Canadian Integrated Program For Antimicrobial Resistance Surveillance Research Articles

Comparison of antimicrobial resistance among Salmonella enterica serovars isolated from Canadian turkey flocks, 2013 to 2021

The emergence of antimicrobial resistance (AMR) in Salmonella from turkeys has raised a food safety concern in Canada as certain serovars have been implicated in human salmonellosis outbreaks in recent years. While several studies evaluated AMR in broiler chickens in Canada, there are limited studies that assess AMR in turkey flocks. This study analyzed data collected between 2013 and 2021 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) farm turkey surveillance program to determine the prevalence of AMR and differences in resistance patterns among Salmonella serovars recovered from turkey flocks. Salmonella isolates were tested for susceptibility to 14 antimicrobials using a microbroth dilution method. Hierarchical clustering dendrograms were constructed to compare the individual AMR status of Salmonella serovars. Differences in the probability of resistance between Salmonella serovars were determined using generalized estimating equation logistic regression models to account for farm-level clustering. Of the 1,367 Salmonella isolates detected, 55.3% were resistant to at least one antimicrobial and 25.3% were multidrug resistant (MDR) (resistant to ≥3 antimicrobial classes). The Salmonella isolates exhibited high resistance to tetracycline (43.3%), streptomycin (47.2%), and sulfisoxazole (29.1%). The 3 most frequently occurring serovars were S. Uganda (22.9%), S. Hadar (13.5%), and S. Reading (12.0%). Streptomycin-sulfisoxazole-tetracycline (n=204) was the most frequent MDR pattern identified. Heatmaps showed that S. Reading exhibited coresistance to the quinolone class antimicrobials, ciprofloxacin, and nalidixic acid; S. Heidelberg to gentamicin and sulfisoxazole; and S. Agona to ampicillin and ceftriaxone. Salmonella Hadar isolates had higher odds of resistance to tetracycline (OR: 152.1, 95% CI: 70.6-327.4) while the probability of being resistant to gentamicin and ampicillin was significantly higher in S. Senftenberg than in all the other serovars. Moreover, S. Uganda had the highest odds of being MDR (OR: 4.7, 95% CI: 3.7-6.1). The high resistance observed warrants a reassessment of the drivers for AMR, including AMU strategies and other production factors. Differences in AMR patterns highlight the need to implement serovar-specific mitigation strategies.

Integrated surveillance of extended-spectrum beta-lactamase (ESBL)-producing Salmonella and Escherichia coli from humans and animal species raised for human consumption in Canada from 2012 to 2017.

Resistance to beta-lactam antimicrobials caused by extended-spectrum beta-lactamase (ESBL)-producing organisms is a global health concern. The objectives of this study were to (1) summarise the prevalence of potential ESBL-producing Escherichia coli (ESBL-EC) and Salmonella spp. (ESBL-SA) isolates from agrifood and human sources in Canada from 2012 to 2017, and (2) describe the distribution of ESBL genotypes among these isolates. All data were obtained from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). CIPARS analysed samples for the presence of ESBLs through phenotypic classification and identified beta-lactamase genes (blaTEM, blaSHV, blaCTX, blaOXA, blaCMY-2) using polymerase chain reaction (PCR) and whole genome sequencing (WGS). The prevalence of PCR-confirmed ESBL-EC in agrifood samples ranged from 0.5% to 3% across the surveillance years, and was detected most frequently in samples from broiler chicken farms. The overall prevalence of PCR-confirmed ESBL-SA varied between 1% and 4% between 2012 and 2017, and was most frequently detected in clinical isolates from domestic cattle. The TEM-CMY2 gene combination was the most frequently detected genotype for both ESBL-EC and ESBL-SA. The data suggest that the prevalence of ESBL-EC and ESBL-SA in Canada was low (i.e. <5%), but ongoing surveillance is needed to detect emerging or changing trends.

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.

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.

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.

