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Effect of orally administered food-grade carrageenans on antibody-mediated and cell-mediated immunity in the inbred rat.

Experiments were performed to investigate the immunological consequences associated with the persorption of poorly degradable carregeenans from the diet. Using an inbred strain of rat it was demonstrated histochemically, by the carrageenan-specific Alcian blue staining technique, that small quantities of food-grade carrageenans given at 0.5% in drinking-water for 90 days could penetrate the intestinal barrier of adult animals. This apparently occurred via an intact mucosa in the absence of inflammatory or pathological lesions. The carrageenan was demonstrated in macrophage-like cells present within the villi and lamina propria of the small intestine. The oral administration of kappa, lambda or iota food-grade carrageenans did not affect local (biliary) or systemic antibody responses to gut commensal microorganisms, or to orally-administered sheep erythrocytes. However, when sheep red blood cells were administered parenterally the ensuing anti-sheep red blood cell haemagglutinating antibody response was temporarily suppressed in carrageenan-fed rats. lambda-Carrageenan and iota-carrageenan both significantly (P less than or equal to 0.01 and P less than or equal to 0.05, respectively) reduced the mid-phase (14-28 days) haemagglutinin response; kappa-carrageenan (L100) was less effective but caused significant depression at day 21 (P less than or equal to 0.01). Individual responses were, however, within the control range 35 days after sheep erythrocyte administration, thus indicating the temporary nature of this effect. Although carrageenan administration depressed the anti-sheep erythrocyte antibody response, it did not affect T-cell immune competence as measured by the popliteal lymph node assay for graft-versus-host reactivity.

Administration, Oral↗

Selective binding and transcytosis of Ulex europaeus 1 lectin by mouse Peyer's patch M-cells in vivo.

The in vivo interaction of the lectin Ulex europaeus agglutinin 1 with mouse Peyer's patch follicle-associated epithelial cells was studied in the mouse Peyer's patch gut loop model by immunofluorescence and electron microscopy. The lectin targets to mouse Peyer's patch M-cells and is rapidly endocytosed and transcytosed. These processes are accompanied by morphological changes in the M-cell microvilli and by redistribution of polymerised actin. The demonstration of selective binding and uptake of a lectin by intestinal M-cells in vivo suggests that M-cell-specific surface glycoconjugates might act as receptors for the selective adhesion/uptake of microorganisms.

Actins↗

Characterisation of Pseudomonas aeruginosa isolated from freshwater culture systems.

Members of the genus Pseudomonas are important phytopathogens and agents of human infections, while other strains and species exhibit bioremediation and biocontrol activities. Species-specific detection of Pseudomonas species in the environment may help to gain a more complete understanding of the ecological significance of these microorganisms. The objective of present study was comparative analysis of biochemically and PCR based confirmed 10 isolates of Pseudomonas aeruginosa (6 from fish intestine and 4 from pond sediment). PCR-ribotyping and PAGE revealed that there was extensive heterogeneity at the genetic and protein levels. Both genetic and phenotypic heterogeneity were more in the sediment isolates compared to the fish isolates. SDS-PAGE clearly demonstrated the differences between fish and sediment isolates as evident from the higher range of protein profiling. In antibiotic sensitivity test no habitat specific antibiogram was obtained. Zinc adversely affected the DNA of all the isolates to be amplified by PCR as DNA banding pattern was different from normal DNA in stressed DNA. Thus stress, particularly, zinc may interfere monitoring of Pseudomonas by PCR.

Animals↗

Immunonutrition: role of biosurfactants, fiber, and probiotic bacteria.

