Review of antimicrobial therapy of selected bacterial diseases in broiler chickens in Canada

This paper reviews common therapeutic applications of antimicrobials in broiler chicken production in relation to Canadian guidelines, surveillance data, and emerging public health concerns about antimicrobial use (AMU). Escherichia coli, Clostridium perfringens, and Staphylococcus spp., were reviewed because of their animal health and economic significance. Enterococcus cecorum and Salmonella were included because of their importance in antimicrobial resistance (AMR) surveillance. This review identified that i) antimicrobials are available in Canada to treat infections by these agents, but may be through over the counter or extra-label use, ii) prevalence rates for these diseases are unknown, iii) antimicrobial use estimates in broilers are lacking, and iv) AMR has emerged in clinical isolates, though data are very sparse. This review highlights the need for surveillance of AMU and AMR in broiler chickens in Canada.     

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.     

Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) Farm Program: results from finisher pig surveillance

In 2006, the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) Farm Program was implemented in sentinel grower-finisher swine herds in Québec, Ontario, Manitoba, Saskatchewan and Alberta. Herds were visited 1-3 times annually. Faecal samples were collected from pens of close-to-market (CTM) weight (>80 kg) pigs and antimicrobial use (AMU) data were collected via questionnaires. Samples were cultured for generic Escherichia coli and Salmonella and tested for antimicrobial susceptibility. This paper describes the findings of this program between 2006 and 2008. Eighty-nine, 115 and 96 herds participated in this program in 2006, 2007 and 2008 respectively. Over the 3 years, antimicrobial resistance (AMR) levels remained consistent. During this period, resistance to one or more antimicrobials was detected in 56-63% of the Salmonella spp. isolates and 84-86% of E. coli isolates. Resistance to five or more antimicrobials was detected in 13-23% of Salmonella and 12-13% of E. coli. Resistance to drugs classified as very important to human health (Category I) by the Veterinary Drug Directorate (VDD), Health Canada, was less than or equal to 1% in both organisms. AMU data were provided by 100 herds in 2007 and 95 herds in 2008. Nine herds in 2007 and five herds in 2008 reported no AMU. The most common route of antimicrobial administration (75-79% of herds) was via feed, predominantly macrolides/lincosamides (66-68% of herds). In both 2007 and 2008, the primary reasons given for macrolide/lincosamide use were disease prevention, growth promotion and treatment of enteric disease. The Category I antimicrobials, ceftiofur and virginiamycin were not used in feed or water in any herds in 2008, but virginiamycin was used in feed in two herds in 2007. Parenteral ceftiofur was used in 29 herds (29%) in 2007 and 20 herds (21%) in 2008. The reasons for ceftiofur use included treatment of lameness, respiratory disease and enteric disease.     

Antimicrobial therapy of selected diseases in turkeys,laying hens,and minor poultry species in Canada

This paper identifies common poultry diseases requiring antimicrobial therapy, antimicrobials deemed efficacious to treat these diseases, and antimicrobial resistance (AMR) in these commodity-pathogen combinations, and describes current residue issues and minor use minor species (MUMS) guidelines. Veterinarians with turkey/layer expertise and diagnosticians were surveyed to determine the bacterial and protozoal diseases diagnosed in the last 5 years. Avian pathogenic Escherichia coli, Staphylococcus aureus, and Ornithobacterium rhinotracheale were the 3 most frequently diagnosed pathogens of turkeys. In layers, E. coli-peritonitis, and Clostridium perfringens/Eimeria spp. infections were the most common diagnoses. A literature review identified 32 antimicrobials as efficacious and/or recommended for treating these diseases. Surveillance and monitoring indicate the presence of enteric resistant organisms from some of these avian species (including resistance to antimicrobials of very high importance to human medicine). This paper highlights the need for surveillance of pathogen frequency, antimicrobial use (AMU), and AMR particularly in turkeys.     

