Investigation of antimicrobial resistance and antimicrobial use in western Canadian cow-calf herds

This thesis summarizes an investigation of antimicrobial resistance (AMR) and antimicrobial use (AMU) in cow-calf herds. The specific objectives of this project were to describe common reasons for treatment and the types of antimicrobials used in cow-calf herds, to describe the frequency of AMR in generic fecal Escherichia coli isolated from various age groups commonly found on cow-calf farms, to determine risk factors associated with the occurrence of AMR, and finally to investigate the underlying molecular mechanisms of AMR in cow-calf herds. At least 86% of the herds treated one or more calves or cows during the study period; however, the overall proportion of both calves and cows reported as treated was less than 14% for calves and 3% for cows. The majority of antimicrobials reported as used in cow-calf operations were for individual therapeutic use rather than prophylaxis, metaphylaxis, or growth promotion. Injectable formulations were the most commonly reported method of antimicrobial administration on cow-calf farms. Cow-calf herds in Wetern Canada are not a significant reservoir for resistance to antimicrobials classified as very important to human medicine such as ciprofloxacin and ceftiofur. The three most common resistances detected were to tetracycline, sulphamethoxazole, and streptomycin regardless of age group. Young calves sampled in the spring of the year were more likely to be shedding AMR E. coli than older calves sampled in the fall of the year or than cows sampled in the spring of the year. The cow-calf pair relationship was not an important factor in transfer of AMR from the individual cow to her calf, but the presence of AMR in the general cow herd was associated with AMR in the calf population. The potential importance of co-selection for AMR at the molecular level was demonstrated by both the risk factor analysis and the molecular work. Phenotypic resistance to streptomycin, tetracycline, and sulphamethoxazole were each associated with the presence of resistance genes from all six families of antimicrobials examined in this study. Several statistically significant associations were also detected between the resistance genes considered. No significant associations were detected between any of the AMR phenotypes or genotypes and the STEC virulence factors stx1, stx2 and eae

Clostridium difficile and methicillin-resistant Staphylococcus aureus shedding by slaughter-age pigs

<p>Abstract</p> <p>Background</p> <p><it>Clostridium difficile </it>and methicillin-resistant <it>Staphylococcus aureus </it>are critical human pathogens and of increasing concern in food animals. Because of the apparent impact of age on prevalence of these organisms, studies of slaughter age pigs are important when considering the potential for contamination of food. This study evaluated <it>C. difficile </it>and MRSA shedding by slaughter age pigs from farms across Canada.</p> <p>Results</p> <p><it>Clostridium difficile </it>was isolated from 30/436 (6.9%) samples from 15/45 (33%) farms. After adjusting for clustering at the herd level, the prevalence was 3.4%. Ribotype 078 (toxinotype V, North American Pulsotype 7) was the most common strain, accounting for 67% of isolates. MRSA was isolated from 21/460 (4.6%) pigs from 5/46 (11%) farms. The prevalence in pigs after adjusting for clustering at the herd level was 0.2%. Seven different spa types were identified, with 3 related spa types (t011, t034, new) accounting for 16 (76%) consistent with ST398 predominating.</p> <p>Both MRSA and <it>C. difficile </it>samples were collected from 45 farms. Both MRSA and <it>C. difficile </it>were detected on 2 (4.4%), with <it>C. difficile </it>only on 13 (29%), MRSA only on 3 (6.7%) and neither on 27 (60%).</p> <p>Conclusions</p> <p>The prevalence of <it>C. difficile </it>and MRSA in slaughter age pigs was relatively low, particularly in comparison with studies involving younger pigs. The predominance of <it>C. difficile </it>ribotype 078 and MRSA ST398 was not surprising, but there was diversity in strain types and the majority of isolates of both organisms were strains that can be found in humans. While the prevalence of <it>C. difficile </it>and MRSA in slaughter age pigs was relatively low, there is clearly potential for contamination of meat from healthy pigs carrying this pathogen into slaughterhouses.</p

