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Microflora of the honeybee gastrointestinal tract.

Microorganisms in the midgut and rectum of the honeybee were enumerated and characterized. Counts of aerobic microorganisms were distinctly lower than counts of anaerobes (10(5)-10(6) viable cells per g of intestinal content vs. 10(8)-10(9) per g). Total numbers of anaerobic microorganisms were almost identical with the count of anaerobic Gram-positive acid resistant rods. A higher number of coliform bacteria and Bacillus spp. was detected in the rectum (10(5) per g). Anaerobic and aerobic microorganisms, coliforms, enterococci, Bacillus spp., Pseudomonas spp. and yeasts were found in all bees; lactobacilli, staphylococci and moulds were not found.

Animals↗

Isolation and identification of intestinal bacteria from Japanese tree frog (Hlya japonica) with the special reference to anaerobic bacteria.

The bacteria in the large intestines of eight Japanese tree frogs (Hlya japonica) were enumerated by using an anaerobic culture system. The microorganisms at approximately 3.1 x 10(9) bacteria per g (wet weight) of intestinal contents were present in the intestine of all the frogs tested. No difference of the total bacteria in the frog intestine was observed between two different incubation-temperatures (room temperature and 37 degrees C). Eleven genera and 16 species were isolated from the frog intestine. In most frogs, Bacteroides (B.) caccae and B. vulgatus were detected as the predominant organisms. Escherichia coli was also present in greater numbers in the intestine. Other bacteria isolated at high dilutions were strict anaerobes, including Fusobacterium and Clostridium. Enterococcus faecalis was frequently isolated from the frog intestine. However, four genera of Bifidobacterium, Eubacterium, Peptostreptococcus, and Lactobacillus were not isolated from the frog intestine.

Animals↗

[Consequences of antibiotic therapy to the intestinal ecosystem].

Ecological impact of antibiotherapy results from the interaction between microorganisms in the ecosystems and antibiotics at which they are exposed. The amount of antibiotics use in the world is continuously increasing. The fraction devoted to human care is only about half the total amount. There are multiple other fields of usage, in agriculture, breeding and veterinary medicine. Bacterial ecosystems exposed at antibiotherapy in man are mainly the skin and the gastrointestinal and respiratory tracts. The gastrointestinal system is quantitatively predominant and the consequences of the bacterial imbalance induced by antibiotics are potentially severe. It is the reason why it is the most extensively studied, in the literature and in the present review. The origin of resistant bacteria will be briefly discussed.

Animals↗

The role of probiotics in gastrointestinal surgery.

PURPOSE OF REVIEW: Postoperative complications in gastrointestinal elective surgery have been described as low. In certain circumstances, however, such as those including high-risk patients, the incidence is higher. In such situations, bacteremia and infectious complications may be caused by translocation of either gastrointestinal bacteria or its toxins as the result of bacterial overgrowth, loss of intestinal epithelial integrity, or immunologic compromising of the host. Attempts should be made to reduce the impact of these factors. Probiotics, which confer to the host health benefits similar to those offered by a balanced indigenous microbiota, are thought to act through a variety of mechanisms including the competition with potential pathogens for nutrients or enterocyte adhesion sites, degradation of toxins, production of antimicrobial factors, and local and systemic immunomodulation. RECENT FINDINGS: Recent studies have shown that probiotics play a role in decreasing postoperative complications in patients undergoing major gastrointestinal operations. SUMMARY: Considering the mechanisms of action of probiotics in the gastrointestinal tract, mainly in the intestine, evidence strongly suggests that the use of these microorganisms might positively affect the complication rate in patients undergoing surgery on the gastrointestinal tract.

Bacterial Translocation↗

Comparison of the concentrations of phenolic compounds in olive oils and other plant oils: correlation with antimicrobial activity.

The antimicrobial activity of different edible vegetable oils was studied. In vitro results revealed that the oils from olive fruits had a strong bactericidal action against a broad spectrum of microorganisms, this effect being higher in general against Gram-positive than Gram-negative bacteria. Thus, olive oils showed bactericidal activity not only against harmful bacteria of the intestinal microbiota (Clostridium perfringens and Escherichia coli) also against beneficial microorganisms such as Lactobacillus acidophilus and Bifidobacterium bifidum. Otherwise, most of the foodborne pathogens tested (Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica, Yersinia sp., and Shigella sonnei) did not survive after 1 h of contact with olive oils. The dialdehydic form of decarboxymethyl oleuropein and ligstroside aglycons, hydroxytyrosol and tyrosol, were the phenolic compounds that statistically correlated with bacterial survival. These findings were confirmed by testing each individual phenolic compound, isolated by HPLC, against L. monocytogenes. In particular, the dialdehydic form of decarboxymethyl ligstroside aglycon showed a potent antimicrobial activity. These results indicate that not all oils classified as "olive oil" had similar bactericidal effects and that this bioactivity depended on their content of certain phenolic compounds.