Salmonella spp. prevalence and antimicrobial resistance in broiler chicken and turkey flocks in Canada from 2013 to 2018

Salmonella infections are a major human health concern. In the elderly and immunocompromised, infections can be life-threatening and may require antibiotic therapy. Where antibiotic therapy is required, antimicrobials of choice include fluoroquinolones and extended-spectrum cephalosporins (ESC). The aim of this study is to utilize data from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) to compare the prevalence of Salmonella serovars between broiler chicken and turkey flocks across Canada, and to gain an understanding of the prevalence of resistance to antimicrobials categorized as important to human health. There were 1,596 Salmonella isolates obtained from 514 broiler chicken flocks, and 659 Salmonella isolates obtained from 217 turkey flocks (2013-2018). All isolates were obtained from pooled faecal samples. Among broiler chicken flocks, the top three serovars were Kentucky (n=573, 36%), Enteritidis (n=314, 20%) and Heidelberg (n=127, 8%). Resistance to ceftriaxone among Salmonella ser. Kentucky decreased from 27% in 2013 to 22% in 2018. There was no resistance among Salmonella ser. Enteritidis reported until 2018 when one isolate from British Columbia was resistant to ampicillin, streptomycin, sulphisoxazole and tetracycline. Salmonella ser. Heidelberg resistance to ceftriaxone decreased from 19% in 2013 to 14% in 2018. Among turkey flocks the top three serovars were Uganda (n=109, 16.5%), Hadar (n=85, 12%) and Muenchen (n=66, 10%). No isolates of Salmonella ser. Uganda or Salmonella ser. Muenchen were resistant to any β-lactams. Salmonella ser. Hadar (34/81, 42%) exhibited resistance to ampicillin. There was no resistance to quinolones among turkey isolates. Emerging resistance among Salmonella ser. Enteritidis, and resistance to β-lactams and fluoroquinolones among Salmonella ser. Kentucky from broilers are cause for concern as these classes of antimicrobials are important for treatment of salmonellosis.

Evaluating the Integration of One Health in Surveillance Systems for Antimicrobial Use and Resistance: A Conceptual Framework.

It is now widely acknowledged that surveillance of antimicrobial resistance (AMR) must adopt a “One Health” (OH) approach to successfully address the significant threats this global public health issue poses to humans, animals, and the environment. While many protocols exist for the evaluation of surveillance, the specific aspect of the integration of a OH approach into surveillance systems for AMR and antimicrobial Use (AMU), suffers from a lack of common and accepted guidelines and metrics for its monitoring and evaluation functions. This article presents a conceptual framework to evaluate the integration of OH in surveillance systems for AMR and AMU, named the Integrated Surveillance System Evaluation framework (ISSE framework). The ISSE framework aims to assist stakeholders and researchers who design an overall evaluation plan to select the relevant evaluation questions and tools. The framework was developed in partnership with the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). It consists of five evaluation components, which consider the capacity of the system to: [1] integrate a OH approach, [2] produce OH information and expertise, [3] generate actionable knowledge, [4] influence decision-making, and [5] positively impact outcomes. For each component, a set of evaluation questions is defined, and links to other available evaluation tools are shown. The ISSE framework helps evaluators to systematically assess the different OH aspects of a surveillance system, to gain comprehensive information on the performance and value of these integrated efforts, and to use the evaluation results to refine and improve the surveillance of AMR and AMU globally.

A cross-sectional study of the prevalence factors associated with fluoroquinolone resistant Campylobacter jejuni in broiler flocks in Canada

Campylobacter infections in humans are usually self-limiting; however, antibiotic intervention may be necessary in the case of severe infection. Fluoroquinolones are often the drug of choice for treatment of campylobacteriosis; however, resistance to these drugs can develop rapidly, complicating treatment protocols. Increasing resistance to fluoroquinolones in human infections has coincided with approval of use of fluoroquinolones in animals, therefore, isolation of fluoroquinolone resistant (FQr) Campylobacter in broiler flocks is concerning. This cross-sectional study utilized data collected from 2013-2018 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) on-farm surveillance program to investigate prevalence factors associated with the isolation of FQr C. jejuni from broiler faecal samples. Mixed effects logistic regression models accounting for clustering of flocks within hatcheries, with and without a fixed effect for the presence of flock level tetracycline resistance were used to assess prevalence factors among 536 C. jejuni isolates from 158 flocks. Both models indicated that the type of bird used (Ross versus Cobb or mixed), the use of virginiamycin as a feed additive, the use of traps to control rodent populations in the barn, and the total number of birds in the barn were significant prevalence factors for increased FQr C. jejuni in a flock. In the model where flock level tetracycline resistance was included as a fixed effect, the odds of FQr C. jejuni increased by 16 (95% CI: 3.74, 68), and the magnitude of the effect of each of the identified prevalence factors was larger. Both models indicated that methods of disinfection of water lines between production cycles is important, with the use of chlorine being protective in the model where tetracycline resistance was included as a fixed effect, and the use of hydrogen peroxide being a risk factor in the model where tetracycline resistance was not included as a fixed effect. The use of hot water to wash the barn between production cycles was also a significant protective factor in the model where tetracycline resistance was not included as a fixed effect. These results indicate that biosecurity and sanitation procedures play a role in the dissemination of FQr C. jejuni in broiler flocks. Future analysis should seek to understand the effect of different disinfectant products on the isolation of FQr C. jejuni. Gaining a better understanding of the management of these critical practices may allow for the reduction of this enteric pathogen in broiler flocks in Canada.

