of new IMIs were found, where administration of TSs was most effective against S. dysgalactiae (OR=0.47; 95% CI=0.23-0.98), non-dysgalactiae streptococci (OR=0.60; 95% CI=0.49-0.74), E. coli (OR=0.62; 95% CI=0.50-0.77), Corynebacterium spp. (OR=0.68; 95% CI=0.52-0.90) and coagulase-negative staphylococci (OR=0.85; 95% CI=0.76-0.94). However, additional TS infusion did not significantly reduce new IMIs in the remaining pathogen groups. The current meta-analytic evidence supports the efficacy of using TS add-on infusions in dairy cows at dry-off for reducing the incidence and prevalence of IMIs post-calving; however, pathogen group differences should be considered.

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.

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.

Quantifying antimicrobial use on Canadian dairy farms using garbage can audits.

Antimicrobial resistance in pathogenic bacteria is one of the preeminent concerns for the future of global health. There is a dose-dependent relationship between antimicrobial use (AMU) and the prevalence of antimicrobial-resistant pathogens. As most AMU in Canada is related to animal agriculture, there is a need to reduce overall AMU, which could be accomplished through surveillance of AMU in animal agriculture, including the dairy industry. The objective of this study was to quantify AMU on dairy farms across Canada. This study had two parts: a description of data collected in 2019-2020, and a meta-analysis comparing this data to previous estimates of AMU in the Canadian dairy industry. The first included a garbage can audit (GCA) on 107 farms in four Canadian provinces (British Columbia, Alberta, Ontario, and Nova Scotia) in 2020; AMU data were converted to the dose-based metrics of defined course doses (DCD) and defined daily doses (DDD). Mixed-effect linear models were fit to determine the relationship between province and use of different classes of antimicrobials. On average, for every 100 animals on the farm, 117 DCD of antimicrobials were administered per year (IQR: 55, 158). These treatments amounted to 623 DDD / 100 animal-yr (IQR: 302, 677 DDD/100 animal-years). Penicillins were the most used class of antimicrobials, followed by first-and third-generation cephalosporins. Farms in Ontario used more third-generation cephalosporins than other provinces. The second part of this study compared AMU in 2020 to previously reported Canadian studies through a meta-analysis. A GCA was conducted in 2007-2008 in Alberta, Ontario, Québec, and the Maritime provinces (Prince Edward Island, New Brunswick and Nova Scotia); another GCA was conducted in Québec in 2018. Overall, AMU was lower in 2018-2020 than in 2007-2008, with the exception of third-generation cephalosporin use, which increased.

Evaluation of antibody and antigen cross-reaction in Kenyan dairy cattle naturally infected with two pestiviruses: Bovine viral diarrhea virus and classical swine fever virus.

Background and Aim: Bovine viral diarrhea virus (BVDV) and classical swine fever virus (CSFV) are important pathogens of cattle and pigs, respectively, and belong to the genus Pestivirus. As CSFV has been shown to infect cattle, it can create diagnostic challenges of BVDV results through possible cross-reactivity where cattle could be exposed to pigs and CSFV. This study aimed to determine the possible cross-reactivity of BVDV and CSFV enzyme-linked immunosorbent assay (ELISA) results for antigen (Ag) and antibody (Ab) among smallholder dairy cattle in Kenya. Materials and Methods: This was a cross-sectional study based on a single visit to farms to collect serum samples and other descriptive farm-level and animal-level information. Testing for BVDV Ag and Ab was conducted on serum samples from 320 dairy cows and heifers, with CSFV Ag and Ab testing conducted on a subset of 133 and 74 serum samples, respectively. CSFV testing was based on BVDV test results and the availability of enough sample volume from farms that kept pigs. The Ag and Ab tests utilized IDEXX ELISA for both BVDV and CSFV. Results: For the 74 samples with Ab tests for both viruses, 40 (54.0%) were BVDV Ab positive, while 63 (85.1%) were CSFV Ab positive. Of the 40 BVDV Ab positive samples, 36 cattle (90.0%) tested positive for CSFV Ab. However, of the 34 BVDV Ab negative samples, 27 (79.4%) were CSFV Ab test-positive. For the 133 samples with Ag tests for both viruses, 125 (94.0%) were BVDV Ag positive, while 2 (1.5%) samples were CSFV Ag positive. None of the eight BVDV Ag negative samples was positive for CSFV Ag and only two (1.6%) of the 125 BVDV Ag positive samples were positive for CSFV Ag. Conclusion: The results indicate either substantial cross-reactivity of the two Ab ELISA tests, or reactivity with some other protein in the samples that led to the positive Ab test results. There was only limited evidence for cross-reactivity of the two Ag ELISA tests. We recommend that Pestivirus genus cross-reactivity be considered when interpreting BVDV ELISA results in cattle, more for Ab than Ag tests. Further research is needed to clarify the levels of cross-reactivity between BVDV and other Pestivirus Ag and Ab tests from animals on mixed-species farms.