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 and Resistance in Aquaculture: Findings of a Globally Administered Survey of Aquaculture‐Allied Professionals

There is limited published information regarding antimicrobial use (AMU) and antimicrobial resistance (AMR) in aquaculture. Our objective was to determine the opinions of aquaculture‐allied professionals around the world on the frequency of AMU and AMR in common aquatic species. The study questionnaire included five sections: respondent demographics, extent of AMU in aquaculture, frequency of observations of AMR in aquaculture, AMR monitoring and surveillance and antimicrobial susceptibility testing in various jurisdictions. It was administered in English and Spanish to 604 professionals in 25 countries and with varying expertise in aquaculture. The response rate was 33% (199/604). Over half of the participants had >10 years of experience in aquaculture: 70% (140/199) were involved in fish health/clinical work and their primary experience was with salmon, tilapia, trout, shrimp (including prawn) and/or catfish. Tetracycline use was reported by 28%, 46%, 18%, 37% and 9% of respondents working with catfish, salmon, tilapia, trout and shrimp, respectively. Resistance to tetracycline in one or more species of bacteria was reported as ‘frequent‐to‐almost always’ for the same aquaculture species by 39%, 28%, 17%, 52% and 36% of respondents, respectively. ‘Frequent‐to‐almost always’ use of quinolone was reported by 70% (32/46) and 67% (8/12) of respondents from the United States and Canada, respectively, where quinolone products are not approved for aquaculture, and extra‐label fluoroquinolone use is either prohibited (United States) or discouraged (Canada). Similar frequencies of quinolone use were also reported by the majority of respondents from Europe [70% (7/10)] and Asia [90% (9/10)] where labelled indications exist. This baseline information can be used to prioritize research or surveillance for AMU and AMR in aquaculture.     

Antimicrobial resistance in Campylobacter spp. isolated from Ontario sheep flocks and associations between antimicrobial use and antimicrobial resistance

The objectives of this study were to determine the prevalence of antimicrobial resistance (AMR) in faecal Campylobacter spp. from lambs and adult sheep and associations between antimicrobial use (AMU) and AMR. A total of 275 faecal samples collected during initial and final visits from 51 sheep flocks, including one feedlot, across southern Ontario were tested for the presence of Campylobacter spp. Campylobacter jejuni was detected in 52% (143/275) of the faecal samples, Campylobacter coli in 7% (19/275), Campylobacter lari in 1% (2/275) and 2% (4/275) were non-speciated Campylobacter. Broth microdilution was used to test antimicrobial susceptibility of 162 isolates to nine antimicrobials. Campylobacter jejuni isolates (n = 142) were resistant to tetracycline (39%), ciprofloxacin (4%), nalidixic acid (4%) and telithromycin (1%). C. coli isolates (n = 19) were resistant to tetracycline (74%), and azithromycin, clindamycin, erythromycin, and telithromycin (5%). The C. lari isolate displayed resistance to nalidixic acid. No statistically significant associations were found between AMU and AMR during multivariate modelling in this study.     

Evaluation of selective media in antimicrobial surveillance programs capturing broad-spectrum β-lactamase producing Escherichia coli from chickens at slaughter

Antimicrobial resistance surveillance targeting agricultural animals is practiced in many countries but does not often include media selective for cephalosporin resistance. Here, we compared the frequency of recovery of resistant Escherichia coli using selective and non-selective media from the cecal contents of 116 chickens collected by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS). Third generation cephalosporin resistance was detected in 24 samples including 12, 10, and 2 on selective, non-selective, and both media, respectively. Isolates producing the CTX-M-1 ESBL were grown from 11 samples, 10 on selective medium only. Our results suggest that current surveillance approaches underestimate the true prevalence of resistance to critically important antimicrobials.     

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 through feed, water, and injection in 20 swine farms in Alberta and Saskatchewan

Antimicrobial resistance (AMR) is an emerging animal welfare and public health issue linked to antimicrobial use (AMU) in livestock. This study was conducted in 2004 on 20 swine farms in Alberta and Saskatchewan. On-farm records and questionnaires were used to retrospectively describe the antimicrobial exposures of pigs through feed, water, and injection. Antimicrobial use in all production categories was described over 12 months. On-farm records and questionnaires provided sufficient data to describe antimicrobial exposure rates through feed and water. In contrast, on-farm records did not supply sufficient data to describe parenteral antimicrobial exposure rates. Records lacked data on the number of exposures per treatment, therefore parenteral AMU was described as an exposure incidence. Parenteral exposure records were often unavailable for pigs less than 22 kg, in which case questionnaires were used. The incidence of parenteral AMU was significantly higher in herds reporting exposure by questionnaire compared with existing records, suggesting that on-farm records did not reliably describe parenteral AMU. However, because antimicrobial exposures in feed and water were markedly more common than through injection, it was concluded that existing on-farm data would be a valuable resource for investigating AMU and AMR in pigs.     