Antimicrobial Resistance of Campylobacter in Broiler Chicken Along the Food Chain in Canada

Antimicrobial resistance (AMR) is a major public health threat worldwide. The main objective of this study was to compare AMR in Campylobacter from broiler chickens raised on Canadian farms and their products in different geographical regions of Canada. To do this, antimicrobial susceptibility results from isolates of Campylobacter recovered from a national microbiological baseline study conducted in federally registered establishments and in the retail marketplace were analyzed. Among 1460 isolates tested, 774 (53%) were resistant to at least one antimicrobial, with a predominance of three profiles: tetracycline (39%), quinolone–tetracycline (6.6%), and quinolones only (3.5%). The results showed no significant difference in the frequency of resistant profiles (p ≥ 0.05) among the isolates originating from different points in the food processing chain at slaughterhouses and in retail establishments. This suggests that AMR observed in Campylobacter isolates from raw chicken at retail originated further upstream in the system. A difference in the frequency of certain resistance profiles was observed between the regions of Canada. For instance, in British Columbia, there was more resistance to quinolones, while in Ontario and Quebec, Campylobacter isolates were more resistant to tetracyclines, macrolides, ketolides, and lincosamides. Comparison of AMR data from this study with those from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) did not show any significant difference and provides evidence that CIPARS produces nationally representative resistance results.

Antimicrobial resistance of Salmonella and generic Escherichia coli isolated from surface water samples used for recreation and a source of drinking water in southwestern Ontario, Canada.

Antimicrobial resistance (AMR) in the aquatic environment represents an important means of introduction and dissemination of resistance genes, and presence of resistant pathogens in surface waters may pose a public health concern to recreational and drinking water users. The purpose of this study was to explore antimicrobial resistance patterns in water samples collected from the Grand River watershed (southwestern Ontario, Canada) to describe the composition, trends and potential risks of AMR in the aquatic environment. As part of FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), stream water samples were collected bi-weekly from sampling sites within the Grand River watershed in the Waterloo, Ontario sentinel site and tested for the presence and antimicrobial susceptibility of Salmonella spp. (2005-2013) and generic Escherichia coli (2012-2013). Of all samples tested, 16% of Salmonella and 22% of E.coli isolates were resistant to at least one antimicrobial, including three Salmonella isolates and two E.coli isolates that were resistant to Category I antimicrobials, which are classified as very high importance for the treatment of serious bacterial infections in humans. The greatest proportion of resistant E.coli isolates were observed from the river site upstream of the drinking water intake, while the greatest proportion of resistant Salmonella isolates were from sites upstream in the watershed, and at one recreational water site. Salmonella resistance trends remained fairly stable between 2007 and 2013, with the exception of streptomycin and tetracycline which increased in 2010 and 2013. Continued surveillance of antimicrobial resistance patterns and exploration of risk factor data will allow for a better understanding of resistance transmission in the aquatic environment.

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.

Antimicrobials Used for the Therapy of Necrotic Enteritis and Coccidiosis in Broiler Chickens and Turkeys in Canada, Farm Surveillance Results (2013-2017).

