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Microbial perspective on fiber utilization by swine.

Dietary fiber may contribute up to 30% of the maintenance energy needs of growing pigs. Higher energy contributions may be obtained from dietary fiber fed to sows, along with some improvements in reproduction, health, and well-being. As long as cereal grain supplies and high-quality protein supplements are abundant, the use of fibrous feeds for swine most likely will be limited. However, as the human demand for cereal grains increases, swine producers, especially those with reproductive animals, may be economically forced to incorporate alternative feedstuffs. These feedstuffs might include lignified plant cell wall material such as grasses and legumes, and feed-milling and distillery by-products that contain a high level of fiber residues. The microflora in swine large intestine will be able to adapt to these lignified forages and by-product feeds much better than the microflora in humans. Swine microflora contain highly active ruminal cellulolytic and hemicellulolytic bacterial species, which include Fibrobacter succinogenes (intestinalis), Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio spp., and Prevotella ruminicola. Additionally, a new highly active cellulolytic bacterium, Clostridium herbivorans, has been recently isolated from pig large intestine. The populations of these microorganisms are known to increase in response to the ingestion of diets high in plant cell wall material. The numbers of cellulolytic bacteria from adult animals are approximately 6.7 times greater than those found in growing pigs. None of these highly active cellulolytic bacterial species are found in the human large intestine. Thus, the pig large intestinal fermentation of fiber seems to more closely resemble that of ruminants than that of humans.

Animals↗

Candida albicans and Saccharomyces cerevisiae induce interleukin-8 production from intestinal epithelial-like Caco-2 cells in the presence of butyric acid.

Intestinal epithelial cells (IEC) are important in initiation and regulation of immune responses against numerous foreign substances including food, microorganisms and their metabolites in the intestine. Since the responses of IEC against yeasts have not yet been well understood, we investigated the effects of Candida albicans, Saccharomyces cerevisiae, and their cell wall components on interleukin-8 (IL-8) secretion by the IEC-like Caco-2 cells. Live cells of both yeast species stimulated Caco-2 cells to produce IL-8 only in the presence of butyric acid, which is a metabolite produced by intestinal bacteria. S. cerevisiae zymosan and glucan also enhanced IL-8 secretion. Treatment of Caco-2 cells with butyric acid increased the expression of mRNAs coding for Toll-like receptor 1 (TLR1), TLR6 and dectin-1, which recognize zymosan. C. albicans induced more IL-8 secretion and also decreased transepithelial electrical resistance more rapidly than S. cerevisiae. These results suggest that both yeasts in the intestine stimulate the host's mucosal immune systems by interacting with IEC.

Butyric Acid↗

[Ecologic impact of antibiotherapy. Role of substitution microorganisms in the control of antibiotic-related diarrhea and colitis].

Mild or severe episodes of diarrhoea are the main side effects of antibiotic therapy. The major form of intestinal disorders is pseudomembranous colitis due to Cl. difficile. Among ecological means for control of these diarrhoea, which result from disruption of the intestinal ecosystem, several non-pathogenic microorganisms can be used in order to establish a new equilibrium; among the available microorganisms, S. boulardii is a non-pathogenic yeast, which has given rise to many experimental and clinical studies. Moreover, results of an in vitro study of susceptibilities to antibiotics of the available microorganisms are reported; for S. boulardii, these in vitro data confirm the bases of persistence of the organism in the gut during antibiotic therapy. Besides these in vitro data, experimental studies in animal models (gnotobiotic mice and hamsters with Cl. difficile colitis) have confirmed the potential effectiveness of S. boulardii in preventing the pathogenic effect of toxins A and B and in decreasing numbers of Cl. difficile colonies in the gut. Similarly, in intensive care unit patients, controlled prospective studies have shown that administration of S. boulardii prevented diarrhoea, pseudo-membranous colitis and intestinal translocation of gram-negative bacilli when combined with selective digestive decontamination.

Animals↗

Intestinal microbiota of patients with bacterial infection of the respiratory tract treated with amoxicillin.

