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A Strong Impact of Genetic Background on Gut Microflora in Mice.

Esworthy RS, Smith DD, Chu FF - Int J Inflam (2010)

Bottom Line: We used diet to modulate ileocolitis; a casein-based defined diet with AIN76A micronutrients (AIN) attenuates inflammation compared to conventional LabDiets.We found that mouse strain had the strongest impact on the composition of microbiota than diet and GPx genotypes.From 129 DKO strictly, we found overgrowth of Escherichia coli.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Beckman Research Institute of the City of Hope, 1500 Duarte Road, Duarte, CA 91010-3000, USA.

ABSTRACT
Genetic background affects susceptibility to ileocolitis in mice deficient in two intracellular glutathione peroxidases, GPx1 and GPx2. The C57BL/6 (B6) GPx1/2 double-knockout (DKO) mice have mild ileocolitis, and 129S1/Sv (129) DKO mice have severe inflammation. We used diet to modulate ileocolitis; a casein-based defined diet with AIN76A micronutrients (AIN) attenuates inflammation compared to conventional LabDiets. Because luminal microbiota induce DKO ileocolitis, we assessed bacterial composition with automated ribosomal intergenic-spacer analysis (ARISA) on cecal DNA. We found that mouse strain had the strongest impact on the composition of microbiota than diet and GPx genotypes. In comparing AIN and LabDiet, DKO mice were more resistant to change than the non-DKO or WT mice. However, supplementing yeast and inulin to AIN diet greatly altered microflora profiles in the DKO mice. From 129 DKO strictly, we found overgrowth of Escherichia coli. We conclude that genetic background predisposes mice to colonization of potentially pathogenic E. coli.

No MeSH data available.


Related in: MedlinePlus

Box and whisker plots of the levels of cecal E. coli (large colonies on LB plates under aerobic conditions) and Enterococcus sp. (small colonies on LB plates; E. faecalis,  E. hirae, or E. gallinarum) for mice on AIN diet. Bacteria counts are based on colony forming units per gram of cecal contents (CFU/g) at 24 hours, 37°C. Representative colonies were stabbed with a micropipet tip and transferred to ARISA PCR mix. The PCR products were resolved on agarose gels for comparison to laboratory E. coli and the PCR products were further isolated and sequenced for final identification of species.
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Related In: Results  -  Collection


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fig9: Box and whisker plots of the levels of cecal E. coli (large colonies on LB plates under aerobic conditions) and Enterococcus sp. (small colonies on LB plates; E. faecalis, E. hirae, or E. gallinarum) for mice on AIN diet. Bacteria counts are based on colony forming units per gram of cecal contents (CFU/g) at 24 hours, 37°C. Representative colonies were stabbed with a micropipet tip and transferred to ARISA PCR mix. The PCR products were resolved on agarose gels for comparison to laboratory E. coli and the PCR products were further isolated and sequenced for final identification of species.

Mentions: The prediction that the 129 N10 DKO mice would show overgrowth of E. coli was substantiated by counts of cultures from cecal contents on LB plates (Figure 9). E. coli was detected in 28 of 31 mice on AIN diet at an average of ~1 × 109 CFU/g cecal contents. On LabDiet, 6 of 6 DKO mice had detectable E. coli at an average ~1.5 × 109 CFU/g. WT mice had average E. coli levels of ~1 × 106 CFU/g, detected in 3 of 12 mice on AIN diet, which are at the lower limit of detection.


A Strong Impact of Genetic Background on Gut Microflora in Mice.

Esworthy RS, Smith DD, Chu FF - Int J Inflam (2010)

Box and whisker plots of the levels of cecal E. coli (large colonies on LB plates under aerobic conditions) and Enterococcus sp. (small colonies on LB plates; E. faecalis,  E. hirae, or E. gallinarum) for mice on AIN diet. Bacteria counts are based on colony forming units per gram of cecal contents (CFU/g) at 24 hours, 37°C. Representative colonies were stabbed with a micropipet tip and transferred to ARISA PCR mix. The PCR products were resolved on agarose gels for comparison to laboratory E. coli and the PCR products were further isolated and sequenced for final identification of species.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2957666&req=5

fig9: Box and whisker plots of the levels of cecal E. coli (large colonies on LB plates under aerobic conditions) and Enterococcus sp. (small colonies on LB plates; E. faecalis, E. hirae, or E. gallinarum) for mice on AIN diet. Bacteria counts are based on colony forming units per gram of cecal contents (CFU/g) at 24 hours, 37°C. Representative colonies were stabbed with a micropipet tip and transferred to ARISA PCR mix. The PCR products were resolved on agarose gels for comparison to laboratory E. coli and the PCR products were further isolated and sequenced for final identification of species.
Mentions: The prediction that the 129 N10 DKO mice would show overgrowth of E. coli was substantiated by counts of cultures from cecal contents on LB plates (Figure 9). E. coli was detected in 28 of 31 mice on AIN diet at an average of ~1 × 109 CFU/g cecal contents. On LabDiet, 6 of 6 DKO mice had detectable E. coli at an average ~1.5 × 109 CFU/g. WT mice had average E. coli levels of ~1 × 106 CFU/g, detected in 3 of 12 mice on AIN diet, which are at the lower limit of detection.

Bottom Line: We used diet to modulate ileocolitis; a casein-based defined diet with AIN76A micronutrients (AIN) attenuates inflammation compared to conventional LabDiets.We found that mouse strain had the strongest impact on the composition of microbiota than diet and GPx genotypes.From 129 DKO strictly, we found overgrowth of Escherichia coli.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Beckman Research Institute of the City of Hope, 1500 Duarte Road, Duarte, CA 91010-3000, USA.

ABSTRACT
Genetic background affects susceptibility to ileocolitis in mice deficient in two intracellular glutathione peroxidases, GPx1 and GPx2. The C57BL/6 (B6) GPx1/2 double-knockout (DKO) mice have mild ileocolitis, and 129S1/Sv (129) DKO mice have severe inflammation. We used diet to modulate ileocolitis; a casein-based defined diet with AIN76A micronutrients (AIN) attenuates inflammation compared to conventional LabDiets. Because luminal microbiota induce DKO ileocolitis, we assessed bacterial composition with automated ribosomal intergenic-spacer analysis (ARISA) on cecal DNA. We found that mouse strain had the strongest impact on the composition of microbiota than diet and GPx genotypes. In comparing AIN and LabDiet, DKO mice were more resistant to change than the non-DKO or WT mice. However, supplementing yeast and inulin to AIN diet greatly altered microflora profiles in the DKO mice. From 129 DKO strictly, we found overgrowth of Escherichia coli. We conclude that genetic background predisposes mice to colonization of potentially pathogenic E. coli.

No MeSH data available.


Related in: MedlinePlus