Surveillance of antimicrobial resistance in Escherichia coli, Salmonella, and Campylobacter recovered from laying hens, their environment and products in Canada indicated a stable level of resistance to critically important antimicrobials, in varying time periods between 2007 and 2021.

The study objective was to determine the occurrence and antimicrobial resistance (AMR) attributes of select foodborne bacteria recovered from egg-producing (layer) chickens between 2007 and 2021 using different sample matrices (Study 1: liquid whole eggs, Study 2: spent hen cecal samples, Study 3: environmental sponge swabs and fecal samples from layer chicken barns, and Study 4: fecal samples from layer chicken barns). Samples from each study were submitted for the culture of Escherichia coli and Salmonella. In addition, samples from layer chicken barns were submitted for the culture of Campylobacter. Isolates were tested by microbroth dilution and interpreted using both clinical breakpoints and epidemiological cut-offs (ECOFFs). The ECOFFs were applied to detect non-wild type (NWT) strains. The proportion of resistant, NWT, and distribution of minimum inhibitory concentrations (MIC) were assessed. Ceftriaxone resistance was detected at a low-level in E. coli (< 2 %, all studies) and Salmonella (4.3 %, Study 2). Very low-level ciprofloxacin resistance was detected in E. coli (<1 %, Study 1) but a slightly elevated ciprofloxacin NWT E. coli (1 % to 6 %) observed. Only the farm fecal samples in Studies 2 and 3 were tested for Campylobacter as part of its study design, and moderate level ciprofloxacin resistance (<15 %) was observed. The MIC distribution patterns were similar across the organisms tested/studies and no substantial shifts in the distributions were detected. This analysis demonstrated that resistance to very important antimicrobials in bacteria from layers in Canada remains low, however, the detection of ciprofloxacin-resistant Campylobacter and the implications of this observation to the safety of egg products, and the role of laying flocks (i.e., as reservoir for resistant organisms) needs to be investigated.

Decrease in the prevalence of antimicrobial resistance in Escherichia coli isolates of Canadian turkey flocks driven by the implementation of an antimicrobial stewardship program.

The emergence of antimicrobial-resistant organisms at the human-animal-environment interface has raised global concern prompting governments and various stakeholders to take action. As a part of the stewardship initiative, Canadian turkey producers have implemented an antimicrobial use (AMU) strategy to manage antimicrobial resistance (AMR) in their sector. This study evaluated farm-level AMU and AMR data collected between 2016 and 2021 in major turkey-producing provinces/regions through the Canadian Integrated Program for Antimicrobial Resistance Surveillance to assess the progress of the strategy by characterizing the prevalence of homologous and multidrug resistance (MDR) in Escherichia coli isolated from turkeys. Multivariable mixed-effect logistic regression models assessed temporal and provincial/regional variations in AMR and MDR. Negative binomial regression models examined the temporal and regional variations in the total AMU. The total AMU (measured in mg/kg turkey biomass) significantly decreased in all provinces/regions in 2020 and 2021. Escherichia coli isolates from turkey flocks showed a significant decrease in resistance to gentamicin, sulfisoxazole, and tetracyclines during the six-year study period, consistent with the timing of the AMU reduction strategy. The prevalence of MDR isolates was significantly lower in 2020 and 2021 compared to 2016. Higher prevalence was observed in the Western region compared to Québec and Ontario. Two common AMR patterns were identified: ampicillin-streptomycin-tetracyclines and streptomycin-sulfisoxazole-tetracyclines. These AMR patterns indicate possible cross-resistances (same class), co-selection (unrelated classes) for resistance, or potential carryover of resistance determinants from previous production cycles. The decreasing prevalence of resistance to homologous antimicrobials, MDR, and AMU quantity are suggestive that the turkey sector's AMU strategy is achieving its desired impact. However, antimicrobials previously eliminated for preventive use in turkey flocks and the use of highly important antimicrobials in human medicine suggest that the AMU reduction strategy should be monitored and re-evaluated periodically to mitigate the emergence of MDR bacteria and safeguard animal and public health.

A One Health Genomic Investigation of Gentamicin Resistance in Escherichia coli from Human and Chicken Sources in Canada, 2014 to 2017.

