藏鸡鸡毒支原体的分离鉴定及对常用抗菌药物的敏感性试验
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摘要
为了探究藏鸡鸡毒支原体(MG)的生物学特性,采用分离培养法对疑似慢性呼吸道病(CRD)藏鸡气管进行了MG分离纯化,根据菌落形态、鸡红细胞吸附试验、生长抑制试验和PCR进行鉴定,利用微量法测定了分离株对临床常用抗菌药物的敏感性。结果表明;从30份样本中仅分离到3株符合MG特征,分别命名为ZJ-1、ZJ-2和ZJ-3;通过MG 16srRNA序列比对,证实3株分离株均为与S6和HS株亲缘关系较近的较强毒力致病性MG;对试验用抗菌药物的敏感性较为一致,即对泰妙菌素、泰乐菌素、氟苯尼考、强力霉素、氧氟沙星、恩诺沙星、环丙沙星、林可霉素、庆大霉素均高度敏感;对红霉素中度敏感;对土霉素不敏感,但未发现高度耐药现象。证实了藏鸡群中存在致病性MG流行,且毒力具有增强趋势,临床中可使用泰乐菌素、强力霉素等高度敏感药物进行防治。
In order to explore the biological characteristics of Mycoplasma gallisepticam(MG)from Tibetan chicken.MG was separated and purified from trachea samples of Tibetan chickens with chronic respiratory disease(CRD).All the isolates were further identified by using colony morphology,absorption test for chicken red blood cell,growth inhibition test and polymerase chain reaction.The sensitivity of isolates to common antibacterial agent drug was investigate by micro method.The results showed that three isolates named as ZJ-1,ZJ-2,and ZJ-3 were separated from thirty specimens.By 16 SrRNA sequence alignment,the three isolates are highly pathogenic with close relationship to S6 and HS strains,and highly sensitive to tiamulin,tylosin,florfenicol,doxycycline,ofloxacin,enrofloxacin,ciprofloxacin,lincomycin and gentamicin,and are moderately sensitive to erythromycin,but not sensitive to oxytetracycline,drug resistant isolates was not investigated.The examined Tibetan chickens were infected by pathogenic MG with increasing virulence tendency,such highly sensitive drugs as Tiamulin,TYlosin and so on can be clinically used to prevent and cure MG infection.
In order to explore the biological characteristics of Mycoplasma gallisepticam(MG)from Tibetan chicken.MG was separated and purified from trachea samples of Tibetan chickens with chronic respiratory disease(CRD).All the isolates were further identified by using colony morphology,absorption test for chicken red blood cell,growth inhibition test and polymerase chain reaction.The sensitivity of isolates to common antibacterial agent drug was investigate by micro method.The results showed that three isolates named as ZJ-1,ZJ-2,and ZJ-3 were separated from thirty specimens.By 16 SrRNA sequence alignment,the three isolates are highly pathogenic with close relationship to S6 and HS strains,and highly sensitive to tiamulin,tylosin,florfenicol,doxycycline,ofloxacin,enrofloxacin,ciprofloxacin,lincomycin and gentamicin,and are moderately sensitive to erythromycin,but not sensitive to oxytetracycline,drug resistant isolates was not investigated.The examined Tibetan chickens were infected by pathogenic MG with increasing virulence tendency,such highly sensitive drugs as Tiamulin,TYlosin and so on can be clinically used to prevent and cure MG infection.
引文
[1]GROSS W B.Factors affecting the development of respiratory disease complex in chickens[J].Avian Dis,1990,34(3):607-61.
[2]GHARAIBEH S,Al-RASHDAN M.Change in antimicrobial susceptibility of Mycoplasma gallisepticum field isolates[J].Vet Microbial,2011,150(3):379-383.
[3]FAN H H,KLEVEN S H,JACKWOOD M W.Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum[J].Avian Diseases,1995,39(4):729-735
[4]毕丁仁.禽源霉形体的分离和鉴定[J].中国兽医科技,1985(12):49-57.
[5]毕丁仁,吴世钦.用血细胞吸附抑制试验快速鉴定鸡毒霉形体[J].中国兽医科技,1990(1):29-30.
[6]GHARAIBEH S,Al ROUSSAN D.The use of molecular techniques in isolation and characterization of Mycoplasma gallisepticumfrom commercial chickens in Jordan[J].Int J Poult Sci,2008,7(1):28-35.
[7]丁铲,于圣青,刘晓文,等.败血支原体16SrRNA基因的克隆与核酸序列分析[J].中国预防兽医学报,1999,21(2):134-136.
[8]毕丁仁,屈小玲.鸡毒霉形体人工感染肉用仔鸡后呼吸系统的病理形态学观察[J].中国兽医科技,1999,29(6):7-10.