Antimicrobial resistance in fecal generic Escherichia coli and Salmonella spp. obtained from Ontario sheep flocks and associations between antimicrobial use and resistance

The purpose of this study was to examine the prevalence and patterns of antimicrobial resistance (AMR) in enteric bacteria obtained from Ontario sheep flocks, and associations between antimicrobial use (AMU) and AMR. Forty-nine sheep producers participated for a 1-year interval between 2006 and 2008. Two-hundred and eighty-three pooled fecal samples were collected from the flocks during initial and final visits. Up to 3 isolates of Salmonella spp. and generic E. coli per pooled fecal sample were tested for susceptibility to 15 antimicrobials. Resistance was infrequent among Salmonella (0%, n = 7 isolates) and low among E. coli (13.1%; n = 849) isolates. A small number of isolates were resistant to antimicrobials classified as being of very high importance to human health. Tetracycline resistance was most frequently observed (12.0%). Logistic regression was used to model potential AMU (qualitative and quantitative) risk factors for tetracycline resistance in generic E. coli from final visits. Qualitative analysis indicated that the use of injectable sulfonamides [including trimethoprim-sulfonamide combinations (TMS)] and tetracycline in the feed and water were significantly associated with tetracycline resistance (OR = 2.6, P = 0.01; and OR = 4.8, P ≤ 0.01, respectively). Quantitative analysis also indicated that TMS exposure rate was significantly associated with tetracycline resistance, which varied depending on the exposure rate. The exposure rate of tetracycline in the feed and water was only significant after the removal of one influential flock, warranting further research examining flocks with higher tetracycline exposure rates. Although the prevalence of AMR in participating flocks was relatively low, risk factors for resistance were identified.     

Surveillance of antimicrobial resistance in clinical isolates of Pasteurella multocida and Streptococcus suis from Ontario swine

Susceptibility results for Pasteurella multocida and Streptococcus suis isolated from swine clinical samples were obtained from January 1998 to October 2010 from the Animal Health Laboratory at the University of Guelph, Guelph, Ontario, and used to describe variation in antimicrobial resistance (AMR) to 4 drugs of importance in the Ontario swine industry: ampicillin, tetracycline, tiamulin, and trimethoprim–sulfamethoxazole. Four temporal data-analysis options were used: visualization of trends in 12-month rolling averages, logistic-regression modeling, temporal-scan statistics, and a scan with the “What’s strange about recent events?” (WSARE) algorithm. The AMR trends varied among the antimicrobial drugs for a single pathogen and between pathogens for a single antimicrobial, suggesting that pathogen-specific AMR surveillance may be preferable to indicator data. The 4 methods provided complementary and, at times, redundant results. The most appropriate combination of analysis methods for surveillance using these data included temporal-scan statistics with a visualization method (rolling-average or predicted-probability plots following logistic-regression models). The WSARE algorithm provided interesting results for quality control and has the potential to detect new resistance patterns; however, missing data created problems for displaying the results in a way that would be meaningful to all surveillance stakeholders.     

Antimicrobial resistance and recovery of Salmonella,Campylobacter, and Escherichia coli from chicken egg layer flocks in Canadian sentinel surveillance sites using 2 types of sample matrices

