Analysis of the large bowel microbiota of colitic mice using PCR/DGGE

Bibiloni, Rodrigo; Simon, Mary Alice; Albright, Carol A.; Sartor, Balfour R.; Tannock, Gerald W.

doi:10.1111/j.1472-765x.2005.01720.x

Cited by 53 publications

(43 citation statements)

References 36 publications

(72 reference statements)

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“…There was no correlation between the Stx2 concentration in vivo and the severity of disease in mice (Table 2). (4,21,31). When conventional or SPF mice are inoculated with live EHEC, bacteria are detectable only briefly in the feces, and bacterial counts fall to undetectable levels within a few days (7,24,29,40).…”

Section: %)mentioning

confidence: 99%

Pathogenesis of Renal Disease Due to Enterohemorrhagic Escherichia coli in Germ-Free Mice

et al. 2008

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Enterohemorrhagic Escherichia coli (EHEC) is a food-borne pathogen that causes hemorrhagic colitis and acute renal failure. We used a germ-free mouse model to investigate the role of host factors, Shiga toxin 2 (Stx2), and bacterial strain in disease due to EHEC. Germ-free male and female Swiss-Webster mice that were 3 days to 12 weeks old were orally inoculated with 1 of 10 EHEC strains or derivatives of two of these strains with Stx2 deleted. All inoculated mice became infected regardless of the inoculum dose. All bacterial strains colonized the intestines, reaching levels of 10 9 to 10 12 CFU/g of feces by 4 days after inoculation. Seven of the 10 wild-type strains caused disease. However, the two Stx2 deletion mutants, unlike the Stx2 ؉ parental strains, did not cause disease. The clinical signs of disease in mice included lethargy, dehydration, polyuria, polydypsia, and death. Postmortem examination of affected mice revealed dehydration and luminal cecal fluid accumulation. Histologic examination revealed close adherence of bacteria to the intestinal epithelium in the ileum and cecum but not in the colon. Other lesions included progressive renal tubular necrosis, glomerular fibrin thrombosis, and red blood cell sludging. The severity of disease varied according to the bacterial strain and age, but not sex, of the host. This study demonstrated that EHEC colonizes germ-free mice in large numbers, adheres to the intestinal epithelium, and causes luminal cecal fluid accumulation and progressive renal failure. The disease in mice was Stx2 and bacterial strain dependent. This animal model should be a useful tool for studying the pathogenesis of renal disease secondary to EHEC infection.Enterohemorrhagic Escherichia coli (EHEC), a food-borne bacterial pathogen of humans, is a normal inhabitant of the ruminant intestine. This organism is most often transmitted to humans in undercooked meat, but it has also caused outbreaks associated with contaminated vegetables (5, 28), fruit juices (6), and drinking water (41). The disease manifestations include watery or bloody diarrhea and can be severe enough to cause hospitalization and even death (18). In addition, some affected children (and rarely adults) develop hemolytic-uremic syndrome (HUS), a rapidly progressive, often fatal form of renal failure that is attributed to expression of Shiga toxins (Stx) by pathogenic bacteria (2, 18). HUS is defined by a triad of symptoms that includes hemolysis, uremia, and thrombocytopenia. It is a rare complication of disease due to EHEC, but its importance is amplified by its severity, tendency to affect young children, and the absence of any specific treatment or preventative measure. Thus, the significance of disease due to EHEC is derived from its food-borne nature and its potentially severe consequences.Both HUS and hemorrhagic colitis caused by EHEC have been attributed to the production of Stx by pathogenic bacteria (2, 50) referred to as Stx-producing E. coli (STEC). The Stxs are macromolecular cytotoxins that are phage encode...

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Section: %)mentioning

confidence: 99%

Pathogenesis of Renal Disease Due to Enterohemorrhagic Escherichia coli in Germ-Free Mice

et al. 2008

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“…Wohlgemuth et al (2009) found no correlation between increasing E. coli cell numbers in Il10 -/-mice and colitis severity. Another study showed no increase in E. coli numbers in germ-free Il10 -/-mice colonized with faecal bacteria from healthy wild-type mice (Bibiloni et al, 2005).…”

Section: Effect Of Host Genotype On Caecal Bacteriamentioning

confidence: 99%

“…Both human clinical studies and animal studies have shown differences in the bacterial communities of the intestine between disease and control subjects, which suggest the involvement of luminal bacteria and bacterial products in the initiation and progression of intestinal inflammation (Mangin et al, 2004;Bibiloni et al, 2005). Antibiotic treatment decreased concentrations of intestinal bacteria and attenuated colitis in CD patients (Colombel et al, 1997;Gui et al, 1997) …”

