Emergence of Cfr-Mediated Linezolid Resistance among Livestock-Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) from Healthy Pigs in Portugal.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST398 is mainly found in Europe and North America, colonizing the nasal cavity of pigs. This study characterized the MRSA isolates recovered from pig nasal swabs (n = 171) by evaluating the antimicrobial susceptibility profile by broth microdilution and characterizing the genetic lineages by spa-typing. Three linezolid-resistant isolates were subjected to Whole-Genome Sequencing (WGS). All strains harbored the mecA gene and were resistant to tetracycline and susceptible to vancomycin. A high frequency of multidrug resistance (97.6%) was evidenced, with 55 different multidrug resistance profiles identified. The MRSA strains were found to belong to 17 spa-types, three being novel. The linezolid-resistant strains appeared to belong to the ST398 type, spa-type t011, and SCCmec_type_Vc and to harbor the cfr, fexA, blaZ, mecA, tetM, and tetK genes. The cfr gene was predicted to be carried in the plasmid, flanked by ISSau9 and the transposon TnpR. MRSA from Portuguese fattening pigs present a high diversity of genetic lineages. The presence of cfr-positive LA-MRSA may represent a risk of transmission to humans, mainly to those in contact with livestock.

Assessing antimicrobial resistance occurrence in the Portuguese food system: Poultry, pigs and derived food, 2014-2018.

Antimicrobial resistance (AMR) spread in the food system is a major threat to public and animal health. We describe AMR trends in zoonotic bacteria and identify risk factors associated with AMR occurrence in animals and derived food in Portugal. Data from the Portuguese AMR surveillance programme on food-producing animals, 2014-2018, were used. AMR frequencies were calculated for Escherichia coli, Campylobacter and Salmonella in broilers, turkeys, pig populations and their derived food products. AMR-associated factors were studied for Salmonella isolates: population, sampling stage (farm, slaughterhouse and processing plant), sample type (environmental, carcase and food), sampler (HACCP, industry, official and official and industry), sample context (control and eradication programmes and monitoring), year (2014-2018) and season. Logistic regression was applied to estimate crude odds ratio and adjusted odds ratio (aOR) with 95% confidence intervals (95% CI). The final models were obtained using a backward stepwise method. This study included 2,157 Escherichia coli, 561 Campylobacter and 1,071 Salmonella isolates. The highest prevalence estimates amongst tested antimicrobials for each bacterial species in 2014-2018 had the following ranges: (i) (fluoro)quinolones: E. coli: 84%-93%, Campylobacter: 94%-98%; (ii) tetracyclines: E. coli: 68%-91%, Campylobacter: 87%-91%, Salmonella: 72%; (iii) penicillins: E. coli: 82%-100%; (iv) sulphonamides: E. coli: 68%-82%. Compared with the reference categories for host (broiler), year (2014), season (winter) and sampler (HACCP own checks), resistance to at least one antimicrobial in Salmonella was significantly less likely in laying hens (aOR 0.28; 95% CI: 0.18-0.42), 2016 (aOR 0.56; 95% CI: 0.33-0.93), 2017 (aOR 0.29; 95% CI: 0.17-0.51) and 2018 (aOR 0.35; 95% CI: 0.20-0.61), autumn (aOR 0.63; 95% CI: 0.40-0.97) and more likely to occur in broiler products (aOR 5.14; 95% CI: 2.61-10.54), pork products (aOR 6.84; 95% CI: 3.74-12.98) and official and industry combined sampling (aOR 2.16; 95% CI: 1.06-4.47). This study reveals a high prevalence of Salmonella resistance, especially during the summer and in post-farm stages of the Portuguese food system to nearly all antimicrobials and in the summer in farms to (fluoro)quinolones. Measures to tackle resistance are required.

Multi-Drug and ß-Lactam Resistance in Escherichia coli and Food-Borne Pathogens from Animals and Food in Portugal, 2014-2019.

Animal and food sources are seen as a potential transmission pathway of antimicrobial resistance (AMR) to humans. The aim of this study is to describe Campylobacter, Salmonella, and commensal Escherichia coli multi-drug resistance (MDR) in the food chain between 2014 and 2019 in Portugal. AMR surveillance data from food-producing animals and food were assessed. MDR relative frequencies were estimated by bacterial genus and year. AMR profiles were created using observations of resistance to antimicrobial classes from each isolate. Antimicrobial susceptibility testing results were clustered using k-modes. Clusters were described by population, AMR classification, ß-lactamases, sample stage, sample type, season, and year. Overall, MDR was more prevalent for E. coli, ranging from 74-90% in animal and 94-100% in food samples. MDR was found to be more widespread in resistance profiles that were common among E. coli and Salmonella isolates and in those exclusively observed for E. coli, frequently including (fluoro)quinolones and cephalosporins resistance. ß-lactam resistance was observed around 75% to 3rd/4th-generation cephalosporins in E. coli. Clusters suggest an escalating MDR behaviour from farm to post-farm stages in all bacteria and that Salmonella (fluoro)quinolones resistance may be associated with broilers. These findings support policy and decision making to tackle MDR in farm and post-farm stages.