Vaccine use in Canadian cow-calf herds and opportunities for improvement

Vaccinations are one of the most impactful tools available to cow-calf producers to control within herd disease and later, in feedlots. While vaccine use has been studied across Canada, inconsistent and variable regional data makes analysis and interpretation difficult. The objective of this study was to describe vaccination protocols and factors associated with vaccine use in Canadian cow-calf herds and define associations between vaccine use and productivity outcomes. Surveys describing vaccine use in 2020 were collected from 131 cow-calf herds (40 eastern, 91 western), recruited through a national beef cattle surveillance program. Ninety-two percent of cows and replacement heifers, and 72% of bulls were vaccinated with Infectious Bovine Rhinotracheitis (IBR), Bovine Viral Diarrhea Virus (BVDV), Parainfluenza3 Virus (PI3), and Bovine Respiratory Syncytial Virus (BRSV). At least half of cows and bulls were vaccinated for clostridial pathogens and cows and heifers for viral calf scours. Clostridial vaccines were significantly more likely to be used in western Canada compared to eastern Canada. While 92% of producers vaccinated suckling calves against IBR/BRSV/PI3, only 47% provided a second vaccine prior to weaning; 78% of calves were also vaccinated at least once for BVDV before weaning. Producers who vaccinated calves against IBR/BRSV/PI3 before 3 months of age provided a second dose prior to weaning more often than producers who administer the first IBR/BRSV/PI3 vaccine later. Vaccine use has increased across Canada, particularly in calves, prior to weaning. Relative to label recommendations for annual vaccination, clostridial vaccines were generally underutilized in cows and bulls, and by producers in eastern Canada as compared to western Canada. Opportunities also exist to improve adherence to label recommendations for the booster dose of scours vaccine when used in bred replacement heifers. Protocols including product choices, the timing and boosting of respiratory vaccines in nursing calves vary widely across herds. Use of intranasal vaccines in neonatal calves less than 2 weeks old has increased in western Canada compared to previous reports. There is a need to better understand how timing of vaccination in nursing calves contributes to effectiveness, for respiratory disease in nursing and weaned calves

Prevalence and molecular characterization of Clostridium difficile isolated from feedlot beef cattle upon arrival and mid-feeding period

<p>Abstract</p> <p>Background</p> <p>The presence of indistinguishable strains of <it>Clostridium difficile </it>in humans, food animals and food, as well as the apparent emergence of the food-animal-associated ribotype 078/toxinotype V as a cause of community-associated <it>C. difficile </it>infection have created concerns about the potential for foodborne infection. While studies have reported <it>C. difficile </it>in calves, studies of cattle closer to the age of harvest are required. Four commercial feedlots in Alberta (Canada) were enrolled for this study. Fecal samples were collected at the time of arrival and after acclimation (< 62, 62-71 or > 71 days on feed). Selective culture for <it>Clostridium difficile </it>was performed, and isolates were characterized by ribotyping and pulsed-field gel electrophoresis. A logistic regression model was built to investigate the effect of exposure to antimicrobial drugs on the presence of <it>C. difficile</it>.</p> <p>Results</p> <p><it>Clostridium difficile </it>was isolated from 18 of 539 animals at the time of feedlot arrival (CI = 2.3-6.1) and from 18 of 335 cattle at mid-feeding period (CI = 2.9-13.1). Overall, there was no significant difference in the prevalence of <it>C. difficile </it>shedding on arrival versus mid-feeding period (<it>P </it>= 0.47). No association between shedding of the bacterium and antimicrobial administration was found (<it>P </it>= 0.33). All the isolates recovered were ribotype 078, a toxinotype V strain with genes encoding toxins A, B and CDT. In addition, all strains were classified as NAP7 by pulsed field gel electrophoresis (PFGE) and had the characteristic 39 base pairs deletion and upstream truncating mutation on the <it>tcd</it>C gene.</p> <p>Conclusions</p> <p>It is apparent that <it>C. difficile </it>is carried in the intestinal tracts of a small percentage of feedlot cattle arriving and later in the feeding period and that ribotype 078/NAP7 is the dominant strain in these animals. Herd management practices associated with <it>C. difficile </it>shedding were not identified, however further studies of the potential role of antimicrobials on <it>C. difficile </it>acquisition and shedding are required.</p

Associations between antimicrobial resistance in fecal Escherichia coli isolates and antimicrobial use in Canadian turkey flocks