We investigated whether gentamicin resistance (Genr) in Escherichia coli isolates from human infections was related to Genr E. coli in chicken and whether resistance may be due to coselection from use of lincomycin-spectinomycin in chickens on farms. Whole-genome sequencing was performed on 483 Genr E. coli isolates isolated between 2014 and 2017. These included 205 human-source isolates collected by the Canadian Ward (CANWARD) program and 278 chicken-source isolates: 167 from live/recently slaughtered chickens (animals) and 111 from retail chicken meat collected by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). The predominant Genr gene was different in human and chicken sources; however, both sources carried aac(3)-IId, aac(3)-VIa, and aac(3)-IVa. Forty-one percent of human clinical isolates of Genr E. coli contained a blaCTX-M extended-spectrum beta-lactamase (ESBL) gene (84/205), and 53% of these were sequence type 131 (ST131). Phylogenomic analysis revealed a high diversity of Genr isolates; however, there were three small clusters of closely related isolates from human and chicken sources. Genr and spectinomycin resistance (Specr) genes were colocated in 148/167 (89%) chicken animal isolates, 94/111 (85%) chicken retail meat isolates, and 137/205 (67%) human-source isolates. Long-read sequencing of 23 isolates showed linkage of the Genr and Specr genes on the same plasmid in 14/15 (93%) isolates from chicken(s) and 6/8 (75%) isolates from humans. The use of lincomycin-spectinomycin on farms may be coselecting for gentamicin-resistant plasmids in E. coli in broiler chickens; however, Genr isolates and plasmids were mostly different in chickens and humans.

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 (nCR E. 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-nCR E. coli linkages, the most robust association was between the parenteral aminoglycosides use and nCR E. 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.

One Health Genomic Analysis of Extended-Spectrum ß-Lactamase‒Producing Salmonella enterica, Canada, 2012‒2016.

Extended-spectrum ß-lactamases (ESBLs) confer resistance to extended-spectrum cephalosporins, a major class of clinical antimicrobial drugs. We used genomic analysis to investigate whether domestic food animals, retail meat, and pets were reservoirs of ESBL-producing Salmonella for human infection in Canada. Of 30,303 Salmonella isolates tested during 2012-2016, we detected 95 ESBL producers. ESBL serotypes and alleles were mostly different between humans (n = 54) and animals/meat (n = 41). Two exceptions were blaSHV-2 and blaCTX-M-1 IncI1 plasmids, which were found in both sources. A subclade of S. enterica serovar Heidelberg isolates carrying the same IncI1-blaSHV-2 plasmid differed by only 1-7 single nucleotide variants. The most common ESBL producer in humans was Salmonella Infantis carrying blaCTX-M-65, which has since emerged in poultry in other countries. There were few instances of similar isolates and plasmids, suggesting that domestic animals and retail meat might have been minor reservoirs of ESBL-producing Salmonella for human infection.

Antimicrobial use in lactating sows, piglets, nursery, and grower-finisher pigs on swine farms in Ontario, Canada during 2017 and 2018.

BACKGROUND: Data on antimicrobial use (AMU) in pig production are needed for the development of good antimicrobial stewardship practices to reduce the risk of antimicrobial resistance in bacteria that can cause illness in animals and humans. In Canada, there is a lack of quantitative data on AMU in the farrowing and nursery stages of pig production. This study aimed to determine which antimicrobial active ingredients are currently used in farrowing, nursery, and grower-finisher herds in the province of Ontario, Canada, and to quantify AMU using various metrics. We collected data on herd demographics, biosecurity, health status, and AMU during one production cycle from 25 farrowing and 25 nursery herds in Ontario, between May 2017 and April 2018, and obtained data from 23 Ontario grower-finisher herds during the same time frame from the Public Health Agency's Canadian Integrated Program for Antimicrobial Resistance Surveillance. We applied frequency measures, and weight-, and dose-based metrics to the data. RESULTS: In all pigs, the highest quantity of AMU was administered in-feed. By all routes of administration and compared to other production stages, nursery pigs used more antimicrobials in mg/kg biomass and the number of Canadian defined daily doses per 1000 pig-days (doseCA rate), while grower-finisher pigs used more antimicrobials in total kilograms and the number of Canadian defined daily doses per pig. In suckling pigs in some herds, there was routine disease prevention use of ceftiofur, an antimicrobial active ingredient categorized as very highly important in human medicine by Health Canada. The top antimicrobial used in each stage of pig production often varied by the metric used. There was producer-reported growth promotion use of antimicrobials in suckling and grower-finisher feed. CONCLUSIONS: The results of this study provide a current picture of AMU in pigs in Ontario and can be used as a basis for further research on AMU in farrowing and nursery herds in Canada. Our findings confirm that it would be useful to include farrowing and nursery herds in routine AMU surveillance in Canada. A future analysis using data from this project will examine factors that affect the quantity of AMU.