Since 2013, the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) has collected antimicrobial use (AMU) and antimicrobial resistance data from sentinel broiler chicken flocks (Br, five provinces) and sentinel turkey flocks (Tk, one province 2013-2015, three provinces 2016-2017). The objectives of this paper were to describe various preventive strategies aimed at controlling necrotic enteritis (NE) and coccidiosis in the broiler chicken and turkey flocks participating in CIPARS and FoodNet Canada Farm Surveillance Program between 2013 and 2017, to quantify and identify trends in antimicrobials used in feed, and to describe temporal changes in the diagnoses of bacterial and protozoal diseases in relation to antimicrobial use in feed. Comprehensive data were collected (by questionnaire) enabling AMU assessment by various count-based metrics (i.e., frequency and number of medicated rations), weight-based metrics (i.e., inclusion rate in feed and kilograms consumed), and technical indicators (i.e., milligrams per population correction unit [mg/PCU]). Qualitative information such as reasons for use and frequency of diagnosed diseases provided context to the trends in AMU. Between 2013 and 2017, 646 broiler flocks (14.9 million kg biomass) and 234 turkey flocks (12.4 million kg biomass) were surveyed. Overall, antimicrobials used for the prevention of Clostridium perfringens infections (NE) contributed to 85% (109/128 mg/PCUBr) and 95% (59/62 mg/PCUTk) of the quantity of antimicrobials administered via feed in broiler chickens and turkeys, respectively. Three NE programs were used: either 1, 2, or ≥3 antimicrobials administered throughout the production cycle. The treatment protocol in which a single antimicrobial was used throughout the cycle was the most frequent NE preventive program for broiler chickens (58%) and turkeys (76%). Bacitracin and virginiamycin were the top two most frequently used antimicrobials in both species for NE. For coccidiosis control, ionophores and chemical coccidiostats contributed to 66% (3091 kg) and 68% (1561 kg) of the total feed antimicrobial exposures in broiler chickens and turkeys, respectively. Documented coccidiosis programs included continuous or straight (1 drug/cycle), shuttle or dual control (≥2 drugs/cycle), and vaccination. Variations in coccidiosis programs between species were noted: broiler chickens frequently used a shuttle or dual-control program (68%), whereas turkey flocks used primarily a continuous or straight program (74%). Flocks raised without antibiotics and organic farms (10.3% of broiler chickens and 9.8% turkey flocks) used vaccines to prevent coccidiosis. A small number of broiler flocks (n = 6) used a combination of a vaccination and a coccidiostat during the cycle. During the surveillance timeframe used for this paper, the total feed AMU decreased over time in broiler chickens from 136 to 120 mg/PCUBr and in turkeys from 85 to 62 mg/ PCUTk, with no remarkable changes in the frequency of flocks diagnosed with bacterial and protozoal diseases. Surveillance findings such as these will be used as valid reference points in light of the upcoming changes in Canadian federal AMU regulations and industry-led initiatives aimed at reducing AMU.

A comparison of modelling options to assess annual variation in susceptibility of generic Escherichia coli isolates to ceftiofur, ampicillin and nalidixic acid from retail chicken meat in Canada

Statistical modelling of antimicrobial resistance (AMR) data is an important aspect of AMR surveillance programs; however, minimum inhibitory concentration (MIC) data can be challenging to model. The conventional approach is to dichotomize data using established breakpoints, then use logistic regression modelling for analysis. A disadvantage of this approach is a loss of information created by dichotomizing the data. The objectives of the study were to compare the performance and results of different regression models for the analysis of annual variation in susceptibility of generic Escherichia coli (E. coli) isolates to ceftiofur, ampicillin and nalidixic acid from retail chicken meat surveillance samples. E. coli susceptibility data for the three antimicrobials from retail chicken samples from 2007 to 2014 were obtained from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). Annual variation in susceptibility for each antimicrobial was evaluated using multivariable linear, tobit, logistic, multinomial, ordinal and complementary log-log regression models (clog-log). MIC (log2), censored MIC (log2), resistant/susceptible, and categorized MIC (3 or 4 categories) data were used as outcome variables for the appropriate statistical models. Year and region were modelled as categorical predictor variables. Random intercepts were included in the ceftiofur and ampicillin models to account for clustering by retail establishment. The model assumptions evaluated for the mixed models included homoscedasticity and normality of residuals (linear and tobit), homoscedasticity and normality of best linear unbiased predictions (all models), proportional odds (ordinal), and proportional hazards (clog-log). Fixed effects models were used for the nalidixic acid models. The model assumptions evaluated for the fixed effects models included homoscedasticity and normality of residuals (linear and tobit), goodness-of-fit test (logistic and multinomial), proportional odds (ordinal), and proportional hazards (clog-log).Only logistic and multinomial models met model assumptions. Significant annual variation in susceptibility to all three antimicrobials was identified by the multinomial regression models, whereas the logistic regression models only identified significant annual variation in susceptibility to ceftiofur. The multinomial regression model consistently identified additional significant annual variation in susceptibility compared to the logistic regression model. The multinomial modelling approach was able to identify differences between MIC categories within susceptible MIC values, which were below the breakpoint (R) detection level. Given the convention of dichotomizing susceptibility data, the logistic regression approach is likely to remain the standard method of analysis for AMR surveillance data; however, the results of this study demonstrate that multinomial regression should be considered for the analysis of AMR surveillance data.

