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PubMed2002

Developmental regulation of intestinal angiogenesis by indigenous microbes via Paneth cells.

The adult mouse intestine contains an intricate vascular network. The factors that control development of this network are poorly understood. Quantitative three-dimensional imaging studies revealed that a plexus of branched interconnected vessels developed in small intestinal villi during the period of postnatal development that coincides with assembly of a complex society of indigenous gut microorganisms (microbiota). To investigate the impact of this environmental transition on vascular development, we compared the capillary networks of germ-free mice with those of ex-germ-free animals colonized during or after completion of postnatal gut development. Adult germ-free mice had arrested capillary network formation. The developmental program can be restarted and completed within 10 days after colonization with a complete microbiota harvested from conventionally raised mice, or with Bacteroides thetaiotaomicron, a prominent inhabitant of the normal mouse/human gut. Paneth cells in the intestinal epithelium secrete antibacterial peptides that affect luminal microbial ecology. Comparisons of germ-free and B. thetaiotaomicron-colonized transgenic mice lacking Paneth cells established that microbial regulation of angiogenesis depends on this lineage. These findings reveal a previously unappreciated mechanism of postnatal animal development, where microbes colonizing a mucosal surface are assigned responsibility for regulating elaboration of the underlying microvasculature by signaling through a bacteria-sensing epithelial cell.

Animals↗
PubMed1980

[Phenomenon of intestinal microfloral increase after stopping the use of poorly adsorbed antibiotics].

The results of studying intestine microflora in conventional mice (CBA X C57 BL/F1) during enteral treatment and after discontinuation of low-adsorbing antibiotics (gentamicin, restomycin and nystatin) are presented. The average daily doses of the antibiotics were 2.4, 3 mg and 110000 units respectively. For determination of the intestine microflora composition various media were used. It was found that a 7-day exposure of the mice to the above antibiotic combination resulted in total decontamination of their intestine. After discontinuation of the antibiotic there was a marked increase in the number of Coli bacteria, i. e. 10(7) times higher than the initial level. The number of other microorganisms returned to normal without any significant increase. It was shown that when the normal Coli microflore of the mice contained equal numbers of lactose-positive and lactose-negative strains, the increase after discontinuation of the antibiotic was mainly due to the lactose-positive organisms. Reduction of the initial quantitative and qualitative composition of the Coli flora was observed 7--8 days after discontinuation of the antibiotic. The analysis of the experimental and literature data demonstrated that the marked increase in the number of the Coli forms in the intestine microflora resulted from discontinuation of antibiotic. Its universal character, connection with antagonistic relations of some microorganisms, the role of exogenic contamination and means for its decreasing are discussed.

Animals↗
PubMed1984

Impact of antimicrobial agents on the gastrointestinal microflora and the risk of infections.

The most common and significant cause of disturbances in the normal gastrointestinal microflora is the administration of antimicrobial agents. The microflora can be influenced by antimicrobial agents because of incomplete absorption of any orally administered antimicrobial agent, secretion of an antimicrobial agent by the salivary glands and in the bile, or secretion from the intestinal mucosa. In most cases the influence is not beneficial to the patient because suppression of the indigenous microorganisms often permits potential pathogens to overgrow and cause septic conditions, diarrhea, or colitis. Antimicrobial agents that influence the normal microflora also promote the emergence of antimicrobial-resistant strains. The authors' experience on the impact of different beta-lactams, erythromycin, clindamycin, tetracycline, and nitroimidazoles on the gastrointestinal microflora and the risk of infections when these agents are used is reviewed.

Anti-Bacterial Agents↗
PubMed1999

Vibrio parahaemolyticus thermostable direct hemolysin modulates cytoskeletal organization and calcium homeostasis in intestinal cultured cells.

Vibrio parahaemolyticus is a marine bacterium known to be the leading cause of seafood gastroenteritis worldwide. A 46-kDa homodimer protein secreted by this microorganism, the thermostable direct hemolysin (TDH), is considered a major virulence factor involved in bacterial pathogenesis since a high percentage of strains of clinical origin are positive for TDH production. TDH is a pore-forming toxin, and its most extensively studied effect is the ability to cause hemolysis of erythrocytes from different mammalian species. Moreover, TDH induces in a variety of cells cytotoxic effects consisting mainly of cell degeneration which often leads to loss of viability. In this work, we examined the cellular changes induced by TDH in monolayers of IEC-6 cells (derived from the rat crypt small intestine), which represent a useful cell model for studying toxins from enteric bacteria. In experimental conditions allowing cell survival, TDH induces a rapid transient increase in intracellular calcium as well as a significant though reversible decreased rate of progression through the cell cycle. The morphological changes seem to be dependent on the organization of the microtubular network, which appears to be the preferential cytoskeletal element involved in the cellular response to the toxin.