Canadian Collaboration to Identify a Minimum Dataset for Antimicrobial Use Surveillance for Policy and Intervention Development across Food Animal Sectors.

Surveillance of antimicrobial use (AMU) and antimicrobial resistance (AMR) is a core component of the 2017 Pan-Canadian Framework for Action. There are existing AMU and AMR surveillance systems in Canada, but some stakeholders are interested in developing their own AMU monitoring/surveillance systems. It was recognized that the establishment of core (minimum) AMU data elements, as is necessary for policy or intervention development, would inform the development of practical and sustainable AMU surveillance capacity across food animal sectors in Canada. The Canadian Animal Health Surveillance System (CAHSS) AMU Network was established as a multisectoral working group to explore the possibility of harmonizing data inputs and outputs. There was a consensus that a minimum AMU dataset for AMU surveillance (MDS-AMU-surv) should be developed to guide interested parties in initiating AMU data collection. This multisectoral collaboration is an example of how consultative consensus building across relevant sectors can contribute to the development of harmonized approaches to AMU data collection and reporting and ultimately improve AMU stewardship. The MDS-AMU-surv could be used as a starting point for the progressive development or strengthening of AMU surveillance programs, and the collaborative work could serve as a model for addressing AMR and other shared threats at the human-animal-environment interface.

Informing Stewardship Measures in Canadian Food Animal Species through Integrated Reporting of Antimicrobial Use and Antimicrobial Resistance Surveillance Data-Part II, Application.

Using the methodology developed for integrated analysis and reporting of antimicrobial use (AMU) and antimicrobial resistance (AMR) data, farm-level surveillance data were synthesized and integrated to assess trends and explore potential AMU and AMR associations. Data from broiler chicken flocks (n = 656), grower-finisher pig herds (n = 462) and turkey flocks (n = 339) surveyed by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) at the farm-level (2015-2019) were used. The analyses showed a reduction in mean flock/herd level number of defined daily doses using Canadian standards (nDDDvetCA) adjusted for kg animal biomass that coincided with the decline in % resistance in the three species. This was noted in most AMU-AMR pairs studied except for ciprofloxacin resistant Campylobacter where resistance continued to be detected (moderate to high levels) despite limited fluoroquinolone use. Noteworthy was the significantly negative association between the nDDDvetCA/kg animal biomass and susceptible Escherichia coli (multispecies data), an early indication that AMU stewardship actions are having an impact. However, an increase in the reporting of diseases in recent years was observed. This study highlighted the value of collecting high-resolution AMU surveillance data with animal health context at the farm-level to understand AMR trends, enable data integration and measure the impact of AMU stewardship actions.

Informing Stewardship Measures in Canadian Food Animal Species through Integrated Reporting of Antimicrobial Use and Antimicrobial Resistance Surveillance Data-Part I, Methodology Development.

This study explores methodologies for the data integration of antimicrobial use (AMU) and antimicrobial resistance (AMR) results within and across three food animal species, surveyed at the farm-level by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). The approach builds upon existing CIPARS methodology and principles from other AMU and AMR surveillance systems. Species level data integration involved: (1) standard CIPARS descriptive and temporal analysis of AMU/AMR, (2) synthesis of results, (3) selection of AMU and AMR outcomes for integration, (4) selection of candidate AMU indicators to enable comparisons of AMU levels between species and simultaneous assessment of AMU and AMR trends, (5) exploration of analytic options for studying associations between AMU and AMR, and (6) interpretation and visualization. The multi-species integration was also completed using the above approach. In addition, summarized reporting of internationally-recognized indicators of AMR (i.e., AMR adjusted for animal biomass) and AMU (mg/population correction unit, mg/kg animal biomass) is explored. It is envisaged that this approach for species and multi-species AMU-AMR data integration will be applied to the annual CIPARS farm-level data and progressively developed over time to inform AMU-AMR integrated surveillance best practices for further enhancement of AMU stewardship actions.

A One-Health Genomic Investigation of Gentamicin Resistance in Salmonella from Human and Chicken Sources in Canada, 2014 to 2017.

