广东规模化鸡场死鸡胚中鸡毒支原体的分离鉴定、致病性及药物敏感性

doi:10.11843/j.issn.0366-6964.2024.01.027

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (1): 290-299.doi: 10.11843/j.issn.0366-6964.2024.01.027

广东规模化鸡场死鸡胚中鸡毒支原体的分离鉴定、致病性及药物敏感性

陈玥彤^1,2, 刘晓涵^1,3, 王芷洋¹, 赵宇馨¹, 周铁忠², 胡增金¹, 朱悦¹, 王少辉¹, 田明星¹, 丁思羽⁴, 祁晶晶^1*, 于圣青^1*

1. 中国农业科学院上海兽医研究所, 上海 200241;
2. 锦州医科大学畜牧兽医学院, 锦州 121000;
3. 山东畜牧兽医职业学院, 潍坊 261000;
4. 扬州优佳创生物科技有限公司, 扬州 225261

收稿日期:2023-04-19 出版日期:2024-01-23 发布日期:2024-01-24
通讯作者: 于圣青,主要从事水禽重要病原细菌的分子致病机制以及疫苗研究,E-mail:yus@shvri.ac.cn;祁晶晶,主要从事畜禽细菌性传染病的诊断、疫苗、致病机制研究,E-mail:qijingjing@shvri.ac.cn
作者简介:陈玥彤(1997-),女,辽宁营口人,硕士生,主要从事禽支原体疾病研究,E-mail:c18342632780@163.com;刘晓涵(1996-),男,山东烟台人,硕士生,主要从事禽支原体疾病研究,E-mail:2454605207@qq.com。陈玥彤和刘晓涵为同等贡献作者
基金资助:
国家自然科学基金（31902244）；中央级公益性科研院所基本科研业务费专项资金（2022JB05）；辽宁省教育厅基本科研项目（LJKMZ20221254）；扬州市市级计划-现代农业项目（YZ2022046）

Isolation, Identification, Pathogenicity and Drug Susceptibility of Mycoplasma gallisepticum from Dead Chicken Embryos in Large-scale Chicken Farms in Guangdong Province

CHEN Yuetong^1,2, LIU Xiaohan^1,3, WANG Zhiyang¹, ZHAO Yuxin¹, ZHOU Tiezhong², HU Zengjin¹, ZHU Yue¹, WANG Shaohui¹, TIAN Mingxing¹, DING Siyu⁴, QI Jingjing^1*, YU Shengqing^1*

1. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China;
2. College of Animal Husbandry and Veterinary Medicine, Jinzhou Medicine University, Jinzhou 121000, China;
3. Shandong Vocational Animal Science and Veterinary College, Weifang 261000, China;
4. Yangzhou Youjiachuang Biotechnology Co., Ltd, Yangzhou 225261, China

Received:2023-04-19 Online:2024-01-23 Published:2024-01-24

摘要/Abstract

摘要： 旨在从广东某规模化鸡场死鸡胚进行鸡毒支原体（Mycoplasma gallisepticum，MG）的分离，并进行遗传进化分析、致病性及药物敏感性研究。本研究从广东某规模化鸡场的带菌死胚卵黄组织中分离禽支原体，通过菌落观察、血清学试验、16S rRNA支原体通用引物序列鉴定等方法，对分离的菌株进行种属鉴定，同时将分离的毒株对鸡胚和SPF鸡进行攻毒试验及抗菌药物敏感性试验。结果显示：显微镜下菌落呈典型的"荷包蛋"状。平板凝集试验显示其与MG阳性血清有凝集反应，与MS阳性血清不反应；16S rRNA序列测序发现各分离株与MG相似性达99.9%，因此确定分离株为MG。将各分离株感染7日龄SPF鸡胚，结果显示感染鸡胚大多于临近出壳时死亡；感染3周龄的SPF鸡，3周后解剖发现鸡气囊发生显著病理变化，说明其具有较强致病力；药物敏感性试验结果显示，5株分离株对盐酸沃尼妙林、多西环素、泰万菌素及泰妙菌素有较高敏感性，对替米考星、泰乐菌素、恩诺沙星、红霉素、吉他霉素、林可霉素呈现出不同程度耐药性。综上，从广东某规模化鸡场死鸡胚中成功分离到5株MG分离株，各分离株均可引起红细胞凝集，导致鸡胚死亡，对SPF鸡可引起典型的气囊炎症状，且对多种药物存在不同程度耐药性。本研究为临床MG的分离鉴定及用药选择提供了可参考的技术方法及指导，为MG攻毒模型建立提供了研究基础。

关键词: 鸡毒支原体, 死鸡胚, 分离鉴定, 致病性, 药物敏感性

Abstract: The aim of this study was to isolate Mycoplasma gallisepticum (MG) from dead chicken embryos in a large-scale chicken farm in Guangdong, and genetic evolution analysis, pathogenicity and drug susceptibility tests were carried out. Isolation of MG were conducted, then the isolated strains were identified by colony observation, serological test and 16S rRNA sequencing. At the same time, the isolated strains were used to infect chicken embryos and SPF chickens, and were subjected to antimicrobial susceptibility tests. Under the microscope, the colonies showed typical ‘poached egg’. Plate agglutination test showed that it had agglutination reaction with MG positive serum and did not react with MS positive serum. The 16S rRNA sequencing showed that the homology of each isolate with MG was 99.9%, so the isolates were identified as MG strains. Each isolate was used to infect 7-day-old SPF chicken embryos, and the results showed that most of the infected chicken embryos died near hatching. After 3 weeks of infection in 3-week-old SPF chickens, significant pathological changes were found in the air sac of chickens, indicating that it had strong pathogenicity. The drug sensitivity test showed that the 5 isolates were highly sensitive to warnimulin hydrochloride, doxycycline, tywanmycin and tymectin, and were resistant to tilmicosin, tylosin, enrofloxacin, erythromycin, kitasamycin and lincomycin to varying degrees. Five MG isolates were successfully isolated from dead chicken embryos in a large-scale chicken farm in Guangdong province. All of the isolates could cause chicken embryo death and cause typical air sac inflammatory symptoms to SPF chickens, and they had varying degrees of resistance to multiple drugs. This study provides reference technical methods and guidance for the isolation, identification and drug selection for clinical MG, and also provides a research basis for the establishment of MG challenge model.

Key words: Mycoplasma galliscepticum, dead chicken embryos, isolation and identification, pathogenicity, drug susceptibility

中图分类号:

S852.62

陈玥彤, 刘晓涵, 王芷洋, 赵宇馨, 周铁忠, 胡增金, 朱悦, 王少辉, 田明星, 丁思羽, 祁晶晶, 于圣青. 广东规模化鸡场死鸡胚中鸡毒支原体的分离鉴定、致病性及药物敏感性[J]. 畜牧兽医学报, 2024, 55(1): 290-299.

CHEN Yuetong, LIU Xiaohan, WANG Zhiyang, ZHAO Yuxin, ZHOU Tiezhong, HU Zengjin, ZHU Yue, WANG Shaohui, TIAN Mingxing, DING Siyu, QI Jingjing, YU Shengqing. Isolation, Identification, Pathogenicity and Drug Susceptibility of Mycoplasma gallisepticum from Dead Chicken Embryos in Large-scale Chicken Farms in Guangdong Province[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 290-299.

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广东规模化鸡场死鸡胚中鸡毒支原体的分离鉴定、致病性及药物敏感性

Isolation, Identification, Pathogenicity and Drug Susceptibility of Mycoplasma gallisepticum from Dead Chicken Embryos in Large-scale Chicken Farms in Guangdong Province

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