Antimicrobial resistance (AMR) in enteric bacteria continues to be detected in turkey flocks and retail products worldwide, including in Canada. However, studies assessing linkages between on-farm antimicrobial use (AMU) and the development of AMR are lacking. This study aims to identify AMU characteristics that impact the development of AMR in the indicator bacteria Escherichia coli in turkey flocks, building on the Canadian Integrated Program for Antimicrobial Resistance Surveillance methodology for farm-level AMU and AMR data integration. Two analytic approaches were used: (1) multivariable mixed-effects logistic regression models examined associations between AMU (any route, route-specific, and route-disease-specific indication) summarized as the number of defined daily doses in animals using Canadian standards ([nDDDvetCA]/1,000 kg-animal-days at risk) and AMR and (2) multivariable mixed-effects Poisson regression models studied the linkages between AMU and the number of classes to which an E. coli isolate was resistant (nCRE. coli). A total of 1,317 E. coli isolates from a network of 16 veterinarians and 334 turkey producers across the five major turkey-producing provinces in Canada between 2016 and 2019 were used. Analysis indicated that AMR emerged with the use of related antimicrobials (e.g., tetracycline use-tetracycline resistance), however, the use of unrelated antimicrobial classes was also impacting AMR (e.g., aminoglycosides/streptogramins use-tetracycline resistance). As for studying AMU-nCRE. coli linkages, the most robust association was between the parenteral aminoglycosides use and nCRE. coli, though in-feed uses of four unrelated classes (bacitracin, folate pathway inhibitors, streptogramins, and tetracyclines) appear to be important, indicating that ongoing uses of these classes may slow down the succession from multidrug-resistant to a more susceptible E. coli populations. The analysis of AMU (route and disease-specific)-AMR linkages complemented the above findings, suggesting that treatment of certain diseases (enteric, late-stage septicemic conditions, and colibacillosis) are influential in the development of resistance to certain antimicrobial classes. The highest variances were at the flock level indicating that stewardship actions should focus on flock-level infection prevention practices. This study added new insights to our understanding of AMU-AMR linkages in turkeys and is useful in informing AMU stewardship in the turkey sector. Enhanced surveillance using sequencing technologies are warranted to explain molecular-level determinants of AMR

Does swab type matter? Comparing methods for \u3ci\u3eMannheimia haemolytica\u3c/i\u3e recovery and upper respiratory microbiome characterization in feedlot cattle

Background: Bovine respiratory disease (BRD) is caused by interactions among host, environment, and pathogens. One standard method for antemortem pathogen identification in cattle with BRD is deep-guarded nasopharyngeal swabbing, which is challenging, costly, and waste generating. The objective was to compare the ability to recover Mannheimia haemolytica and compare microbial community structure using 29.5 inch (74.9 cm) deep-guarded nasopharyngeal swabs, 16 inch (40.6 cm) unguarded proctology swabs, or 6 inch (15.2 cm) unguarded nasal swabs when characterized using culture, real time-qPCR, and 16S rRNA gene sequencing. Samples for aerobic culture, qPCR, and 16S rRNA gene sequencing were collected from the upper respiratory tract of cattle 2 weeks after feedlot arrival. Results: There was high concordance of culture and qPCR results for all swab types (results for 77% and 81% of sampled animals completely across all 3 swab types for culture and qPCR respectively). Microbial communities were highly similar among samples collected with different swab types, and differences identified relative to treatment for BRD were also similar. Positive qPCR results for M. haemolytica were highly concordant (81% agreed completely), but samples collected by deep-guarded swabbing had lower amounts of Mh DNA identified (Kruskal–Wallis analysis of variance on ranks, P \u3c 0.05; Dunn-test for pairwise comparison with Benjamini–Hochberg correction, P \u3c 0.05) and lower frequency of positive compared to nasal and proctology swabs (McNemar’s Chi-square test, P \u3c 0.05). Conclusions: Though differences existed among different types of swabs collected from individual cattle, nasal swabs and proctology swabs offer comparable results to deep-guarded nasopharyngeal swabs when identifying and characterizing M. haemolytica by culture, 16S rRNA gene sequencing, and qPCR

Antimicrobial Use and Antimicrobial Resistance Indicators—Integration of Farm-Level Surveillance Data From Broiler Chickens and Turkeys in British Columbia, Canada