We investigated whether the increased prevalence of gentamicin resistance in Salmonella from human infections was related to a similar increased prevalence in isolates from broiler chickens and whether this increase may have been due to coselection from use of lincomycin-spectinomycin in chickens on farms. Whole-genome sequencing was performed on gentamicin-resistant (Genr) Salmonella isolates from human and chicken sources collected from 2014 to 2017 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). We determined the genomic relatedness of strains and characterized resistance genes and plasmids. From 2014 to 2017, 247 isolates of Genr Salmonella were identified by CIPARS: 188 were from humans, and 59 were from chicken sources (26 from live animals on farm and 33 from retail meat). The five most common Genr serovars were Salmonella enterica serovars Heidelberg (n = 93; 31.5%), 4,[5],12:i:- (n = 42; 14.2%), Kentucky (n = 37; 12.5%), Infantis (n = 33; 11.2%), and Typhimurium (n = 23; 7.8%). Phylogenomic analysis revealed that for S. Heidelberg and S. Infantis, there were closely related isolates from human and chicken sources. In both sources, resistance to gentamicin and spectinomycin was most frequently conferred by aac(3)-VIa and ant(3'')-Ia, respectively. Plasmid closure confirmed linkages of gentamicin and spectinomycin resistance genes and revealed instances of similar plasmids from both sources. Gentamicin and spectinomycin resistance genes were linked on the same plasmids, and some plasmids and isolates from humans and chickens were genetically similar, suggesting that the use of lincomycin-spectinomycin in chickens may be selecting for gentamicin-resistant Salmonella in broiler chickens and that these resistant strains may be acquired by humans.

Choosing which metrics to use when reporting antimicrobial use information to veterinarians in the Canadian swine industry.

The objective of this study was to evaluate preferences for various metrics and denominators among Canadian swine veterinarians, in order to improve reporting of antimicrobial use (AMU) information to these stakeholders and to facilitate enhanced stewardship decisions. An online survey was made available to swine veterinarians across Canada; 12 responses (estimated response rate 17.6%) were submitted and analyzed. Responses represented veterinarians from every major pig-producing province and from a range of year of graduation from veterinary college. Participants self-evaluated their understanding of dose-based metrics as higher than weight- and frequency-based metrics and interpreted most results of AMU analyses correctly. Participants preferred dose-based metrics over others, and had various objectives for AMU information, including improving AMU on their clients' farms and enabling comparisons with other farms. The results are useful to those making decisions about which AMU metrics to use in reports targeted to swine veterinarians.

Choix des paramètres à utiliser lors de la communication d'informations sur l'utilisation des antimicrobiens aux vétérinaires de l'industrie porcine canadienne. L'objectif de la présente étude était d'évaluer les préférences pour divers paramètres et dénominateurs chez les vétérinaires porcins canadiens, afin d'améliorer la déclaration de l'information sur l'utilisation d'antimicrobiens (UMA) à ces intervenants et de faciliter des décisions de gérance améliorées. Un sondage en ligne a été mis à la disposition des vétérinaires porcins partout au Canada; 12 réponses (taux de réponse estimé à 17,6 %) ont été soumises et analysées. Les réponses représentaient des vétérinaires de toutes les grandes provinces productrices de porcs et d'une gamme d'années d'obtention du diplôme d'un collège vétérinaire. Les participants ont auto-évalué leur compréhension des mesures basées sur la dose comme étant supérieure aux mesures basées sur le poids et la fréquence et ont interprété correctement la plupart des résultats des analyses UMA. Les participants préféraient les mesures basées sur la dose aux autres, et avaient divers objectifs pour l'information sur l'UMA, notamment l'amélioration de l'UMA dans les fermes de leurs clients et la possibilité de comparer avec d'autres fermes. Les résultats sont utiles à ceux qui prennent des décisions sur les paramètres d'UMA à utiliser dans les rapports destinés aux vétérinaires porcins.(Traduit par Dr Serge Messier).

Antimicrobial Use Indices-The Value of Reporting Antimicrobial Use in Multiple Ways Using Data From Canadian Broiler Chicken and Turkey Farms.