The intestinal tract harbors a huge diversity of metabolically-active aerobic and anaerobic bacteria that interact, forming a complex ecosystem. This microbiota has an important role in human metabolism, nutrition, immunity, and protection against colonization by pathogenic microorganisms. Several factors can influence the intestinal microbiota; these include age, diet, inflammatory and infectious processes, and the use of antimicrobials. We investigated the influence of bacterial infection of the respiratory tract and of amoxicillin therapy on the normal intestinal microbiota of patients. Bacterial infectious processes affecting the respiratory tract were found to influence the intestinal microbiota, significantly decreasing the number of colony-forming units (CFUs) of Bacteroides spp. and Lactobacillus spp. per gram of feces. The use of amoxicillin also influenced the intestinal microbiota, significantly decreasing the CFU of Bifidobacterium spp. and Lactobacillus spp./g of feces. Changes in the composition of the intestinal microbiota need to be observed, since a decrease in the normal microorganisms can pose a number of hazards for hosts, including decreased resistance to colonization. With proper follow-up, health-care teams can minimize such hazards by implementing suitable therapy- and diet-related measures, thus reducing the occurrence of detrimental effects on the gastrointestinal ecosystem.

Acute Disease↗

[Routes of E. coli 055 bacterial penetration through the intestinal wall in gnotobiotic and ordinary animals].

Peculiarities of permeability of intestinal barrier in germfree animals for enteropathogenic E. coli 055 were studied. Both germfree and conventional guinea pigs and rats were used. An increase of bacteriemia was revealed in gnotobiotes during the first day of oral E. coli 055 monocontamination; bacteriemia was transient in conventional animals. Under electron microscopy alterations of intercellular contacts and formation of spaces between enterocytes containing numerous microorganisms were found in the intestinal mucous membrane in gnotobiotes. Also more pronounced changes of microvessels of the intestinal mucous membrane were discovered in gnotobiotes. The processes of ingestionan and digestion of Escherichia coli by enterocytes and leukocytes were noted in conventional animals. The revealed derangements of the intestinal barrier in gnotobiotes explain the cause of higher bacteriemia in germfree animals. An important role of the microbial factor in the formation of intestinal barrier is indicated by the data obtained.

Animals↗

Reduction of non-digestible oligosaccharides in soymilk: application of engineered lactic acid bacteria that produce alpha-galactosidase.

Human consumption of soy-derived products has been limited by the presence of non-digestible oligosaccharides (NDO), such as the alpha-galactooligosaccharides raffinose and stachyose. Most mammals, including man, lack pancreatic alpha-galactosidase (alpha-Gal), which is necessary for the hydrolysis of these sugars. However, such NDO can be fermented by gas-producing microorganisms present in the cecum and large intestine, which in turn can induce flatulence and other gastrointestinal disorders in sensitive individuals. The use of microorganisms expressing alpha-Gal is a promising solution to the elimination of NDO before they reach the large intestine. In the present study, lactic acid bacteria engineered to degrade NDO have been constructed and are being used as a tool to evaluate this solution. The alpha-Gal structural genes from Lactobacillus plantarum ATCC8014 (previously characterized in our laboratory) and from guar have been cloned and expressed in Lactococcus lactis. The gene products were directed to different bacterial compartments to optimize their possible applications. The alpha-Gal-producing strains are being evaluated for their efficiency in degrading raffinose and stachyose: i) in soymilk fermentation when used as starters and ii) in situ in the upper gastrointestinal tract when administered to animals orally, as probiotic preparations. The expected outcomes and possible complications of this project are discussed.

Animals↗

Enteric immunization: promises and challenges.

Immunization to prevent many intestinal infections is inadequate because most available vaccines are given parenterally, a route that does not effectively stimulate the intestinal immune system. Thus, investigators are pursuing several strategies for achieving enteric protection through oral immunization. The most promising approaches are the incorporation of immunogens into microparticles for protection and enhanced uptake of the immunogen by intestinal lymphoid tissues, the genetic manipulation of microorganisms, and the use of mucosal adjuvants. The achievement of effective oral immunization against intestinal infections could lead to control of serious diarrheal illnesses, which still are a major worldwide health problem.