Comparison of annual and regional variation in multidrug resistance using various classification metrics for generic Escherichia coli isolated from chicken abattoir surveillance samples in Canada

Antimicrobial resistance (AMR) and related multidrug resistance (MDR) are important global public health issues. The Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) conducts surveillance of AMR in enteric bacteria and monitors MDR. However, the analysis of MDR is complicated by the lack of consensus for MDR definitions. The objectives were to describe the most common resistance patterns in generic E. coli isolates from chicken cecal samples and determine the impact of using different MDR metrics for analysis of annual and regional variation in MDR. From 2006 to 2015, 1598 E. coli isolates were collected from chickens at slaughter for CIPARS. Three MDR classification metrics were used: MDR-drug (MDR if the isolate was resistant (R) to ≥3 of the 13 antimicrobials); MDR-cat (MDR if R to ≥3 of the 9 antimicrobials categories); and MDR-class (MDR if R to ≥3 of the 6 antimicrobial classes). The most frequent resistance patterns overall, and by year and region were extracted along with patterns that included resistance to quinolones, and third generation cephalosporins and/or β-lactams with β-lactamase inhibitors. For each MDR metric, mixed logistic regression models, which included random intercepts for abattoir, were fitted to analyze the association between prevalence of MDR, and year and region. Interaction effects between year and region were evaluated. Overall, and in all years and regions, non-resistant was the most common resistance pattern (24.9%, 95% CI 22.8–27.1%). Resistance patterns that included third generation cephalosporins and β-lactams with β-lactamase inhibitors were common. The prevalence of MDR was variable: MDR-class 38.5% (95% CI 36.1–41.0%); MDR-cat 49.4% (95% CI 46.9–51.9%); and MDR-drug 53.3% (95% CI 50.8–55.8%). Based on models fitted with individual fixed effects, significant annual variation in the prevalence of MDR was identified with MDR-drug and MDR-class models. Significant regional variation was identified for all three MDR metric models. Significant interaction effects between year and region were identified with the MDR-drug and MDR-cat multivariable mixed logistic regression models. The interpretation of the association between the prevalence of MDR, and year and region differed depending on the MDR metric used. These results are supportive of the previous concerns that caution must be taken when comparing MDR results between studies. Global consensus is needed for the optimal MDR classification metric for foodborne enteric bacteria AMR surveillance.

Exposure to antimicrobial-resistant Escherichia coli through the consumption of ground beef in Western Canada