We have previously described the importance of using multiple indicators for reporting national farm-level antimicrobial use (AMU) information, but the distribution of flock-level AMU and how these indicators relate to each other has not yet been fully explored. Using farm-level surveillance data (2013-2019), for broiler chickens (n = 947 flocks) and turkeys (n = 427), this study aims to (1) characterize flock-level AMU and identify high users, (2) identify appropriate AMU indicators and biomass denominator [population correction unit (PCU) vs. kg weight at pre-slaughter], and (3) make recommendations on the application to veterinarian-producer and national-level reporting. Diverse AMU patterns were identified in broiler chickens (156 patterns) and turkeys (68 patterns); of these, bacitracin, reported by 25% of broiler chicken and 19% of turkey producers, was the most frequently occurring pattern. Depending on the indicator chosen, variations in reported quantity of use, temporal trends and relative ranking of the antimicrobials changed. Quantitative AMU analysis yielded the following results for broiler chickens: mean 134 mg/PCU; 507 number (n) of Canadian (CA) defined daily doses (DDDvet) per 1,000 chicken-days and 18 nDDDvetCA/PCU. Analysis in turkey flocks yielded the following: mean 64 mg/PCU, 99 nDDDvetCA/1,000 turkey-days at risk and 9 nDDDvetCA/PCU. Flocks were categorized based on the percentiles of the mg/PCU distribution: "medium" to "low" users (≤75th percentile) and "high" users (>75th percentile). The odds of being a high user in both broiler chickens and turkeys were significantly increased: if water medications were used, and if trimethoprim-sulfonamides, bacitracins, and tetracyclines were used. Pairwise correlation analysis showed moderate correlation between mg/PCU and the nDDDvetCA/1,000 animal days at risk and between mg/PCU and nDDDvetCA/PCU. Significantly high correlation between nDDDvetCA/1,000 animal days at risk and nDDDvetCA/PCU was observed, suggestive that either of these could be used for routine monitoring of trends in AMU. One source of discrepancy between the indicators was the antimicrobial. Understanding the choice of parameter input and effects on reporting trends in AMU will inform surveillance reporting best practices to help industry understand the impacts of their AMU reduction strategies and to best communicate the information to veterinarians, their producers, and other stakeholders.

Extended-Spectrum ß-Lactamase and AmpC ß-Lactamase-Producing Escherichia coli Isolates from Chickens Raised in Small Flocks in Ontario, Canada.

Food-producing animals are recognized to play a role in the epidemiology of antimicrobial resistance in Canada. However, the presence of resistant organisms in particular groups of animals, such as chickens raised in small-holder flocks, has not been studied. The purpose of this study was, therefore, to identify and characterize Escherichia coli possessing broad-spectrum ß-lactamase genes among a collection of third-generation cephalosporin-resistant isolates recovered from 205 small flocks in southern Ontario. Extended-spectrum ß-lactamase (ESBL; CTX-M-1) positive strains were isolated from 26 out of 205 flocks (12.7%), whereas 39 strains possessing AmpC (CMY-2) were grown from 31 out of 205 flocks (15.1%). Pulsed-field gel electrophoresis (PFGE) revealed that the isolates were genetically heterogeneous. Further testing by multi-locus sequence typing confirmed that none of the PFGE-defined clusters belonged to ST131. Our results suggest that the dissemination of this resistance in bacteria isolated from chickens in small-holder flocks may be associated with the spread of plasmids rather than particular E. coli clones and that these isolates do not possess the ESBL types most commonly associated with human infections (CTX-M-15).

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.

A method to detect Escherichia coli carrying the colistin-resistance genes mcr-1 and mcr-2 using a single real-time polymerase chain reaction and its application to chicken cecal and porcine fecal samples.

Colistin is one of the last-resort antibiotics for the treatment of multidrug-resistant infections in humans, but transmissible colistin-resistance genes have emerged in bacteria from animals. The rapid and sensitive detection among animals of colonization with bacteria carrying these genes is critical in helping to control further spread. Here we describe a method for broth enrichment of colistin-resistant Escherichia coli from animal fecal and cecal samples followed by real-time polymerase chain reaction (PCR) for the simultaneous detection of two of the main colistin-resistance genes, mcr-1 and mcr-2. The PCR uses a single set of nondegenerative primers, and mcr variants can be differentiated by melt-curve analysis. Overnight culture enrichment was effective for amplifying colistin-resistant E. coli, even when initially present in numbers as low as 10 bacteria per gram of sample. The mcr-1 and mcr-2 genes were not found in any of the Ontario swine and poultry samples investigated.