Eggs are important to the diet of Canadians. This product is one of the supply-managed commodities in Canada, but unlike other commodities, where food safety risks are extensively explored and reported, information on the prevalence of enteric organisms (e.g., Salmonella, Campylobacter) and antimicrobial resistance (AMR) in layers in Canada are limited. This study was conducted to determine the prevalence of select bacteria and the associated AMR patterns in layer flocks using 2 sample matrices. Farms were located within FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance sentinel sites (SS). Fecal samples (Ontario: ON_SS1a, ON_SS1b) and environmental sponge swabs (British Columbia: BC_SS2a) were collected. Salmonella prevalence was 29% and 8% in ON_SS1a and ON_SS1b, respectively, and 7% in BC_SS2a. S. Kentucky and S. Livingstone were the most frequently isolated serovars and no S. Enteritidis was detected. Campylobacter was not detected in the BC sponge swabs but was isolated from 89% and 53% of Ontario fecal samples (ON_SS1a and ON_SS1b, respectively). Seven C. jejuni from Ontario were ciprofloxacin-resistant. Escherichia coli prevalence was high in both sample types (98%). Overall, tetracycline resistance among E. coli ranged from 26% to 69%. Resistance to ceftiofur (n = 2 isolates) and gentamicin (n = 2) was relatively low. There were diverse resistance patterns (excludes susceptible isolates) observed among E. coli in Ontario (10 patterns) and British Columbia (14 patterns). This study revealed that fecal samples are more informative for farm-level monitoring of pathogen and AMR prevalence. Without further validation, sponge swabs are limited in their utility for Campylobacter detection and thus, for public health surveillance.     

Outpacing the resistance tsunami: Antimicrobial stewardship in equine medicine,an overview

Antimicrobial stewardship (AMS) is the term increasingly used to describe the multiple approaches needed to sustain the efficacy of antimicrobial drugs in the face of the increasing development and spread of antimicrobial resistance in bacterial pathogens, and the global crisis in medicine that it is engendering. The concept and the practices associated with AMS continue to evolve but the general approach is a dynamic and multifaceted one of continuous improvement based on reducing, improving, monitoring and evaluating the use of antimicrobials so as to preserve their future efficacy and to protect human and animal health. Using many equine examples, this basic overview discusses the multiple and interacting elements of AMS: Practice guidelines, infection control and prevention, clinical microbiology, resistance and use surveillance, dosage, pharmacokinetics and pharmacodynamics, regulation, education and owner compliance, leadership, coordination and measurement. There have been impressive advances in recent years in reporting and analysis of AMR in horses, in the scrutiny and assessment of how antimicrobial drugs are used in horses and in identification of areas for improvement including dosing, surgical prophylaxis, infection control, development of practice standards and the use of clinical microbiology. Antimicrobial stewardship is taking shape as we start to see the emergence of evidence-based recommendations but far more is required. Containing and even rolling back AMR will need the continued engagement of practitioners, equine national and international practitioner organisations, researchers and educators in the academic community, horse owners, regulators and others.     

Analysis of antimicrobial sales data of the main distributor in Quebec from 2016 to 2019: An estimate of usage in dairy cattle,horses, and small animals

Because antimicrobial usage (AMU) data are crucial in understanding and dealing with the threat that antimicrobial resistance (AMR) poses to global health, data of the sale of antimicrobials from 2016 to 2019 of CDMV Inc. — a major distributor of veterinary products in Canada — were collected and analyzed for the province of Quebec. The primary objective was to describe the evolution of AMU sales data for dairy cattle, small animals, and horses; a secondary objective was to determine effects of a new provincial regulation on antimicrobials of very high importance (in Quebec) on sales for dairy cattle. Results are described in milligrams of antimicrobials per kilogram of animal biomass (mg/PCU) for dairy cattle, small animals, and horses; intramammary products were analyzed for number of treatments per 100 cow-years; and results for dairy cattle were compared in Canadian-defined course doses for cattle (DCDbovCA) per 100 cow-years to a recent study for this species in Quebec. Between 2016 and 2019, there were decreased sales of Category 1 antimicrobials for all species included in the study (Category 1-VI for small animals). This reduction was even more apparent for dairy cattle, for which a 76% decrease occurred from 2018 to 2019 (1.7 to 0.4 mg/PCU). This marked reduction was attributed to the new regulation implemented in February 2019. Since a farm- and clinic-level AMU monitoring system has not yet been implemented in Quebec, analysis of CDMV Inc. sales enabled observations of temporal trends in AMU for dairy cattle, horses, and small animals. These temporal trends based on CDMV Inc. sales will be useful for making comparisons and validating trends derived from farm- and clinic-level data generated by a monitoring system.     