Section: Introductionmentioning

confidence: 99%

Diversity of caecal bacteria is altered in interleukin-10 gene-deficient mice before and after colitis onset and when fed polyunsaturated fatty acids

et al. 2010

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Interleukin-10 gene-deficient (Il10 –/–) mice show a hyper-reaction to normal intestinal bacteria and develop spontaneous colitis similar to that of human Crohn's disease when raised under conventional (but not germ-free) conditions. The lack of IL10 protein in these mice leads to changes in intestinal metabolic and signalling processes. The first aim of this study was to identify changes in the bacterial community of the caeca at 7 weeks of age (preclinical colitis) and at 12 weeks of age (when clinical signs of colitis are present), and establish if there were any changes that could be associated with the mouse genotype. We have previously shown that dietary n-3 and n-6 polyunsaturated fatty acids (PUFA) have anti-inflammatory effects and affect colonic gene expression profiles in Il10 –/– mice; therefore, we also aimed to test the effect of the n-3 PUFA eicosapentaenoic acid (EPA) and the n-6 PUFA arachidonic acid (AA) on the bacterial community of caeca in both Il10 –/– and C57 mice fed these diets. The lower number of caecal bacteria observed before colitis (7 weeks of age) in Il10 –/– compared to C57 mice suggests differences in the intestinal bacteria that might be associated with the genotype, and this could contribute to the development of colitis in this mouse model. The number and diversity of caecal bacteria increased after the onset of colitis (12 weeks of age). The increase in caecal Escherichia coli numbers in both inflamed Il10 –/– and healthy C57 mice might be attributed to the dietary PUFA (especially dietary AA), and thus not be a cause of colitis development. A possible protective effect of E. coli mediated by PUFA supplementation and associated changes in the bacterial environment could be a subject for further investigation to define the mode of action of PUFA in colitis.

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“…PCR-DGGE for 16S rRNA Gene 16S rRNA gene fragments were amplified from stool DNA with the forward primer HDA1-GC clamp and the reverse primer HDA2 21 (Supplementary Table 2). PCR reactions were performed for 4 min at 94 1C, with 30 cycles of 30 s at 94 1C, 30 s at 56 1C, and 1 min at 68 1C, and then a final step for 7 min at 68 1C.…”

Section: Dna Preparations From Mouse Stool Samplesmentioning

confidence: 99%

Butyric acid attenuates intestinal inflammation in murine DSS-induced colitis model via milk fat globule-EGF factor 8

Mishiro

Kusunoki

Otani

et al. 2013

Laboratory Investigation

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Butyric acid, a short-chain fatty acid and one of the main metabolites of intestinal microbial fermentation of dietary fiber, has been shown to have an important role in maintaining the integrity of the intestinal mucosa, while it also has been shown to exert potent anti-inflammatory effects both in vitro and in vivo. However, the precise mechanisms underlying those effects have not been fully identified. We exposed colonic epithelial cells to butyric acid, then extracted total RNA samples, and subsequently hybridized them to microarray chips. Among the upregulated genes, milk fat globuleepidermal growth factor 8 (MFG-E8) was elevated by approximately fivefold. We previously reported that the potential therapeutic benefits of MFG-E8 in intestinal tissue injury were dependent not only on enhanced clearance of apoptotic cells but also required diverse cellular events for maintaining epithelial integrity. The influence of butyric acid on cell function is often attributed to its inhibition of histone deacetylases (HDACs). We found that acetylation on histone 3 lysine 9 (acetyl-H3K9) around the MFG-E8 promoter was significantly increased with butyric acid exposure. Experimental colitis was induced by administration of dextran sodium sulfate (DSS) in C57BL/6N (MFG-E8 þ / þ ) and MFG-E8 À / À mice. Although the colonic bacterial compositions in wild-type (WT) and MFG-E8 À / À mice were not significantly different, intrarectal administration of butyric acid during an acute phase of colitis attenuated intestinal inflammatory parameters and inhibited body weight loss in the WT mice. Our novel findings suggest that butyric acid has significant antiinflammatory effects partly via MFG-E8 on DSS-induced murine experimental colitis.

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Analysis of the large bowel microbiota of colitic mice using PCR/DGGE

Cited by 53 publications

References 36 publications

Pathogenesis of Renal Disease Due to Enterohemorrhagic Escherichia coli in Germ-Free Mice

Pathogenesis of Renal Disease Due to Enterohemorrhagic Escherichia coli in Germ-Free Mice

Diversity of caecal bacteria is altered in interleukin-10 gene-deficient mice before and after colitis onset and when fed polyunsaturated fatty acids

Butyric acid attenuates intestinal inflammation in murine DSS-induced colitis model via milk fat globule-EGF factor 8

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