Anti-Infective Agents↗

[Effect of antibiotics on intestinal microflora and production of metabolites].

Methodical approaches to detection of relation between intestinal microflora and its metabolites are described. The microbial origin of certain compounds can be asserted by a decrease in their production after exposure to antibacterial drugs or the absence of their production in microbe-free animals. The authors consider that parallel investigation of intestinal microflora and its metabolites after exposure to various agents e.g. narrow spectrum antibiotics or specific substrates is the most accurate methodical approach to detection of their interrelations. Data on the effect of four drugs i.e. kanamycin, metronidazole, cefotaxime and bactrim on production of 10 bacterial metabolites: p-cresol, phenol, indican, acetic, propionic, butyric, isobutyric, valeric, isovaleric and caproic acids in rats are presented. Correlation between the metabolites and the intestinal microflora composition was revealed. It is concluded that detection of microorganisms responsible for production of definite metabolites requires at the maximum: (1) exposure to drugs of different spectra, (2) detection of changes in intestinal microflora by biotope++ and (3) investigation of mucosa microflora which more exactly characterizes metabolism of definite biotops.

Animals↗

Characterization of M cell development during indomethacin-induced ileitis in rats.

BACKGROUND: M cells play an important role in the intestinal immune system as they have a high capacity for transcytosis of a wide range of microorganisms and macromolecules. However, little is known about the role of M cells during intestinal inflammation. AIM: We studied M cell development during indomethacin-induced intestinal inflammation in rats. METHODS: Ileitis in rats was induced by two subcutaneous injections with indomethacin (7.5 mg/kg) given 24 h apart. Rats were sacrificed after 14 days and tissue was analysed by fluorescence microscopy and electron microscopy. M cells could be visualized by using the FITC-labelled mAb anti-cytokeratin (CK)-8 (clone 4.1.18), which was recently identified as specific M cell marker in rats. The number of cytokeratin-8 positive M cells was related to the surface of the follicle associated epithelium. For morphological studies, we used both transmission electron microscopy (T.E.M.) and scanning electron microscopy (S.E.M.). RESULTS: In non-inflamed ileum M cells were scarce. Only 4% of the follicle associated epithelium were M cells, whereas an increase of M cells up to 11% was found in inflamed follicle associated epithelium (P < 0.001). The rate of M cell induction depended on the macroscopic degree of inflammation. T.E.M./S.E.M. studies showed that in inflamed tissue most M cells underwent apoptosis with typical morphological signs. In contrast to apoptotic M cells, the neighbouring enterocytes usually appeared intact. The number of mononuclear cells below the follicle associated epithelium was significantly increased. S.E.M. studies revealed that during induced ileitis mononuclear cells migrated from the lamina propria into the gut lumen by passing through apoptotic M cells. CONCLUSIONS: During indomethacin-induced ileitis in rats the increase in M cell number in association with apoptosis of M cells may alter the intestinal barrier function. These observations may play a pivotal role in the pathogenesis of chronic intestinal inflammation, e.g. in inflammatory bowel disease.

Animals↗

Expression of antimicrobial neutrophil defensins in epithelial cells of active inflammatory bowel disease mucosa.

BACKGROUND/AIMS: The normal intestinal epithelium is increasingly being recognised as an important component of the mucosal innate protection against microorganisms. Human neutrophil defensins 1-3 (HNP 1-3) and lysozyme are components of the systemic innate immunity. The aim of this study was to investigate the expression of HNP 1-3 and lysozyme in normal and active inflammatory bowel disease (IBD) mucosa. METHODS: Mucosal tissue sections were studied by immunohistochemistry using antibodies to neutrophil defensins 1-3 and lysozyme. Extracts of purified intestinal epithelial cells were used for immunoblotting studies and antimicrobial activity against the phoP negative strain of Salmonella typhimurium. RESULTS: Surface epithelial cells strongly immunoreactive for neutrophil defensins and lysozyme were seen in active ulcerative colitis and Crohn's disease (but not normal or inactive IBD) mucosal samples. Many of these cells coexpressed both of the antimicrobial proteins. Immunoblotting studies confirmed the expression of neutrophil defensins in extracts of purified ulcerative colitis epithelial cells, which also demonstrated antimicrobial activity. CONCLUSION: HNP 1-3 and lysozyme are expressed in surface enterocytes of mucosa with active IBD and they may play an important role in intestinal host defence against luminal microorganisms.

