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Colonization of C57BL/6 Mice by a Potential Probiotic Bifidobacterium bifidum Strain under Germ-Free and Specific Pathogen-Free Conditions and during Experimental Colitis.

Grimm V, Radulovic K, Riedel CU - PLoS ONE (2015)

Bottom Line: Colonization of and persistence in the gastrointestinal tract is thus contributing to the beneficial effects of these strains.Conversely, B. bifidum S17/pMGC, i.e., a strain of human origin, adhered at significantly higher levels to human compared to murine IECs (p < 0.001).Collectively, these results suggest a selective disadvantage of B. bifidum S17/pMGC in the competition with the normal murine microbiota and an anti-inflammatory effect that requires live, metabolically active bacteria.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology and Biotechnology, University of Ulm, 89068, Ulm, Germany.

ABSTRACT
The effects of at least some probiotics are restricted to live, metabolically active bacteria at their site of action. Colonization of and persistence in the gastrointestinal tract is thus contributing to the beneficial effects of these strains. In the present study, colonization of an anti-inflammatory Bifidobacterium bifidum strain was studied in C57BL/6J mice under germ-free (GF) and specific pathogen-free (SPF) conditions as well as during dextran sulfate sodium (DSS)-induced colitis. B. bifidum S17/pMGC was unable to stably colonize C57BL/6J mice under SPF conditions. Mono-association of GF mice by three doses on consecutive days led to long-term, stable detection of up to 109 colony forming units (CFU) of B. bifidum S17/pMGC per g feces. This stable population was rapidly outcompeted upon transfer of mono-associated animals to SPF conditions. A B. animalis strain was isolated from the microbiota of these re-conventionalized mice. This B. animalis strain displayed significantly higher adhesion to murine CMT-93 intestinal epithelial cells (IECs) than to human Caco-2 IECs (p = 0.018). Conversely, B. bifidum S17/pMGC, i.e., a strain of human origin, adhered at significantly higher levels to human compared to murine IECs (p < 0.001). Disturbance of the gut ecology and induction of colitis by DSS-treatment did not promote colonization of the murine gastrointestinal tract (GIT) by B. bifidum S17/pMGC. Despite its poor colonization of the mouse GIT, B. bifidum S17/pMGC displayed a protective effect on DSS-induced colitis when administered as viable bacteria but not as UV-inactivated preparation. Collectively, these results suggest a selective disadvantage of B. bifidum S17/pMGC in the competition with the normal murine microbiota and an anti-inflammatory effect that requires live, metabolically active bacteria.

No MeSH data available.


Related in: MedlinePlus

Colonization and effect of B. bifidum S17/pMGC in DSS-induced colitis.(A) Fecal counts of B. bifidum S17/pMGC in C57BL/6J mice before during and after administration of DSS. Animals received daily doses of 2×109 CFU/animal of B. bifidum S17/pMGC starting 5 days prior to DSS challenge until to day 6 (i.e. 1 day after DSS treatment was stopped). Values are CFU/g feces and are mean ± standard deviation (n = 7 animals until day 6 and n = 3 thereafter). (B) and (C) Effect of B. bifidum S17/pMGC on DSS-induced weight loss (B) and colonic weight:length ratio (C). Mice of the DSS-challenged and B. bifidum S17-treated group (DSS/S17) are four out of seven animals shown in (A). Control mice received PBS as placebo and water with or without DSS (DSS/PBS and H20/PBS respectively, both n = 4). Values are mean ± standard error of the mean. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test analysis for each day (B) or at the end of the trial (C). Asterisks indicate levels of statistical significance differences for comparison to H2O/PBS group and letter for comparisons to DSS/PBS group (*: p < 0.05; **,a: p<0.01; ***,b: p<0.001).
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pone.0139935.g005: Colonization and effect of B. bifidum S17/pMGC in DSS-induced colitis.(A) Fecal counts of B. bifidum S17/pMGC in C57BL/6J mice before during and after administration of DSS. Animals received daily doses of 2×109 CFU/animal of B. bifidum S17/pMGC starting 5 days prior to DSS challenge until to day 6 (i.e. 1 day after DSS treatment was stopped). Values are CFU/g feces and are mean ± standard deviation (n = 7 animals until day 6 and n = 3 thereafter). (B) and (C) Effect of B. bifidum S17/pMGC on DSS-induced weight loss (B) and colonic weight:length ratio (C). Mice of the DSS-challenged and B. bifidum S17-treated group (DSS/S17) are four out of seven animals shown in (A). Control mice received PBS as placebo and water with or without DSS (DSS/PBS and H20/PBS respectively, both n = 4). Values are mean ± standard error of the mean. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test analysis for each day (B) or at the end of the trial (C). Asterisks indicate levels of statistical significance differences for comparison to H2O/PBS group and letter for comparisons to DSS/PBS group (*: p < 0.05; **,a: p<0.01; ***,b: p<0.001).

Mentions: To further investigate if changes in the microbiota or host physiology during colitis promotes colonization by B. bifidum S17/pMGC, C57BL/6J mice were pre-treated with B. bifidum S17/pMGC for 5 days and then administered DSS in drinking water for another 5 days to induce colitis. Treatment with bifidobacteria was continued until one day after DSS administration was stopped. Fecal shedding of B. bifidum S17/pMGC was monitored before, during and after DSS treatment (Fig 5A, n = 7 animals). This revealed that fecal levels of the strain were between 1 × 104 and 1 × 105 CFU/g during the pre-treatment and DSS phase of the experiment. However, numbers of fecal B. bifidum S17/pMGC quickly dropped and were below the limit of detection three days after treatment with bifidobacteria was stopped.


