Within-farm dynamics of ESBL/AmpC-producing Escherichia coli in veal calves: a longitudinal approach.

OBJECTIVES: To assess the within-farm dynamics of extended-spectrum ß-lactamase (ESBL)/AmpC-producing Escherichia coli in veal calves. METHODS: Three veal-calf fattening farms were screened. Faecal samples from all calves within a compartment (109-150 per farm) were taken upon arrival on the farm (T0) and after 3, 6, 8 and 10 weeks (T3-T10). ESBL/AmpC genes were characterized by PCR and sequencing. Plasmids were characterized by transformation, PCR-based replicon typing and plasmid multilocus sequence typing (MLST). E. coli genotypes were analysed by MLST. RESULTS: At T0 the prevalence of ESBL/AmpC-producing E. coli ranged from 18% to 26%. These were predominantly isolates carrying blaCTX-M-1 and blaCTX-M-15 genes, located on various plasmids and E. coli sequence types (STs). Farm 1 was negative for ESBL/AmpC-producing E. coli after T0. Farm 2 showed an increase up to 37% at T3, which subsequently decreased gradually to 0% at T10. The presence from T3 to T10 on farm 2 was mainly caused by the clonal spread of a multiresistant E. coli ST57 harbouring blaCTX-M-14 on an IncF F2:A-:B- plasmid. Farm 3 showed a gradual decrease in prevalence to 1.4% at T10, with a relative increase of the identical clonal variant as shown for farm 2. A second clonal variant found in farm 3 was a multiresistant E. coli ST10 harbouring blaCTX-M-14 on an IncK plasmid. CONCLUSIONS: The prevalence of ESBL/AmpC-producing E. coli decreased over time. A clonal spread was observed on farm 2 and farm 3, illustrative of the complex dynamics probably associated with the use of antimicrobials.

Quantifying antimicrobial resistance at veal calf farms.

This study was performed to determine a sampling strategy to quantify the prevalence of antimicrobial resistance on veal calf farms, based on the variation in antimicrobial resistance within and between calves on five farms. Faecal samples from 50 healthy calves (10 calves/farm) were collected. From each individual sample and one pooled faecal sample per farm, 90 selected Escherichia coli isolates were tested for their resistance against 25 mg/L amoxicillin, 25 mg/L tetracycline, 0.5 mg/L cefotaxime, 0.125 mg/L ciprofloxacin and 8/152 mg/L trimethoprim/sulfamethoxazole (tmp/s) by replica plating. From each faecal sample another 10 selected E. coli isolates were tested for their resistance by broth microdilution as a reference. Logistic regression analysis was performed to compare the odds of testing an isolate resistant between both test methods (replica plating vs. broth microdilution) and to evaluate the effect of pooling faecal samples. Bootstrap analysis was used to investigate the precision of the estimated prevalence of resistance to each antimicrobial obtained by several simulated sampling strategies. Replica plating showed similar odds of E. coli isolates tested resistant compared to broth microdilution, except for ciprofloxacin (OR 0.29, p ≤ 0.05). Pooled samples showed in general lower odds of an isolate being resistant compared to individual samples, although these differences were not significant. Bootstrap analysis showed that within each antimicrobial the various compositions of a pooled sample provided consistent estimates for the mean proportion of resistant isolates. Sampling strategies should be based on the variation in resistance among isolates within faecal samples and between faecal samples, which may vary by antimicrobial. In our study, the optimal sampling strategy from the perspective of precision of the estimated levels of resistance and practicality consists of a pooled faecal sample from 20 individual animals, of which 90 isolates are tested for their susceptibility by replica plating.