畜牧兽医学报 ›› 2008, Vol. 39 ›› Issue (7): 955-961.doi:

• 预防兽医 • 上一篇    下一篇

采用PCR-DGGE技术分析蛋鸡肠道细菌种群结构及多样性

倪学勤1*,Joshua Gong2*,Hai Yu2,曾东1,Shayan Sharif3,周小秋4   

  1. 1.四川农业大学动物医学院 动物疫病与人类健康四川省重点实验室,雅安 625014;2.Food Research Program, Agriculture and AgriFood Canada, Guelph, Ontario, N1G 2W1,Canada;3.Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada;4.四川农业大学动物营养研究所,雅安 625014
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-07-23 发布日期:2008-07-23

The Bacterial Community and Diversity in the Layer Gastrointestinal Tract: From Crop to Cecum Analyzed by PCR-DGGE

NI Xue-qin1*, GONG Joshua2*, YU Hai2, ZENG Dong1, SHARIF Shayan3, ZHOU Xiao-qiu4   

  1. 1.College of Animal Medicine, Sichuan Agricultural University, Ya’an 625014, China; 2.Food Research Program, Agriculture and AgriFood Canada, Guelph, Ontario, N1G 2W1,Canada; 3. Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada; 4.Institute of Animal Nutrition, Sichuan Agricultural University, Ya’an 625014, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-07-23 Published:2008-07-23

摘要: 利用基于16S rDNA的PCRDGGE(变性梯度凝胶电泳)图谱技术结合特异性和共性条带割胶回收DNA进行克隆和测序,对2、4、6和8周龄蛋鸡嗉囊、十二指肠、空肠、回肠和盲肠内容物细菌群落的结构和多样性进行了比较,并鉴定了8周龄蛋鸡部分特异性和共性群落成员,分析蛋鸡年龄和肠段部位对微生物群落结构和多样性的影响。肠道菌群DGGE图谱显示肠道部位对细菌种群结构影响很大,盲肠内微生物的多样性最高,其次是回肠和空肠,嗉囊和十二指肠的微生物多样性比较低。十二指肠内细菌与其它肠段相比差异最大,其次是盲肠与其它肠段细菌组成的差异。随着蛋鸡周龄的增加,消化道微生物经历了由简单(2周)到复杂(4周),再回复简单(6周)到复杂(8周)的变化过程。DGGE图谱中共性条带序列分析表明8周龄蛋鸡消化道前段的优势细菌是三得利乳杆菌(Lactobacillus suntoryeus)、索氏梭菌(Clostridium sordellii)和大肠杆菌,盲肠中的特异性条带主要是各种不可培养的细菌以及超巨巨单胞菌(Megamonas hypermegale)和梭菌属细菌(Clostridium spp)。该研究结果表明蛋鸡肠道部位决定细菌群落的结构和多样性,蛋鸡的周龄影响肠道细菌多样性。

Abstract: The bacterial community and diversity in the layer whole gastrointestinal tract were studied by using 16S rDNA-based denaturing gradient gel electrophoresis (DGGE) and some common and special bands were identified, in order to investigate the effect of compartment and age on the predominant eubacteria in the crop, duodenal, jejunal, ileal and ceacal digesta at the age of 2, 4, 6 and 8 weeks. The DGGE profiles provided a snapshot of the bacterial population and allowed calculation of the degree of similarity in the predominant intestinal microflora of commercial layers. PCRDGGE DNA profiles of the V3 region gene of 16S rDNA of microbiota in crop, duodenal, jejunal, ileal and ceacal digesta showed different profiles, the highest levels of diversity were found in cecum, followed by ileum and jejunum, while lowest levels of diversity were found in crop and duodenum. Similarity coefficients between banding patterns of duodenum and the other compartments were significantly different, followed by that of cecum. The number of bands in all intestinal compartants increased when layers grew from 2 to 4 weeks, then decreased in 6 weeks and increased in 8 weeks. 4 common and 9 special 16S rDNA amplicons in the DGGE pattern of layers at 8 weeks were identified by cloning and sequence analysis. Lactobacillus suntoryeus, Clostridium sordellii and E. coli were predominant in the upper GI tract. The special bacterial in ceacal digesta of commercial layers were Clostridium spp,Megamonas hypermegale and plenty of uncultured bacterial. The results of this study indicated that compartmentspecific factors play an important role in the composition and diversity of the dominant eubacterial population in every region of gastrointestinal tract of layers at different age. Furthermore the age of layers also plays an important role in the diversity of the intestinal bacterial community.