The study aimed to investigate the prevalence of antibiotic resistance genes (ARGs) within the ecosystem of natural plague foci, assessing their potential impact on the efficacy of plague treatments. Employing 16S rRNA gene sequencing and high-throughput quantitative PCR, microbial communities and ARGs were detected, with subsequent analysis of interactions among ARGs, mobile genetic elements (MGEs), environmental factors, and microbial species. Tetracycline resistance genes were found to be dominant, with multidrug and tetracycline resistance ARGs primarily associated with marmots and ecological soil, while pikas predominantly harbored β-lactam resistance ARGs. High detection rates were observed for resistance genes rpsl and sul1, which are relevant to streptomycin and sulfonamides, antibiotics commonly used in plague treatment. The total dissolved solids (TDS) in soil significantly promoted the presence of tetR-02, and Ni was found to inhibit vanHB. The tnpA-03 MGE was identified as a significant contributor to the dissemination of the aadE gene. The high prevalence of ARGs, particularly rpsl and sul1, poses a potential risk to the efficacy of main antibiotic treatments for plague. The study suggests that environmental microbiomes may be the greatest risk factor for the emergence of drug-resistant Yersinia pestis, given the low misuse of antibiotics in animals within natural plague foci. Monitoring the risk of drug-resistant strain emergence and preparing alternative antibiotic or combination therapy strategies based on ARG pollution levels in plague-affected areas is deemed necessary.
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