| 摘要: |
| 研究依莎褐蛋鸡和艾维茵肉鸡发育正常及迟缓鸡群的盲肠细菌种群结构和多样性变化。使用基于16S rDNA的PCR-DGGE技术,结合割胶回收DNA进行克隆和测序,分别对4、6、10、16、20和40周龄蛋鸡及1、2、4、6、7和8周龄肉鸡发育正常、迟缓鸡群盲肠内容物的细菌群落进行特异性引物图谱指纹和聚类分析,鉴定特异性和共性群落。结果表明,Lactobacillus属在两品种发育正常鸡群盲肠内容物细菌的相似性均高于迟缓鸡群,两品种各周龄和发育正常、迟缓鸡群间指纹图谱平均条带数差异显著(P<0.05)。而Bacteroides属在发育正常鸡群盲肠内容物细菌的相似性与迟缓鸡群较为相近;发育正常、迟缓鸡群间平均条带数差异显著(P<0.05)。Clostridium属在蛋鸡20、40周龄的平均条带数差异不显著(P>0.05),但肉鸡各周龄和发育正常、迟缓鸡群间的平均条带数差异显著(P<0.05)。序列测序结果,Lactobacillus属在蛋鸡产蛋期发育正常、迟缓鸡群均定居于Lactobacillus agilis;而育雏和育成期定居于Lactobacillus aviaries和不可培养的细菌。两品种发育正常、迟缓鸡群均定居于Bacteroides属的Bacteroides acidifaciens、Uncultured bacterium,而发育正常、迟缓蛋鸡群存在Clostridium属Uncultured proteobacterium,发育正常肉鸡群定居Shigella sonnei,而两品种发育迟缓鸡群均缺乏此类菌种。结果显示,依莎褐蛋鸡和艾维茵肉鸡群各饲养阶段盲肠细菌群落差异显著,对鸡群正常生长发育影响较大。 |
| 关键词: 鸡 盲肠 菌落 变性梯度凝胶电泳(DGGE) |
| DOI：10.11841/j.issn.1007-4333.2011.04.019 |
| 投稿时间：2011-02-10 |
| 基金项目:山东省教育厅科技计划项目(J06K58); 临沂大学博士科研启动基金(BS07011); 日本学生支援机构归国外国留学生短期研究制度基金项目(JP22GR037) |
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| Analysis of chicken cecal microbial flora the structure growthand development of PCR-DGGE |
|
LI Yong-zhu1, Yongquan_Cui2
|
| (1.College of Life Science,Linyi University,Linyi 276005,China;2.Laboratory of Animal Genetics,Okayama University,Okayama 700-8530,Japan) |
| Abstract: |
| This research is to study the effects of Normal,slow development layers and broiler on cecal bacterial population structure and diversity.To identify specific and common community members,cecal bacterial population structure and diversity in 4-,6-,10-,16-,20-,and 40-week-old commercial layers and 1-,2-,4-,6-,7-,and 8-week-old broiler chickens of Normal development,slow comparatively studied using PCR-DGGE of 16S rDNA,combined with cloning and sequencing of DNA recovered from specific and common tapping bands of PCR production with specific primers.Lactobacillus-specific primers and cluster analysis showed more similarity of cecal bacteria in the chicken of both varieties of Normal than in those of slow.The average band number of PCR-DGGE fingerprints of 16S rDNA from the chicken of the same variety at different weeks and Normal development,slow of age was significantly different (P<0.05).Bacteroides-specific primers and cluster analysis exhibited more common bands and closer similarity of cecal bacteria in the two varieties of Normal,slow development.The average band numbers of PCR-DGGE fingerprints of 16S rDNA from the two varieties at different weeks of age did not display significant difference (P>0.05),but the difference between chickens of Normal,slow development was significant (P<0.05).Clostridium specific primers and cluster analysis revealed that the difference between the average fingerprint band number from the commercial layers 20- and 40-week old was not significant (P>0.05),but the difference between that from health and poor broiler chickens at different weeks of age was significantly different (P<0.05).Sequence analysis of DNA recovered from Lactobacillus-specific tapping bands of PCR production showed that cecal Lactobacillus agilis abounded in commercial layers of both Normal development,slow during laying period,Lactobacillus aviaries and unculturable bacteria were abundant at the brooding and rearing periods.Unculturable Weissella sp.and Lactobacillus animalis were plentiful in chickens of the two varieties of Normal,slow development.Sequence analysis of DNA recovered from Bacteroides-specific tapping bands of PCR production exhibited that cecal Bacteroides acidifaciens and uncultured bacterium were abundant in the two varieties of Normal development,slow.Sequence analysis of DNA recovered from Clostridium specific tapping bands of PCR production displayed that the cecum of the two varieties of Normal,slow development were plentiful of unculturable bacterium.Unculturable proteobacterium were abundant in commercial layers.Shigella sonnei existed in large quantities in healthy broiler chickens,but were absent in the two species of poor health.The results of this study showed that cecal bacterial community structure in chicken at different feeding stages was affected by chicken species and their health conditions. |
| Key words: laying hen cecal bacterial flora denaturing gradient gel electrophoresis |
