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

曾东; 颜宗凯; 曾燕; 倪学勤; 王印; 李涛

doi:10.3969/j.issn.1006-267x.2014.06.012

动物营养学报 >

2014 , Vol. 26 >Issue 6: 1521 - 1526

DOI: https://doi.org/10.3969/j.issn.1006-267x.2014.06.012

分子营养 Molecular Nutrition

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

曾东 ,
颜宗凯 ,
曾燕 ,
倪学勤 ,
王印 ,
李涛

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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

收稿日期: 2013-12-30

网络出版日期: 2014-05-30

基金资助

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

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

ZENG Dong ,
YAN Zongkai ,
ZENG Yan ,
NI Xueqin ,
WANG Yin ,
LI Tao

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1. Animal Micro-Ecology Institute, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China;
2. Animal Biotechnology Center, Sichuan Agricultural University, Ya'an 625014, China

Received date: 2013-12-30

Online published: 2014-05-30

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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。

关键词： 猪; 唾液; 细菌; PCR-DGGE; 聚类分析; 主成分分析

本文引用格式

曾东 , 颜宗凯 , 曾燕 , 倪学勤 , 王印 , 李涛 . 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[J]. 动物营养学报, 2014 , 26(6) : 1521 -1526 . DOI: 10.3969/j.issn.1006-267x.2014.06.012

Abstract

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.

Key words： pig; saliva; bacteria; PCR-DGGE; cluster analysis; principal component analysis

参考文献

[1] KAZOR C E,MITCHELL P M,LEE A M,et al.Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients[J].Journal of Clinical Microbiology,2003,41(2):558-563.

[2] PASTER B J,BOCHES S K,GALVIN J L,et al.Bacterial diversity in human subgingival plaque[J].Journal of Bacteriology,2001,183(12):3770-3783.

[3] BONETTA S,BONETTA S,CARRARO E,et al.Microbiological characterization of Robiola di Roccaverano cheese using PCR-DGGE[J].Food Microbiology,2008,25(6):786-792.

[4] KOLLS J K,LINDN A.Interleukin-17 family members and inflammation[J].Immunity,2004,21(4):467-476.

[5] 吴明桃,徐晓,陈慧,等.运用DGGE分析慢性牙周炎患者唾液微生物群落结构[J].口腔医学,2012,32(2):65-68.
[6] EDWARDS J E,MCEWAN N R,TRAVIS A J,et al.16S rDNA library-based analysis of ruminal bacterial diversity[J].Antonie Van Leeuwenhoek,2004,86(3):263-281.

[7] MUYZER G,DE WAAL E C,UITTERLINDEN A G.Profiling of complex microbial population by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes encoding for 16S rRNA[J].Applied and Environmental Microbiology,1993,59(3):695-700.
[8] 刘怡然,王伟健,沈家平,等.PCR-DGGE检测早期儿童龋治疗前后口腔菌群多样性的初步研究[J].口腔生物医学,2012,3(4):201-204.
[9] 田晓娟,张志红,黄江丽,等.体外培养瘤胃微生物总DNA的提取与纯化[J].江西科学,2012,30(1):44-46,102.
[10] WALTER J,HERTEL C,TANNOCK G W,et al.Detection of Lactobacillus,Pediococcus,Leuconostoc and Weissellaspecies in human feces by using group-specific PCR primers and denaturing gradient gel electrophoresis[J].Applied and Environmental Microbiology,2001,67(6):2578-2585.

[11] 倪学勤,GONG J,YU H,等.采用PCR-DGGE技术分析蛋鸡肠道细菌种群结构及多样性[J].畜牧兽医学报,2008,39(7):955-961.
[12] 张柏林,秦贵信,孙泽威,等.仔猪胃肠道微生物菌群定植规律及其功能的研究进展[J].中国畜牧杂志,2009,45(19):66-69.
[13] 林均德,鄂禄祥,宋连喜,等.猪和兔胚胎胃肠道菌群的初步研究[J].中国畜牧杂志,2007,43(10):24-26.
[14] 高侃,汪海峰,章文明,等.益生菌调节肠道上皮屏障功能及作用机制[J].动物营养学报,2013,25(9):1936-1945.
[15] GOLDSTEIN E J C,TYRRELL K L,CITRON D M,et al.Anaerobic and aerobic bacteriology of the saliva and gingiva from 16 captive Komodo dragons (Varanus komodoensis):new implications for the "bacteria as venom" model[J].Journal of Zoo and Wildlife Medicine,2013,44(2):262-272.

[16] RUBY J D,SHIREY R E,GERENCSER V F,et al.Extracellular iodophilic polysaccharide synthesized by Neisseria in human dental plaque[J].Journal of Dental Research,1982,61(5):627-631.

[17] 于淼,仪红,孟玲娜,等.不同龋状况母子间口腔微生物群落结构分析[J].口腔医学研究,2010,26(3):387-390.
[18] 禹慧明,廖玲,陈平洁,等.断奶仔猪肠道菌群的研究[J].中国微生态学杂志,2000,12(2):81-82.
[19] JAMI E,MIZRAHI I.Composition and similarity of bovine rumen microbiota across individual animals[J].PloS One,2012,7(3):e33306.

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