[1]周 姣,刘宝生,韩春生,等.江西地区仔猪肠道A型产气荚膜梭菌的分离及其β2毒素的研究[J].中国预防兽医学报,2018,(02):101-105.[doi:0.3969/j.issn.1008-0589.201708015]
 ZHOU Jiao,LIU Bao-sheng,HAN Chun-sheng,et al.CPA isolation and its β2 toxin investigation from the intestinal tract of piglets in Jiangxi province[J].Chinese journal of preventive veterinary medicine,2018,(02):101-105.[doi:0.3969/j.issn.1008-0589.201708015]





CPA isolation and its β2 toxin investigation from the intestinal tract of piglets in Jiangxi province

周 姣刘宝生韩春生李 昆戴益民张锦华*

 (江西农业大学 动物科学技术学院,江西 南昌 330045)

ZHOU Jiao LIU Bao-sheng HAN Chun-sheng LI Kun DAI Yi-min ZHANG Jin-hua*

 (College of Animal Science and Technology Jiangxi Agricultural University, Nanchang 330045, China)
为研究A型产气荚膜梭菌(CpA)的β2毒素对仔猪坏死性肠炎(NEC)的影响,本试验采用改良FTG 培养基从江西地区健康与腹泻哺乳仔猪新鲜粪便中初分离Cp,再对所分离Cp菌株进行生化和16S rDNA分子鉴定,统计江西省7个地区健康和腹泻仔猪肠道Cp分离率;通过多重PCR技术对CpA菌株及CpA β2+毒素基因携带率进行测定;将CpA (β2+/β2-)分离株进行小鼠毒性试验。结果显示:255份哺乳仔猪粪便样品中,腹泻样品Cp分离率为89.6 % (138/154),健康样品Cp分离率为90.1 % (91/101),其中CpA占分离菌株的89.5 % (205/229);CpA 菌株的β2毒素携带率为99.5 % (204/205),表明江西地区仔猪肠道CpA携带率及CpA (β2+)比例均较高;所分离的CpA菌株通常携带2个以上质粒,且β2毒素基因位于CpA菌其中的质粒中;来源于腹泻仔猪的CpA (β2+)、CpA (β2-)和健康仔猪的CpA (β2+)培养上清分别以腹腔接种小鼠,均可引起小鼠不同程度死亡。从小鼠毒性试验结果表明CpA与菌株来源健康或腹泻仔猪、是否携带β2毒素没有直接关系,其毒性作用可能主要与宿主肠道环境有关。本实验为研究CpA (β2+)在NEC中的影响提供初步实验依据。
To study the effect of β2 toxin produced by Clostridium perfringens type A (CpA) on the Necrotizing enterocolitis (NEC) of piglets, C.perfringens (Cp) was isolated from the feces of piglets with or without NEC using the modifying FTG (Fluid Thioglycollate) medium and further confirmed by biochemical and 16S rDNA molecular indetification. The speration rate of Cp carrying by piglets in seven regions of Jiangxi province was statistically analyzed. CpA and β2 toxin gene carried by CpA were analyzed using a multiplex PCR method. Finally, we established infection model with mice using the isolates CpA (β2+/β2-). The results demonstrated that the speration rates of Cp in diarrhea and healthy piglets were 89.6% (138/154) and 90.1% (91/101) respectively. The speration rate of CpA was 89.5% (205/229) among the all isolates, of which 99.5% (204/205) were CpA (β2+). These results showed that the incidence of CpA and β2 toxin were at a relatively high percentage in piglets of Jiangxi province. The culture supernatants of CpA (β2+) and CpA (β2-) from NEC piglets, CpA (β2+) from healthy piglets respectively injected intraperitoneally mice all could induce a different level of death in mice. Results of plasmid extraction also confirmed that there