Phospholipids constitute an important part of cellular membranes, and membrane fluidity and permeability are dependent on the fatty acid composition of the phospholipid. The composition, which changes with aging and disease is, to a large degree, influenced by nutrient supply. Phospholipids have been effective in protecting cellular membranes such as those of the gastrointestinal tract to an extent not much different from that observed with external supply of established mucosa-protective drugs such as misoprostol and sucralfate. Polar lipids have also been shown to be effective in preventing microbial translocation. The effect is further potentiated by an external supply of probiotic fibers such as pectin, guar gum, and oat gum. These and many other fibers also have documented strong mucosa preventive effects. Prebiotic bacteria such as Lactobacillus plantarum have demonstrated a strong ability to preserve food and prevent spoilage. In addition, L. plantarum seems to not only preserve key nutrients such as omega-3 fatty acids, but also increases its content during storage conditions. L. plantarum alone or in combination with various fibers has demonstrated a strong ability to reduce and eliminate potentially pathogenic microorganisms both in vitro and in vivo. It has recently been shown that L. plantarum possesses the ability to adhere to and colonize intestinal mucosa. It seems unique among the lactobacilli for L. plantarum to use mannose-specific adhesins, uncommon among gram-positive, but common among gram-negative bacteria, which makes it possible that L. plantarum competes with gram-negative other potential pathogens for receptor sites at the mucosal cell surfaces. Additionally, L. plantarum seems to be effective in eliminating nitrate and producing nitric oxide. These functions of L. plantarum are among the reasons why it has been used in combination with various fibers and polar lipids to recondition the gastrointestinal mucosa. For the purpose of a L. plantarum-containing formula being produced and tried, a treatment policy is regarded as an extension of the immunonutrition program and called ecoimmunonutrition.

Bacteria↗

Modulation of gut physiology through enteric toxins.

Diarrheal diseases caused by microorganisms and their toxins are a major cause of mortality and morbidity throughout the world. Acute diarrhea is mainly caused due to increased intestinal secretion, commonly as a result of infection with enterotoxin producing organisms (enterotoxigenic Escherichia coli, Vibrio cholera) or due to decreased intestinal absorption from infection with organisms that damage the intestinal epithelium (enteropathogenic E. coli sp., Shigella sp., Salmonella sp.) The studies of the impact of enteric pathogens and their virulence factors exert their effect by producing toxins, called bacterial toxins. The protein toxins are produced by diverse group of bacteria. Most of the bacterial toxins exert their effect through involvement of ADP-ribosylation proteins; otherwise essential for several cellular functions while other toxins involve guanylate cyclase systems or calcium and protein kinases for their ultimate action.

Animals↗

Rumen metabolism and absorption of a 14C-labelled elastomeric copolymer and its value as a roughage substitute for cattle.

Several synthetic elastomeric and plastomeric polymers were tested for suitability as artificial roughages. They were fed to rumenfistulated cattle fed grain only. Several of the polymers were regurgitated, remasticated and reswallowed, and they formed thin strands of intermeshed fiber that produced a large, loosely woven hay-like mass that floated on the rumen contents. An elastomeric polymer consisting of copolymers of 80 to 90% ethylene and 10 to 20% propylene, with a tensile strength at yield of 45.7 kg/cm2, a hardness of 30 units (Shore D hardness scale) and a tensile strength at 300% elongation of 51.0 kg/cm2, was selected for further testing. The copolymer was fed at about 90 g/head daily for 127 days to cattle fed grain only. At slaughter, rumens contained an average of 8.0 kg copolymer (dry basis). Cattle fed the copolymer had healthier rumen papillae and epithelia of the abomasum and small intestines than did control animals fed grain only. Using 14C-labeled copolymer, we found that the copolymer was not degraded by rumen microorganisms or acid-pepsin solution. When 14C-labeled copolymer was fed to milking cows, no 14C activity was found in milk, blood or urine. Upon slaughter, about 100% of the 14C activity was recovered from digesta and feces. We concluded that the copolymer was not absorbed from the digestive tract.

Absorption↗

Spontaneous bacterial peritonitis: pathogenesis, diagnosis, treatment.