Animals↗
PubMed1992

Microbial transformation of deoxynivalenol (vomitoxin).

Microbial inocula from rumen fluid, soil, and contents of the large intestines of chickens (CLIC) and of swine (SLIC) were tested for their ability to transform deoxynivalenol (vomitoxin) in vitro. Microorganisms in (CLIC) completely transformed pure vomitoxin, and this activity was retained through six serial subcultures. No alteration of the toxin by incubation with SLIC was detected, whereas 35% of the vomitoxin was metabolized in the original culture of rumen fluid and 50% was metabolized by the soil sample, though metabolism was decreased in subsequent subcultures of either sample. A single metabolite was isolated and identified as deepoxy vomitoxin. The increase in concentration of deepoxy vomitoxin in the culture medium corresponded with the decrease in vomitoxin concentration. The vomitoxin transformation rate was not affected by either the ratio of CLIC to vomitoxin (5 to 0.2 g of CLIC per mg of vomitoxin) or the initial concentration of vomitoxin (14 to 1,400 ppm) in the medium. Biotransformation of vomitoxin was completely inhibited when the pH in the medium was lowered to 5.20. Sodium azide at a 0.1% (wt/vol) concentration in the medium blocked the transformation of vomitoxin, suggesting that the deepoxidation of vomitoxin is an energy-dependent process. About 50% of the vomitoxin in moldy corn in culture medium was transformed by microorganisms from CLIC. The vomitoxin transformation rate in moldy corn was not affected when the concentration of CLIC changed from 0.2 to 0.8 g/ml of medium. Vomitoxin in the moldy corn was not transformed when CLIC were added to corn without culture medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2005

Prevention of febrile neutropenia in cancer patients by probiotic strain Enterococcus faecium M-74. Phase II study.

Febrile neutropenia (FN) remains a potentially life-threatening complication of anticancer chemotherapy. Bacterial translocation via intestinal mucosa is a significant mechanism of FN development. Competitive inhibition of bowel colonization by pathogenic microorganisms by lactic acid bacteria could be a useful prevention of FN. The aim of the study was the prevention of FN by probiotic strain Enterococcus faecium M-74 enriched with selenium in leukemic patients. Fourteen (six males/eight females) patients with myelogenous leukemia treated by induction or consolidation chemotherapy were included in the study. Patients received prophylaxis with E. faecium M-74 during one cycle of chemotherapy. The daily dose was 36 x 10(9) CFU tid. Prophylaxis started between day -2 and day +2 of chemotherapy and continued until the absolute neutrophile count (ANC) was >1,000/microl. All patients experienced febrile neutropenia. During 231 days of severe neutropenia, 30 febrile episodes occurred. No any febrile episode or infection provoked by the strain tested was noticed. Tolerance of therapy was excellent without significant adverse effects. Our results demonstrate the safety of the probiotic strain E. faecium M-74 enriched with selenium in leukemic patients with severe neutropenia. However, its administration was not effective in the prevention of febrile neutropenia, but this does not preclude the protective effect of other probiotic strains.

Adult↗
PubMed1999

Colonization of microflora in mice: mucosal defense against luminal bacteria.

To investigate the pathogenesis of inflammatory bowel disease, it is critical to develop a system that uses simple and reproducible models for analyzing the "normal" mucosal defense mechanism. In the present study, germ-free mice were conventionalized by the oral administration of microorganisms prepared from the feces of genetically identical mice. Histological assessment and mucin characterization of small intestine and colon were then carried out. Histological findings in the gut were site-dependent and clearly time-dependent. Acute inflammation was most evident in the cecum. The cecal mucosa exhibited hyperplastic changes in epithelial cells, infiltration of polymorphonuclear cells, crypt abscesses, and epithelial projections on the epithelial surface 7 days after conventionalization. Some of the changes were similar to those seen in human ulcerative colitis. The histological findings in the conventionalized mice were comparable to those in specific pathogen-free mice after 28 days. Mucin histochemistry revealed that bacterial colonization altered the number of rectal goblet cells and the mucin composition in a time-dependent fashion. Although this model shares only some characteristics of human inflammatory bowel disease, it is unique in demonstrating the acquisition of mucosal defense. Understanding of this process is critical for the elucidation of inflammatory bowel disease pathogenesis.