[9]SENTERFITL B,TAYLOR-ROBINSON D,ROBERTSON J A,et al.Antibiotic sensitivity testing of mycoplasmas[J].Meth Mycoplasmol,1986,2:397-401.
[10]孔意端,林居纯,陈继荣,等.鸡毒支原体不同地区分离株对常用抗菌药物的敏感性试验[J].动物医学进展,2008,29(6):35-38.
[11] REINHARDT A K,KEMPF I,KOBISCH M,et al.Fluoroquinolone resistance in Mycoplasma gallisepticum:DNA gyrase as primary target of enrofloxacin and impact of mutations in topoisomerases on resistance level[J].J Antimicrob Chemoth,2002,50(4):589-592.
[12] GERCHMAN I,LEVISOHN S,MIKULA I,et al.Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticumstrains isolated from poultry[J].Res Vet Sci,2011,42(1):90.
[13] LEVISOHN S.Antibiotic sensitivity patterns in field isolates of Mycoplasma gallisepticum as a guide to chemotherapy[J].Israel J Med Sci,1981,17(7):661-666.
[14] GAUTIER-BOUCHARDON A V,REINHARDT A K,KOBISCH M,et al.In vitro development of resistance to enrofloxacin,erythromycin,tylosin,tiamulin and oxytetracycline in Mycoplasma gallisepticum,Mycoplasma iowae and Mycoplasma synoviae[J].Vet Microbiol,2002,88(1):47-58.
[15]李建,高航飞,安维雪,等.牛支原体TaqMan实时荧光定量PCR检测方法的建立[J].中国兽医学报,2017,37(6):1059-1064.
[16]倪博,白方方,刘茂军,等.支原体脂蛋白及其变异与宿主互作研究进展[J].生物技术通报,2013,1(2):49-54.
[2]GHARAIBEH S,Al-RASHDAN M.Change in antimicrobial susceptibility of Mycoplasma gallisepticum field isolates[J].Vet Microbial,2011,150(3):379-383.
[3]FAN H H,KLEVEN S H,JACKWOOD M W.Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum[J].Avian Diseases,1995,39(4):729-735
[4]毕丁仁.禽源霉形体的分离和鉴定[J].中国兽医科技,1985(12):49-57.
[5]毕丁仁,吴世钦.用血细胞吸附抑制试验快速鉴定鸡毒霉形体[J].中国兽医科技,1990(1):29-30.
[6]GHARAIBEH S,Al ROUSSAN D.The use of molecular techniques in isolation and characterization of Mycoplasma gallisepticumfrom commercial chickens in Jordan[J].Int J Poult Sci,2008,7(1):28-35.
[7]丁铲,于圣青,刘晓文,等.败血支原体16SrRNA基因的克隆与核酸序列分析[J].中国预防兽医学报,1999,21(2):134-136.
[8]毕丁仁,屈小玲.鸡毒霉形体人工感染肉用仔鸡后呼吸系统的病理形态学观察[J].中国兽医科技,1999,29(6):7-10.
[9]SENTERFITL B,TAYLOR-ROBINSON D,ROBERTSON J A,et al.Antibiotic sensitivity testing of mycoplasmas[J].Meth Mycoplasmol,1986,2:397-401.
[10]孔意端,林居纯,陈继荣,等.鸡毒支原体不同地区分离株对常用抗菌药物的敏感性试验[J].动物医学进展,2008,29(6):35-38.
[11] REINHARDT A K,KEMPF I,KOBISCH M,et al.Fluoroquinolone resistance in Mycoplasma gallisepticum:DNA gyrase as primary target of enrofloxacin and impact of mutations in topoisomerases on resistance level[J].J Antimicrob Chemoth,2002,50(4):589-592.
[12] GERCHMAN I,LEVISOHN S,MIKULA I,et al.Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticumstrains isolated from poultry[J].Res Vet Sci,2011,42(1):90.
[13] LEVISOHN S.Antibiotic sensitivity patterns in field isolates of Mycoplasma gallisepticum as a guide to chemotherapy[J].Israel J Med Sci,1981,17(7):661-666.
[14] GAUTIER-BOUCHARDON A V,REINHARDT A K,KOBISCH M,et al.In vitro development of resistance to enrofloxacin,erythromycin,tylosin,tiamulin and oxytetracycline in Mycoplasma gallisepticum,Mycoplasma iowae and Mycoplasma synoviae[J].Vet Microbiol,2002,88(1):47-58.
[15]李建,高航飞,安维雪,等.牛支原体TaqMan实时荧光定量PCR检测方法的建立[J].中国兽医学报,2017,37(6):1059-1064.
[16]倪博,白方方,刘茂军,等.支原体脂蛋白及其变异与宿主互作研究进展[J].生物技术通报,2013,1(2):49-54.