东北民猪源停乳链球菌的分离鉴定与药敏试验
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摘要
为分离东北民猪源停乳链球菌(Streptococcus dysgalactiae)并鉴定其生物学特性,从齐齐哈尔市某东北民猪场死亡仔猪的肝脏及其关节液样品中分离病原菌,通过生理生化试验和分子生物学试验鉴定分离菌,研究了分离菌的致病性和药物敏感性。结果表明,分离到革兰阳性球菌1株,生化指标与停乳链球菌停乳亚种相符。系统进化树分析表明,分离菌株与停乳链球菌停乳亚种参考菌株的同源性为97%～98%,其中与AB537921.1(Japan)同源性最高,达到98.99%,进一步从分子水平鉴定该菌为停乳链球菌停乳亚种。以0.2 mL/只(3.2×10~9CFU/mL)接种量感染小鼠,发病率和病死率均为100%。分离菌对多黏菌素B、阿莫西林、氨苄西林、环丙沙星、氧氟沙星、庆大霉素、氟苯尼考、林可霉素、新霉素均敏感,仅对罗红霉素耐药。
In order to isolate and identify Streptococcus dysgalactiae from northeastern min pigs,a bacterial strain was isolated from the liver and joint fluid of dead piglets and identified by biological characteristics and 16 S rDNA gene analysis.At the same time,the pathogenicity and drug sensitivity of isolate were studied.The results showed that a strain of Gram positive cocci was isolated,and the biochemical indexes were consistent with the subspecies of Streptococcus dysgalactiae.Alignment analysis showed that the gene shared more than 97%-98% homology with the sequences of Streptococcus dysgalactiae subsp. dysgalactiae strain.And the homology with the strain JF789447.1(Canada)was up to 98.99%,which confirmed the isolate was as Streptococcus dysgalactiae subsp. dysgalactiae strain.Furthermore,the isolate had strong pathogenicity to mice and the mortality was 100% when inoculated with 3.2×10~9 CFU/mL.While,the isolate was sensitive to polymyxin B,amoxicillin,ampicillin,ciprofloxacin,ofloxacin,gentamicin,florfenicol,lincomycin,neomycin,and resistant to roxithromycin only.
In order to isolate and identify Streptococcus dysgalactiae from northeastern min pigs,a bacterial strain was isolated from the liver and joint fluid of dead piglets and identified by biological characteristics and 16 S rDNA gene analysis.At the same time,the pathogenicity and drug sensitivity of isolate were studied.The results showed that a strain of Gram positive cocci was isolated,and the biochemical indexes were consistent with the subspecies of Streptococcus dysgalactiae.Alignment analysis showed that the gene shared more than 97%-98% homology with the sequences of Streptococcus dysgalactiae subsp. dysgalactiae strain.And the homology with the strain JF789447.1(Canada)was up to 98.99%,which confirmed the isolate was as Streptococcus dysgalactiae subsp. dysgalactiae strain.Furthermore,the isolate had strong pathogenicity to mice and the mortality was 100% when inoculated with 3.2×10~9 CFU/mL.While,the isolate was sensitive to polymyxin B,amoxicillin,ampicillin,ciprofloxacin,ofloxacin,gentamicin,florfenicol,lincomycin,neomycin,and resistant to roxithromycin only.
引文
[1] 杨移斌,杨秋红,刘永涛,等.俄罗斯鲟源停乳链球菌停乳亚种分离、鉴定及药敏特性研究[J].中国预防兽医学报,2017,39(9):717-721.
[2] 王宇婷,王云光,吴丹丹,等.2007-2015年黑龙江省猪链球菌病分子流行病学调查[J].黑龙江畜牧兽医,2016(18):109-111.
[3] 祁金汉,张其祝.猪链球菌病的检疫检验[J].中国畜牧兽医文摘,2016,32(11):123-124.
[4] 王萍.培养方法对土壤可培养细菌多样性的影响及两株新菌的初步鉴定[D].江苏南京:南京农业大学,2009.
[5] Williams A E,Blakemore W F.Pathology of Streptococcal meningitis following intravenous intracisternal and natural routes of infection[J].Neuropathol Appl Neurobiol,1990,16(4):345-356.
[6] 陈雯雯,韦显凯,闭璟珊,等.广西部分地区猪链球菌的分离鉴定及小鼠致病性研究[J].中国畜牧兽医,2017,44(3):861-867.
[7] 张彦丽,刘容珍,张桂玲,等.从一起聚集感染事件中分离培养出停乳链球菌似马亚种[J].中国卫生检验杂志,2014(13):1889-1891.
[8] 董志民,杨春蕾,田向学,等.猪链球菌的分离鉴定及其致病性分子基础研究[J].中国预防兽医学报,2018(2):96-100.
[9] 李善春,周婷婷,冯凯,等.牛源金黄色葡萄球菌和牛源停乳链球菌灭活菌苗对小鼠免疫效果的比较与评估[J].新疆农业大学学报,2016,39(3):218-221.