Animals↗
PubMed1987

Effect of dietary fiber on colonic cell proliferation and its relationship to colon carcinogenesis.

The addition of specific fiber supplements to semipurified diets has been shown to stimulate large bowel cell proliferation in laboratory rodents. Relatively insoluble fibers such as cellulose, which is poorly fermented, the more-soluble oat bran, and inert bulking agents such as kaolin produce little or no effect on cell growth. On the other hand, wheat bran, pectin, guar gum, and degraded carageenan all stimulate large bowel cell proliferation, the greatest growth response tending to occur in the cecum or proximal colon. The proximal large bowel is also the major site for the intestinal fermentation of dietary fiber and any other nonabsorbed carbohydrates. The fermentation of fiber by colonic microorganisms results in the production of short-chain fatty acids and a lower pH of large bowel contents, metabolic events known to be associated with increased epithelial cell growth. In general, factors that stimulate cell growth also enhance tumor development, a concept that holds true in the colon even for dietary fibers such as pectin and guar gum. Wheat bran can also stimulate colon carcinogenesis when fed only during carcinogen exposure. Oat bran and corn bran may stimulate colon carcinogenesis by increasing fecal bile acid excretion, a feature of many soluble fibers, while the acidification of large bowel contents is associated with an increased frequency of chemically induced colonic cancers. A greater understanding of colonic metabolism and cell physiology is needed to define fully the mechanisms by which dietary fibers modify colon cancer development.

Animals↗
PubMed2026

Intestinal infections establish antigen-specific, long-lived memory CD4+ T cells in the brain and meninges.

The meninges form the border between the brain and periphery and house a rich network of immune cells. Here we show that gastrointestinal challenges (intracellular or extracellular bacteria and parasites) reshape the nature of CD4+ T cells in the dura mater, the outer meningeal layer, changing the dominant polarization states to T helper (TH) 1, TH17 and TH2 cells, respectively, with differing cytokine profiles. This occurs via CXCR6-CXCL16-dependent migration of gut-activated CD4+ T cells to the central nervous system, where they establish long-lived memory populations around the dural venous sinuses, within dural lymphoid aggregates and in the brain. Functionally, these orally primed dural CD4+ T were capable of rapid, antigen-specific recall responses, proliferating and producing cytokines upon intravenous rechallenge. Our findings reveal a direct link between intestinal and dural immunity, enabling the central nervous system borders to acquire immunological memory of gut microorganisms, a major source of bloodborne pathogens capable of reaching the brain via fenestrated dural vasculature.

Journal Article↗
PubMed2007

Roundtrip ticket for secretory IgA: role in mucosal homeostasis?

An important activity of mucosal surfaces is the production of Ab referred to as secretory IgA (SIgA). SIgA serves as the first line of defense against microorganisms through a mechanism called immune exclusion. In addition, SIgA adheres selectively to M cells in intestinal Peyer's patches, thus mediating the transepithelial transport of the Ab molecule from the intestinal lumen to underlying gut-associated organized lymphoid tissue. In Peyer's patches, SIgA binds and is internalized by dendritic cells in the subepithelial dome region. When used as carrier for Ags in oral immunization, SIgA induces mucosal and systemic responses associated with production of anti-inflammatory cytokines and limits activation of dendritic cells. In terms of humoral immunity at mucosal surfaces, SIgA appears thus to combine properties of a neutralizing agent (immune exclusion) and of a mucosal immunopotentiator inducing effector immune responses in a noninflammatory context favorable to preserve local homeostasis of the gastrointestinal tract.

Breast Feeding↗
PubMed1979

[Etiology of porcine dysentery].

The etiological agent of swine dysentery is an anaerobic spiralshaped microorganism of the Treponema genus. Examination of a scraping from the mucous membrane of the large intestine or the examination of faeces in a native preparation by phase contrast are the fastest diagnostical methods of determining swine dysentery. Large treponemas, as predominating microflora, 6--9 micrometer long, 0.3--0.4 micrometer in diameter, spiralshaped and moving in a serpentine way are found in faeces and the mucous membrane in the case of acute swine dysentery. These large treponemas were identified in 14% of clinically healthy pigs but in very small numbers. Treponemas are sensitive to their habitat. Treponemas survive for up to five days only in faeces which are suitably reduced. This finding is important both for subduing the infection and for diagnosis.