La colistine est un des antibiotiques de dernier recours pour le traitement d'infections causées par des bactéries multi-résistantes chez l'humain, mais des gènes transmissibles de résistance à la colistine ont émergé chez des bactéries provenant d'animaux. La détection rapide et sensible parmi les animaux de la colonisation par ces bactéries porteuses de ces gènes est critique afin d'aider à limiter la propagation. Nous décrivons ici une méthode pour un enrichissement en bouillon des souches de Escherichia coli résistantes à la colistine provenant d'échantillons de fèces animales et de caecum suivi d'une réaction d'amplification en chaine par la polymérase (ACP) en temps réel pour la détection simultanée des deux gènes principaux de résistance à la colistine, mcr-1 et mcr-2. La réaction d'ACP utilise une seule série d'amorces non-dégénératives, et les variants de mcr peuvent être différenciés par l'analyse de la courbe de fusion. Une culture d'enrichissement d'une nuit était efficace pour amplifier les E. coli résistants à la colistine, même si présents initialement en quantité aussi faible que 10 bactéries par gramme d'échantillon. Les gènes mcr-1 et mcr-2 n'ont pas été trouvés dans aucun des échantillons porcins ou aviaires étudiés.(Traduit par Docteur Serge Messier).

Changes in antimicrobial resistance levels among Escherichia coli, Salmonella, and Campylobacter in Ontario broiler chickens between 2003 and 2015.

Poultry has been identified as a reservoir of foodborne enteric pathogens and antimicrobial resistant bacteria. The objective of this study was to describe and compare antimicrobial resistant isolates from an Ontario broiler chicken farm-level baseline project (2003 to 2004) to the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) Ontario abattoir and retail surveillance data from 2003, and to the most recent (2015) CIPARS Ontario chicken surveillance data in order to assess the impact of an industry-wide policy change in antimicrobial use. Ceftiofur resistance (TIO-R) prevalence in Salmonella decreased by 7% on farm between 2003 and 2004 and 2015. During the same timeframe, TIO-R E. coli prevalence decreased significantly by 16%, 11%, and 8% in farm, abattoir, and retail samples, respectively. Gentamicin resistant (GEN-R) E. coli, however, increased by 10% in farm and 15% in retail-derived isolates, and trimethoprim-sulfamethoxazole resistant (TMSm-R) E. coli increased significantly by 20%, 18%, and 5% in farm, abattoir, and retail isolates, respectively. Similarly, ciprofloxacin-resistant (CIP-R) Campylobacter spp. significantly increased in retail isolates by 11% and increased in farm (33%) and abattoir isolates (7%). The decrease in TIO-R Salmonella/E. coli in recent years is consistent with the timing of an industry-led intervention eliminating the preventive use of ceftiofur, a third generation cephalosporin and class of antimicrobials deemed critically important to human medicine. The rise in GEN-R and TMSm-R prevalence is indicative of recent shifts in antimicrobial use. Our study highlights the importance of integrated surveillance in detecting emerging trends and determining the efficacy of interventions to improve food safety.

La volaille a été identifiée comme étant un réservoir d'agents pathogènes entériques d'origine alimentaire et de bactéries résistantes aux antimicrobiens. L'objectif de la présente étude était de décrire et comparer des isolats résistants aux antimicrobiens provenant d'une ferme ontarienne de poulets à griller obtenus dans le cadre d'un projet de base (2003 à 2004) aux données de surveillance de 2003 du Programme intégré canadien de surveillance de la résistance aux antimicrobiens (PICRA) d'abattoir et de ventes au détail en Ontario, et aux plus récentes données de surveillance (2015) du PICRA Ontario pour la volaille afin d'évaluer l'impact d'un changement à l'ensemble de l'industrie dans l'utilisation des antimicrobiens. La prévalence de la résistance au cefiofur (TIO-R) de Salmonella a diminué de 7 % sur la ferme entre 2003 à 2004 et 2015. Durant ce même intervalle de temps, la prévalence de TIO-R de E. coli diminua de manière significative de 16 %, 11 %, et 8 % dans les échantillons provenant de la ferme, de l'abattoir et de la vente au détail, respectivement. Toutefois, les E. coli résistant à la gentamicine (GEN-R) ont augmenté de 10 % et 15 % dans les échantillons pris à la ferme et de la vente au détail, respectivement. Les E. coli résistants au trimethoprime-sulfaméthoxazole (TMSm-R) ont augmenté de manière significative par 20 %, 18 %, et 5 % dans les isolats de la ferme, de l'abattoir et de la vente au détail, respectivement. Les isolats de Campylobacter spp. résistants au ciprofloxacin (CIP-R) augmentèrent de manière significative dans les échantillons de vente au détail (11 %), ceux de la ferme (33 %) ainsi que ceux de l'abattoir (7 %). La diminution de la TIO-R chez Salmonella/E. coli au cours des dernières années concorde avec une intervention menée par l'industrie d'éliminer l'utilisation en prévention du ceftiofur, une céphalosporine de troisième génération qui est une classe d'antimicrobiens considérée d'importance critique en médecine humaine. L'augmentation de la prévalence de GEN-R et de TMSm-R est indicative d'un changement récent dans l'utilisation des antimicrobiens. Notre étude fait ressortir l'importance d'un programme intégré de surveillance pour détecter les tendances émergentes et déterminer l'efficacité des interventions pour améliorer la salubrité alimentaire.(Traduit par Docteur Serge Messier).

