| 符德佳 1,2，康桦华 2，曾宪军 3,4，樊志红 3,4，陈 杰 3,4，黄炜乾 5，蒋顺进 5，彭新宇 2.鸡场饮水中肺炎克雷伯氏菌的分离鉴定及耐药性研究[J].广东农业科学,2024,51(2):124-138 |
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鸡场饮水中肺炎克雷伯氏菌的分离鉴定及耐药性研究 |
| Isolation, Identification and Drug Resistance Research of Klebsiella pneumoniae in Drinking Water from a Chicken Farm |
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| DOI：10.16768/j.issn.1004-874X.2024.02.012 |
| 中文关键词: 鸡 饮水 肺炎克雷伯氏菌 多重耐药性 耐药基因 |
| 英文关键词: chicken drinking water Klebsiella pneumoniae multiple drug resistance resistance gene |
| 基金项目:清远市 2023 年省科技创新战略专项（大专项 + 任务清单）项目（2023DZX012）；科技创新战略专项资金（高水平农科院建设）（R2020PY-JC001）；广东省农业科学院驻镇帮扶村农村科技特派员项目（KTP20210010）；广东省畜禽疫病防治研究重点实验室项目（2023B1212060040） |
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| 中文摘要: |
| 【目的】研究养殖场鸡饮水中肺炎克雷伯氏菌（Klebsiella pneumoniae）的耐药表型和耐药基因的携带情况，并分析其相关性，为有效预防该病提供参考。【方法】从养殖场鸡饮水系统中分离、鉴定肺炎克雷伯氏菌，采用药敏纸片扩散法检测分离菌株对 25 种常用抗菌药的耐药性，通过 PCR 检测菌株携带的常见耐药基因。对其中 4 株肺炎克雷伯氏菌进行全基因组测序，利用 CARD 抗性基因数据库预测耐药基因的携带情况。【结果】从养殖场鸡饮水系统中共分离到 9 株肺炎克雷伯氏菌，药敏试验表明这些菌株呈现出多重耐药，仅对多粘菌素敏感。PCR 检测发现，9 株肺炎克雷伯氏菌共携带 13 种常见的耐药基因，其中，β- 内酰胺酶类耐药基因 CMY-1、BlaSHV 和 BlaCTX-M，碳青霉烯类耐药基因 KPC，喹诺酮类耐药基因 QnrB、QnrS 和 QnrA，氨基糖苷类耐药基因Aph(3')-Ia，磺胺类耐药基因 Sul2 等 12 种耐药基因与其耐药表型表现一致，而 Mcr-1 基因与其表型相反。全基因组扫描分析结果显示，4 株肺炎克雷伯氏菌共携带了 59 种耐药基因，涉及 31 类抗生素药物，其中四环素类抗生素的耐药基因最多，其次是大环内酯类抗生素和氟喹诺酮类抗生素耐药基因；预测到覆盖度和同源性均在 98% 以上的耐药基因共有 23 种，分别为 FosA6、CRP、KpnF、KpnFG、KpnE、OqxA、OqsB、SHV-11、Tet(D)、CTX-M-27、QnrB65、DHA-1、Aph(4')-Ia、MsrE、QnrB2、Aph(3')-Ia、Sul1、CmlA6、FloR、AadA12、Ant(3')-IIa、ArmA、Sul2。【结论】在养殖场鸡饮水中分离到的肺炎克雷伯氏菌耐药性严重，携带多种耐药基因，其中在分离菌株上均检测到 Mcr-1 基因，但分离菌株均对多粘菌素敏感，且均未检测到 D 类碳青霉烯类酶 OXA-48 基因。 |
| 英文摘要: |
| 【Objective】It aims to investigate resistance phenotypes and resistance genes of Klebsiella pneumoniae in drinking water of chicken farm, and analyze the correlation between them, to provide references for effective prevention of the disease.【Method】Strains of K. pneumoniae were isolated and identified from the drinking water system of a chicken farm. The resistances of the isolated strains to 25 antibiotics were detected by the drug-sensitive disk diffusion method. The presence of common drug-resistance genes in all these strains was detected by PCR. The whole genomes of four strains of K. pneumoniae were sequenced to analyze and predict the carrying of drug resistance genes by the Comprehensive Antibiotic Research Database (CARD).【Result】Nine strains of K. pneumoniae were isolated from the drinking water system of the chicken farm. The results of drug sensitivity testing revealed that these strains were multiple drug resistant, and only susceptive to colistin sulfate. Thirteen common drug resistance genes carried by all nine strains were confirmed by PCR, and consistency between drug resistance phenotypes and genes was observed in 12 resistant genes including β-lactamase resistance genes CMY-1, BlaSHV and BlaCTX-M; carbapenem resistance gene KPC; quinolone resistance genes QnrB, QnrS and QnrA; aminoglycoside resistance gene Aph(3')-Ia; sulfa resistance gene Sul2 and etc; while inconsistency between the presence of Mcr-1 and its corresponding phenotype was observed. A total of 59 resistance genes, involving 31 classes of antibiotics, were carried by four strains of K. pneumoniae through whole genome scanning analysis. Among these, the number of genes resistant to tetracycline antibiotics was maximum, followed by genes resistant to macrolide and fluoroquinolone antibiotics. Additionally, a total of 23 drug-resistant genes were predicted when the coverage and homology were both set at a rate of higher than 98%, including FosA6, CRP, KpnF, KpnFG, KpnE, OqxA, OqsB, SHV-11, Tet(D), CTX-M-27, QnrB65, DHA-1, Aph(4')-Ia, MsrE, QnrB2, Aph(3')-Ia, Sul1, CmlA6, FloR, AadA12, Ant(3')-IIa, ArmA and Sul2.【Conclusion】Strains of K. pneumoniae isolated from the drinking water of the chicken farm show serious drug resistance and carried multiple drug resistance genes, among which Mcr-1 gene is detected on the isolated strains. However, the isolated strains are sensitive to polymyxin, and the D-type carbapenemase OXA-48 gene was not detected in the isolated strains. |
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