Due to inadequate defence mechanisms, cirrhotic patients with ascites have an increased susceptibility to infections, the most frequent and the most severe one being spontaneous bacterial peritonitis (SBP). SBP diagnosis is based on testing of the ascitic fluid obtained by paracentesis. A polymorphonuclear cell count of more than 250 cells/mm3 of ascitic fluid is considered diagnostic and from cultures of ascitic fluid only one germ should be isolated. 60% of the SBP episodes are produced by gram negative enteric bacilli - E. coli and Klebsiella spp. being the most frequent isolated microorganisms. The most important pathogenic mechanism for SBP is bacterial translocation. In liver cirrhosis, three mechanisms are proposed for the pathogenesis of SBP: intestinal bacterial overgrowth, the alterations (structural and functional) of the intestinal mucosal barrier and the deficiencies of the local immune response. The most appropriate antibiotic treatment is a third generation cephalosporin (Cefotaxim or Ceftriaxon) which should be administrated for 5 days. With early start of the antibiotic treatment, the short-term prognosis of cirrhotic patients with SBP has improved significantly. Unfortunately, the long term prognosis remains extremely poor due to the severity of subjacent liver disease.

Anti-Bacterial Agents↗

Manipulation of the crop and intestinal flora of the newly hatched chick.

The ceca of newly hatched chicks obtained either from commercial hatcheries or from a carefully controlled experimental hatchery have been shown to contain high numbers of a variety of microorganisms. Fecal streptococci, clostridia, enterobacteria, pediococci, and occasionally Pseudomonas aeruginosa have all been isolated, but never lactobacilli. After the chick has been on feed for 1 day, the numbers of lactobacilli in the crops and ceca are quite variable; by the 3rd day, however, large numbers are present throughout the alimentary tract. Lactobacilli could be established after 1 day by incorporating them in the feed or by treating the chicks with 0.5 ml of a broth culture. Spraying the eggs before hatching did not lead to their early establishment. Lactobacilli given alone to the newly hatched chick failed to prevent the establishment of Salmonella. However, when the chicks were given a complex mixture of facultative anaerobes and anaerobes which had been isolated from an adult bird, cecal colonization with Salmonella was prevented. The possible importance of the facultative anaerobes, particularly Streptococcus faecalis, in lowering the redox potential to encourage the growth of the anaerobes is discussed.

Animal Feed↗

Chymotrypsinlike activity of some intestinal bacteria.

Some tests of exocrine pancreatic function are based on the estimation of intestinal chymotryptic activity by using specific substrates such as N-benzoyl-L-tyrosyl-PABA (BT-P). In this study the action of a number of bacteria isolated from human or monkey intestines on BT-P was investigated. The monkeys studied had an intestinal flora closely resembling that of man. Except for Bacteroides, Proteus vulgaris No. 73, Proteus morgani No. 202, and Pseudomonas aeruginosa No. 76, significant splitting of BT-P did not take place when the common enteric microorganisms were tested. It was concluded that with the possible exception of bacterial overgrowth of the small bowel, microbial chymotryptic activity is not of importance.

4-Aminobenzoic Acid↗

[Chronic enteropathy of unknown etiology in patients with AIDS. An analysis of 40 cases].

BACKGROUND: Data about the etiology of chronic enteropathy in AIDS patients are scarce and are very dependent upon the geographical area. The aim of this study was to detect microorganisms potentially associated with chronic enteropathy in AIDS patients with diarrhoea for more than one month, and initial negative routine stool bacterial cultures and examinations for ova and parasites. The degrees of associated intestinal malabsorption and immunodeficiency were also analysed. PATIENTS AND METHODS: Forty consecutive patients were recruited from January 1993 to December 1994. The following studies were performed: Intestinal absorption tests (d-xylose and 14C-triolein), CD4/CD8 cell counts, microbiological studies (standard stool cultures for detection of bacteria and examinations for ova and parasites including the detection of Enterocitozoon bieneusi spores by the Weber's stain), upper gastrointestinal endoscopy or colonoscopy with intestinal biopsies and blood cultures for CMV and mycobacteria. RESULTS: The median duration of diarrhoea was 4 months and the mean weight loss was 8.4 kg. Ninety percent of patients had less than 0.1 x 10(9) CD4+ cells/l, with a mean CD4+ cell count of 0.035 x 10(9)/l. Malabsorption was found in 84% of patients. An etiological diagnosis of chronic enteropathy was reached in 60% of the patients. The yield of pathological examination was 37% and the microbiological test using samples of faeces and blood were positive in 45% and 20% of cases respectively. The most frequently identified microorganisms were CMV (10 cases), E. bieneusi (9), enterobacteria (8), Cryptosporidium parvum (5), Leishmania donovani (2). Patients with enteropathy caused by E. bieneusi had lower count of CD4 cells (p = 0.005) and with higher serum levels of alkaline phosphatase (p = 0.02) than patients with CMV enteropathy. CONCLUSIONS: Stool Weber's stain and CMV and mycobacterial blood cultures should be added to the standard work-up diagnosis in patients with chronic diarrhoea and a CD4+cells count below 0.1 x 10(9) l. Upper and/or lower gastrointestinal endoscopies with intestinal biopsies should be performed only in patients with persistent diarrhea without microbiological diagnosis or a lack of response to treatment.

