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Evaluation of the components of a commercial probiotic in gnotobiotic mice experimentally challenged with Salmonella enterica subsp. enterica ser. Typhimurium.

Vitacanis((R)), a probiotic preparation containing a Lactobacillus acidophilus, an Enterococcus faecium and a Saccharomyces cerevisiae, has been developed for the prevention of intestinal disorders in dogs and cats. In the present study, these microorganisms were tested jointly or singly during experimental infection of gnotobiotic mice with Salmonella Typhimurium. Four experimental groups consisting of animals given probiotics jointly or singly and a control group consisting of germfree mice were used. The groups were treated with one or three of the microorganisms (experimental) or PBS (control) 10 days before intragastric challenge with a suspension containing about 10(2) cells of the bacterial pathogen. A higher survival (P<0.05) was observed in gnotobiotic mice given E. faecium (82%). All the animals in the other groups died after the challenge but the survival time was longer (P<0.05) for groups given all three of the microorganisms (7.4+/-2.4 days) or given only L. acidophilus (7.2+/-2.9 days) than for the control mice (4.4+/-1.1 days) and the mice that received S. cerevisiae (4.9+/-1.6 days) mice. The survival data agreed with the histopathological findings which showed more severe liver and intestinal lesions in control mice and in mice given Saccharomyces. In vitro antagonistic assays showed inhibition growth of E. faecium and S. Typhimurium around the colonies of L. acidophilus and for S. Typhimurium around the colonies of E. faecium. However, in vivo, S. Typhimurium became similarly established in the digestive tract of gnotobiotic mice at levels ranging from 10(8) to 10(10)CFU/g of feces and remained at these high levels until the animals died or were sacrificed. Among the three probiotic components of the commercial product Vitacanis((R)), E. faecium was the only one that provided protection against challenge with S. Typhimurium. Protection was not due to the reduction of the intestinal populations of the pathogenic bacteria.

Animals↗

Defensins and mucosal protection.

The gastrointestinal tract is constantly exposed to microorganisms, mainly a large and complex population of bacteria resident in the colon and distal small intestine. Although the normal host relationship with the resident luminal bacteria is often mutually beneficial, the host also requires protection against these microorganisms. Epithelial cells play a critical role in mediating these protective responses and there is increasing appreciation of the likely importance of antimicrobial peptides of the defensin family that they express. The enteric alpha-defensins (human defensins [HD]5 and 6) are expressed by Paneth cells, which are normally confined to the small intestine, but are also seen in the colon in patients with inflammatory bowel disease. Studies have shown that HD5 is stored in Paneth cell granules in precursor form and requires processing to the mature peptide. Human beta-defensin (HBD)1 is constitutively expressed in intestinal epithelial cells, whereas the expression of HBD2 is induced in inflammatory bowel disease. HBDs have also been shown to be chemotactic for immature dendritic cells and memory T cells. Thus, they may not only mediate innate immunity, but also regulate adaptive immune responses in inflammatory bowel disease.

Animals↗

Effect of fermented soy milk on the intestinal bacterial ecosystem.

AIM: To investigate the effect of fermented soy milk on human ecosystem in the intestinal tract by way of examining the population of different microorganisms isolated from fecal samples. METHODS: A crossover experimental design was applied. Twenty-eight healthy adults completed this experiment. Each subject consumed 250 mL, twice a day between meals, of either fermented soy milk or regular soy milk first for 2 wk, then switched to the other drink after 2 wk. Fecal samples were collected from all subjects every week starting from the second week to the end of the experiment. The microorganisms analyzed were Bifidobacterium spp., Lactobacillus spp., Clostridium perfringens, coliform organisms, and total anaerobic organisms. RESULTS: In the period of fermented soy milk consumption, the populations of Bifidobacterium spp. and Lactobacillus spp. increased (P<0.05) as well as the ratios of Bifidobacterium spp. and Lactobacillus spp. to Clostridium perfringens (P<0.05). The population of coliform organisms decreased (P<0.05) when subjects were in the period of fermented soy milk consumption. CONCLUSION: Intake of fermented soy milk significantly improved the ecosystem of the intestinal tract in the body by increasing the amount of probiotics.

Adult↗

Lectin histochemical investigations of the distal gut of chicks with special emphasis on the follicle-associated epithelium.

