Abstract
The overall complexity of the microbial communities in the gastrointestinal (GI) tracts of mammals has hindered observations of dynamics and interactions of individual bacterial populations. However, such information is crucial for understanding the diverse disease-causing and protective roles that gut microbiota play in their hosts. Here, we determine the spatial distribution, interanimal variation, and persistence of bacteria in the most complex defined-flora (gnotobiotic) model system to date, viz., mice colonized with the eight strains of the altered Schaedler flora (ASF). Quantitative PCR protocols based on the 16S rRNA sequence of each ASF strain were developed and optimized to specifically detect as few as 10 copies of each target. Total numbers of the ASF strains were determined in the different regions of the GI tracts of three C.B-17 SCID mice. Individual strain abundance was dependent on oxygen sensitivity, with microaerotolerant Lactobacillus murinus ASF361 present at 10(5) to 10(7) cells/g of tissue in the upper GI tract and obligate anaerobic ASF strains being predominant in the cecal and colonic flora at 10(8) to 10(10) cells/g of tissue. The variation between the three mice was small for most ASF strains, except for Clostridium sp. strain ASF502 and Bacteroides sp. strain ASF519 in the cecum. A comparison of the relative distribution of the ASF strains in feces and the colon indicated large differences, suggesting that fecal bacterial levels may provide a poor approximation of colonic bacterial levels. All ASF strains were detected by PCR in the feces of C57BL/6 restricted flora mice, which had been maintained in an isolator without sterile food, water, or bedding for several generations, providing evidence for the stability of these strains in the face of potential competition by bacteria introduced into the gut.
MeSH Terms
Animals
Bacteria/classification,genetics,growth & development
Digestive System/microbiology
Ecosystem
Feces/microbiology
Female
Genes, rRNA
Germ-Free Life
Mice
Mice, Inbred C57BL
Mice, SCID
Models, Animal
Molecular Sequence Data
RNA, Ribosomal, 16S/genetics
Sequence Analysis, DNA
Chemicals
RNA, Ribosomal, 16S
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sarma-Rupavtarm Ramahi B
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Ge Zhongming
Schauer David B
Fox James G
Polz Martin F
References (31)
31 references, click to expand
-
Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut.
Appl Environ Microbiol. 1999 Nov;65(11):4799-807
PMID: 10543789
-
ASSOCIATION OF GERMFREE MICE WITH BACTERIA ISOLATED FROM NORMAL MICE.
J Exp Med. 1965 Jul 1;122:77-82
PMID: 14325475
-
rrndb: the Ribosomal RNA Operon Copy Number Database.
Nucleic Acids Res. 2001 Jan 1;29(1):181-4
PMID: 11125085
-
Molecular analysis of commensal host-microbial relationships in the intestine.
Science. 2001 Feb 2;291(5505):881-4
PMID: 11157169
-
Commensal host-bacterial relationships in the gut.
Science. 2001 May 11;292(5519):1115-8
PMID: 11352068
-
Escherichia coli strains colonising the gastrointestinal tract protect germfree mice against Salmonella typhimurium infection.
Gut. 2001 Jul;49(1):47-55
PMID: 11413110
-
DNA isolation protocols affect the detection limit of PCR approaches of bacteria in samples from the human gastrointestinal tract.
Syst Appl Microbiol. 2001 Nov;24(3):405-10
PMID: 11822677
-
Culture-independent analysis of gut bacteria: the pig gastrointestinal tract microbiota revisited.
Appl Environ Microbiol. 2002 Feb;68(2):673-90
PMID: 11823207
-
Mucosa-associated bacteria in the human gastrointestinal tract are uniformly distributed along the colon and differ from the community recovered from feces.
Appl Environ Microbiol. 2002 Jul;68(7):3401-7
PMID: 12089021
-
Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria.
Microbiology. 2002 Nov;148(Pt 11):3651-60
PMID: 12427955
-
The indigenous flora of the gastrointestinal tract.
Dis Colon Rectum. 1967 Jan-Feb;10(1):23-34
PMID: 6018347
-
The gastrointestinal epithelium and its autochthonous bacterial flora.
J Exp Med. 1968 Jan 1;127(1):67-76
PMID: 4169441
-
The gnotobiotic animal as a tool in the study of host microbial relationships.
Bacteriol Rev. 1971 Dec;35(4):390-429
PMID: 4945725
-
Human fecal flora: the normal flora of 20 Japanese-Hawaiians.
Appl Microbiol. 1974 May;27(5):961-79
PMID: 4598229
-
Microbial ecology of the gastrointestinal tract.
Annu Rev Microbiol. 1977;31:107-33
PMID: 334036
-
Protective role of intestinal flora against infection with Pseudomonas aeruginosa in mice: influence of antibiotics on colonization resistance.
Infect Immun. 1985 Jan;47(1):118-22
PMID: 2856912
-
Ecology of Candida albicans gut colonization: inhibition of Candida adhesion, colonization, and dissemination from the gastrointestinal tract by bacterial antagonism.
Infect Immun. 1985 Sep;49(3):654-63
PMID: 3897061
-
Gastrointestinal microflora in mammalian nutrition.
Annu Rev Nutr. 1986;6:155-78
PMID: 3524615
-
Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology.
Appl Environ Microbiol. 1988 May;54(5):1079-84
PMID: 3389805
-
Basic local alignment search tool.
J Mol Biol. 1990 Oct 5;215(3):403-10
PMID: 2231712
-
In vivo and in vitro models of the human colonic flora.
Crit Rev Food Sci Nutr. 1992;31(4):299-331
PMID: 1581008
-
Colonization resistance.
Antimicrob Agents Chemother. 1994 Mar;38(3):409-14
PMID: 8203832
-
Phylogenetic identification and in situ detection of individual microbial cells without cultivation.
Microbiol Rev. 1995 Mar;59(1):143-69
PMID: 7535888
-
Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples.
Appl Environ Microbiol. 1995 Aug;61(8):3069-75
PMID: 7487040
-
Inflammatory bowel disease: an immunity-mediated condition triggered by bacterial infection with Helicobacter hepaticus.
Infect Immun. 1997 Aug;65(8):3126-31
PMID: 9234764
-
The influence of normal microbial flora on the development of chronic mucosal inflammation.
Res Immunol. 1997 Oct-Dec;148(8-9):567-76
PMID: 9588836
-
Impact of culture-independent studies on the emerging phylogenetic view of bacterial diversity.
J Bacteriol. 1998 Sep;180(18):4765-74
PMID: 9733676
-
Creating and maintaining the gastrointestinal ecosystem: what we know and need to know from gnotobiology.
Microbiol Mol Biol Rev. 1998 Dec;62(4):1157-70
PMID: 9841668
-
Noninvasive measurement of anatomic structure and intraluminal oxygenation in the gastrointestinal tract of living mice with spatial and spectral EPR imaging.
Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4586-91
PMID: 10200306
-
Phylogeny of the defined murine microbiota: altered Schaedler flora.
Appl Environ Microbiol. 1999 Aug;65(8):3287-92
PMID: 10427008
-
The role of normal flora in Giardia lamblia infections in mice.
J Infect Dis. 2000 Apr;181(4):1510-2
PMID: 10751141