Animals↗
PubMed2002

Diagnosis of paratuberculosis in naturally infected goats.

A survey was carried out to verify if an immunohistochemical method associated with agar gel immunodiffusion (AGID) will establish a firm diagnosis of caprine paratuberculosis. One hundred and thirty-six goats were tested by AGID for antibodies against Mycobacterium avium subsp. paratuberculosis at two different times: the first time 22 (19.1%) were positive and the second time 25 (18%). One seronegative goat with severe diarrhea and 5 seropositive goats, two of which showing similar clinical signs, were sacrificed and necropsied. Samples were taken from small intestine, liver, spleen, mesenteric lymph nodes for bacteriological, histological and immunohistochemical examinations. M.a. paratuberculosis was isolated from intestine samples of 4 seropositive goats and from mesenteric lymph nodes of one seropositive goat; the microorganism was not isolated from samples of one seropositive and the seronegative animals. Ziehl Neelsen staining showed acid-fast bacilli in macrophages of the 5 seropositive animals and the immunohistochemical method for M. a. paratuberculosis detected bacterial antigen in the same samples.

Animals↗
PubMed2002

[The probiotic Lactobacillus acidophilus--an alternative treatment of bacterial vaginosis].

The probiotics or biotherapeutical agents are microorganisms that have an antagonistic activity regarding pathogens in vivo. Therapeutic application of these microorganisms is mostly in cases of infections of non-sterile mucous surfaces--vagina and large intestines. With the help of the probiotic L. acidophilus bacterial vaginosis can be treated successfully in a natural way. The lack of systemic side effects makes it a drug of choice in the treatment of pregnant women. More studies in this direction and development of new strategies for treatment of bacterial vaginosis in pregnant and non-pregnant women are needed.

Clinical Trials as Topic↗
PubMed1985

[Experimental diarrhea in mice induced by oral infection with an enterotoxigenic strain of E. coli].

It has been shown in experiments on adult random-bred and BALB/c Sto mice that oral infection with human strain of E. coli N-10407 induces enterocolitis characterized by marked diarrheal syndrome but without lethal outcome. The action of the strain not colonizing the intestinal mucosa of mice may be accounted for by a weak cytopathic effect of the infecting microorganism and its toxin. The data obtained indicate that adult mice may serve as a convenient model for investigation of colibacillary diarrhea induced by enterotoxigenic strain of E. coli, BALB/c Sto mice being a more sensitive model than random-bred animals.

Animals↗
PubMed1993

[Epidemiology of bacterial endocarditis in France in 1991].

In order to update our information about the incidence and demographic, microbiological and clinical characteristics of infective endocarditis (IE) in France, a 12 month long study was carried out in three regions: Ile de France, Rhône-Alpes and Lorraine. Four hundred and fifteen cases of IE were recensed: certain (32%), probable (53%) or possible (15%). The annual incidence was 24.3 per million. The average age was 56 +/- 19 years. There was no past history of cardiac disease in 34% of cases; 33% had native valvular heart disease and 22% had one or more valvular prostheses. The site of IE was mitral in 39%, aortic in 36%, tricuspid in 6% and other or multivalvular in 19% of cases. The causal microorganism was isolated in 92% of cases. It was a streptococcus in 58% of cases (S. viridans in 27%; group D streptococcus + enterococcus in 23%); a staphylococcus was isolated in 23% of cases (Staphylococcus Aureus in 18%) and another microorganism in 11% of cases. The presumed portal of entry was dental in 24%, gastro-intestinal in 13%, cutaneous in 6% and urinary in 4% of cases. Twenty patients were intravenous drug addicts. Forty-five patients had medical or surgical procedure. Twenty-four per cent of patients were operated during the first two months, 17% died during this period (15% of operated and 18% of non-operated patients). Despite the advances in antibiotic therapy and in cardiac surgical techniques, IE seems to be as common and as severe as ten years ago.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2005

Niche heterogeneity determines bacterial community structure in the termite gut (Reticulitermes santonensis).