Emergence and dissemination of antimicrobial resistance (AMR) in food-borne bacteria is a public health issue. Retail meat is considered an important carrier for antimicrobial-resistant Escherichia coli. Currently, resistance of E. coli strains to third generation cephalosporins are of particular concern, with significant potential animal and public health consequences. Resistance to tetracyclines is also a concern, due to high prevalence of this resistance and important co-resistance patterns. However, the actual likelihood of exposure to antimicrobial-resistant bacteria via the consumption of retail meats, beyond the simple frequencies of resistance found in samples of meat at the grocery store, in Canada remains to be investigated. This study was conducted to estimate the potential human exposure to ceftriaxone- and tetracycline-resistant E.coli (CREC and TREC; the hazards of interest) through the consumption of ground beef in Western Canada.Our exposure assessment simulation model included five modules: 1) estimation of prevalence and concentration of the hazards of interest in retail ground beef samples collected by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), 2011–2014; 2) potential growth of the hazards during transportation from retail to home; 3) potential growth or reduction of the hazards during home storage; 4) thermal inactivation of the hazards due to cooking; and 5) consumption. The outputs of the model were presented as the probability of consumers' exposure to various threshold levels of the hazards [10 to 106 colony forming units (CFU)] in one serving of cooked ground beef.Overall, the probabilities of exposure to high threshold levels of the hazards (>4 log10 CFU/serving) were estimated to be lower than 0.12% in the study population (2011–2014). The mean probabilities of exposure to at least 1 log10 CFU CREC and TREC in a single meal containing cooked ground beef were 0.067% (SD: 0.001%) and 1.20% (SD: 0.02%), respectively. This probability substantially decreased as the threshold level of exposure increased to ≥6 log10 CFU. The probability of exposure to TREC was consistently greater than that for CREC. Cooking led to a prominent drop in the mean concentration of the hazards (4.7 log10 CFU/g). The findings from this research could inform the policy-making process and provide suggestions for adjustments in future retail surveillance plans. In addition, important knowledge gaps in this area have been highlighted.

Risk factors associated with the A2C resistance pattern among E. coli isolates from broiler flocks in Canada

The extra-label use of ceftiofur in Canadian hatcheries was cause for concern due to an increased prevalence of ceftiofur resistant Salmonella Heidelberg in chickens and humans in Québec. Due to on-going concerns related to human health the use of ceftiofur was eventually phased out of the poultry production industry in 2014–2015. Simultaneous resistance to amoxicillin-clavulanic acid, ceftiofur and cefoxitin, a pattern known as A2C, caused by the presence of blaCMY-2 has become increasingly prevalent in broiler flocks worldwide. This study used data from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) broiler farm surveillance program to examine the risk factors for A2C resistant Escherichia coli isolates (A2C-REI) from 1482 samples taken from 371 broiler flocks in Canada, between 2013 and 2015. Results of a mixed effects Poisson model with fixed effects at the flock level and random effects at the hatchery level indicated that the in ovo use of ceftiofur (Prevalence Ratio (PR): 1.91, 95% CI: 1.43–2.57), and the use of hydrogen peroxide to disinfect water lines during the growing period (PR: 1.52, 95% CI: 1.11–2.07), were risk factors for increased expected prevalence of A2C-REI, compared to farms not using these strategies. Controlling for wild birds (PR: 0.64, 95% CI: 0.48–0.84), use of avilamycin in feed (PR: 0.63, 95% CI: 0.41–0.98), and storage of manure on the farm (PR: 0.67, 95% CI: 0.51–0.89) were associated with a lower expected prevalence of A2C-REI, than farms that did not use these strategies. This study identified important management factors that were associated with the prevalence of A2C-REI on Canadian broiler farms. The findings of this study can be used as a baseline for the monitoring of the effects of the removal of ceftiofur from poultry production in Canada.

Prevalence and antimicrobial resistance among Escherichia coli and Salmonella in Ontario smallholder chicken flocks.

Surveillance is an important component of an overall strategy to address antimicrobial resistant bacteria in food animals and the food chain. The poultry market has many points of entry into the Canadian food chain, and some production practices are underrepresented in terms of surveillance. For example, pathogen carriage and antimicrobial resistance surveillance data are limited in smallholder chicken flocks raised for slaughter at provincially inspected abattoirs. In Canada, antimicrobial resistance in Escherichia coli and Salmonella isolated from commercial broiler chicken flocks, slaughtered at federally inspected abattoirs, is monitored by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). The objective of this study was to establish baseline information of antimicrobial resistance presence in E.coli and Salmonella isolated from smallholder flocks in Ontario, utilizing CIPARS collection and isolation methodologies, and to compare findings with CIPARS federally inspected abattoir data from Ontario, Canada. Five chickens per flock were sampled from 205 smallholder flocks. Of 1,025 samples, the E.coli prevalence was 99% (1,022/1,025), and 47% (483/1,022) of positive E.coli isolates were resistant to one or more of the 14 antimicrobials. Furthermore, as compared to results reported for the CIPARS commercial flocks, E.coli isolates from smallholder flocks had significantly lower resistance prevalence to six of 14 individual antimicrobials. Recovery of E.coli did not differ between federally inspected and provincially inspected flocks. Salmonella prevalence at the bird level in smallholder flocks was 0.3% (3/1,025), significantly lower (p≪0.0001, 95% CI 0.080%-0.86%) than federally inspected commercial flocks. The overall differences found between the commercial and smallholder flocks may be explained by differences in poultry husbandry practices and hatchery sources.