Cross-sectional study of antimicrobial use and treatment decision for preweaning Canadian dairy calves.

Antimicrobials should be used prudently in farm animals to prevent the development of resistant bacteria in both humans and animals. The objective of this study was to investigate Canadian dairy producers' practices for antimicrobial use in the treatment of disease in preweaning dairy calves. In-person questionnaires were administered to 144 dairy producers across 5 provinces in Canada between July 2019 and August 2020. Almost all (96%) producers used antimicrobials to treat calves with respiratory disease, but only 27% indicated they had a written treatment protocol for respiratory disease. Most (95%) of these protocols for respiratory disease were developed with input from the herd veterinarian. Seventy-four percent of producers used antimicrobials to treat calf diarrhea, with 37% of producers having a written treatment protocol for calf diarrhea with input from the herd veterinarian. The combinations of signs adopted by the producers for antimicrobial treatment in calf respiratory disease and diarrhea were evaluated based on findings from other studies. More than half (56%) of producers who used antimicrobials for calf respiratory disease decided to use antimicrobials by evaluating multiple clinical signs. Eighty-two percent of producers who used antimicrobials for calf diarrhea made decisions based on systemic signs of disease, presence of bloody stool, no response to previous treatment, or on the recommendation from the herd veterinarian. Producers with a written treatment protocol had 3 to 7 times greater odds of using antimicrobials based on multiple signs or systemic signs of disease compared with those without a protocol. Further research may investigate other calf management practices related to decision-making by producers in using antimicrobials to improve antimicrobial stewardship on dairy farms.

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.

Genetic and phenotypic characterization of the bla(CMY) gene from Escherichia coli and Salmonella enterica isolated from food-producing animals, humans, the environment, and retail meat.

The bla(CMY-2) family of the ampC beta-lactamase genes confer broad-spectrum resistance to beta-lactam antimicrobials, including ceftriaxone and ceftiofur, as well as to beta-lactamase inhibitors, such as clavulanic acid. Organisms with the bla(CMY-2) phenotype have been recovered from the environment and from retail meat products, posing a potential public health risk. The objectives of this study were to sequence the bla(CMY-2) gene from Escherichia coli and Salmonella enterica from multiple sources that had a reduced susceptibility to ceftriaxone and to determine the effect of observed mutations in the bla(CMY-2) gene on the antimicrobial resistance phenotype (spectrum and minimum inhibitory concentration/susceptibility patterns) of the isolates. The bla(CMY-2) genes from 52 bacterial isolates were sequenced for this study. Sixty-two percent (32/52) were E. coli and 38% (20/52) were S. enterica. Of the 32 E. coli isolates, 30 were found to carry a beta-lactamase gene that was 100% homologous to bla(CMY-2). One of the E. coli isolates was found to contain a gene that was 90% homologous to bla(CMY-2). This isolate also had lower minimum inhibitory concentrations to tetracyclines, streptomycin, and the sulfonamide antimicrobials than are commonly expected for isolates containing the bla(CMY-2). Of the 20 genes obtained from Salmonella isolates, 8 (40%) were found to be homologous to bla(CMY-2), with no altered susceptibility phenotypes observed.