应用聚合酶链式反应-变性梯度凝胶电泳技术分析健康猪唾液细菌结构及多样性  

Analysis of Structure and Diversity of Salivary Bacteria of Healthy Pigs Using Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis

作　　者：曾东[1] 颜宗凯 曾燕[1] 倪学勤[1] 王印[2] 李涛[1] 

机构地区：[1]四川农业大学动物医学院动物微生态研究中心,雅安625014 [2]四川农业大学动物生物技术中心,雅安625014

出　　处：《动物营养学报》2014年第6期1521-1526,共6页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：教育部留学回国人员科研启动基金资助项目(200835-1);四川省学术带头人培养基金资助

摘　　要：本试验旨在研究健康猪唾液细菌结构及多样性。试验采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术结合PCR-DGGE图谱及特异性条带的克隆和测序,同时采用主成分分析和聚类分析方法分析试验结果。结果表明:PCR-DGGE图谱中仔猪唾液细菌平均条带数为23条,而育肥猪为28条;育肥猪唾液细菌种群的多样性指数、均匀度和丰富度均较高,分别为3.34、0.877和28.40,而仔猪分别为3.17、0.834和23.20;聚类分析显示,虽然不同生长阶段猪唾液细菌群落具有一定差异性,但所有样品间相似性系数均高于0.66。PCR-DGGE图谱中共性条带属于发酵乳杆菌(Lactobacillus fermentum)、大肠杆菌(Escherichia coli)、Mycobacterium arosiense、厚壁菌(Firmicutes)和猪链球菌(Streptococcus suis),它们是猪口腔中的优势菌群;仔猪的特异性条带属于Neisseria dentiae和Neisseria shayeganii,而育肥猪的特异性条带主要是葡萄球菌亚种(Staphylococcus sp.)。结果提示,健康仔猪和育肥猪唾液细菌结构存在明显差异,随着猪的生长发育,唾液细菌多样性增加且趋于稳定。This experiment was conducted to study the structure and diversity of salivary bacteria of healthy pigs. The bacterial diversity was studied using polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE）, cluster analysis and principal component analysis. Some common and special bands were identified by clone and sequencing at the same time. The results showed that in PCR-DGGE profile, the average band number of piglets was 23 and the average band number of finishing pigs was 28; the diversity index, richness and evenness of finishing pigs were higher, and they were 3.34, 0.877 and 28.40, respectively, while the diversity index, richness and evenness of piglets were 3.17, 0.834 and 23.20, respectively; the cluster analysis suggested that there were some differences in salivary bacterial population during different growth stages of pigs, but the similarity coefficient in all the samples was higher than 0.66. In PCR-DGGE profile, Lactobacillus fermentum, Escherichia coli, Mycobacterium arosiense, Firmicutes and Streptococcus suis were predominant in mouth cavity of pigs; the special bacteria in saliva of piglets were Neisseria dentiae and Neisseria shayeganii, and the special bacteria in saliva of finishing pigs was Staphylococcus sp.. There is a significant difference in the diversity of salivary bacteria between piglets and finishing pigs. The diversity of salivary bacteria is increasing and gradually stabilized with the growth of pigs.

关 键 词：猪 唾液 细菌 PCR—DGGE 聚类分析 主成分分析 

分 类 号：Q938[生物学—微生物学]