Administration, Oral↗

[Gastrointestinal manifestations of HIV infection].

The intestinal (in particular rectal) mucosa is the main portal of entry for HIV in homosexual men, who represent the vast majority of HIV-infected patients in Europe and North America. There are several possibilities for HIV to reach the CD4-positive T cells, macrophages and follicular-dendritic cells in the intestinal mucosa. HIV may be transported through M-cells directly to mucosal lymph follicles. Alternatively HIV may infect enterocytes via Fc-receptor by antibody-bound HIV or via a CD4-independent receptor. By successive budding on the basolateral side of the enterocytes HIV may be released into the lamina propria. The loss and functional impairment of activated CD4-positive lamina propria T-cells could be responsible for both the decreased immune defense and altered structure and function of the mucosa. The common intestinal symptoms in HIV-infected patients may be caused by a variety of mechanisms. The high number of secondary opportunistic or non-opportunistic infections and secondary malignancies of the gut may be responsible for the observed symptoms. However, the pathogenic relevance of some of these pathogens is questionable since there is often no correlation between symptoms and presence of the pathogen. In addition, there is a considerable percentage of symptomatic patients without identifiable microorganisms. Yet unidentified pathogens, small intestinal bacterial overgrowth, damage of intestinal nerve fibres, or secretory diarrhea may contribute to the pathogenesis of gastrointestinal symptoms. The findings of a pathogen-negative diarrhea, of HIV-infected mononuclear cells in the gut, and of epithelial hypoproliferation and enterocyte dysmaturation is in agreement with the hypothesis that there is an enteropathy caused by HIV itself.

AIDS-Related Opportunistic Infections↗

Adherence to lipids and intestinal mucin by a recently recognized human pathogen, Campylobacter upsaliensis.

Campylobacter upsaliensis is a recently recognized human enteric pathogen associated with enteritis, colitis, bacteremia, and sepsis. Very little is known about the mechanisms of pathogenesis of this organism. The goals of this study were to determine whether C. upsaliensis binds to epithelial cells and whether there are specific lipid molecules that might serve as cell membrane receptors. In addition, we also explored C. upsaliensis binding to purified human small-intestinal mucin, since the mucus gel overlying the epithelium provides an initial contact surface for the bacteria and must be penetrated for the organisms to reach their cell receptors. Binding of C. upsaliensis to model epithelial cells was shown by microscopy adhesion assays, and binding to lipids was detected by thin-layer chromatography-overlay assays. Bacteria bound to phosphatidylethanolamine (PE), gangliotetraosylceramide (Gg4), and, more weakly, to phosphatidylserine (PS). There was no binding to ceramide, cholesterol, phosphatidylcholine, and globosides. Using receptor-based microtiter well immunoassays, we observed binding to be equal, specific, and saturable for PE and Gg 4 but low and nonspecific for PS. At least five bacterial surface proteins (50 to 90 kDa) capable of PE binding were identified by a lipid-silica affinity column technique. In slot blot overlay assays, biotin-labeled C. upsaliensis also bound in a concentration-dependent fashion to purified human small-intestinal mucin, implying that these microorganisms also express an adhesin(s) recognizing a specific mucin epitope(s). We speculate that binding to mucins may influence access of the bacteria to cell membrane receptors and thereby influence host resistance to infection.

Adhesins, Bacterial↗

[Effect of the lavage of the digestive tract on microflora in patients with polyps in the large intestine].

The microbial status of the intestine and the influence of lavage with polyethylene glycol and balanced electrolyte solution (PEG + E), used in the process of the preparation of patients to polypectomy, on this status were evaluated. The study of microflora was made before oral lavage after, and 48-72 hours later its completion. For control, a group of healthy volunteers, also subjected to oral lavage with PEG + E, was used. The lavage of the digestive tract with PEG + E led to a sharp change in the microbial status in both groups. Some microorganisms, previously absent in the intestine, were found after lavage. The processes of the restoration of intestinal microflora after lavage in healthy volunteers and in patients with polyps had certain differences. In healthy volunteers intestinal microflora was completely restored, and even improved, 48-72 hours after lavage with PEG + E, while at the expiration of this time intestinal microflora in the patients with polyps could be characterized as dysbiotic.

