Azithromycin resistance in Escherichia coli and Salmonella from food-producing animals and meat in Europe.

OBJECTIVES: To characterize the genetic basis of azithromycin resistance in Escherichia coli and Salmonella collected within the EU harmonized antimicrobial resistance (AMR) surveillance programme in 2014-18 and the Danish AMR surveillance programme in 2016-19. METHODS: WGS data of 1007 E. coli [165 azithromycin resistant (MIC > 16 mg/L)] and 269 Salmonella [29 azithromycin resistant (MIC > 16 mg/L)] were screened for acquired macrolide resistance genes and mutations in rplDV, 23S rRNA and acrB genes using ResFinder v4.0, AMRFinder Plus and custom scripts. Genotype-phenotype concordance was determined for all isolates. Transferability of mef(C)-mph(G)-carrying plasmids was assessed by conjugation experiments. RESULTS: mph(A), mph(B), mef(B), erm(B) and mef(C)-mph(G) were detected in E. coli and Salmonella, whereas erm(C), erm(42), ere(A) and mph(E)-msr(E) were detected in E. coli only. The presence of macrolide resistance genes, alone or in combination, was concordant with the azithromycin-resistant phenotype in 69% of isolates. Distinct mph(A) operon structures were observed in azithromycin-susceptible (n = 50) and -resistant (n = 136) isolates. mef(C)-mph(G) were detected in porcine and bovine E. coli and in porcine Salmonella enterica serovar Derby and Salmonella enterica 1,4, [5],12:i:-, flanked downstream by ISCR2 or TnAs1 and associated with IncIγ and IncFII plasmids. CONCLUSIONS: Diverse azithromycin resistance genes were detected in E. coli and Salmonella from food-producing animals and meat in Europe. Azithromycin resistance genes mef(C)-mph(G) and erm(42) appear to be emerging primarily in porcine E. coli isolates. The identification of distinct mph(A) operon structures in susceptible and resistant isolates increases the predictive power of WGS-based methods for in silico detection of azithromycin resistance in Enterobacterales.

Analysis of Antimicrobial Use and the Presence of Antimicrobial-Resistant Bacteria on Austrian Dairy Farms-A Pilot Study.

The assumed link between high levels of antimicrobial use on farms and selection for antimicrobial-resistant (AMR) bacteria on that farm remains difficult to prove. In the pilot study presented here, we analysed total antimicrobial use on 50 dairy farms in Austria and also collected environmental samples to ascertain whether specific AMR bacteria were present. Antimicrobial use (AMU) analysis was based on electronic veterinary treatment records over a one-year period. Faecal samples for the assessment of extended-spectrum beta-lactamase (ESBL)-producing E. coli were collected from cowsheds, calf pens, and youngstock housing areas, as well as dust samples from barns, to isolate methicillin-resistant Staphylococcus aureus (MRSA). Bacteriological cultures were carried out on selective agar. Farms were split into groups of 25 of the highest antimicrobial users and 25 of the lowest users. Overall, samples from 13/50 (26.0%) farms were found to be positive for the presence of ESBL-producing E. coli. Of these, eight farms were in the low user group and five were in the high user group. Only one farm was confirmed to harbour MRSA. Statistical analyses demonstrated that there was no significant difference in this study population between high or low antimicrobial use with respect to the presence of ESBL-producing E. coli on farms (p = 0.33). In conclusion, the presence of specific AMR bacteria on farms in this study population was not found to have a statistically proven relationship with their level of antimicrobial use.

Differences in antimicrobial resistance of commensal Escherichia coli isolated from caecal contents of organically and conventionally raised broilers in Austria, 2010-2014 and 2016.

The prevalence of antimicrobial resistance in commensal Escherichia coli isolated from organically raised broiler flocks was compared to the prevalence in isolates from conventional flocks. From 2010 to 2014, and in 2016, resistance trends and multidrug resistance in isolates from the caecal contents of flocks from both broiler production forms were analyzed. Samples were taken in four abattoirs accounting for at least 90% of the national slaughtered broiler population. In total, 962 commensal E. coli were obtained from organically raised broiler flocks (n = 142) and from conventionally raised broiler flocks (n = 820). The mean prevalence of commensal E. coli isolates, which were fully susceptible to the antimicrobials tested, was 43.3% in organically raised broiler flocks and thus significantly higher (P < 0.001) compared to 16.7% in conventionally operated flocks. During the study period, the proportion of fully susceptible isolates increased significantly in both broiler populations. Antimicrobial resistance rates were significantly lower in commensal E. coli isolated from organic compared to conventional production regarding ciprofloxacin (33.3% versus 69.1%), nalidixic acid (33.7% versus 67.4%), sulfamethoxazole (26.7% versus 39.9%), ampicillin (19.0% versus 33.8%) and trimethoprim (12.8% versus 24.9%). Regarding tetracycline, tigecycline and ceftazidime resistance rates were slightly but not significantly higher in isolates from organic flocks (27.6% versus 25.9%; 4.0% versus 1.4%; 2.0% versus 1.9%). This fact is surprising for tetracycline, as none of the investigated organic flocks had been treated with this antimicrobial during their lifetime. No resistances were found in isolates from both production forms against colistin and meropenem, and from organic flocks against azithromycin. The annual prevalence of resistance against ciprofloxacin and nalidixic acid decreased significantly in isolates from both broiler production forms. In isolates from organic flocks, it also decreased regarding ampicillin and sulfamethoxazole. Significant increasing trends were observed in the resistance prevalence against trimethoprim and borderline significantly for ampicillin in commensal E. coli from conventional flocks. Multidrug resistance was detected at a significantly higher prevalence in isolates from conventionally raised flocks (35.1%) compared to organic flocks (22.7%). Findings from this study clearly indicate the influences of organic compared to conventional broiler production practices on the prevalence of antimicrobial resistance in commensal E. coli from broiler flocks.