Acquired Immunodeficiency Syndrome↗

[Opsonizing activity of the serum of gnotobiotic guinea pigs contaminated with individual representatives of intestinal microflora].

The influence of contamination of germfree guinea pigs with individual representatives of the intestinal microflora (Bac. mesentericus, Bac. subtilis, S. albus, and S. faecalis) on the formation of the serum opsonic activity was studied. An increase of the opsonic activity to all the microorganisms on the 11th day after a corresponding monocontamination and a stimulating influence of the serum on the intracellular digestion of Bac. mesentericus and Bac. subtilis microbes was noted. As to the pathogenic microorganisms (E. coli 055), S. Faecalis only were capable of stimulating the serum opsonic activity. The results indicated the presence of an association between the microflora composition and the opsonic activity of the animal blood serum. The value of this index also depended on the properties of the phagocytosis object.

Animals↗

Reovirus type I infection of small intestinal epithelium in suckling mice and its effect on M cells.

In 10-day-old mice, reovirus serotype I (reo I) selectively adheres to the apical surface of M cells and penetrates the intestinal epithelium via M cells overlying Peyer's patches before causing disseminated infection. Recently, reo I enteritis has been described in adult mice. We wished to determine if reo I enteritis also occurs in suckling mice and, if so, to determine which major epithelial cell types become infected and where the virus enters epithelial cells other than M cells. Transmission electron microscopy revealed that after oral inoculation of 10-day-old mice, peak infection of M cells preceded that of absorptive and undifferentiated crypt cells. The percentage of M cells in the dome epithelial cell population was reduced more than 4-fold between 4 and 72 hours after reo I inoculation compared with saline-inoculated mice. By 6 days after inoculation, reo I replication was no longer observed and there was a more than 2-fold increase in M cells overlying Peyer's patches domes of reo I-inoculated mice compared with saline controls. By 13 days, control and infected mice had similar percentages of M cells. When incubated with isolated intestinal epithelial sheets, reo I adhered selectively to and was endocytosed via the basal plasma membrane of absorptive cells. Thus, reo I initially penetrates the intestinal epithelium via the apical surface of M cells which become infected. Virions subsequently enter absorptive and crypt cells via their basal surfaces. During the first 3 days of enteritis, the M cell population becomes markedly depleted which may affect the permeability of the mucosal barrier to microorganisms and other antigens as well as influence the host immune response.

Animals↗

[Mycoplasmas and AIDS].

AIDS is a complex illness due to HIV type 1 and 2 infection. It is characterized by an important immunodeficiency mainly caused by depletion of CD4+ T lymphocytes. The reasons for this depletion have not been sufficiently clarified yet. In 1986, Shy Ching Lo astonished the scientific community with reported evidence concerning the direct role played by mycoplasma in the etiopathology of AIDS. Since then, different theories have pointed to mycoplasma as cofactors, commensals or opportunistic agents. Although in vivo and in vitro experiments are controversial they suggest a possible mechanism that would explain the synergism between both agents: the mycoplasma belonging to normal intestinal flora could move to urethra, oropharynx or blood due to high risk sexual practice. There it would proliferate favoured by early immunological disorders related to HIV. It has been speculated that several microorganisms including mycoplasma, acting as superantigens, could induce a chronic CD4+ and CD8+ T lymphocytes activation resulting in apoptosis of the infected lymphocytes. The release of cytokines induced by mycoplasma could influence the progression of the disease.

