摘要
为了探究江苏省部分地区鸡滑液囊支原体(MS)的菌株基因分型及致病性和药物敏感性情况,从江苏省部分地区分离鸡滑液囊支原体,通过菌落观察、玻片凝集试验、16S rRNA测序、vlhA基因PRR区分型等方法,对分离的菌株进行种属及分型鉴定,同时进行SPF鸡攻毒试验及药敏试验.结果显示,所采集样品中分离出3株疑似MS菌株,均可在固体培养基上生成典型的"油煎蛋"样菌落;经16SrRNA序列分析发现各分离株与MS相似性达99.9％;玻片凝集试验显示其与MS标准阳性血清、MS感染血清有凝集反应,与SPF鸡阴性血清无反应,因此将分离株确定为MS.SPF鸡攻毒试验显示,3株分离株均可引起鸡只明显的关节、足垫肿大以及轻微气囊炎症状,其中关节攻毒组症状最为明显.MIC测定结果显示,与标准株MS WVU1853相比,这3株分离株对替米考星和恩诺沙星的敏感性显著降低,对泰乐菌素、吉他霉素、泰妙菌素、林可霉素、氟苯尼考及大观霉素敏感性出现不同程度降低;对盐酸沃尼妙林、多西环素、土霉素和泰万菌素具有较高敏感性.本研究为当地防控MS感染提供了科学依据,为临床MS的分离鉴定及用药选择提供了方法指导,为MS疫苗评估中攻毒模型的建立奠定了研究基础.
Abstract
In order to explore the genotyping,pathogenicity,and aitibiotics susceptibility of My-coplasma synoviae(MS)in selected areas of Jiangsu Province,this study isolated and characterized MS strains.The isolates were identified through observation of colony morphology,serological testing,16S rRNA sequencing,and vlhA-PRR typing.Additionally,SPF chicken challenge tests and antimicrobial susceptibility tests were performed.The results showed that three suspected MS strains were isolated from the collected samples,exhibiting characteristic'fried egg'colonies on solid medium.The 16S rRNA sequence analysis showed that each isolate had 99.9％similarity with MS.Furthermore,plate agglutina-tion test showed that it had agglutination reaction with MS-positive serum and MS-infected serum,but did not react with negative serum of SPF chicken,so it was confirmed that the isolates were MS.The SPF chicken challenge test showed that the three isolates caused obvious joint,foot pad swelling and mild air sac inflammation in chickens,and the symptoms of the joint challenge group were the most obvious.The MIC test showed that,compared to the standard strain MS WVU1853,the three isolates displayed a sig-nificantly reduced sensitivity to tilmicosin and enrofloxacin,as well as varying degrees of reduced sensitivity to tylosin,kitasamycin,tiamulin,lincomycin,florfenicol,and spectinomycin.However,they retained a high sensitivity to valnemulin hydrochloride,doxycycline,oxytetracycline,and tyl-valosin.This study provides scientific basis for local prevention and control of MS infections,offers methodological guidance for the isolation,identification,and aitibiotics selection of clinical MS cases,and lays a research foundation for the establishment of a challenge model in MS vaccine evaluation.
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