Effects of orally administered tetracycline on the intestinal community structure of chickens and on tet determinant carriage by commensal bacteria and Campylobacter jejuni

Abstract

There is a growing concern that antibiotic usage in animal production has selected for resistant food-borne bacteria. Since tetracyclines are common therapeutic antibiotics used in poultry production, we sought to evaluate the effects of oral administration on the resistance of poultry commensal bacteria and the intestinal bacterial community structure. The diversity indices calculated from terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA amplicons did not indicate significant changes in the cecal bacterial community in response to oxytetracycline. To evaluate its effects on cultivable commensals, Enterococcus spp., Escherichia coli, and Campylobacter spp. were isolated from the cecal droppings of broiler chickens. Enterococcus spp. and E. coli expressed tetracycline MICs of >8 microg/ml and harbored a variety of tet resistance determinants regardless of the tetracycline exposure history of the birds. The enterococcal isolates possessed tetM (61%), tetL (25.4%), and tetK (1.3%), as well as tetO (52.5%), the determinant known to confer a tetracycline resistance phenotype in Campylobacter jejuni. E. coli isolates harbored tetA (32.2%) or tetB (30.5%). Tetracycline MICs remained at <2 microg/ml for Campylobacter isolates before and after tetracycline treatment of the chickens, even though isolates expressing MICs of >16 mug/ml were commonly cultured from flocks that did not receive oxytetracycline. The results imply that complex ecological and genetic factors contribute to the prevalence of antibiotic resistance arising from resistance gene transfer in the production environment.

FIG. 1.

PFGE strain typing of tetracycline-susceptible (MIC = 1 μg/ml) C. jejuni isolates cultured from oxytetracycline-treated commercial broiler chickens in flock 1 and their relationship to the tetracycline-resistant (MIC = 64 μg/ml) isolates from an untreated sequential flock raised in the same house on farm CF-2. Lane 1 contains the S. cerevisiae molecular weight marker; lane 2 shows the SmaI pattern common to isolates cultured from flock 1; lane 3 represents the SmaI pattern detected in isolates cultured from flock 2; lane 4 contains the λ molecular weight marker, lane 5 shows the KpnI pattern for flock 1; lane 6 shows the KpnI pattern for flock 2.