[1] Uzal F A, Freedman J C, Shrestha A, et al. Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease [J]. Future Microbiol, 2014, 9(3): 361-377.
[2]Schotte U, Truyen U, Neubauer H. Significance of beta 2-toxigenic Clostridium perfringens infections in animals and their predisposing factors--a review [J]. J Vet Med B Infect Dis Vet Public Health, 2004, 51(10): 423-426.
[3]屠伟英,蒋玉文,赵代文,等. 仔猪产气荚膜梭菌肠毒血症病原诊断 [J]. 中国兽药杂志,2001,35(6):10-13.
[4]Kongsted H, Jonach B, Haugegaard S, et al. Microbiological, pathological and histological findings in four Danish pig herds  affected by a new neonatal diarrhoea syndrome [J]. BMC Vet Res, 2013, 9: 206.
[5]Bueschel D M, Helen Jost B, Billington S J, et al. Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringens field isolates: correlation of genotype with phenotype [J]. Vet Microbiol, 2003, 94(2): 121-129.
[6]Herholz C, Miserez R, Nicolet J, et al. Prevalence of b2-Toxigenic Clostridium perfringens in horses with intestinal disorders [J]. J Clin Microbiol,1999, 37(2): 358-361.
[7]Freedman J C, Theoret J R, Wisniewski J A, et al. Clostridium perfringens type A-E toxin plasmids [J]. Res Microbiol, 2015, 166: 264-279.
[8]Marchesi J R, Sato T, Welghtman A J, et al.  Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA [J]. Appl Environ Microb, 64(2): 795-799.
[9]Garmory H S, Chanter N, French N P, et al. Occurrence of Clostridium perfringens β2-toxin amongst animals, determined using genotyping and subtyping PCR assays [J]. Epidemiol Infect, 2000, 124(1): 61-67.
[10]Songer J G, Uzal F A, Clostridial enteric infections in pigs [J]. J Vet Diagn Invest, 2005, 17(6): 528-536.
[11]Afshari A, Jamshidi A, Razmyar J, et al. Genotyping of Clostridium perfringens isolated from broiler meat in northeastern of Iran [J]. Vet Res Forum, 2015, 6(4): 279-284.
[12]Chan G, Farzan A, Soltes G, et al. The epidemiology of Clostridium perfringens type A on Ontario swine farms, with special reference to cpb2-positive isolates [J]. BMC Vet Res, 2012, 8: 156.
[13]Waters M, Savoie A, Garmory H S, et al. Genotyping and phenotyping of beta2-toxigenic Clostridium perfringens fecal isolates associated with gastrointestinal diseases in piglets [J]. J Clin Microbiol, 2003, 41(8): 3584-3591.
[14]Farzan A, Kircanski J, DeLay J, et al. An investigation into the association between cpb2-encoding Clostridium perfringens type A and diarrhea in neonatal piglets [J]. Can J Vet Res, 2013, 77(1): 45-53.
[15]Fisher D J, Miyamoto K, Harrison B, et al. Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene [J]. Mol Microbiol, 2005, 56(3): 747-762.
[16]Azcarate-Peril M A, Foster D M, Cadenas M B, et al. Acute necrotizing enterocolitis of preterm piglets is characterized by dysbiosis of ileal mucosa-associated bacteria [J]. Gut Microbes, 2014, 2(4): 234-243.
[17]Allaart J G, van Asten A J, Vernooij J C,et al. Effect of Lactobacillus fermentum on beta2 toxin production by Clostridium perfringens [J]. Appl Environ Microb, 2011, 77(13): 4406-4411.



 WU Sheng-long,YUAN Zhi-wei,JU Hui-ping,et al.Polymorphisms of the FUT1 gene M307 locus in post-weaning Sutai breed piglet and resistance to F18 fimbrial Escherichia coli in vitro[J].Chinese journal of preventive veterinary medicine,2007,(02):783.
 ZHANG Hong-bing*.The enhancement of the transfer factors on immunization of inactivated porcine circovirus type 2 vaccine in pigs[J].Chinese journal of preventive veterinary medicine,2013,(02):920.[doi:10.3969/j.issn.1008-0589.2013.11.14]
 WANG Xiao-bo,XU Jin-jun,WU Li-li,et al.Isolation and identification of Toxoplasma gondii strain from piglets[J].Chinese journal of preventive veterinary medicine,2013,(02):940.[doi:10.3969/j.issn.1008-0589.2013.11.19]
 ZHANG Hong-bing. The effect of porcine interleukin-2,4 as immunopotentiator for PCV inactivated vaccine to enhanceimmune responses in piglets[J].Chinese journal of preventive veterinary medicine,2014,(02):395.[doi:10.3969/j.issn.1008-0589.2014.05.15]
 CHEN Zhang-hua*,DENG Jun-liang,CHEN Cang-liang.Effect of "Zhukangsan" on the immune function of pig erythrocytes[J].Chinese journal of preventive veterinary medicine,2015,(02):970.[doi:10.3969/j.issn.1008-0589.2015.12.17]
[7]孙加节*,蒋 勇*,习欠云,等.人参复合多糖提高猪圆环病毒疫苗免疫效果的研究[J].中国预防兽医学报,2016,(09):734.[doi:0.3969/j.issn.1008-0425.2016.09.13]
 SUN Jia-jie*,JIANG Yong*,XI Qian-yun,et al.Ginseng polysaccharides complex enhances immune response to porcine circovirus in piglets[J].Chinese journal of preventive veterinary medicine,2016,(02):734.[doi:0.3969/j.issn.1008-0425.2016.09.13]
[8]马 琰,宗秋芳,权国梅,等.F4大肠杆菌黏附素受体易感性仔猪的体外鉴定研究[J].中国预防兽医学报,2017,(05):366.[doi:10.3969/j.issn.1008-0425.2017.05.07]
 MA Yan,ZONG Qiu-fang,QUAN Guo-mei,et al. Identification of genetically susceptible piglets with the small intestinal epithelia receptors to F4 fimbriated Escherichia coli in vitro[J].Chinese journal of preventive veterinary medicine,2017,(02):366.[doi:10.3969/j.issn.1008-0425.2017.05.07]

更新日期/Last Update: 2018-03-09