Cell Extracts↗

Alterations in the proliferating compartment of gastric mucosa during Helicobacter pylori infection: the putative role of epithelial cells expressing p27(kip1).

The proliferating zone contains stem cells that give rise to all epithelial cells of the gastric mucosa. In the present study, we investigated the turnover of gastric epithelial cells in the proliferating zone of Helicobacter pylori-infected mucosa, with or without intestinal metaplasia, before and after eradication of the microorganism. In addition, we studied the topographical distribution of the cyclin dependent kinase inhibitor p27(Kip1), which plays a critical role in cell cycle progression and differentiation programs. Twenty-eight patients (22 male), aged 32-78 years and with dyspeptic symptoms, were endoscoped, and gastric biopsies were obtained from antrum and corpus for histopathological examination and the Campylobacter-like organisms test; eradication therapy was given to infected patients, and all patients were re-endoscoped after 105 +/- 33 days (mean +/- SD). The kinetics of gastric epithelial cells and p27(Kip1) status was assessed by means of immunohistochemistry and TUNEL (Tdt-mediated dUTP-biotin nick end labeling) assay. Twenty-one (21) of 28 patients were H. pylori positive, and 7 were found H. pylori negative and served as controls. In antrum, intestinal metaplasia was detected in 7/21 (33.3%). In H. pylori gastritis, Ki67 expression was found increased in the proliferating zone, compared with normal (P =.03); analogous results were obtained with the other proliferation markers, namely retinoblastoma protein and topoisomerase IIalpha. An inverse relationship between proliferation index and atrophy was disclosed (P =.02). A reduction in the proliferation index was observed after eradication, albeit not significant. Apoptotic epithelial cells were found significantly increased (P <.01) in H. pylori gastritis, and a significant reduction was observed after eradication (P <.01). In addition, apoptotic index was found to correlate with H. pylori density. The topographical study of p27(Kip1) revealed a p27(kip1)-positive epithelial cell population that resided deep in the proliferating zone; these cells were considered to be stem cells and were found significantly increased in areas with intestinal metaplasia (P <.05); in H. pylori gastritis, there was also an increase that did not reach statistical significance. H. pylori infection induces apoptosis and increases proliferation in the proliferating zone. The increased cellular turnover, together with the increased number of putative p27(Kip1)-positive stem cells in the context of intestinal metaplasia, provides further evidence for the role of H. pylori infection in gastric carcinogenesis.

Adult↗

Infection and mucosal injury in cancer treatment.

The oral and gastrointestinal mucosa acts as an important mechanical barrier that prevents local or systemic invasion by microorganisms. Cytotoxic chemotherapy-induced mucosal injury (MI) of oral cavity and intestinal epithelium occurs in many patients treated for malignancy. Compromise of the mucosal barrier can contribute to local invasion by colonizing microorganisms and, subsequently, to systemic infection. Historically, gram-negative bacteremia has been the most problematic bacterial infection in neutropenic patients, but its incidence has reduced over time because of the use of prophylactic antibiotics. There has been a shift in the type of infecting organisms responsible for bacteremia in these patients, from predominantly gram-negative organisms to gram-positive cocci. The viridans group of streptococci is composed of the most frequent bacterial pathogens associated with MI. When speciated, oral colonizers such as Streptococcus mitis, Streptococcus oralis, and Streptococcus sangulis II are the most frequently identified pathogens. Other systemic infections caused by vancomycin-resistant enterococci, Stenotrophomonas maltophilia, and Candida species have also been associated with MI after cancer treatment. Infection can also exacerbate MI after cancer treatment. The best recognized example is herpes simplex virus type 1 (HSV-1). Latent virus is frequently reactivated in HSV-seropositive patients; this reactivation leads to stomatitis, which can be indistinguishable from MI caused by cytoreductive therapies. Antiviral prophylaxis or treatment can control the virus-induced MI and bring about overall amelioration of MI. Recognition of this infectious cause of MI is important in order for clinicians to anticipate and minimize oral toxicity and to facilitate optimal delivery of the antineoplastic regimen.

Antineoplastic Agents↗

Events at the host-microbial interface of the gastrointestinal tract. II. Role of the intestinal epithelium in pathogen-induced inflammation.

