猪源荚膜血清F型多杀性巴氏杆菌的分离鉴定、毒力基因检测与致病性研究
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摘要
为调查多杀性巴氏杆菌(P.multocida)在湖南猪群中的流行情况,本实验室于2015年6月~2016年3月在湖南大型屠宰场采集猪肺脏222份和猪扁桃体180份临床样品,采用血琼脂培养和PCR方法进行P.multocida的分离鉴定,得到4株F型P.multocida。对分离的4株F型P.multocida进行毒力基因检测以及小鼠的致病性研究,结果显示4株菌均含有ptf A、pfhA、fim A、sodA、sodC、ompA、ompH、oma87、plpB、exbB、exbD、tonB、hgbB、Fur、nan H、hsf-1和hsf-2毒力基因,但tadD、toxA、hgbA、nanB、pm HAS、tbpA毒力基因均为阴性;4株菌致病性试验结果显示,感染小鼠的剂量分别为4 cfu/只、5 cfu/只、11 cfu/只和17 cfu/只时可导致小鼠在60 h内全部死亡,表明这4株菌对小鼠有强致病力。病理切片显示,感染小鼠有明显的肺泡充血、脾淋巴细胞凋亡、肾小管和肾小球萎缩等病理变化。本实验首次在湖南地区分离到猪源荚膜血清F型P.multocida,并进行了较为详细的生物学特性研究,为国内P.multocida的流行情况和生猪多杀性巴氏杆菌病的防控补充了参考资料。
Four Pasteurella multocida capsule serotype F strains were isolated in this lab by using blood agar and PCR method from 222 lungs and 180 tonsils of slaughtered pigs in Hunan province during June 2015 to March 2016. 23 virulence-associated genes harboured in those 4 isolates were positive of virulence-associated genes of ptf A, pfhA, fimA, sodA, sodC, ompA, ompH,oma87, plpB, exbB, exbD, tonB, hgbA, Fur, nanH, hsf-1 and hsf-2 detected by PCR. Pathogenic test showed that all of the mice died after 60 hrs when inoculated intraperitoneally with the isolates at dose of 4 cfu, 5 cfu, 11 cfu and 17 cfu for each mouse,respectively, indicating these isolates were high pathogenic to mice. The alveolar hyperaemia, splenic lymphocyte apoptosis,glomerular and renal tubular atrophy of mice were revealed by histopathological examination. These results provid a basis for further study prevention and epidemiological surveillance P.multocida infection in pigs.
Four Pasteurella multocida capsule serotype F strains were isolated in this lab by using blood agar and PCR method from 222 lungs and 180 tonsils of slaughtered pigs in Hunan province during June 2015 to March 2016. 23 virulence-associated genes harboured in those 4 isolates were positive of virulence-associated genes of ptf A, pfhA, fimA, sodA, sodC, ompA, ompH,oma87, plpB, exbB, exbD, tonB, hgbA, Fur, nanH, hsf-1 and hsf-2 detected by PCR. Pathogenic test showed that all of the mice died after 60 hrs when inoculated intraperitoneally with the isolates at dose of 4 cfu, 5 cfu, 11 cfu and 17 cfu for each mouse,respectively, indicating these isolates were high pathogenic to mice. The alveolar hyperaemia, splenic lymphocyte apoptosis,glomerular and renal tubular atrophy of mice were revealed by histopathological examination. These results provid a basis for further study prevention and epidemiological surveillance P.multocida infection in pigs.
引文
[1]Wilson B A,Ho M.Pasteurella multocida:from Zoonosis to Cellular Microbiology[J].Clin Microbiol Rev,2013,26(3):631-655.
[2]彭忠,王庚,吴广敬,等.猪源多杀性巴氏杆菌的分离与鉴定[J].中国兽医科学,2015,(09):943-951.
[3]汤细彪.猪源多杀性巴氏杆菌的分子流行病学与致病性研究[D].华中农业大学,2010.
[4]林星宇,王印,杨泽晓,等.猪源荚膜血清F型多杀性巴氏杆菌的分离鉴定[J].中国兽医科学,2015,(06):567-571.
[5]Townsend K M,Boyce J D,Chung J Y,et al.Genetic organization of Pasteurella multocida cap Loci and development of a multiplex capsular PCR typing system[J].J Clin Microbiol,2001,39(3):924-929.
[6]Khamesipour F,Momtaz H,Azhdary M M.Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran[J].Front Microbiol,2014,5:536.
[7]Rimler R B,Rhoades K R.Serogroup F.a new capsule serogroup of Pasteurella multocida[J].J Clin Microbiol,1987,25(4):615-618.
[8]Rhoades K R,Rimler R B.Capsular groups of Pasteurella multocida isolated from avian hosts[J].Avian Dis,1987:895-898.