Differences in microenvironment and interactions of microorganisms within and across habitat boundaries should influence structure and diversity of the microbial communities within an ecosystem. We tested this hypothesis using the well characterized gut tract of the European subterranean termite Reticulitermes santonensis as a model. By cloning and sequencing analysis and molecular fingerprinting (terminal restriction fragment length polymorphism), we characterized the bacterial microbiota in the major intestinal habitats - the midgut, the wall of the hindgut paunch, the hindgut fluid and the intestinal protozoa. The bacterial community was very diverse (> 200 ribotypes) and comprised representatives of several phyla, including Firmicutes (mainly clostridia, streptococci and Mycoplasmatales-related clones), Bacteroidetes, Spirochaetes and a number of Proteobacteria, all of which were unevenly distributed among the four habitats. The largest group of clones fell into the so-called Termite group 1 (TG-1) phylum, which has no cultivated representatives. The majority of the TG-1 clones were associated with the protozoa and formed two phylogenetically distinct clusters, which consisted exclusively of clones previously retrieved from the gut of this and other Reticulitermes species. Also the other clones represented lineages of microorganisms that were exclusively recovered from the intestinal tract of termites. The termite specificity of these lineages was underscored by the finding that the closest relatives of the bacterial clones obtained from R. santonensis were usually derived also from the most closely related termites. Overall, differences in diversity between the different gut habitats and the uneven distribution of individual phylotypes support conclusively that niche heterogeneity is a strong determinant of the structure and spatial organization of the microbial community in the termite gut.

Animals↗
PubMed1998

Molecular phylogenetic identification of the intestinal anaerobic microbial community in the hindgut of the termite, Reticulitermes speratus, without cultivation.

A termite maintains an anaerobic microbial community in its hindgut, which seems to be the minimum size of an anaerobic habitat. This microbial community consists of bacteria and various anaerobic flagellates, and it is established that termites are totally dependent on the microbes for the utilization of their food. The molecular phylogenetic diversity of the intestinal microflora of a lower termite, Reticulitermes speratus, was examined by a strategy that does not rely on cultivation of the resident microorganisms. Small subunit ribosomal RNA (ssrRNA) genes were directly amplified from the mixed-population DNA of the termite gut by polymerase chain reaction (PCR) and clonally isolated. Most sequenced clones were phylogenetically affiliated with the four major groups of the domain Bacteria: the Proteobacteria group, the Spirochete group, the Bacteroides group, and the Low G + C gram-positive bacteria. The 16S rRNA sequence data show that the majority of the intestinal microflora of the termite consists of new species that are yet to be cultured. The phylogeny of a symbiotic methanogen inhabiting the gut of a lower termite (R. speratus) was analyzed without cultivation. The nucleotide sequence of the ssrDNA and the predicted amino acid sequence of the mcrA product were compared with those of the known methanogens. Both comparisons indicated that the termite symbiotic methanogen belonged to the order Methanobacteriales but was distinct from the known members of this order. The diversity of nitrogen-fixing organisms was also investigated without culturing the resident microorganisms. Fragments of the nifH gene, which encodes the dinitrogenase reductase, were directly amplified from the mixed-population DNA of the termite gut and were clonally isolated. The phylogenetic analysis of the nifH amino acid sequences showed that there was a remarkable diversity of nitrogenase genes in the termite gut. The molecular phylogeny of a symbiotic hypermastigote Trichonympha agilis (class Parabasalia; order Hypermastigida) in the hindgut of R. speratus was also examined by the same strategy. The whole-cell hybridization experiments indicated that the sequence originated from a large hypermastigote in the termite hindgut, Trichonympha agilis. According to the phylogenetic trees constructed, the hypermastigote represented one of the deepest branches of eukaryotes. The hypermastigote along with members of the order Trichomonadida formed a monophyletic lineage, indicating that the hypermastigote and trichomonads shared a recent common ancestry.

Animals↗
PubMed1998

Cloning of the novel gene intelectin, which is expressed in intestinal paneth cells in mice.

Using a large-scale in situ hybridization screening method, we isolated the cDNA gene intelectin, whose mRNA is expressed in small intestinal paneth cells in mice. Northern blot analysis revealed that the mRNA corresponding to the cDNA was 1.2 kp in length, and expression was specific to the tissue in the small intestine. We termed this gene intelectin because the deduced amino acid sequence is similar to the previously cloned oocyte lectin gene of Xenopus laevis. The function of intelectin may be involved in the defence of microorganisms.

Amino Acid Sequence↗