新疆驼乳源克雷伯菌分离鉴定及耐药性与耐药基因分析

doi:10.16431/j.cnki.1671-7236.2025.10.039

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (10): 4941-4951.doi: 10.16431/j.cnki.1671-7236.2025.10.039

• 基础兽医 • 上一篇

新疆驼乳源克雷伯菌分离鉴定及耐药性与耐药基因分析

徐晓炜^1,2,4, 高姣姣^1,2, 马宪兰^1,2, 阿丽亚·吐尔迪^1,2, 邵伟², 郑楠³, 赵艳坤^1,3

1. 新疆维吾尔自治区农业科学院农业质量标准与检测技术研究所, 农业农村部农产品质量安全风险评估实验室 (乌鲁木齐), 新疆农产品质量安全实验室, 乌鲁木齐 830091;
2. 新疆农业大学动物科学学院, 乌鲁木齐 830052;
3. 中国农业科学院北京畜牧兽医研究所, 农业农村部奶及奶制品质量安全控制重点实验室, 北京 100193;
4. 伊犁职业技术学院动物科学学院, 伊犁 835000

收稿日期:2024-12-31 发布日期:2025-09-30
通讯作者: 赵艳坤 E-mail:yankunzhao90@163.com
作者简介:徐晓炜，E-mail：1967649437@qq.com。
基金资助:
国家自然科学基金(32460881)；自治区杰出青年基金(2024D01E12)；自治区天山英才项目(2023TSYCCX0034)；新疆农业科学院农业科技创新稳定支撑项目(xjnkywdzc-2024003-51)

Isolation,Identification and Analysis of Drug Resistance and Drug Resistance Genes of Klebsiella from Camel Milk in Xinjiang

XU Xiaowei^1,2,4, GAO Jiaojiao^1,2, MA Xianlan^1,2, ALIA Tuerdi^1,2, SHAO Wei², ZHENG Nan³, ZHAO Yankun^1,3

1. Xinjiang Agricultural Product Quality and Safety Laboratory, Agricultural Product Quality and Safety Risk Assessment Laboratory of the Ministry of Agriculture and Rural Affairs (Urumqi), Institute of Agricultural Quality Standards and Testing Technology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China;
2. College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China;
3. Key Laboratory for Quality & Safety Control for Milk and Dairy Products of Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
4. College of Animal Science, Yili Vocational and Technical College, Yili 835000, China

Received:2024-12-31 Published:2025-09-30

摘要/Abstract

摘要： 【目的】探明新疆地区生鲜驼乳中克雷伯菌流行情况及耐药风险隐患，为该地区生鲜驼乳中克雷伯菌的监测提供理论依据。【方法】采用平板划线法对新疆地区生鲜驼乳进行细菌分离纯化，通过形态学观察、革兰染色镜检、生化鉴定及PCR技术对分离株进行鉴定，并利用微量肉汤稀释法对其耐药性进行分析，采用PCR方法对分离菌进行17种关键耐药基因检测。【结果】从新疆地区采集的95份生鲜驼乳样本中共分离出58株克雷伯菌，分离率为61.05%。所有分离株均呈隆起的粉红色圆形菌落，革兰染色镜检呈红色、短杆的革兰阴性菌。生化鉴定结果显示，分离菌尿素酶、枸橼酸盐、葡萄糖、乳糖、甘氨酸、鸟氨酸及赖氨酸检测均呈现阳性；硫化氢、甲基红试验及V-P试验结果均为阴性。PCR扩增结果显示，分离株khe特异性基因、16S rDNA PCR扩增分别获得大小为428和1 476 bp目的条带，测序结果显示分离菌16S rDNA序列与NCBI数据库中克雷伯菌的参考序列相似性均≥99.0%，综合判定分离菌均为克雷伯菌。药敏试验结果显示，分离菌对氨苄西林、磺胺异噁唑耐药率分别为27.59%和15.52%，对阿莫西林/克拉维酸、美罗培南、链霉素、庆大霉素、四环素、多四环素、氟苯尼考、环丙沙星、复方新诺明敏感率为100%，对卡那霉素敏感率为98.28%。其中有1株克雷伯菌为多重耐药菌。耐药基因检测结果显示，β-内酰胺类耐药基因中，bla_CTX-M-14、bla_SHV、bla_CTX-M-123、bla_CTX-M阳性率分别为28.0%、24.0%、24.0%和4.0%。未检出多肽类耐药基因mcr-1、mcr-2、mcr-3和磺胺类耐药基因sul1、sul2、sul3。【结论】新疆地区生鲜驼乳中克雷伯菌流行情况较严重，且存在部分耐药现象，因此，对生鲜驼乳中克雷伯菌耐药性的监测具有重要意义。