Carbohydrates on epithelial cell surfaces play an important role as attachment sites for different microorganisms like bacteria, viruses and protozoa. To obtain more information about the distribution of carbohydrates on the luminal surface along the intestine, lectin histochemical studies on different gut segments of chicks of different age groups were carried out using a panel of 13 lectins with specificities for Man, Glc, Gal, GalNAc, GlcNAc or GlcNAc oligosaccharides and Sia. Furthermore, we tried to find out whether previously reported specificities of certain lectins for M cells (membranous or multifold cells) in the bursa of Fabricius (BF) can be observed also on M cells of the intestine. As a result we were able to demonstrate binding of all lectins employed in these studies in all investigated gut segments. In some cases, the application of the same lectin led to varying staining intensities of the same histological structures in different age-groups (e.g. staining of the brush border with WGA, LEA, MAA or Conarva) or different gut segments (e.g. staining of goblet cells with CMA II, LEA and MPA). Hence, terminal carbohydrate residues of glycoconjugates on the intestinal epithelium vary depending on age and organ site. As glycoconjugates can act as attachment sites for microorganisms, these differences in the distribution of sugar residues may be one explanation for the site-specificity of certain pathogens. Furthermore, the binding of lectins to the follicle-associated epithelium (FAE) of the BF differs from that to the FAE of the intestine again stressing the site specificity of lectin binding. Thus, up to now no universal M-cell marker along the chicken intestine exists.

Animals↗

Potential role of epithelial cell-derived histone H1 proteins in innate antimicrobial defense in the human gastrointestinal tract.

In the human gastrointestinal tract, microorganisms are present in large numbers in the colon but are sparse in the proximal small intestine. In this study, we have shown that acid extracts of fresh human terminal ileal mucosal samples mediate antimicrobial activity. Following cation-exchange chromatography, one of the eluted fractions demonstrated antibacterial activity against bacteria normally resident in the human colonic lumen. This activity was further fractionated by reverse-phase high-performance liquid chromatography and identified as histone H1 and its fragments. We have also shown that in tissue sections, immunoreactive histone H1 is present in the cytoplasm of villus epithelial cells. In vitro culturing of detached (from the basement membrane) villus epithelial cells led to the release of antimicrobial histone H1 proteins, while the cells demonstrated ultrastructural features of programmed cell death. Our studies suggest that cytoplasmic histone H1 may provide protection against penetration by microorganisms into villus epithelial cells. Moreover, intestinal epithelial cells released into the lumen may mediate antimicrobial activity by releasing histone H1 proteins and their fragments.

Amino Acid Sequence↗

Topical antimicrobial prophylaxis of nosocomial pneumonia in mechanically ventilated patients. Microbiological observations.

Generally, reduction of colonization and infection with potentially pathogenic microorganisms in intensive care units (ICU) is attempted by a combination of antimicrobial agents administered topically in the digestive tract and systematically. We tested the efficacy of topical antimicrobial prophylaxis of the oropharynx and stomach administered in combination with sucralfate without systemic prophylaxis in 25 mechanically ventilated ICU patients. The regimen successfully reduced colonization with potentially pathogenic microorganisms in the oropharynx and trachea without modifying the intestinal flora. However, colonization and infections with gram-positive cocci and gram-negative rods other than Enterobacteriaceae and Pseudomonadaceae and resistant to one or both the antimicrobial agents used were observed.

Administration, Intranasal↗

[Bacteria and dietary fiber].

The presence of cellulose particles, so in-vitro studies have shown, influences the growth rate and metabolic activity of the bacteria Escherichia coli, Bifidobacterium adolescentis and Chromobacterium violaceum. After the homogenization in Waring Blendor, the growth rate has been found to be highest in a medium containing 10% cellulose particles and to show a decreasing tendency in media containing 5%, 1% and 0% cellulose. The biochemical intensity, judged by glucose consumption and intensity of nitrate reduction, showed an inverse tendency. It seems necessary to add also the factor "biochemical activity" of the microorganisms to the effect of dietary fibres in the intestinal tract.

Bifidobacterium↗

Regional community-acquired urinary tract infections in Israel: diagnosis, pathogens, and antibiotic guidelines adherence: a prospective study.