Antimicrobial stewardship in the dairy industry: responding to the threat of antimicrobial resistance

Dairy Australia is the national service body for the Australian dairy industry. Its role is to help farmers adapt to a changing operating environment and achieve a profitable, sustainable dairy industry. Although the use of antibiotics in Australian agriculture is relatively low in global terms, Dairy Australia recognises important drivers for continuous improvement in antimicrobial stewardship (AMS). Dairy Australia’s first strategic priority is to support profitable farms. This priority has driven the development of a range of on‐farm change management programs in the animal health and welfare fields to optimise the unit cost of production and dairy cattle welfare. Dairy Australia’s third strategic priority is to further develop a ‘trusted dairy industry’. Previous and current work under these two strategies position the dairy industry favourably with respect to confronting the challenge of antimicrobial resistance (AMR) and supporting sound AMS with the guiding principle of ‘as little as possible, as much as necessary’. However, given an incomplete but ongoing threat of AMR, more work is needed. Supported by Dairy Australia, the dairy industry has developed an antimicrobial use strategy aligning with the Australian Animal Sector National AMR Plan 2018.     

Rates and determinants of antimicrobial use, including extra-label, on Ontario sheep farms

Few antimicrobials are licensed for use in sheep in Canada, and the range of indications is narrow. Treatment in an "extra-label" manner may be ineffective. In addition, potentially harmful drug residues in food-animal products and antimicrobial resistance in bacteria may be associated with extra-label drug use (ELDU). No data had been documented on drug use, specifically antimicrobial use (AMU), in Ontario sheep, although it was thought that much use was extra-label. This study investigated AMU and ELDU on 49 lamb-producing Ontario sheep farms. Data were prospectively collected over 12 months from the participating farms, and farm-level practices were ascertained with a questionnaire. Treatment-level and farm-level variables were investigated for associations with rates of AMU by means of Poisson rate regression models fit with a generalized estimating equation to control for clustering at the farm level. Antimicrobials with high mean exposure rates included chlortetracycline (in feed), penicillins, and oxytetracycline. The exposure rate in lambs was significantly lower (P < 0.01) with antimicrobial treatment of systemic signs, respiratory disease, or wound or injury than with treatment of other reported diseases or conditions; it was also significantly lower with 3 or more lambing periods per year (α = 0.05). The exposure rate in adult sheep was significantly lower with treatment of 5 of the 6 most prevalent diseases or conditions (α = 0.05) and significantly higher with producer decision to treat and producer experience of 20 y or greater. Rates of using antimicrobials not licensed for use in sheep were high, as was extra-label use of licensed antimicrobials. Diseases reportedly treated most often with antimicrobials (e.g., systemic signs, mastitis) were significantly associated with lower rates of ELDU (α = 0.05). Compared with the rates in adult sheep, the mean rate of use of nonlicensed antimicrobials was similar in the lambs, whereas the mean rate of extra-label use of licensed antimicrobials was lower among the lambs. The results are useful in determining if public health concerns about antimicrobial use in Ontario sheep are warranted and in creating drug use and licensure strategies for the Canadian sheep industry.     

Use of a bacterial antimicrobial resistance gene microarray for the identification of resistant Staphylococcus aureus

As diagnostic and surveillance activities are vital to determine measures needed to control antimicrobial resistance (AMR), new and rapid laboratory methods are necessary to facilitate this important effort. DNA microarray technology allows the detection of a large number of genes in a single reaction. This technology is simple, specific and high-throughput. We have developed a bacterial antimicrobial resistance gene DNA microarray that will allow rapid antimicrobial resistance gene screening for all Gram-positive and Gram-negative bacteria. A prototype microarray was designed using a 70-mer based oligonucleotide set targeting AMR genes of Gram-negative and Gram-positive bacteria. In the present version, the microarray consists of 182 oligonucleotides corresponding to 166 different acquired AMR gene targets, covering most of the resistance genes found in both Gram-negative and -positive bacteria. A test study was performed on a collection of Staphylococcus aureus isolates from milk samples from dairy farms in Québec, Canada. The reproducibility of the hybridizations was determined, and the microarray results were compared with those obtained by phenotypic resistance tests (either MIC or Kirby-Bauer). The microarray genotyping demonstrated a correlation between penicillin, tetracycline and erythromycin resistance phenotypes with the corresponding acquired resistance genes. The hybridizations showed that the 38 antimicrobial resistant S. aureus isolates possessed at least one AMR gene.