Antimicrobial use surveillance in broiler chicken flocks in Canada, 2013-2015.

There is a paucity of data on the reason for and the quantity of antimicrobials used in broiler chickens in Canada. To address this, the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) implemented surveillance of antimicrobial use (AMU) and antimicrobial resistance (AMR) in broiler chicken flocks in 2013. Shortly after this (2014), the poultry industry banned the preventive use of ceftiofur in broiler chickens. The objectives of this analysis were to describe antimicrobial use (AMU) in Canadian broiler chickens between 2013 and 2015 (n = 378 flocks), compare these results to other animal species in Canada, to highlight the utility of farm surveillance data to evaluate the impact of a policy change, and to explore how different antimicrobial use metrics might affect data interpretation and communication. The surveillance data indicated that the poultry industry policy resulted in lower antimicrobial use and resistance, and they successfully captured information on when, where, why, and how much antimicrobials were being used. The majority of antimicrobials were administered via the feed (95%). The relative frequency of antimicrobial classes used in broiler chickens differed from those used in swine or in food animal production in general. Coccidiostats were the most frequently used antimicrobial classes (53% of total kg). Excluding coccidiostats, the top three most frequently used antimicrobial classes were bacitracin (53% of flocks), virginiamycin (25%) and avilamycin (21%), mainly used for the prevention of necrotic enteritis. Depending on the AMU metric utilized, the relative rankings of the top antimicrobials changed; hence the choice of the AMU metric is an important consideration for any AMU reporting. When using milligrams/Population Correction Unit (mg/PCU) the top three antimicrobial classes used were bacitracins (76 mg/PCU), trimethoprim-sulfonamides (24 mg/PCU), and penicillins (15 mg/PCU), whereas when using a number of Defined Daily Doses in animals using Canadian standards /1,000 chicken-days at risk (nDDDvetCA/1,000 CD) the ranking was bacitracins (223 nDDDvetCA/1,000 CD), streptogramins (118 nDDDvetCA/1,000 CD), and trimethoprim-sulfonamides (87 nDDDvetCA/1,000 CD). The median animal treatment days in feed for one cycle (ATD/cycle) during the three-year study were 34 ATD/cycle; this was equal to the mean age of the flocks at pre-harvest sampling day (days at risk), indicating that the studied flocks except those that were raised without antibiotics and organic, were fed with medicated rations throughout the observation period. Overall, more than half (59%) of antimicrobials used in broiler chickens were in classes not used in human medicine, such as ionophores and chemical coccidiostats aimed to prevent coccidiosis. Compared to grower-finisher pigs and in production animal species (national sales data), the mg/PCU of antimicrobials used in broiler chickens was relatively lower. The findings of this paper highlighted the importance of farm-level AMU surveillance in measuring the impact of interventions to reduce antimicrobials in poultry.

A Whole-Genome Sequencing Approach To Study Cefoxitin-Resistant Salmonella enterica Serovar Heidelberg Isolates from Various Sources.