Colonization of C57BL/6 Mice by a Potential Probiotic Bifidobacterium bifidum Strain under Germ-Free and Specific Pathogen-Free Conditions and during Experimental Colitis.

Grimm V, Radulovic K, Riedel CU - PLoS ONE (2015)

Colonization and effect of B. bifidum S17/pMGC in DSS-induced colitis.(A) Fecal counts of B. bifidum S17/pMGC in C57BL/6J mice before during and after administration of DSS. Animals received daily doses of 2×109 CFU/animal of B. bifidum S17/pMGC starting 5 days prior to DSS challenge until to day 6 (i.e. 1 day after DSS treatment was stopped). Values are CFU/g feces and are mean ± standard deviation (n = 7 animals until day 6 and n = 3 thereafter). (B) and (C) Effect of B. bifidum S17/pMGC on DSS-induced weight loss (B) and colonic weight:length ratio (C). Mice of the DSS-challenged and B. bifidum S17-treated group (DSS/S17) are four out of seven animals shown in (A). Control mice received PBS as placebo and water with or without DSS (DSS/PBS and H20/PBS respectively, both n = 4). Values are mean ± standard error of the mean. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test analysis for each day (B) or at the end of the trial (C). Asterisks indicate levels of statistical significance differences for comparison to H2O/PBS group and letter for comparisons to DSS/PBS group (*: p < 0.05; **,a: p<0.01; ***,b: p<0.001).
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pone.0139935.g005: Colonization and effect of B. bifidum S17/pMGC in DSS-induced colitis.(A) Fecal counts of B. bifidum S17/pMGC in C57BL/6J mice before during and after administration of DSS. Animals received daily doses of 2×109 CFU/animal of B. bifidum S17/pMGC starting 5 days prior to DSS challenge until to day 6 (i.e. 1 day after DSS treatment was stopped). Values are CFU/g feces and are mean ± standard deviation (n = 7 animals until day 6 and n = 3 thereafter). (B) and (C) Effect of B. bifidum S17/pMGC on DSS-induced weight loss (B) and colonic weight:length ratio (C). Mice of the DSS-challenged and B. bifidum S17-treated group (DSS/S17) are four out of seven animals shown in (A). Control mice received PBS as placebo and water with or without DSS (DSS/PBS and H20/PBS respectively, both n = 4). Values are mean ± standard error of the mean. Statistical analysis was performed by one-way ANOVA with Bonferroni post-test analysis for each day (B) or at the end of the trial (C). Asterisks indicate levels of statistical significance differences for comparison to H2O/PBS group and letter for comparisons to DSS/PBS group (*: p < 0.05; **,a: p<0.01; ***,b: p<0.001).
Mentions: To further investigate if changes in the microbiota or host physiology during colitis promotes colonization by B. bifidum S17/pMGC, C57BL/6J mice were pre-treated with B. bifidum S17/pMGC for 5 days and then administered DSS in drinking water for another 5 days to induce colitis. Treatment with bifidobacteria was continued until one day after DSS administration was stopped. Fecal shedding of B. bifidum S17/pMGC was monitored before, during and after DSS treatment (Fig 5A, n = 7 animals). This revealed that fecal levels of the strain were between 1 × 104 and 1 × 105 CFU/g during the pre-treatment and DSS phase of the experiment. However, numbers of fecal B. bifidum S17/pMGC quickly dropped and were below the limit of detection three days after treatment with bifidobacteria was stopped.

Bottom Line: Colonization of and persistence in the gastrointestinal tract is thus contributing to the beneficial effects of these strains.Conversely, B. bifidum S17/pMGC, i.e., a strain of human origin, adhered at significantly higher levels to human compared to murine IECs (p < 0.001).Collectively, these results suggest a selective disadvantage of B. bifidum S17/pMGC in the competition with the normal murine microbiota and an anti-inflammatory effect that requires live, metabolically active bacteria.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microbiology and Biotechnology, University of Ulm, 89068, Ulm, Germany.

ABSTRACT
The effects of at least some probiotics are restricted to live, metabolically active bacteria at their site of action. Colonization of and persistence in the gastrointestinal tract is thus contributing to the beneficial effects of these strains. In the present study, colonization of an anti-inflammatory Bifidobacterium bifidum strain was studied in C57BL/6J mice under germ-free (GF) and specific pathogen-free (SPF) conditions as well as during dextran sulfate sodium (DSS)-induced colitis. B. bifidum S17/pMGC was unable to stably colonize C57BL/6J mice under SPF conditions. Mono-association of GF mice by three doses on consecutive days led to long-term, stable detection of up to 109 colony forming units (CFU) of B. bifidum S17/pMGC per g feces. This stable population was rapidly outcompeted upon transfer of mono-associated animals to SPF conditions. A B. animalis strain was isolated from the microbiota of these re-conventionalized mice. This B. animalis strain displayed significantly higher adhesion to murine CMT-93 intestinal epithelial cells (IECs) than to human Caco-2 IECs (p = 0.018). Conversely, B. bifidum S17/pMGC, i.e., a strain of human origin, adhered at significantly higher levels to human compared to murine IECs (p < 0.001). Disturbance of the gut ecology and induction of colitis by DSS-treatment did not promote colonization of the murine gastrointestinal tract (GIT) by B. bifidum S17/pMGC. Despite its poor colonization of the mouse GIT, B. bifidum S17/pMGC displayed a protective effect on DSS-induced colitis when administered as viable bacteria but not as UV-inactivated preparation. Collectively, these results suggest a selective disadvantage of B. bifidum S17/pMGC in the competition with the normal murine microbiota and an anti-inflammatory effect that requires live, metabolically active bacteria.

No MeSH data available.


Related in: MedlinePlus