Administration, Oral↗

Lactobacilli, anticarcinogenic activities and human intestinal microflora.

Lactobacilli belong to the normal oropharyngeal and intestinal microflora in humans. These microorganisms contribute to the stabilization of the microflora and maintain the colonization resistance against pathogens. Lactobacilli have been used as dietary supplements in order to prevent gastrointestinal disturbances. Claims have been made that certain strains of lactobacilli possibly exert anticarcinogenic activities. The activity of bacterial enzymes, implicated in colon carcinogenesis may be elevated by a high meat, Western-type diet. Supplements of Lactobacillus acidophilus decreased these levels in both rats and humans. Colon cancer patients given L. acidophilus fermented milk showed a significant increase both in numbers of intestinal lactobacilli and dietary calcium intake, while decreasing trends in levels of both soluble faecal bile acids and faecal bacterial enzymes, two risk makers for colon cancer, were observed. In vitro studies have revealed that lactobacilli and other lactic acid bacteria have the ability to absorb cooked food mutagens. Recent studies in humans have shown that intake of L. acidophilus significantly reduced the mutagen excretion after consumption of fried meat. Several mechanisms by which lactobacilli might exert anticarcinogenic effects are discussed. Thus, certain strains of lactobacilli might lower the colon cancer risk in humans.

Bacterial Physiological Phenomena↗

[Characteristics of experimental antibiotic-induced dysbacteriosis].

Changes in the microflora of the large and small intestines in mice and guinea pigs after the oral administration of canamycin (a hardly absorbable antibiotic) and ampiox (an easily absorbable antibiotic) in different doses. The administration of these antibiotics in different doses (therapeutic, subtherapeutic and over therapeutic) led to an increase in the number of opportunistic microorganisms and the contamination of the small intestine by these organisms. These changes were also well pronounced in guinea pigs, normally having no enterobacteria. After the administration of the antibiotics was stopped, opportunistic microorganisms were gradually eliminated from the small intestine. The rate of decontamination depended on the administered dose of the antibiotic: the higher the dose was the longer the process of the decontamination of the small intestine lasted. An increase in the amount of opportunistic microbes in the large intestine and the decontamination of the small intestine occurred simultaneously with the decrease in the amount of lactobacilli and bifidobacteria in both the small and large intestines.

Ampicillin↗

[Inflammatory bowel disease--do microorganisms play a role?].

This review focuses on the potential pathogenic role of microorganisms in relation to inflammatory bowel diseases, i.e. Crohn's disease and ulcerative colitis. Pathogenic microorganism such as Mycobacterium paratuberculosis, measles and mumps viruses, Epstein-Barr virus, and Listeria monocytogenes are discussed, as well as involvement of the normal intestinal flora. Furthermore, the influence of microorganisms in experimental animal colitis models is discussed. The available results are inconclusive, but there seems to be basis for proposing the hypothesis that the inflammation in inflammatory bowel disease reflects an immune imbalance with loss of tolerance for normally harmless antigens in the mucosal microflora.

Animals↗

Interaction of bifidobacteria with the gut and their influence in the immune function.

Bifidobacteria are predominant in the lumen of the large intestine and confer various health benefits on the host. They are also used in the preparation of new fermented milks (bioyogurts) or added to conventional yogurt to generate probiotic effects. The colonization of the gut by bacteria tends to be host specific due partly to the way in which bacteria adhere to the intestinal wall. Using a homologous strain of Bifidobacterium animalis in an experimental mouse model, we analyzed by immunofluorescence labelled-bacteria and transmission electronic microscopy the importance of the bacterial interaction with epithelial an immune cells associated to the gut, and the effect of feeding of B. animalis in the immune response. It was able to adhere and interact with both small and large intestine. In spite of this interaction with the gut, no modifications in the immune state (secretory or systemic response) were observed. A heterologous strain of Bifidobacterium adolescentis from human faeces, was neither incapable of binding to the intestine, nor influence the immune system activation, when it was administered during 2, 5 or 7 consecutive days; we believe that using a homologous strain, oral tolerance is developed even when the microorganism interacts with the immune cells associated with the intestine. However, we cannot ignore the beneficial effect of these microorganisms, especially in the prevention of intestinal infections. We think that this property exerted by bifidobacteria is more related to other mechanisms such as competitive inhibition, acid production or others, than enhancement of the immune state.