An immense number of bacteria reside within the intestinal lumen. The task of appropriately identifying and responding to microbial threats lies primarily with the single layer of cells that line the intestinal tract. Intestinal epithelial cells have developed a number of strategies aimed at identifying microorganisms and eliciting the appropriate inflammatory response. The pathogen recognition mechanisms and the signaling and inflammatory events that ensue within the intestine are the focus of this review.

Animals↗

[Digestive and cutaneous colonization of Staphylococcus epidermidis in hairless axenic mice].

In order to determine the cutaneous and digestive colonization of S. epidermidis in germ-free HRS mice, several groups of animals were contaminated with this strain according to different methods. The bacterial cutaneous enumerations were performed after preliminary crushing of the cutaneous biopsy. In this study, whatever the method of contamination of germ-free HRS mice, the kinetics of S. epidermidis cutaneous colonization was parallel to the kinetics of intestinal colonization. The isolation of bacteria on skin was possible only when microorganisms were eliminated in faeces (6 h after contamination). During a single experiment, there was only slight variability in colonization, whether intestinal or cutaneous. Some differences could be found from one experiment to another, but all animals in the same trial showed the same cutaneous bacterial count. This experimental model is characterized by an interrelation between the intestinal and cutaneous ecosystems.

Animals↗

Microorganisms associated with abscesses of sheep and goats in the south of Iran.

A total of 86 abscesses (45 and 41) abscesses of sheep and goats, respectively) were examined for their causal agents; 44 of these abscesses were located subcutaneously, and the remaining 42 were in lungs, livers, intestines, and udders. A total of 23 different types of microorganisms were isolated from 78 abscesses; bacteria were not detected in the remaining eight abscesses. Microorganisms isolated were: species of the genera Corynebacterium, Staphylococcus, and Streptococcus and Pasteurella, Escherichia coli, and other gram-negative rods. Peptostreptococcus anaerobius and Eubacterium tortuosum were isolated in pure culture from one abscess each in goats. Two to four different microorganisms were associated with 25 of the 86 abscesses. It was determined that 11 isolates of Corynebacterium pyogenes, 2 isolates of Pasteurella haemolytica, 1 of P multocida, and 1 of C pseudotuberculosis were lethal to mice. Two of the C pyogenes isolates from subcutaneous spreading abscesses of goats were pathogenic for rabbits; these two isolates produced similar suppurative inflammation in goats experimentally, but did not cause death. As determined by experimental inoculation, goats were more susceptible than sheep to the two isolates. In nature, subcutaneous abscesses of goats associated with C pyogenes were of a more diffusive type which resulted in generalization of the infection and death of the animals. Postmortem examination of ten goats dying of field infection showed the presence of larvae of the warble-fly Przhevalskiana silenus at the site of infection.

Abscess↗

Intestinal colonisation, microbiota and future probiotics?

The human intestine is colonized by a large number of microorganisms, collectively termed microbiota, which support a variety of physiological functions. As the major part of the microbiota has not yet been cultured, molecular methods are required to determine microbial composition and the impact of specific dietary components including probiotics. Probiotics are viable microbial food supplements, which have a beneficial impact on human health. Health-promoting properties have been demonstrated for specific probiotic products. The most significant demonstrations for probiotic efficacy include prevention and treatment of antibiotic associated diarrhea, rotavirus diarrhea and allergy prevention. Lactobacillus rhamnosus GG (=ATCC 53103) and Bifidobacterium lactis Bb12 are the among the best-characterized and most studied probiotic strains with demonstrated impact on human health. New complex targets for probiotics include irritable bowel syndrome and Helicobacter pylori infection. For future probiotics the most important target is a demonstrated clinical benefit supported by knowledge on the mechanistic actions in the microbiota of the target population. Molecular and genomics-based knowledge of the composition and functions of the microbiota, as well as deviations from the balanced microbiota, will advance the selection of new and specific probiotics. Potential combinations of specific probiotics may prove to be the next step to reduce the risk on intestinal diseases and reconstruct specific microbial deviations.

Bifidobacterium↗

Lectin histochemical characterisation of the porcine small intestine around weaning.