Acquired Immunodeficiency Syndrome↗

A review on the impact of 4-quinolones on the normal oropharyngeal and intestinal human microflora.

During the last few years the impact of the newer 4-quinolones, ciprofloxacin, enoxacin, norfloxacin, ofloxacin and pefloxacin, on the human microflora has been studied by several investigators. This review article summarizes the published data concerning these studies. The results show that the oropharyngeal flora is affected only slightly or not at all by the 4-quinolones. All the newer 4-quinolones have a similar effect on the normal intestinal flora. The gram-negative aerobic flora is strongly suppressed during administration of 4-quinolones, while the gram-positive flora is only slightly affected. The anaerobic microflora is hardly affected at all. The emergence of resistant bacterial strains is uncommon, although one study shows increased MIC-values for anaerobes during ciprofloxacin administration. Replacement by yeasts or other inherently resistant microorganisms does not often seem to be a problem. High concentrations of the 4-quinolones are reached in faeces, values between 100-2,200 mg/kg being reported. Since the 4-quinolones do not cause marked ecological disturbances in the intestinal microflora, they may be suitable for selective decontamination in immunocompromised patients, for prophylaxis of urinary tract infections and for treatment of bacterial intestinal infections.

Ciprofloxacin↗

Anti-inflammatory potential of the probiotic dietary supplement Lactibiane Tolérance: in vitro and in vivo considerations.

BACKGROUND & AIMS: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. In inflammatory bowel disease (IBD), where major modifications of the intestinal microflora have been reported, there is an increasing interest in modulating the flora with probiotic products. This work addresses the anti-inflammatory potential of Lactibiane Tolérance, a probiotic dietary supplement (mixture of four strains) using in vitro and in vivo approaches. METHODS: Comparison of the four individual strains and the commercial product reconstituted from them was conducted by in vitro tests (cytokine release after 24h stimulation of human peripheral blood mononuclear cells (PBMC)). The potential immunomodulatory characteristics of Lactibiane Tolérance were determined in vivo in an acute mice model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Assessment of colitis included blinded histological and macroscopic scores. RESULTS: We showed that Lactibiane Tolérance has anti-inflammatory properties in vitro by stimulating IL-10 production and in vivo by conferring a significant protective effect in the TNBS-induced colitis model (more than 50% decrease of colitis symptoms, P<0.01). CONCLUSIONS: These results demonstrate that a probiotic dietary supplement, Lactibiane Tolérance, can significantly prevent the initial injury of TNBS and could stimulate the initiation of clinical trials in IBD.

Animals↗

Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens.

The gastrointestinal tract is a complex ecosystem that associates a resident microbiota and cells of various phenotypes lining the epithelial wall expressing complex metabolic activities. The resident microbiota in the digestive tract is a heterogeneous microbial ecosystem containing up to 1 x 10(14) colony-forming units (CFUs) of bacteria. The intestinal microbiota plays an important role in normal gut function and maintaining host health. The host is protected from attack by potentially harmful microbial microorganisms by the physical and chemical barriers created by the gastrointestinal epithelium. The cells lining the gastrointestinal epithelium and the resident microbiota are two partners that properly and/or synergistically function to promote an efficient host system of defence. The gastrointestinal cells that make up the epithelium, provide a physical barrier that protects the host against the unwanted intrusion of microorganisms into the gastrointestinal microbiota, and against the penetration of harmful microorganisms which usurp the cellular molecules and signalling pathways of the host to become pathogenic. One of the basic physiological functions of the resident microbiota is that it functions as a microbial barrier against microbial pathogens. The mechanisms by which the species of the microbiota exert this barrier effect remain largely to be determined. There is increasing evidence that lactobacilli and bifidobacteria, which inhabit the gastrointestinal microbiota, develop antimicrobial activities that participate in the host's gastrointestinal system of defence. The objective of this review is to analyze the in vitro and in vivo experimental and clinical studies in which the antimicrobial activities of selected lactobacilli and bifidobacteria strains have been documented.

Animals↗