Molecular epidemiology of Campylobacter jejuni human and chicken isolates from two health units.

A study was conducted over a 2-year period in the Perth District and Wellington-Dufferin-Guelph health units in Ontario, with an objective of using comparative genomic fingerprinting (CGF) with a 40-gene assay (CGF40) to investigate the association between human cases of campylobacteriosis and spatially and temporally related Campylobacter isolates from retail chicken. CGF results were available for isolates from 115 human cases and 718 retail chicken samples. These data were combined with CGF results from a large reference database of Campylobacter isolates. Isolates were categorized into types based on >90% CGF40 fingerprint similarity (CGF-90%). CGF-90% types were categorized as chicken associated (CA90) when the proportion of animal isolates in the given type that originated from chicken was at least 80% and was statistically significant. Risk factor data were collected from cases by questionnaire. Urban cases were significantly more likely than rural cases to be CA90 and there were significantly fewer CA90 cases in the second year of the study. Due to the population distribution in Canada and most industrialized countries, the majority of campylobacteriosis cases are urban dwellers. Therefore, the association between urban cases and chicken-associated types of Campylobacter emphasizes the importance of educational and food safety efforts to reduce the impact of Campylobacter from retail chicken on public health. Sources other than chicken may be more important for rural dwellers.

Antimicrobial resistance and antimicrobial use associated with laboratory-confirmed cases of Campylobacter infection in two health units in Ontario.

AIM: A population-based study was conducted over a two-year period in the Perth District (PD) and Wellington-Dufferin-Guelph (WDG) health units in Ontario to document antimicrobial resistance and antimicrobial use associated with clinical cases of laboratory-confirmed campylobacteriosis. METHODS: Etest (bioMérieux SA, France) was used to determine the minimum inhibitory concentration of amoxicillin/clavulanic acid, ampicillin, chloramphenicol, ciprofloxacin (CIP), clindamycin, erythromycin (ERY), gentamicin, nalidixic acid and tetracycline. Data regarding antimicrobial use were collected from 250 cases. RESULTS: Of the 250 cases, 165 (65.7%) reported staying home or being hospitalized due to campylobacteriosis. Fifty-four per cent of cases (135 of 249) reported taking antimicrobials to treat campylobacteriosis. In 115 cases (51.1%), fecal culture results were not used for treatment decisions because they were not available before the initiation of antimicrobial treatment and/or they were not available before the cessation of symptoms. Of the 250 cases, 124 (49.6%) had available Campylobacter isolates, of which 66 (53.2%) were resistant to at least one of the antimicrobials tested. No resistance to ampicillin, chloramphenicol or gentamicin was found in these isolates. Six isolates (4.8%) were resistant to CIP. Two isolates (1.6%) were resistant to ERY; however, no isolates were resistant to both CIP and ERY. CONCLUSION: Prudent use practices should be promoted among physicians to reduce the use of antimicrobials for the treatment of gastroenteritis in general and campylobacteriosis in particular, as well as to minimize the future development of resistance to these antimicrobials in Campylobacter species.

