【摘要】 为探究网络交易的粪肥是否成为抗性基因的传播载体,采用高通量荧光定量PCR技术对网上购买的4种粪肥（牛粪肥、鸡粪肥、羊粪肥和鸡羊混合肥）中抗生素抗性基因的多样性和丰度进行了比较分析,并对其潜在的传播风险进行了评估。结果表明:4种网络交易的粪肥中共检测出165种抗生素抗性基因和10种可移动遗传元件,检测到抗性基因的个数表现为:羊粪肥（130种）>鸡粪肥（98种）>鸡羊混合肥=牛粪肥（89种）;4种网络交易的粪肥中抗生素抗性基因的绝对丰度为7.18×10⁷～3.99×10¹¹copies/g（干重）,相对拷贝数为0.013～0.727copies/bacterialcell,抗生素抗性基因的丰度均表现为羊粪肥>鸡羊混合肥>鸡粪肥>牛粪肥;4种网络交易的粪肥主导的抗生素抗性基因主要是氨基糖苷类、多重耐药类和磺胺类抗性基因,而不同粪肥中抗生素抗性基因多样性的差异显著,且都具有其独有的抗性基因;4种网络交易的粪肥中都检测到intI-1LC和tnpA-04等4种可移动遗传元件,且总抗性基因的相对拷贝数与总可移动遗传元件的相对拷贝数呈极显著相关（p<0.01）,表明抗生素抗性基因的水平转移可能加剧其迁移和传播的风险。综上,网络交易的粪肥可能是抗生素抗性基因迁移和传播的潜在载体,该研究对健全我国网络交易中粪肥的微生物安全标准具有重要的参考价值。更多还原

【Abstract】 In order to explore whether the online-trading manure is the potential carrier of Antibiotic Resistance Genes（ARGs）,this paper applies high-throughput real-time fluorescence quantitative PCR（qPCR） to analyzing the diversity and abundance of ARGs in the four manure, including cow manure, chicken manure, sheep manure and chicken and sheep mixed manure, which were purchased in the online shops.The paper also evaluates the transmission risk of ARGs.The results show that a total of 165 ARGs and 10 Mobile Genetic Elements（MGEs） are detected in the four types of manure.The number of detected ARGs types are as follows: sheep manure（130 types）,chicken manure（98 types）,and chicken and sheep mixed manure=earthworm manure（89 types）.The absolute abundance of ARGs in the four manure ranges from 7.18×10⁷ to 3.99×10¹¹ copies/g（dry weight）,and the relative gene copy number ranges from 0.013 to 0.727 copies/bacterialcell.The order of both the absolute and relative abundance of ARGs is as follows: sheep manure> chicken and sheep mixed manure> chicken manure> earthworm manure.The dominant ARGs types in the four manure are aminoglycoside resistance gene, multi-drug resistance genes and sulfonamide resistance genes.The composition of ARGs subtypes significantly differs among the four manure, and each manure has its own unique ARGs subtypes.Four MGEs, such as intI-1LC（integrant） and tnpA-04（transposase）,are detected in all four types of manure.The relative copy number of total ARGs significantly and positively correlates with the relative copy number of total MGEs（p<0.01）,indicating that the horizontal transfer of resistance genes may exacerbate the migration and spread of ARGs.To sum up, online trading manure could be a potential carrier for ARGs.The study provides valuable data for the establishment of microbial safety standards for online trading manure.更多还原