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福建白羽肉鸡鸡毒支原体的分离鉴定及最小抑菌浓度测定
引用本文:吴春琳,黄宝钦,蓝天韵,林琳,张立根,李健,罗忠宝,吴异健.福建白羽肉鸡鸡毒支原体的分离鉴定及最小抑菌浓度测定[J].中国畜牧兽医,2021,48(4):1489-1497.
作者姓名:吴春琳  黄宝钦  蓝天韵  林琳  张立根  李健  罗忠宝  吴异健
作者单位:1. 福建农林大学动物科学学院(蜂学学院), 福州 350002;2. 福建省兽医中药与动物保健重点实验室, 福州 350002;3. 福建圣农发展股份有限公司, 南平 354100
基金项目:福建省自然科学基金项目(2017J01597);校科技创新专项基金项目(CXZX2017054)。
摘    要:为了解福建白羽肉鸡中鸡毒支原体(MG)对临床常用药物的敏感性,本研究从2个商品代白羽肉鸡养殖场采集了疑似患慢性呼吸道病(CRD)的鸡喉拭子40份,接种于适宜的支原体培养基上进行纯化。通过对菌株生物学特性分析、细菌形态学观察、PCR测序及动物回归试验进行鉴定,并利用最低抑菌浓度(MIC)测定了分离株对临床常用抗菌药的敏感性。结果显示,本试验成功获得2株MG,命名为MX和HY。将分离株接种于含有酚红的支原体液体培养基中能分解葡萄糖产酸使培养基颜色由红变黄;固体培养基于低倍镜下观察到典型的"荷包蛋"状菌落;分离株经过Dienes染色后的菌落中心呈致密深蓝色,边缘淡蓝,且30 min后不褪色,并具有较强吸附红细胞的能力,呈现典型MG培养特征;动物回归试验和分离株pvpA基因测序结果显示,2株MG分离株均具有较强致病性,证实其与R株的亲缘关系最近;药敏试验结果显示,MX和HY对泰妙菌素、氟苯尼考和泰乐霉素表现出了高度敏感性,其次是盐酸恩诺沙星、盐酸大观霉素,而对具有与泰乐霉素类似分子结构和抗菌谱的畜禽专用抗生素替米考星敏感性较差,此差异可能与临床上长期应用该药有关。研究结果表明,福建地区商品代白羽肉鸡中存在致病性MG流行,泰妙菌素可以作为临床防治MG的首选药物。

关 键 词:鸡毒支原体(MG)  分离鉴定  生物学特性  最低抑菌浓度(MIC)  
收稿时间:2020-09-24

Isolation,Identification and Minimum Inhibitory Concentration Determination of Mycoplasma gallisepticum from White-feathered Broilers in Fujian
WU Chunlin,HUANG Baoqin,LAN Tianyun,LIN Lin,ZHANG Ligen,LI Jian,LUO Zhongbao,WU Yijian.Isolation,Identification and Minimum Inhibitory Concentration Determination of Mycoplasma gallisepticum from White-feathered Broilers in Fujian[J].China Animal Husbandry & Veterinary Medicine,2021,48(4):1489-1497.
Authors:WU Chunlin  HUANG Baoqin  LAN Tianyun  LIN Lin  ZHANG Ligen  LI Jian  LUO Zhongbao  WU Yijian
Institution:1. College of Animal Science(Bee Academy), Fujian Agriculture and Forestry University, Fuzhou 350002, China;2. Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fuzhou 350002, China;3. Fujian Sunner Development Co., Ltd., Nanping 354100, China
Abstract:To understand the sensitivity of Mycoplasma gallisepticum(MG)to commonly used clinical drugs in Fujian White-feathered broilers,in this study,40 chicken throat swabs suspected of suffering from chronic respiratory disease(CRD)were collected from two commercial White-feathered broiler farms and inoculated on appropriate mycoplasma medium for purification.Through the analysis of the biological characteristics of the strains,bacterial morphology observation,PCR sequencing and animal regression test for identification,and the use of minimum inhibitory concentration(MIC)to determine the sensitivity of isolates to commonly used clinical antibacterial drugs.The results showed that two MG strains,named MX and HY,were successfully isolated and identified.Inoculating the isolated strain in the liquid medium of mycoplasma containing phenol red could decompose glucose to produce acid and change the color of the medium from red to yellow.A typical“poached egg”colony was observed in solid culture based on the observation of low power microscope.The colony center of the isolate after Dienes staining was dense dark blue,the edge was light blue,and did not fade after 30 minutes,and had the ability to adsorb red blood cells,with typical MG culture characteristics.The results of animal regression test and pvpA gene sequencing analysis showed that the two MG isolates had strong pathogenicity,which confirmed that they had the closest genetic relationship with R strain.The results of drug sensitivity test showed that MX and HY were highly sensitive to tiamulin,florfenicol and telithromycin,followed by enofloxacin hydrochloride and spectinomycin hydrochloride,but poor to tilmicosin,a special antibiotic for livestock and poultry with similar molecular structure and antibacterial spectrum.This difference might be related to the long-term clinical use of the drugs.The results showed that there was an epidemic of pathogenic MG in commercial White-feathered broilers in Fujian,and tiamulin could be used as the first choice for clinical prevention and treatment of MG.
Keywords:Mycoplasma gallisepticum(MG)  isolation and identification  biological properties  minimal inhibitory concentration(MIC)
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