Using data from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), we aimed to describe trends in antimicrobial use (AMU) in broiler chickens and turkeys, to compare AMU across species, to compare with trends in antimicrobial resistance (AMR), and to assess the effects of various AMU/AMR units of measurement (metrics and indicators) on data integration. Data on AMU and AMR in enteric bacteria, collected from 2013 to 2017 from broiler chickens (n = 143 flocks) and turkeys (n = 145) were used. In broiler chickens, the total AMU in milligrams/population correction unit (mg/PCUBr) decreased by 6%, the number (n) of defined daily doses for animals using Canadian standards (nDDDvetCA) per 1,000 broiler chicken-days decreased by 12%, and nDDDvetCA/PCU decreased by 6%. In turkeys, the mg/PCUTk decreased by 1%, whereas the nDDDvetCA/1,000 turkey-days and the nDDDvetCA/PCU increased by 1 and 5%, respectively. The types of antimicrobial classes used in both species were similar. Using the frequency of flocks reporting use (i.e., number of flocks reporting use/number of flocks participating) as a measurement, the use of certain antimicrobials changed over time (e.g., Broilers, decreased cephalosporin use, virginiamycin use, emerging use of lincomycin-spectinomycin, and avilamycin; Turkeys: increased trimethoprim-sulfonamides and macrolide use). The trends in resistance to specific antimicrobials paralleled the frequency and quantity of use (e.g., ceftriaxone use decreased—ceftriaxone resistance decreased, and gentamicin use increased—gentamicin resistance increased) in some situations, but not others (decreased fluoroquinolone use—increased ciprofloxacin resistance). AMR data were summarized using the AMR indicator index (AMR Ix). The most notable AMR Ix trend was the decrease in ceftriaxone AMR Ix among Escherichia coli (0.19 to 0.07); indicative of the success of the poultry industry action to eliminate the preventive use of third generation cephalosporins. Other trends observed were the increase in ciprofloxacin AMR Ix among Campylobacter from 0.23 to 0.41 and gentamicin AMR Ix among E. coli from 0.11 to 0.22, suggestive of the persistence/emergence of resistance related to previous and current AMU not captured in our surveillance timeframe. These data highlight the necessity of multiple AMU and AMR indicators for monitoring the impact of stewardship activities and interventions

Evaluating the effects of antimicrobial drug use on the ecology of antimicrobial resistance and microbial community structure in beef feedlot cattle

IntroductionUse of antimicrobial drugs (AMDs) in food producing animals has received increasing scrutiny because of concerns about antimicrobial resistance (AMR) that might affect consumers. Previously, investigations regarding AMR have focused largely on phenotypes of selected pathogens and indicator bacteria, such as Salmonella enterica or Escherichia coli. However, genes conferring AMR are known to be distributed and shared throughout microbial communities. The objectives of this study were to employ target-enriched metagenomic sequencing and 16S rRNA gene amplicon sequencing to investigate the effects of AMD use, in the context of other management and environmental factors, on the resistome and microbiome in beef feedlot cattle.MethodsThis study leveraged samples collected during a previous longitudinal study of cattle at beef feedlots in Canada. This included fecal samples collected from randomly selected individual cattle, as well as composite-fecal samples from randomly selected pens of cattle. All AMD use was recorded and characterized across different drug classes using animal defined daily dose (ADD) metrics.ResultsOverall, fecal resistome composition was dominated by genes conferring resistance to tetracycline and macrolide-lincosamide-streptogramin (MLS) drug classes. The diversity of bacterial phyla was greater early in the feeding period and decreased over time in the feedlot. This decrease in diversity occurred concurrently as the microbiome represented in different individuals and different pens shifted toward a similar composition dominated by Proteobacteria and Firmicutes. Some antimicrobial drug exposures in individuals and groups were associated with explaining a statistically significant proportion of the variance in the resistome, but the amount of variance explained by these important factors was very small (&lt;0.6% variance each), and smaller than associations with other factors measured in this study such as time and feedlot ID. Time in the feedlot was associated with greater changes in the resistome for both individual animals and composite pen-floor samples, although the proportion of the variance associated with this factor was small (2.4% and 1.2%, respectively).DiscussionResults of this study are consistent with other investigations showing that, compared to other factors, AMD exposures did not have strong effects on antimicrobial resistance or the fecal microbial ecology of beef cattle