Development of a real-time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs

被引:459
作者
Guo, X. [1 ]
Xia, X. [2 ]
Tang, R. [1 ]
Zhou, J. [2 ]
Zhao, H. [2 ]
Wang, K. [1 ]
机构
[1] Sichuan Agr Univ, Inst Anim Nutr, Yaan 625014, Sichuan, Peoples R China
[2] Sichuan Agr Univ, Int Ctr Future Agr Human Hlth, Yaan 625014, Sichuan, Peoples R China
关键词
Bacteroidetes; Firmicutes; lean pigs; obese pigs; real-time PCR;
D O I
10.1111/j.1472-765X.2008.02408.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aims: To investigate whether the relative abundance of the Bacteroidetes and Firmicutes divisions in pigs is different between obese and lean animals. Methods and Results: Group-specific primers were designed to target the 16S rRNA genes of Bacteroidetes and Firmicutes present in the gut. After the validation of their specificity, these primers were used in the real-time PCR quantification of all Bacteria, Firmicutes division, Bacteroidetes division and Bacteroides spp. in the faecal samples of obese and lean pigs from Banna mini-pig inbred line. The obese pigs had a similar to 61% fewer percentage (based on all Bacteria) of Bacteroidetes division (P = 0.033) and a similar to 56% fewer proportion of Bacteroides spp. (P = 0.047) than the lean pigs. The proportions of both Bacteroidetes and Bacteroides had a negative correlation (P < 0.01) with the body weight. Conclusion: The results suggested that the fat storage might affect the proportion of Bacteroidetes division in the gut. Significance and Impact of the Study: The real-time PCR assays developed for Firmicutes and Bacteroidetes will be useful for investigating the composition of gut microbiota.
引用
收藏
页码:367 / 373
页数:7
相关论文
共 23 条
[1]   Host-bacterial mutualism in the human intestine [J].
Bäckhed, F ;
Ley, RE ;
Sonnenburg, JL ;
Peterson, DA ;
Gordon, JI .
SCIENCE, 2005, 307 (5717) :1915-1920
[2]   The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis [J].
Cole, JR ;
Chai, B ;
Farris, RJ ;
Wang, Q ;
Kulam, SA ;
McGarrell, DM ;
Garrity, GM ;
Tiedje, JM .
NUCLEIC ACIDS RESEARCH, 2005, 33 :D294-D296
[3]   Reduced dietary intake of carbohydrates by obese subjects results in decreased concentrations of butyrate and butyrate-producing bacteria in feces [J].
Duncan, Sylvia H. ;
Belenguer, Alvaro ;
Holtrop, Grietje ;
Johnstone, Alexandra M. ;
Flint, Harry J. ;
Lobley, Gerald E. .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (04) :1073-1078
[4]   Diversity of the human intestinal microbial flora [J].
Eckburg, PB ;
Bik, EM ;
Bernstein, CN ;
Purdom, E ;
Dethlefsen, L ;
Sargent, M ;
Gill, SR ;
Nelson, KE ;
Relman, DA .
SCIENCE, 2005, 308 (5728) :1635-1638
[5]   Assessment of soil microbial community structure by use of taxon-specific quantitative PCR assays [J].
Fierer, N ;
Jackson, JA ;
Vilgalys, R ;
Jackson, RB .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (07) :4117-4120
[6]   DIETARY MODULATION OF THE HUMAN COLONIC MICROBIOTA - INTRODUCING THE CONCEPT OF PREBIOTICS [J].
GIBSON, GR ;
ROBERFROID, MB .
JOURNAL OF NUTRITION, 1995, 125 (06) :1401-1412
[7]  
KLEESEN B, 2001, BR J NUTR, V86, P375
[8]   Development of Bacteroides 16S rRNA gene TaqMan-based real-time PCR assays for estimation of total, human, and bovine fecal pollution in water [J].
Layton, Alice ;
McKay, Larry ;
Williams, Dan ;
Garrett, Victoria ;
Gentry, Randall ;
Sayler, Gary .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (06) :4214-4224
[9]   Culture-independent analysis of gut bacteria: the pig gastrointestinal tract microbiota revisited [J].
Leser, TD ;
Amenuvor, JZ ;
Jensen, TK ;
Lindecrona, RH ;
Boye, M ;
Moller, K .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (02) :673-690
[10]   Ecological and evolutionary forces shaping microbial diversity in the human intestine [J].
Ley, RE ;
Peterson, DA ;
Gordon, JI .
CELL, 2006, 124 (04) :837-848