ABSTRACTThis study characterized cefoxitin-resistant and -susceptible Salmonella enterica serovar Heidelberg strains from humans, abattoir poultry, and retail poultry to assess the molecular relationships of isolates from these sources in Québec in 2012. Isolates were collected as part of the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). All isolates were subjected to antimicrobial susceptibility testing, PCR for CMY-2, pulsed-field gel electrophoresis (PFGE), and whole-genome sequencing (WGS). A total of 113 S. Heidelberg isolates from humans (n = 51), abattoir poultry (n = 18), and retail poultry (n = 44) were studied. All cefoxitin-resistant isolates (n = 65) were also resistant to amoxicillin-clavulanic acid, ampicillin, ceftiofur, and ceftriaxone, and all contained the CMY-2 gene. PFGE analysis showed that 111/113 (98.2%) isolates clustered together with ≥90% similarity. Core genome analysis using WGS identified 13 small clusters of isolates with 0 to 4 single nucleotide variations (SNVs), consisting of cefoxitin-resistant and -susceptible human, abattoir poultry, and retail poultry isolates. CMY-2 plasmids from cefoxitin-resistant isolates all belonged to incompatibility group I1. Analysis of IncI1 plasmid sequences revealed high identity (95 to 99%) to a previously described plasmid (pCVM29188_101) found in Salmonella Kentucky. When compared to pCVM29188_101, all sequenced cefoxitin-resistant isolates were found to carry 1 of 10 possible variant plasmids. Transmission of S. Heidelberg may be occurring between human, abattoir poultry, and retail poultry sources, and transmission of a common CMY-2 plasmid may be occurring among S. Heidelberg strains with variable genetic backgrounds.

Antimicrobial use and antimicrobial resistance trends in Canada: 2014.

There is a global concern that the emergence of antimicrobial resistance (AMR) threatens our ability to treat infectious diseases. The Canadian Antimicrobial Resistance Surveillance System (CARSS) was created in response to the Government of Canada's commitment to addressing AMR. CARSS integrates information from nine different national surveillance systems for tracking antimicrobial use (AMU) and AMR in both humans and animals to inform AMU/AMR research and policy. To provide highlights of CARSS data on antimicrobial use in humans and animals, AMR trends in human infections in both hospital and community settings and AMR bacteria found in food production animals. Information on AMU in animals and humans is purchased and additional information on AMU in animals is collected through the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). AMR data in humans focuses on first priority organisms. Data on priority organisms for hospital-based AMR is collected through Canadian Nosocomial Infection Surveillance Program (CNISP), Canadian Tuberculosis Laboratory Surveillance System (CTBLSS), Canadian Tuberculosis Laboratory Surveillance System (CTBRS), Canadian Tuberculosis Reporting System (CTBRS) and CIPARS. Data on community-based AMR is collected through CTBLSS, CTBRS, CIPARS, the Antimicrobial-resistant Neisseria gonorrhoeae Surveillance System (ARNGSS) and the National Surveillance of Invasive Streptococcal Disease (NSISD). AMR data on animals is collected through CIPARS. In terms of antibiotic usage in 2014, approximately 82% of antimicrobials were directed to food production animals, 18% to humans and less than one percent each to companion animals (e.g., pets) and crops. Over the past five years, 73% of antimicrobials distributed to food production animals belonged to the same classes as those used in human medicine. Antibiotic usage in humans has remained relatively stable. Trends in 2014 for AMR in hospitals include declining rates of hospital-acquired Clostridium difficile to 3.4 cases per 1,000 patient admissions, methicillin-resistant Staphylococcus aureus (MRSA) infections to 2.89 cases per 10,000 patient days and vancomycin-resistant Enterococci (VRE) to 0.45 cases per 10,000 patient days. Resistance to a number of antimicrobials used to treat Streptococcus pneumoniae has decreased since the introduction of pneumococcal vaccine in 2010. In contrast, trends in 2014 for AMR in the community included increasing rates of community-acquired N. gonorrhoeae - 52.4% of isolates were resistant to at least one antibiotic. Trends for carbapenem-resistant Enterobacteriaceae (CRE) were stable at 0.22 cases per 10,000 patient days. Also, between 2004 and 2014, nine percent of tuberculosis (TB) culture positive cases were resistant to at least one first line anti-tuberculosis drug and this has remained relatively stable over that time. Trends in 2014 for AMR in food production animals showed decreasing resistance of Escherichia coli and Salmonella species to third-generation cephalosporins (ceftriaxone) in poultry associated with a decrease in cephalosporin use on chicken farms, but resistance to ciprofloxacin in Campylobacter species in chicken and cattle has been increasing. Overall, antibiotic use in humans has not declined despite concerns about overuse. Although resistance rates of C. difficile, VRE, MRSA and AMR S. pneumoniae have been gradually decreasing and drug-resistant tuberculosis and CRE have remained stable, community-associated drug-resistant N. gonorrhoeae has been increasing. Although efforts to decrease antibiotic use in animals have been met with some success, AMR continues to occur in fairly high levels in food production animals.