Antibiotic resistance gene abundance in feces of calves fed pirlimycin-dosed whole milk

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Virginia Tech


Exposure to antibiotics has the potential to increase the incidence and proliferation of antibiotic resistance genes (ARG) in the gut and fecal microbiome. Non-saleable, antibiotic-containing milk from cows treated with antibiotics (waste milk) is commonly fed to dairy calves but the effects of ingestion of antibiotics at an early age on the gut microbiome and the development of ARG in the naive gut are not well understood. Pirlimycin, a lincosamide antibiotic acting against Gram positive bacteria through inhibiting protein synthesis by binding to the 50S ribosome, is commonly used as mastitis therapy. Lincosamides are also considered highly important in human medicine, often used against Staphylococcus aureus and Clostridium difficile infections. Emerging microbial resistance to pirlimycin is of concern for both animal and human health. The objective of this study was to determine the effect of early lincosamide antibiotic exposure on the abundance of ARG in feces of milk-fed calves. Eight female Holstein calves were blocked by age, paired by block, and randomly assigned to pasteurized whole milk (control; n = 4) or milk containing 0.2 mg/L of pirlimycin (treatment; n = 4). Calves were enrolled after receiving two colostrum feedings and were fed 5.68 L of pasteurized whole milk, treatment, or control, divided into two daily feedings, from d 1 to d 50 of age. After weaning calves were fed non-medicated starter grain ad libitum. Fecal samples were collected weekly until 85 d of age and freeze-dried. DNA was extracted using QiaAmp® Fast DNA Stool Mini Kit and qPCR was used to quantify the absolute abundance (gene copies/g of wet feces) and relative abundance (gene copies/copies of 16S rRNA genes) of erm(B), tet(O), tet(W) and 16S rRNA genes. Data was analyzed using PROC GLIMMIX in SAS. Abundance of 16S rRNA genes, tet(O) and tet(W) were not different between control and pirlimycin-fed calves nor were the relative abundance of tet(O) (mean = 0.050 tet(O) copies/16S rRNA genes) or tet(W) (0.561 tet(W) copies/16S rRNA genes). While abundance of erm(B) was higher in pirlimycin-fed calves compared to control calves (6.46 and 6.04 log gene copies/g wet feces; P = 0.04) the relative abundance of erm(B) (0.273 gene copies/16S rRNA genes) in feces of calves was not influenced by treatment. There was an effect of day (P < 0.10) for absolute abundance of tet(O), tet(W), and erm(B) indicating that the levels change with time as the fecal microbiome develops. This study suggests that feeding pirlimycin-containing non-saleable milk to growing calves may increase environmental loading of erm(B), which codes for resistance to highly important macrolide and lincosamide antibiotics. Additional research is needed on effects of feeding waste milk to calves on other fecal ARG and on the post-excretion and post-application fate of these genes.



antibiotic resistance, waste milk, dairy calf