OBJECTIF: Les chercheurs ont mené une étude en population pendant une période de deux ans dans les unités sanitaires du district de Perth et de Wellington-Dufferin-Guelph en Ontario, afin d'étayer la résistance antimicrobienne et l'utilisation d'antimicrobiens associées à des cas cliniques de campylobactériose confirmés en laboratoire. MÉTHODOLOGIE: Les chercheurs ont utilisé l'Etest (bioMérieux SA, France) pour déterminer la concentration inhibitrice minimale d'amoxicilline-acide clavulanique, d'ampicilline, de chloramphénicol, de ciprofloxacine (CIP), de clindamycine, d'érythromycine (ERY), de gentamicine, d'acide nalidixique et de tétracycline. Ils ont colligé les données sur l'utilisation d'antimicrobiens auprès de 250 cas. RÉSULTATS: Sur les 250 cas, 165 (65,74 %) ont déclaré être restés à la maison ou avoir été hospitalisés en raison de leur campylobactériose. Cinquante-quatre pour cent des cas (135 sur 249) ont déclaré avoir pris des antimicrobiens pour traiter leur campylobactériose. Dans 115 cas (51,1 %), les résultats de la coproculture n'ont pas été utilisés pour prendre les décisions thérapeutiques parce qu'ils n'étaient pas disponibles avant l'amorce du traitement antimicrobien ou avant l'arrêt des symptômes. Sur les 250 cas, 124 (49,6 %) disposaient d'isolats du Campylobacter, dont 66 (53,2 %) étaient résistants à au moins un des antimicrobiens testés. Les chercheurs n'ont relevé aucun cas de résistance à l'ampicilline, au chloramphénicol ou à la gentamicine dans ces isolats. Six (4,8 %) étaient résistants à la CIP et deux (1,6 %), à l'ERY, mais aucun n'était résistant à la fois à la CIP et à l'ERY. CONCLUSION: Il faudrait promouvoir des pratiques d'utilisation prudentes chez les médecins pour réduire l'utilisation d'antimicrobiens dans le traitement de la gastroentérite en général et de la campylobactériose en particulier, ainsi que pour limiter le plus possible l'apparition future de résistance à ces antimicrobiens dans les espèces de Campylobacter.

Associations between reported on-farm antimicrobial use practices and observed antimicrobial resistance in generic fecal Escherichia coli isolated from Alberta finishing swine farms.

Multilevel random intercept logistic and Poisson regression were used to model associations between various antimicrobial use practices and resistance to individual and multiple antimicrobials among generic fecal Escherichia coli isolated from Alberta finishing swine. In-feed antimicrobial use in finishers was significantly associated with increased risk of resistance to ampicillin, chloramphenicol, streptomycin, and sulfisoxazole in generic E. coli isolates. Chlortetracycline use in grower rations was associated with ampicillin and tetracycline resistance. Use of in-feed antimicrobials in finishers was significantly associated with increased risk of resistance of generic E. coli to multiple antimicrobials. The study findings suggest that certain farm-level interventions related to antimicrobial use might be beneficial in reducing development and emergence of antimicrobial resistance in swine populations.

Antimicrobial use in the Alberta sheep industry.

Information regarding antimicrobial use in sheep is scarce. In 2001, a scrapie surveillance program was initiated in Alberta that also provided a mechanism for collecting other sheep health data including antimicrobial use information between April 2001 and April 2002. A major objective of this study was to describe antimicrobial use in the Alberta sheep industry. This was done by obtaining qualitative antimicrobial use information from all flocks (n = 212) providing cull ewes to the program using a brief, primarily flock-level, questionnaire. The respondents' flocks represented 13.6% of the total provincial flock in Alberta in 2001. By a substantial amount, the most frequent method of administering antimicrobials was through injection followed by in-feed, oral (liquids, pills, boluses), and in-water routes, respectively. Drug-specific use data were collected for injectable antimicrobials only, with the most commonly used antimicrobial classes being penicillins followed by tetracyclines. Producers rarely treated some or all of their flock with injectable antimicrobials after discovering an individual sick animal. Adult sheep were the most common age group treated with injectable antimicrobials and the most frequent reason for injectable antimicrobial use was mastitis followed by respiratory problems. This study provides some initial insight regarding antimicrobial use in Alberta sheep flocks. However, collection of more drug-specific data (drug type, dose/concentration, duration of treatment) for noninjectable routes of administration should be conducted in future studies. Assessing antimicrobial use in other sectors of the Alberta sheep industry (feedlots) and other provinces across Canada would also be beneficial.