The present study was undertaken to characterise the carbohydrate profile of the porcine small intestine using lectin histochemistry during the period from 3 days prior to weaning to 9 days post-weaning. A total of 56 piglets weaned at 4 weeks of age were included in the experiment. The most prominent changes in the glycosylation pattern were observed in the goblet cells. The highest lectin reactivity of the goblet cells in the crypts was observed 7 days post-weaning which suggests that the protective effect of the mucus layer against pathogenic bacteria is increasing during the postweaning period. The staining pattern of the apical membrane remained unchanged during the experimental period. This indicates that the glycosylation process in the goblet cells is rapidly inducible whereas changes in the glycosylation pattern of the apical membrane requires more time. The glycosylation pattern of both goblet cells and apical membrane differed between the positions of the small intestine. 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.

Animals↗

[Pathogenicity of nonagglutinating vibrios].

In studying the pathogenicity of nonagglutinating vibrios it was established that the majority of the strains isolated from the patients suffering from enteritis possessed enteropathogenic properties which were revealed in the trials on nursling rabbits and on the isolated intestinal loop a of an adult rabbit. In difference to the cholera vibryos, these microorganisms produced no typical cholerogenicity syndrome expressing, however, a number of enteropathogenic properties: caused diarrhea, overfilling of the intestine with fluid, etc. Autopsy showed a typical enterocolitis picture, confirmed by histological studies. Nonagglutinating vibrio cultures isolated from the water and from healthy persons possessed no enteropathogenic properties. An isolated intestinal loop of an adult rabbit proved to be the most sensitive experimental model.

Animals↗

Microorganisms associated with epithelial surfaces and stability of the indigenous gastrointestinal microflora.

Indigenous microorganisms of many genera and species associate with mucosal epithelia in the gastrointestinal tracts of animals and humans. These mechanisms may involve a high degree of specificity for host and surface habitat. They may include a capacity of the microorganisms to adhere to the membranes of substratum epithelial cells, to colonize and utilize as a source of nutrients the mucus overlying epithelial cells, and to be motile and attracted into the mucous layer by chemotaxis. The microbes must be able as well to thrive in the nutritional and environmental conditions prevailing on the epithelial surfaces. Microbial communities associated with epithelial surfaces are critical for maintaining a microflora in areas of the tract (i.e., stomach, small intestine) where the lumenal content moves at a rate exceeding the maximum rates at which indigenous microorganisms can multiply. Such communities even may be important in areas (i.e., the cecum and colon) where the content moves at rates below those at which the microbes can multiply. In such areas, microorganisms colonizing mucus on the epithelium and in the crypts of Lieberkuhn may provide a stable inoculum for the lumenal content which may be altered in composition in times of dietary change. Microorganisms associated with gastric and intestinal surfaces undoubtedly serve in a major way to stabilize the composition of the indigenous gastrointestinal microflora. At the molecular level, however, little is known about the mechanisms stabilizing the composition or the biochemical and genetic activities of the microflora. Such mechanisms are important subjects for research in the future.

Animals↗

Lipopolysaccharide of Escherichia coli, polyamines, and acetic acid stimulate cell proliferation in intestinal epithelial cells.

Our aim was to examine whether lipopolysaccharide of Escherichia coli, polyamines of dietetic and/or bacterial origin, and products of the bacterial metabolism influence cell proliferation in epithelial cells from the colon and small intestine. Lipopolysaccharide of Escherichia coli 0111:B4 was incubated with cultures from human colonic mucosa. The mitoses were arrested with Vincristine and the total number of metaphases per crypt was counted. In addition, lipopolysaccharide was incubated with a human colonic epithelial cell line from adenocarcinoma (LS-123 cells) and with a nontransformed small intestinal cell line from germ-free rats (IEC-6 cells) for 24 h. In the last 4 h, the cells were labeled with tritiated thymidine. The cells were incubated with putrescine, cadaverine, and spermidine at 10(-11)-10(-3) M and with acetic acid (10(-5)-10(-1) M), acetaldehyde (10(-10)-10(-4) M) and ammonium chloride (1-20 mM). Lipopolysaccharide of Escherichia coli increased the number of arrested metaphases in human colonic crypts and DNA synthesis in L-123 and IEC-6 cells (P < 0.001). All polyamines increased DNA synthesis in the colonic and small intestinal cell lines, the effects being more marked for putrescine (P < 0.001). The higher concentrations of acetic acid increased DNA synthesis in both epithelial cell lines (P < 0.001). Acetaldehyde slightly decreased DNA synthesis in LS-123 cells at cytotoxic concentrations. Ammonium chloride did not significantly affect DNA synthesis. The final concentration of nonionized ammonia was less than 3%. It is concluded that lipopolysaccharides of Escherichia coli and intraluminal factors derived from microorganisms increase cell proliferation in human colonic crypts and intestinal epithelial cell lines.

Acetaldehyde↗