Genomic Profiling of Multidrug Efflux Pumps and Heavy Metal Proteins in Multidrug-resistant Campylobacter fetus Isolated from Sheath Wash Samples of Bulls in South Africa

Abstract

A substantial evolution of resistance mechanisms among zoonotic bacteria has resulted from anthropogenic factors related to the application of antibiotics in human and veterinary medicine, particularly in contemporary agriculture. This issue associated with the presence of heavy metal-laced protein in zoonotic bacteria should be taken seriously with regard to the health of animals and the general people. To address this issue, the present study employed whole genome sequencing to identify the antimicrobial resistance patterns of Campylobacter fetus subsp. fetus (Cff) and Campylobacter fetus subsp. venerealis (Cfv), resistance and virulence genes, as well as heavy metal protein. Based on culture method biochemical testing and PCR amplification using particular primer pairs (MG3F-MG4R and VenSF-VenSR), bacteria were isolated and identified as C. fetus subsp. fetus and C.fetus subsp. venerealis. Subsequently, antimicrobial disc diffusion tests and whole genome sequencing were performed. Isolated bacteria were resistant to tetracycline at 65%, amoxicillin, and doxycycline at 60%. The resistance was also observed against neomycin at (55%), streptomycin (60%), and gentamycin (55%). Through comprehensive genome sequencing analysis and PCR, multiple efflux pumps linked to multidrug resistance were identified, including the broad-specificity multidrug efflux pump (YkkD), along with CmeA, CmeB, CmeC, and gryA.The genome sequence also revealed genes associated with the production of Cytotoxin (Cdt A, B, and C), adhesion and colonization (VirB10 and VirB9), and invasion (CiaB). In addition, different genomic features in heavy metal resistance included Cobalt-zinc-cadmium resistance protein (CzcD), Tellurite resistance protein (TehA), and arsenic efflux pump protein. The findings of the current study revealed that the emergence of bacterial multidrug resistance is increasingly associated with the substantial and growing contribution of Multidrug resistance efflux pumps, as evident in Cff and Cfv. Therefore, it is crucial to tighten the control of Cff and Cfv in livestock production to prevent the transfer of genes resistant to humans through the food chain.