[10] 幸文定,曾文斌,刘悦欣,等.猪源停乳链球菌类马亚种的分离与鉴定[J].畜牧与兽医,2016,48(2):106-109.
[11] Nomoto R,Munasinghe L I,Jin D H,et al.Lancefield group C Streptococcus dysgalactiae,infection responsible for fish mortalities in Japan[J].J Fish Dis,2004,27(12):679-686.
[12] 潘厚军,刘晓勇,常藕琴,等.西伯利亚鲟停乳链球菌的分离、鉴定与致病性[J].中国水产科学,2009,16(6):891-904.
[13] 张艳波,付安才,孙立光,等.从血液中分离停乳链球菌似马亚种探讨[J].中外医疗,2009,28(31):157-157.
[14] 唐芹芳,居会祥,金浩,等.从血液中分离出停乳链球菌似马亚种1例[J].江苏大学学报:医学版,2008,18(6):552.
[15] Atalay A,Olcu M,Percin D.Antibiotic susceptibilities and serotyping of clinical Streptococcus agalactiae isolates[J].Med J Trakya University,2010,28(4):362-365.
[16] 余丽阳,詹碧翠,杨玮.92株停乳链球菌似马亚种临床感染分布及其药敏分析[J].中国卫生检验杂志,2017(16):151-153.
[17] 杨移斌,杨秋红,刘永涛,等.俄罗斯鲟源停乳链球菌停乳亚种分离、鉴定及药敏特性研究[J].中国预防兽医学报,2017(9):33-37.
[18] 孟祥庆,吴美芹.生猪养殖中链球菌病的防治措施[J].兽医导刊,2017(20):111-111.
[2] 王宇婷,王云光,吴丹丹,等.2007-2015年黑龙江省猪链球菌病分子流行病学调查[J].黑龙江畜牧兽医,2016(18):109-111.
[3] 祁金汉,张其祝.猪链球菌病的检疫检验[J].中国畜牧兽医文摘,2016,32(11):123-124.
[4] 王萍.培养方法对土壤可培养细菌多样性的影响及两株新菌的初步鉴定[D].江苏南京:南京农业大学,2009.
[5] Williams A E,Blakemore W F.Pathology of Streptococcal meningitis following intravenous intracisternal and natural routes of infection[J].Neuropathol Appl Neurobiol,1990,16(4):345-356.
[6] 陈雯雯,韦显凯,闭璟珊,等.广西部分地区猪链球菌的分离鉴定及小鼠致病性研究[J].中国畜牧兽医,2017,44(3):861-867.
[7] 张彦丽,刘容珍,张桂玲,等.从一起聚集感染事件中分离培养出停乳链球菌似马亚种[J].中国卫生检验杂志,2014(13):1889-1891.
[8] 董志民,杨春蕾,田向学,等.猪链球菌的分离鉴定及其致病性分子基础研究[J].中国预防兽医学报,2018(2):96-100.
[9] 李善春,周婷婷,冯凯,等.牛源金黄色葡萄球菌和牛源停乳链球菌灭活菌苗对小鼠免疫效果的比较与评估[J].新疆农业大学学报,2016,39(3):218-221.
[10] 幸文定,曾文斌,刘悦欣,等.猪源停乳链球菌类马亚种的分离与鉴定[J].畜牧与兽医,2016,48(2):106-109.
[11] Nomoto R,Munasinghe L I,Jin D H,et al.Lancefield group C Streptococcus dysgalactiae,infection responsible for fish mortalities in Japan[J].J Fish Dis,2004,27(12):679-686.
[12] 潘厚军,刘晓勇,常藕琴,等.西伯利亚鲟停乳链球菌的分离、鉴定与致病性[J].中国水产科学,2009,16(6):891-904.
[13] 张艳波,付安才,孙立光,等.从血液中分离停乳链球菌似马亚种探讨[J].中外医疗,2009,28(31):157-157.
[14] 唐芹芳,居会祥,金浩,等.从血液中分离出停乳链球菌似马亚种1例[J].江苏大学学报:医学版,2008,18(6):552.
[15] Atalay A,Olcu M,Percin D.Antibiotic susceptibilities and serotyping of clinical Streptococcus agalactiae isolates[J].Med J Trakya University,2010,28(4):362-365.
[16] 余丽阳,詹碧翠,杨玮.92株停乳链球菌似马亚种临床感染分布及其药敏分析[J].中国卫生检验杂志,2017(16):151-153.
[17] 杨移斌,杨秋红,刘永涛,等.俄罗斯鲟源停乳链球菌停乳亚种分离、鉴定及药敏特性研究[J].中国预防兽医学报,2017(9):33-37.
[18] 孟祥庆,吴美芹.生猪养殖中链球菌病的防治措施[J].兽医导刊,2017(20):111-111.
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