Search

Colistin is extensively used in animal production in many low- and middle-income countries. There is a need to develop methodologies to benchmark and monitor changes in resistance in commensal bacterial populations in farms. We aimed to evaluate the performance of a broth microdilution method based on culturing a pooled Escherichia coli suspension (30-50 organisms) from each sample. In order to confirm the biological basis and sensitivity of the method, we prepared 16 standard suspensions containing variable ratios of colistin-susceptible and mcr-1 encoded colistin-resistant E. coli which were grown in 2mg/L colistin. The optical density (OD600nm) readings over time were used to generate a growth curve, and were adjusted to the values obtained in the absence of colistin. The median limit of detection of the method was 1 colistin-resistant in 104 susceptible colonies [1st - 3rd quartile, 1:102 – 1:105]. We applied this method to 108 pooled faecal samples from 36 chicken flocks in the Mekong Delta (Vietnam) over the production cycle. The correlation between this method and the prevalence of colistin resistance in individual colonies harvested from field samples, determined by the Minimum Inhibitory Concentration (MIC), was established. The overall prevalence of colistin resistance at sample and isolate level was 38.9% and 19.4%, respectively. Increased colistin resistance was associated with recent (2 weeks) use of colistin and other, non-colistin antimicrobials (OR=3.67 and OR=1.84, respectively). Our method is a sensitive and affordable approach to monitor changes in colistin resistance in pooled E. coli populations from faecal samples over time.IMPORTANCEColistin (polymyxin E) is an antimicrobial with poor solubility properties, and therefore broth microdilution is the only appropriate method for testing colistin resistance. However, estimating colistin resistance in commensal mixed Escherichia coli populations is laborious since it requires individual colony isolation, identification and susceptibility testing. We developed a growth-based microdilution method suitable for pooled faecal samples. We validated the method by comparing it with results from individual MIC testing of 909 E. coli isolates. We used the method to investigate phenotypic colistin resistance in 108 pooled faecal samples from 36 healthy chicken flocks, each sampled three times over the production cycle. A higher level of resistance was seen in flocks recently supplemented with colistin in drinking water, although the observed generated resistance was short-lived. Our method is affordable, and may potentially be integrated into surveillance systems aiming at estimating the prevalence of resistance at colony level in flocks/herds. Furthermore, it may also be adapted to other complex biological systems, such as farms and abattoirs.

Indiscriminate antimicrobial use (AMU) in animal production is a driver of antimicrobial resistance globally, with a need to define sustainable AMU-reducing interventions in small-scale farms typical of low- and middle-income countries. We conducted a before-and-after intervention study on a random sample of small-scale chicken farms in the Vietnamese Mekong Delta from 2016 to 2019. A baseline was established before providing farms (n=102) with veterinary advice on chicken health and husbandry, and antimicrobial replacement products. Thirty-five (34.2%) farms entered the intervention phase; the remainder no longer continued raising chickens. The intervention reduced AMU (−66%) (hazard ratio [HR]=0.34; p=0.002) (baseline 343.4 Animal Daily Doses per 1,000 chicken-days) and mortality (−40%) (HR=0.60; p=0.005) (weekly baseline 1.60 per 100). Chicken bodyweight increased by 100g (p=0.002) in intervention flocks. Our findings demonstrate that in the Vietnamese context, AMU can be substantially reduced in small-scale chicken farms without compromising flock health by providing veterinary advice.