| 唐伟欣,孙兴滨,高浩泽,程首涛,仇天雷,王旭明.规模化畜禽养殖场粪便中多重耐药菌分离鉴定及其耐药特征[J].农业环境科学学报,2020,39(1):207-216. |
| 规模化畜禽养殖场粪便中多重耐药菌分离鉴定及其耐药特征 |
| Isolation and identification of multidrug-resistant bacteria in feces from large-scale livestock and poultry farms and their antibiotic-resistance characteristics |
| 投稿时间：2019-07-05 |
| DOI：10.11654/jaes.2019-0749 |
| 中文关键词: 禽畜粪便 多重耐药菌 16S rDNA 药敏试验 整合子 |
| 英文关键词: livestock feces multidrug-resistant bacteria 16S rDNA antibiotic susceptibility testing integron |
| 基金项目:北京市自然科学基金面上项目（6182019）；国家重点研发计划重点专项（2017YFD0801402）；北京市农林科学院创新能力建设专项（KJCX20180407）；家禽产业技术体系北京市创新团队专项（BAIC04-2019）；宁夏自治区重点研发计划项目（2019BFG02015） |
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| 中文摘要: |
| 为了解禽畜粪便中多重耐药菌的污染特征，本研究对鸡粪、牛粪、猪粪和有机肥这4种不同样品中多重耐药菌进行了计数分析，并对不同来源的多重耐药菌株进行了分离和纯化，进一步开展了基于16S rDNA序列比对的分子生物学鉴定以确定其种属地位，以及基于药敏试验的耐药性特征分析。结果表明：不同养殖动物粪便中四环素、恩诺沙星、磺胺甲恶唑和泰乐菌素4种抗生素多重耐药菌的绝对数量和相对数量排序均为鸡粪 > 猪粪 > 牛粪，粪源有机肥中可培养的多重耐药菌的绝对数量和相对数量在堆肥后均有所下降。通过对耐药菌株鉴定和分类学分析，发现禽畜粪便中多耐药菌的菌门集中分布在Proteobacteria、Firmicutes和Actinobacteria，多重耐药菌的优势菌属为Escherichia、Corynebacterium、Kurthia；而有机肥样品中多重耐药菌的优势菌属为Staphylococcus、Glutamicibacter。菌株的药敏试验表明，3类动物粪污中的多重耐药菌都对红霉素、四环素有着较高的耐药率，均高于80%，而对阿米卡星这种抗生素表现出较好的敏感性，其耐药率低于30%。通过对Ⅰ、Ⅱ类整合子基因盒的扩增，发现PCR产物的大小从0.8 kb到1.8 kb不等，基因盒adA2、dfrA17、dfrA1及sat2在养殖场粪污中检出率均较高。 |
| 英文摘要: |
| To investigate the characteristics of multidrug-resistant bacteria in livestock manure, the bacterial counts were determined in poultry manure, cow manure, swine manure, and organic fertilizer. First, the multiantibiotic-resistant bacteria were isolated and purified. Then, 16S rDNA sequence-based identification was performed, and their antibiotic susceptibilities were tested by disk diffusion susceptibility method. The absolute and relative numbers of bacteria resistant to tetracycline, enrofloxacin, sulfamethoxazole, and tylosin in manure were ranked, from highest to lowest, according to their percentage:poultry manure, swine manure and cow manure. The relative number of multidrug-resistant bacteria was lower in organic fertilizer than in manure. By the identification and taxonomic analysis of drug-resistant strains, it was found that most multidrug-resistant bacteria in livestock manure belonged to Proteobacteria, Firmicutes, and Actinobacteria; the dominant genus of multidrug-resistant bacteria were Escherichia, Corynebacterium, and Kurthia. The dominant genus of multidrug-resistant bacteria in organic fertilizer were Staphylococcus and Glutamicibacter. Antibiotic susceptibility testing of the strains showed that the multidrug-resistant bacteria in all the types of animal manure had high resistance rates to erythromycin and tetracycline, with more than 80%. Most of strains showed sensitivity to amikacin; the amikacin-resistant rate in all strains was less than 30%. The results of amplification of class Ⅰ and class Ⅱ integron gene cassettes showed that the size of PCR products ranged from 0.8 kb to 1.8 kb, and adA2, dfrA17, dfrA1, and sat2 were the majority class I integron gene cassettes in farm manure. |
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