Culture-independent identification of gut bacteria correlated with the onset of diabetes in a rat model

Bacteria associated with the onset of type 1 diabetes in a rat model system were identified. In two experiments, stool samples were collected at three time points after birth from bio-breeding diabetes-prone (BB-DP) and bio-breeding diabetes-resistant (BB-DR) rats. DNA was isolated from these sample...

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Veröffentlicht in:The ISME Journal 2009-05, Vol.3 (5), p.536-548
Hauptverfasser: Roesch, Luiz FW, Lorca, Graciela L, Casella, George, Giongo, Adriana, Naranjo, Andres, Pionzio, Arianna M, Li, Nan, Mai, Volker, Wasserfall, Clive H, Schatz, Desmond, Atkinson, Mark A, Neu, Josef, Triplett, Eric W
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container_end_page 548
container_issue 5
container_start_page 536
container_title The ISME Journal
container_volume 3
creator Roesch, Luiz FW
Lorca, Graciela L
Casella, George
Giongo, Adriana
Naranjo, Andres
Pionzio, Arianna M
Li, Nan
Mai, Volker
Wasserfall, Clive H
Schatz, Desmond
Atkinson, Mark A
Neu, Josef
Triplett, Eric W
description Bacteria associated with the onset of type 1 diabetes in a rat model system were identified. In two experiments, stool samples were collected at three time points after birth from bio-breeding diabetes-prone (BB-DP) and bio-breeding diabetes-resistant (BB-DR) rats. DNA was isolated from these samples and the 16S rRNA gene was amplified using universal primer sets. In the first experiment, bands specific to BB-DP and BB-DR genotypes were identified by automated ribosomal intergenic spacer analysis at the time of diabetes onset in BB-DP. Lactobacillus and Bacteroides strains were identified in the BB-DR- and BB-DP-specific bands, respectively. Sanger sequencing showed that the BB-DP and BB-DR bacterial communities differed significantly but too few reads were available to identify significant differences at the genus or species levels. A second experiment confirmed these results using higher throughput pyrosequencing and quantitative PCR of 16S rRNA with more rats per genotype. An average of 4541 and 3381 16S rRNA bacterial reads were obtained from each of the 10 BB-DR and 10 BB-DP samples collected at time of diabetes onset. Nine genera were more abundant in BB-DP whereas another nine genera were more abundant in BB-DR. Thirteen and eleven species were more abundant in BB-DP and BB-DR, respectively. An average of 23% and 10% of all reads could be classified at the genus and species levels, respectively. Quantitative PCR verified the higher abundance of Lactobacillus and Bifidobacterium in the BB-DR samples. Whether these changes are caused by diabetes or are involved in the development of the disease is unknown.
doi_str_mv 10.1038/ismej.2009.5
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In two experiments, stool samples were collected at three time points after birth from bio-breeding diabetes-prone (BB-DP) and bio-breeding diabetes-resistant (BB-DR) rats. DNA was isolated from these samples and the 16S rRNA gene was amplified using universal primer sets. In the first experiment, bands specific to BB-DP and BB-DR genotypes were identified by automated ribosomal intergenic spacer analysis at the time of diabetes onset in BB-DP. Lactobacillus and Bacteroides strains were identified in the BB-DR- and BB-DP-specific bands, respectively. Sanger sequencing showed that the BB-DP and BB-DR bacterial communities differed significantly but too few reads were available to identify significant differences at the genus or species levels. A second experiment confirmed these results using higher throughput pyrosequencing and quantitative PCR of 16S rRNA with more rats per genotype. 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subjects Animals
Bacteria
Bacteria - classification
Bacteria - isolation & purification
Bacteroides
Bifidobacterium
Biodiversity
Biomedical and Life Sciences
Cluster Analysis
Deoxyribonucleic acid
Diabetes Mellitus
DNA
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
DNA, Ribosomal - chemistry
DNA, Ribosomal - genetics
DNA, Ribosomal Spacer - chemistry
DNA, Ribosomal Spacer - genetics
Ecology
Evolutionary Biology
Gastrointestinal Tract - microbiology
Genes, rRNA
Genotypes
Lactobacillus
Life Sciences
Microbial Ecology
Microbial Genetics and Genomics
Microbiology
Molecular Sequence Data
original-article
Phylogeny
Rats
RNA, Bacterial - genetics
RNA, Ribosomal, 16S - genetics
Sequence Analysis, DNA
Sequence Homology, Nucleic Acid
title Culture-independent identification of gut bacteria correlated with the onset of diabetes in a rat model
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