畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (10): 1608-1615.doi: 10.11843/j.issn.0366-6964.2014.10.005

• 遗传繁育 • 上一篇    下一篇

梅山猪E.coli F18菌株攻毒试验及其表型的鉴定分析

吴正常1,董文华1,刘颖1,杨建生2,朱国强3,吴圣龙1*,包文斌1*   

  1. (1.扬州大学动物科学与技术学院,扬州 225009; 2.昆山市梅山猪保种有限公司,昆山 215300;3.扬州大学兽医学院, 扬州 225009)
  • 收稿日期:2014-01-12 出版日期:2014-10-23 发布日期:2014-10-23
  • 通讯作者: 包文斌,博士,研究员,主要从事猪抗病育种研究,E-mail:wbbao@yzu.edu.cn;吴圣龙,博士,研究员,主要从事猪抗病育种研究,E-mail:slwu@yzu.edu.cn
  • 作者简介:吴正常(1987-),男,江苏南京人,博士生,主要从事猪遗传育种研究,E-mail:wuzhengchang@126.com
  • 基金资助:

    国家自然科学基金(31372285;31172183;31472066);江苏省高校自然科学研究重大项目(14KJA230003);江苏省科技支撑计划(农业)(BE2013345;BE2012330;BE2014357)

Attack Experiment and Phenotype Analysis of Meishan Piglets by E.coli F18 Strain

WU Zheng-chang1,DONG Wen-hua1,LIU Ying1,YANG Jian-sheng2,ZHU Guo-qiang3,WU Sheng-long1*,BAO Wen-bin1*   

  1. (1.College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China;2.Conservation Ltd.of Meishan Pig of Kunshan in Jiangsu Province,Kunshan 215300,China;3.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China)
  • Received:2014-01-12 Online:2014-10-23 Published:2014-10-23

摘要:

利用E.coli F18菌株攻毒试验来获得梅山猪断奶仔猪E .coli F18敏感和抗性型个体,可以为今后利用高通量技术研究仔猪抗E .coli F18遗传机理提供宝贵的素材,同时也为中国地方猪品种细菌性疾病模型的建立提供指导和参考。本研究以中国地方品种梅山猪断奶仔猪为研究对象,采用F18ab和F18ac菌株口服攻毒试验,并利用仔猪肠道E.coli F18细菌检测、细菌计数以及小肠上皮细胞黏附试验等进行验证。结果表明:攻毒后梅山仔猪出现腹泻、轻度腹泻和不腹泻3种不同的表型,腹泻仔猪肠道内容物中的细菌数量远多于不腹泻仔猪;同时黏附试验发现腹泻仔猪小肠上皮细胞大量黏附F18大肠杆菌,而不腹泻仔猪小肠上皮细胞与细菌基本不黏附;病理组织切片结果表明,重度腹泻仔猪病变肠道黏膜层绒毛高度均缩短,并且回肠组织间隙内有充血现象,空肠组织黏膜层绒毛脱落,由此可以推测大肠杆菌F18与仔猪小肠上皮细胞受体结合后,主要对小肠绒毛完整性造成了影响,从而定居繁殖造成仔猪腹泻。以上试验结果进一步表明,本试验中仔猪腹泻确实由E.coli F18菌株攻毒造成,并成功获得了E.coli F18抗性型和敏感型仔猪个体,为今后猪细菌性腹泻遗传基础研究素材和抗病育种基础群的获得提供了有效可行的方法和途径。

Abstract:

Attack experiment of Meishan piglets caused by E.coli F18 strain will attain individuals of the resistance and susceptibility to E.coli F18,which will provide good material for analyzing the molecular mechanism of the resistance to E.coli F18 by high throughput technology,and provide theoretical reference for further establishing bacterial disease model in Chinese native pig breeds.In the study,the Meishan piglets were used as model animals to test their susceptibility to E.coli F18 by challenging with the pathogens through feeding F18ab and F18ac strains,which was tested and verified by a series of experiments,such as E.coli F18 bacteria detection,bacteria counting and adhesion test of the pathogens to the epithelial cells of small intestine in vitro.The results showed that there were 3 phenotypes of severe diarrhea,mild diarrhea and normality after artificial attack experiment in Meishan piglets.The number of bacteria in the intestinal contents of diarrhea piglets was more than that of normal piglets.Moreover,the adhesion test further showed that most bacteria had adhered to the epithelial cells of the small intestine of diarrhea piglets.The result of pathological tissue section showed that the villus height of pathological intestinal mucosa became shorter,congestion phenomenon appeared in the clearance of ileum tissue,and the villus of ileum tissue mucous layer fell off.These results speculated that E.coli F18 mainly affected the integrity of small intestinal villus when it combined with intestinal epithelial cell receptors,thus E.coli F18 settled in breeding and caused diarrhea in piglets.The above test results further showed that the piglets diarrhea was indeed caused by E.coli F18 attack,and we obtained several individuals of the resistance and susceptibility to E.coli F18 in Meishan piglets,which provided effective ways and methods for obtaining basic material on the genetic mechanisms of pig bacterial diarrhea and disease-resistant breeding base group.

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