INTRODUCTION: The identification and treatment of hospitalized patients with community-acquired urinary tract infections (CAUTI) may be a challenge. The pathogens causing the infection and their relative proportions vary geographically and with time. This observational prospective study had three primary goals: (1) to estimate the likelihood of diagnosis of CAUTI upon admission; (2) to evaluate adherence to the institutional recommendations; (3) to assess the compatibility of the current local antibiotic recommendations with a pathogen's distribution and with its drug sensitivities. METHODS AND RESULTS: Two hundred and twenty-three patients with positive urinary cultures fulfilling criteria for CAUTI were studied. Only 54 (24.2%) were diagnosed as having a urinary tract infection upon admission. Approximately 90% of the patients, who were correctly diagnosed, received the institutional recommended antibiotic therapy (ofloxacin or cefuroxime). Gram-negative intestinal flora comprised 86.1% (192 patients) of the causative microorganisms. Of these, 20.3% of the pathogens demonstrated resistance to ofloxacin and 19.8% to cefuroxime. The prevalence of Escherichia coli, the most common pathogen of UTI, significantly declined in the current study, from 70.5% in 1991 to 56% in 2000. CONCLUSIONS: We observed a low sensitivity in diagnosing community-acquired urinary tract infections upon admission. In patients correctly diagnosed, the use of recommended antibiotics was high. A substantial percentage of the pathogens were resistant to the recommended antibiotics. This study stresses the need for frequent re-evaluation of the prevalence of pathogens involved in regional community-acquired urinary tract infections and the adjustment of the empirical first-line treatment accordingly.

Aged↗

[Immunological aspects of food intolerance in children during first years of life].

A study was made of the morphofunctional status and local defence of the gastrointestinal tract in 122 children aged 4 months to 6 years, suffering from food intolerance showed up by atopic dermatitis in 52 children and by chronic diarrhea in 70 children. Based on the allergological anamnesis, scarification cutaneous tests with food allergens, detection of antibodies to food antigens (RAST, HAIT) food allergy was revealed in all the children. Chronic gastroduodenitis was identified in all the children suffering from atopic dermatitis and in 95% of the children with chronic diarrhea. It should be mentioned that one-third of that group had a graver illness--diffuse duodenitis with sub-atrophy of the villi. The allergic genesis of the impairment of the gastroduodenal mucosa was confirmed. It was more remarkable in atopic dermatitis (tissue eosinophilia and high content of IgE-plasmacytes in the duodenal mucosa). The decrease of local immune defence of the mucous membrane, lactase deficiency, elevated growth of microorganisms in the duodenal contents promote the rise of intestinal barrier permeability for food antigens and enhancement of sensitization.

Age Factors↗

Colonization resistance of the human intestinal microflora: testing the hypothesis in normal volunteers.

Colonization resistance is the mechanism whereby the intestinal microflora protects itself against incursion by new and often harmful microorganisms. Some authors have claimed that colonization resistance is related to the integrity of the anaerobic flora, but this point has not been established in humans. In previous studies in our laboratory cefoxitin, piperacillin, cefoperazone or aztreonam were administered intravenously to healthy volunteers in order to study changes in the intestinal flora and acquisition of new strains. Seven of 16 antibiotic-treated subjects were colonized with gram-negative bacilli, but no correlation was observed between this colonization and the suppression of either anaerobes or any other component of the fecal flora. Marked strains of Escherichia coli and Pseudomonas aeruginosa were also administered by mouth in order to test acquisition of new bacteria. The fed bacteria were found in the stools of both antibiotic-treated and control subjects; the antibiotics had no apparent influence on the ability of these strains to colonize the intestinal tract. Our work, along with findings of others, supports the concept that colonization resistance occurs in humans and is diminished by antibiotic administration. However, it does not support the hypothesis that colonization resistance is related to the anaerobic microflora.

Anti-Bacterial Agents↗

Chronic ethanol feeding affects intestinal mucus lipid composition and glycosylation in rats.

Effect of chronic ethanol feeding on mucus lipid composition and glycosylation was investigated in rat intestine. Ethanol feeding to rats daily for 40 days enhanced cholesterol, phospholipids, triglyceride and cerebroside contents of intestinal mucus. Ethanol feeding exhibited a differential effect on various sugar contents of the mucus. Total hexose content was increased (42%) whereas hexosamines, sialic acid and fucose contents were reduced (p < 0.001). Results on incorporation of labeled sugars into mucus revealed an enhanced (p < 0.01) galactosylation and reduced (p < 0.01) glucosamination of glycoproteins. Separation of glycoproteins on SDS-PAGE showed that proteins of molecular weight >200, 118, 65 and 55 kD were affected. These were highly galactosylated and less glucosaminated in ethanol-fed animals compared to controls. Thus, ethanol feeding markedly alters the lipid composition and glycosylation process of intestinal mucus in rats. These alterations may be of pathological significance in view of the role of mucus in protection and adhesion of microorganisms.

