人、畜粪便中抗生素耐药基因与微生物群落共现分析

Ji-sun Jeong, Aprajita Bhandari, T. Unno
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摘要

背景:畜牧业中用于疾病预防和治疗的抗生素导致抗生素耐药菌的快速发展,这些耐药菌可通过牲畜fe-ce /粪便进入环境,传播抗生素耐药基因(ARGs)。本研究收集济州岛家畜养殖场粪便样本,探讨微生物群落与ARGs之间的关系。方法与结果:应用Illumina MiSeq测序对每个粪便样品中的微生物群落进行表征。采用定量PCR (qPCR)技术,对10个编码四环素耐药(tetB、tetM)、磺胺耐药(sul1、sul2)、氟喹诺酮类耐药(qnrD、qnrS)、氟喹诺酮类和氨基糖苷类耐药(aac(6′)-Ib)、β -内酰胺耐药(bla TEM、bla CTX-M)、大环内酯类耐药(ermC)、1类整合子整合酶基因(intI1)和2类整合子整合酶基因(intI2)的ARGs进行了定量分析。结果表明,人、牛、马和猪组以厚壁菌门和拟杆菌门为主,鸡组以厚壁菌门和放线菌门为主。在ARGs中,检测到的拷贝数最多的是tetM,其次是sul1和sul2。厚壁菌门大多数属与ARGs和整合子基因呈正相关。鸡、牛、猪、人和马分别有97、34、31、25和22个属与ARGs和整合子基因呈正相关。在网络分析中,我们发现了与ARGs和整合子基因相关的微生物群落多样性。结论:本研究确定了人类和牲畜粪便样本中的抗生素耐药模式。
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Co-occurrence Analyses of Antibiotic Resistance Genes and Microbial Community in Human and Livestock Animal Feces
BACKGROUND: Antibiotics used in animal husbandry for disease prevention and treatment have resulted in the rapid progression of antibiotic resistant bacteria which can be introduced into the environment through livestock fe-ces/manure, disseminating antibiotic resistant genes (ARGs). In this study, fecal samples were collected from the livestock farms located in Jeju Island to investigate the relationship between microbial communities and ARGs. METHODS AND RESULTS: Illumina MiSeq sequencing was applied to characterize microbial communities within each fecal sample. Using quantitative PCR (qPCR), ten ARGs encoding tetracycline resistance ( tetB , tetM ), sulfonamide resistance ( sul1 , sul2 ), fluoroquinolone resistance ( qnrD , qnrS ), fluoroquinolone and aminoglyco-side resistance ( aac(6')-Ib ), beta-lactam resistance ( bla TEM , bla CTX-M ), macrolide resistance ( ermC ), a class 1 integrons-integrase gene ( intI1 ), and a class 2 integrons-integrase gene ( intI2 ) were quantified. The results showed that Firmicutes and Bacteroidetes were dominant in human, cow, horse, and pig groups, while Firmicutes and Actinobacteria were dominant in chicken group. Among ARGs, tetM was detected with the highest number of copies, fol-lowed by sul1 and sul2 . Most of the genera belonging to Firmicutes showed positive correlations with ARGs and integron genes. There were 97, 34, 31, 25, and 22 genera in chicken, cow, pig, human, and horse respectively which showed positive correlations with ARGs and integron genes. In network analysis, we identified diversity of microbial communities which correlated with ARGs and integron genes. CONCLUSION(S): In this study, antibiotic resistance pat-terns in human and livestock fecal samples were identified.
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