关键词: 驼乳; 克雷伯菌; 药敏试验; 耐药基因

Abstract: 【Objective】 This study was conducted to explore the prevalence and drug resistance risk of Klebsiella in fresh camel milk in Xinjiang,and provide a theoretical basis for monitoring Klebsiella in fresh camel milk in this area.【Method】 Bacteria were isolated and purified from fresh camel milk in Xinjiang by plate scribing method.The isolates were identified by morphological observation,Gram staining microscopy,biochemical identification and PCR technology.The drug resistance of the isolates was analyzed by broth microdilution method.The isolated bacteria were tested for 17 key resistance genes by PCR.【Result】 A total of 58 strains of Klebsiella were isolated from 95 fresh camel milk samples collected in Xinjiang,and the isolation rate was 61.05%.All isolates were raised pink round colonies,and under Gram-staining microscopy,they showed red,short-stemmed Gram-negative bacteria.The biochemical identification results showed that the isolated bacteria were positive for urease,citrate,glucose,lactose,glycine,ornithine and lysine.The results of hydrogen sulfide test,methyl red test and V-P test were all negative.PCR amplification results showed that 428 and 1 476 bp bands of khe specific gene and 16S rDNA were amplified respectively.The sequencing results showed that the 16S rDNA sequence of the isolated bacteria was ≥99.0% similar to the reference sequence of Klebsiella in the NCBI database,and it was comprehensively determined that all the isolated bacteria were Klebsiella.The results of drug sensitivity test showed that the drug resistance rates of the isolates to ampicillin and sulfamethoxazole were 27.59% and 15.52%,respectively.The sensitivity rates of the isolates to amoxicillin/clavulanic acid,meropenem,streptomycin,gentamicin,tetracycline,dotetracycline,florfenicol,ciprofloxacin,compound sulfamethoxazole were 100%,and the sensitivity rates of the isolates to kanamycin was 98.28%.One strain of Klebsiella was multidrug-resistant.The results of drug resistance gene detection showed that the positive rates of bla_CTX-M-14,bla_SHV,bla_CTX-M-123 and bla_CTX-M were 28.0%,24.0%,24.0% and 4.0%,respectively.No polypeptide resistance genes mcr-1, mcr-2 and mcr-3, and sulfonamide resistance genes sul1,sul2 and sul3 were detected.【Conclusion】 The prevalence of Klebsiella in fresh camel milk in Xinjiang was serious,and there were some drug resistance phenomena.Therefore,it was of great significance to monitor the drug resistance of Klebsiella in fresh camel milk.

Key words: camel milk; Klebsiella; drug susceptibility test; drug resistance genes

中图分类号:

S852.61

徐晓炜, 高姣姣, 马宪兰, 阿丽亚·吐尔迪, 邵伟, 郑楠, 赵艳坤. 新疆驼乳源克雷伯菌分离鉴定及耐药性与耐药基因分析[J]. 中国畜牧兽医, 2025, 52(10): 4941-4951.

XU Xiaowei, GAO Jiaojiao, MA Xianlan, ALIA Tuerdi, SHAO Wei, ZHENG Nan, ZHAO Yankun. Isolation,Identification and Analysis of Drug Resistance and Drug Resistance Genes of Klebsiella from Camel Milk in Xinjiang[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4941-4951.

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新疆驼乳源克雷伯菌分离鉴定及耐药性与耐药基因分析

Isolation,Identification and Analysis of Drug Resistance and Drug Resistance Genes of Klebsiella from Camel Milk in Xinjiang

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