Animals↗

Influence of administered indigenous microorganisms on uptake of [iodine-125] gamma-globulin in vivo by intestinal segments of neonatal calves.

Ten calves less than 14 h of age (average 8.6 h) were anaesthetized, and the intestine was ligated into segments 10 cm in length at 3-cm intervals beginning 1.8 m anterior of the ileocecal junction and proceeding proximally. Seven treatments were assigned in random order to segments in three successive sections of the small intestine. Segments received 1 ml of viable bacteria of intestinal origin, autoclaved bacteria of intestinal origin, or sterile microbiological broth at zero time; then after 4 h they were injected with iodine-125 labeled gamma-globulin. After an additional 1.5 h, the experiment was ended and uptake assayed. Two treatments measured anaerobic microbial growth after 4 h incubation with 1 ml of either sterile broth or live bacteria culture. Residual [iodine-125] gamma-globulin was measured in segments receiving 1 ml of sterile broth or live bacteria culture with 5.5-h incubation followed by 15-s exposure to labeled gamma-globulin. Uptake was lowest in segments receiving live bacteria as compared to segments receiving sterile inocula. Number of bacteria per gram of tissue was correlated negatively with uptake. Low corticosteroids in serum were associated with low uptake of gamma-globulin.

Animals↗

[Characteristics of Escherichia serogroup O132:K isolated in intestinal diseases in monkeys].

The results obtained in the study of the main biological characteristics of 22 Escherichia strains, serogroup O132: K ., isolated from monkeys in the Sukhumi reserve are presented. For the first time Escherichia belonging to serovar O132: K .: H- have been detected; these organisms, in contrast to reference strain O132: K .: H28, possess a number of Shigella-like characteristics and are capable of intraepithelial parasitism in Shereny's test, which impedes their primary identification. The isolation of the above-mentioned cultures from live and dead monkeys with the clinical and pathologic diagnosis of catarrhal enteritis, colitis, enterocolitis, ulcerous colitis with unknown etiology, dysentery, from monkeys having had contacts with sick monkeys at the focus of clinical dysentery and in the process of prophylactic examination, as well as the pathogenicity of these strains for guinea pigs, as determined in Shigella-induced keratoconjunctivitis used as model infection, have allowed the authors to consider Escherichia, serovar O132: K .: H-, to be the etiological factor of E. coli infection in monkeys and to regard them as enteropathogenic. In this connection further research is necessary to find out the role of these microorganisms in the etiology of intestinal infections in animals; for this reason it is expedient to produce diagnostic agglutinating serum on an industrial scale.

Animals↗

Effects of roxithromycin on fecal bacteria in human volunteers and resistance to colonization in gnotobiotic mice.

The ecological impact of roxithromycin given orally at 300 mg/day on the intestinal floras in six human volunteers was studied. The resulting fecal concentrations of active roxithromycin were in the range of 100 to 200 micrograms/g of feces. Consecutive modifications in the composition of the fecal floras were limited to a decrease in counts of total members of the family Enterobacteriaceae. The rest of the intestinal floras, including the predominant anaerobic floras, changed little. No overgrowth of Pseudomonas aeruginosa, staphylococci, fungi, or highly erythromycin-resistant strains of the family Enterobacteriaceae was observed. The strains of Enterobacteriaceae and of anaerobes isolated during treatment were not markedly more resistant to roxithromycin than those isolated before treatment started. Changes in intestinal resistance to colonization by exogenous microorganisms in gnotobiotic mice inoculated with human fecal flora were studied and were also found to be minimal. The impact of oral roxithromycin on the intestinal microbiota appears to be weaker than that previously observed with oral erythromycin, perhaps because the concentrations of roxithromycin in the feces were lower than those previously found for erythromycin.

Administration, Oral↗