Alcoholism↗

[Up-to-date clinical and experimental basis for the use of probiotics]

OBJECTIVE: To evaluate the potential of probiotics or biotherapeutic agents for the prevention and/or treatment of selected intestinal infections. METHODS: Medline database was searched for all relevant articles between 1990 and February 1998. Bibliographies of articles were also used. All animal experiments and placebo-controlled human studies were reviewed in order to provide information on the mechanisms of action, potential efficacy, or adverse effects of these biotherapeutic agents. RESULTS: In the first part of this review, the different mechanisms of action that are effective in the treatment of diarrhea were discussed, and they were well demonstrated in laboratory animals. The most important are: enzymatic induction of disaccharidase activity, trophic effects on the intestinal mucosa, action in blocking bacterial toxins, and also induction of the immunologic response. Therapeutic effects of probiotics in humans, mainly in the gastrointestinal tract, were reported in the second part. Placebo-controlled studies have shown that biotherapeutic agents have been used successfully in the treatment of acute diarrhea in infants, traveler s diarrhea, antibiotic-associated diarrhea, with or without Clostridium difficile-associated enterocolitis (pseudomembranous colitis), and in immunosuppression-associated diarrhea, including AIDS. Lactobacillus, Bifidobacterium and Saccharomyces boulardii were the most important biotherapeutic agents to be considered. CONCLUSIONS: Currently, there is evidence that the administration of selected microorganisms is benefic in the prevention and treatment of certain intestinal infections. According to the literature, Saccharomyces boulardii is the most important probiotic. Possible future indications were discussed, such as the probable synergic effect of many probiotics due to their different and complementary mechanisms of action. The importance of new experimental and clinical studies for the better understanding of actions and the use of probiotics in other clinical situations was emphasized.

Journal Article↗

Laboratory criteria in the diagnostics of intestinal infections of staphylococcal etiology in children.

Faecal samples were investigated from 948 children hospitalized with the diagnose of acute intestinal infection. The authors studied the isolation and properties of a broad spectrum of microorganisms from children's faeces: Salmonella, Shigella, E. coli, S. aureus, rotaviruses, Klebsiella, Morganella, Proteus. As a result, laboratory criteria were formulated of diagnosing in children intestinal infection of staphylococcal etiology.

Acute Disease↗

Effect of ampicillin-induced alterations in murine intestinal microbiota on the survival and competition of environmentally released pseudomonads.

The environmental release of genetically altered microorganisms has prompted the investigation of their potential health effects by the employment of other-than-human models. Although direct health effects are addressed, this investigation examines primarily some potential indirect health effects associated with environmentally released microorganisms. Indirect effects examined include colonization of the gastrointestinal tract, competition with the resident microbiota, and translocation of the dosed microorganisms to other organs. Pseudomonads used in this study were isolated from a commercial product marketed for environmental PCB degradation. When mice were dosed by gavage with approximately 10(9) ampicillin-resistant pseudomonads, an increase in recovery from the intestinal tract, as compared to untreated animals, was observed 48 hr after dosing. Intestinal survival of Pseudomonas aeruginosa strain BC16 was enhanced 1000-fold and that of P. maltophilia strain BC6, 10-fold. Strains BC17 and BC18 were unaffected. Ampicillin treatment had a significant effect on the relative number of microbiota in the intestine, by selecting primarily for the facultative species. The lactose-fermenting enterobacteria, obligately anaerobic predominantly Gram-negative rods, and total aerobic and anaerobic populations were monitored in the presence and absence of the PCB-degrading pseudomonad. P. aeruginosa strain BC17 and P. maltophilia strain BC6 had a dose effect (p less than 0.05) on the total aerobic and anaerobic populations as well as the lactose-fermenting enterobacteria. These results are similar to those for the mouse isolate control, strain PAMG. P. aeruginosa strain BC18 had a dose effect (p less than 0.05) on the total anaerobic population, including the obligately anaerobic Gram-negative bacilli. No translocation of the dosed strains to the liver, spleen, or lung was observed 48 hr after dosing.