[9]Wilson M A,Duncan R M,Nordholm G E,et al.Pasteurella multocida isolated from wild birds of North America:a serotype and DNA fingerprint study of isolates from 1978 to 1993[J].Avian Dis,1995,39(3):587-593.
[10]Ewers C,Lübke-Becker A,Bethe A,et al.Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status[J].Vet Microbiol,2006,114(3):304-317.
[11]Fuller T E,Kennedy M J,Lowery D E.Identification of Pasteurella multocida virulence genes in a septicemic mouse model using signature-tagged mutagenesis[J].Microb Pathog,2000,29(1):25-38.
[12]Jaglic Z,Jeklova E,Leva L,et al.Experimental study of pathogenicity of Pasteurella multocida serogroup F in rabbits[J].Vet Microbiol,2008,126(1):168-177.
[13]Catry B,Chiers K,Schwarz S,et al.Fatal peritonitis caused by Pasteurella multocida capsular type F in calves[J].J Clin Microbiol,2005,43(3):1480-1483.
[14]Davies R L,Watson P J,Caffrey B.Comparative analyses of Pasteurella multocida strains associated with the ovine respiratory and vaginal tracts[J].Vet Rec,2003,152(1):7-10.
[15]May B J,Zhang Q,Li L L,et al.Complete genomic sequence of Pasteurella multocida,Pm70[J].Proceed Nat Aca Sci,2001,98(6):3460-3465.
[16]Johnson T J,Abrahante J E,Hunter S S,et al.Comparative genome analysis of an avirulent and two virulent strains of avian Pasteurella multocida reveals candidate genes involved in fitness and pathogenicity[J].BMC Microbiol,2013,13:106.
[17]Okay S,Kurt K A.Comparative genome analysis of five Pasteurella multocida strains to decipher the diversification in pathogenicity and host specialization[J].Gene,2015,567(1):58-72.
[2]彭忠,王庚,吴广敬,等.猪源多杀性巴氏杆菌的分离与鉴定[J].中国兽医科学,2015,(09):943-951.
[3]汤细彪.猪源多杀性巴氏杆菌的分子流行病学与致病性研究[D].华中农业大学,2010.
[4]林星宇,王印,杨泽晓,等.猪源荚膜血清F型多杀性巴氏杆菌的分离鉴定[J].中国兽医科学,2015,(06):567-571.
[5]Townsend K M,Boyce J D,Chung J Y,et al.Genetic organization of Pasteurella multocida cap Loci and development of a multiplex capsular PCR typing system[J].J Clin Microbiol,2001,39(3):924-929.
[6]Khamesipour F,Momtaz H,Azhdary M M.Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran[J].Front Microbiol,2014,5:536.
[7]Rimler R B,Rhoades K R.Serogroup F.a new capsule serogroup of Pasteurella multocida[J].J Clin Microbiol,1987,25(4):615-618.
[8]Rhoades K R,Rimler R B.Capsular groups of Pasteurella multocida isolated from avian hosts[J].Avian Dis,1987:895-898.
[9]Wilson M A,Duncan R M,Nordholm G E,et al.Pasteurella multocida isolated from wild birds of North America:a serotype and DNA fingerprint study of isolates from 1978 to 1993[J].Avian Dis,1995,39(3):587-593.
[10]Ewers C,Lübke-Becker A,Bethe A,et al.Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status[J].Vet Microbiol,2006,114(3):304-317.
[11]Fuller T E,Kennedy M J,Lowery D E.Identification of Pasteurella multocida virulence genes in a septicemic mouse model using signature-tagged mutagenesis[J].Microb Pathog,2000,29(1):25-38.
[12]Jaglic Z,Jeklova E,Leva L,et al.Experimental study of pathogenicity of Pasteurella multocida serogroup F in rabbits[J].Vet Microbiol,2008,126(1):168-177.
[13]Catry B,Chiers K,Schwarz S,et al.Fatal peritonitis caused by Pasteurella multocida capsular type F in calves[J].J Clin Microbiol,2005,43(3):1480-1483.
[14]Davies R L,Watson P J,Caffrey B.Comparative analyses of Pasteurella multocida strains associated with the ovine respiratory and vaginal tracts[J].Vet Rec,2003,152(1):7-10.
[15]May B J,Zhang Q,Li L L,et al.Complete genomic sequence of Pasteurella multocida,Pm70[J].Proceed Nat Aca Sci,2001,98(6):3460-3465.
[16]Johnson T J,Abrahante J E,Hunter S S,et al.Comparative genome analysis of an avirulent and two virulent strains of avian Pasteurella multocida reveals candidate genes involved in fitness and pathogenicity[J].BMC Microbiol,2013,13:106.
[17]Okay S,Kurt K A.Comparative genome analysis of five Pasteurella multocida strains to decipher the diversification in pathogenicity and host specialization[J].Gene,2015,567(1):58-72.
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