Ampicillin↗

Role of M cells in intestinal barrier function.

M cells are known as specialized epithelial cells of the follicle-associated epithelium of the gastrointestinal tract. As M cells have a high capacity for transcytosis of a wide range of microorganisms and macromolecules, they are believed to act as an antigen sampling system. The primary physiological role of M cells seems to be the rapid uptake and presentation of particular antigens and microorganisms to the immune cells of the lymphoid follicle to induce an effective immune response. In contrast to absorptive enterocytes, M cells do not exert direct defense mechanisms to antigens and pathogens in the gut lumen. Therefore, they provide functional openings of the epithelial barrier. Although M cells represent a weak point of the epithelial barrier, even under noninflamed conditions, there seems to be a balance between antigen uptake and immunological response. The low number of M cells in the gastrointestinal tract and the direct contact to immune cells in the lamina propria usually prevent the occurrence of mucosal inflammation. During chronic intestinal inflammation we observe an increase of M cell number and apoptosis selectively in M cells. M cell damage seems to be responsible for the increase of the uptake of microorganisms that is observed during intestinal inflammation. Under inflammatory conditions in the intestine, the maintenance of the epithelial barrier is broken and M cells seem to play a major role during this process.

Animals↗

Hexamita meleagridis (Spironucleus meleagridis) infection in chukar partridges associated with high mortality and intracellular trophozoites.

An outbreak of infectious catarrhal enteritis, associated with the flagellated protozoan Spironucleus meleagridis (syn. Hexamita meleagridis), is reported from a commercial flock of chukar partridges in California. The disease affected birds between the ages of 4 and 6 wk and resulted in diarrhea, listlessness, depression, and high mortality. Concurrent infection with other intestinal pathogens, including Cryptosporidia, group E Salmonella, long-segmented filamentous microorganisms (LSFMOs), and Rotavirus-like virus particles, was found in some but not all affected birds. Dermatitis of the face, shanks, and feet, suggestive of B-complex vitamin deficiency, was present in most affected birds as well. Flagellated protozoan parasites could be found in the lumen of the duodenum and jejunum and in the intestinal crypts. In some cases the flagellates were wedged between epithelial cells or were located intracellularly within cells of the mucosal epithelium and the intestinal lamina propria.

Animals↗

The influence of a gastrointestinal microflora on natural killer cell activity.

These studies demonstrate that the natural cytotoxicity of BALB/c mouse spleen cells for 51Cr-labeled YAC-1 cells can be significantly enhanced by microorganisms in the alimentary tract. Spleen cells from germfree BALB/c mice, euthymic, athymic, or non-nude background (+/+), had natural cell-mediated cytotoxicity for YAC-1 cells. Intestinal colonization with a few (flora-defined) or many (complex flora-conventionalized) microorganisms significantly enhanced natural cell-mediated cytotoxicity of athymic and euthymic mice over their germfree counterparts. Conversely, colonization of the alimentary tract of athymic and euthymic germfree mice with a pure culture of Candida albicans or colonization with Candida and a Bacillus sp. did not enhance natural cell-mediated cytotoxic activity over germfree levels. Spleen cells from germfree athymic mice were significantly more cytotoxic than spleen cells from germfree BALB/c mice that did not carry the nude gene (ie, +/+). In the germfree or gnotobiotic state, no difference in natural killer cell activity was evident between athymic (nu/nu) and heterozygous (+/nu) littermate mice; however, athymic (nu/nu) flora-defined or conventionalized mouse spleen cells were significantly more cytotoxic for YAC-1 cells than splenocytes from flora-defined or conventionalized heterozygous (+/nu) littermates. Spleen cells from BALB/c mice that were athymic (nu/nu) and colonized with a complex microbial flora (ie, conventionalized) had the highest percentage of cytotoxicity, at three different effector to target ratios, for YAC-1 cells. These studies indicate that the intestinal microflora can alter murine natural cell-mediated cytotoxicity.

Animals↗

Paving the way for pathogens?

Indigenous intestinal microflora are known to afford protection against colonization by pathogenic microorganisms. However, the metabolic activity of at least one species of the indigenous microflora can induce expression of surface glycoconjugates, which may in turn confer susceptibility to infection.

Animals↗