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Appearance of 15N-labeled intestinal microbial amino acids in the venous blood of the pig colon.

Two experiments were done to determine whether pigs possess the ability to absorb amino acids synthesized from urea nitrogen by indigenous microbes in the large intestine. Incorporation of [15N]urea into amino acid fractions of bacterial cells from the rectum and of the deproteinized incubated medium were examined in an experiment in vitro. The isotope was incorporated into 17 amino acids and the ammonia fraction of these samples. The absorption of the microbial amino acids from the colon was investigated by determination of the 15N concentration of the free amino acids in the venous blood of the colon after infusion of the 15N-labeled microorganisms into the cecum. The increase of 15N concentration was also observed in the plasma-free amino acids (threonine, isoleucine, phenylalanine, lysine, histidine, arginine, aspartic acid, serine, alanine, cystine) of the blood from the colic branch of the ileocolic vein. The results of these experiments indicated that pigs have the ability to utilize the microbial amino acids synthesized from urea nitrogen in the large intestine.

Amino Acids↗

Survival of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus in the terminal ileum of fistulated Göttingen minipigs.

The ability of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus administered in yogurt to survive the passage through the upper gastrointestinal tract was investigated with Göttingen minipigs that were fitted with ileum T-cannulas. After ingestion of yogurt containing viable microorganisms, ileostomy samples were collected nearly every hour beginning 3 h after food uptake. Living L. delbrueckii subsp. bulgaricus and S. thermophilus were detected in the magnitude of 10(6) to 10(7) per gram of intestinal contents (wet weight) in all animals under investigation. A calculation of the minimum amount of surviving bacteria that had been administered is presented. Total DNA extracted from ileostomy samples was subjected to PCR, which was species specific for L. delbrueckii and S. thermophilus and subspecies specific for L. delbrueckii subsp. bulgaricus. All three bacterial groups could be detected by PCR after yogurt uptake but not after uptake of a semisynthetic diet. One pig apparently had developed an endogenous L. delbrueckii flora. When heat-treated yogurt was administered, L. delbrueckii was detected in all animals. S. thermophilus or L. delbrueckii subsp. bulgaricus was not detected, indicating that heat-inactivated cells and their DNAs had already been digested and their own L. delbrueckii flora had been stimulated for growth.

Animals↗

[Microbiological aspects of application of fluoroquinolones in surgery].

Antimicrobic activity of fluorochinolons with regard to pathogenic organisms of purulent-septic surgical infection was studied. In comparison with cephalosporins of the third generation, higher activity of fluorochinolons is noted with regard to majority aerobic microorganisms, and to pseudomonads, in particular. Optimal pharmacokinetics properties, good compliance, wide antimicrobic spectrum permits to use fluorochinolons in conduction of empirical antibacterial therapy for the infection occurrence prophylaxis in surgical practice, and for the selective intestinal decontamination, in particular.

Anti-Infective Agents↗

[Ulcerative colitis in childhood and its differential diagnosis].

Ulcerative colitis occurs before the age of 15 years in 20% and before 20 years in 40% of all patients involved. The frequency of this disease in patients at the University Children's Hospital of Munich has clearly risen over the past years. In many patients we found characteristic differences in history, clinical picture and laboratory results as compared with Crohn's disease. The most important diagnostic procedure is endoscopy of the entire colon. The histological examination of the biopsies taken during colonoscopy differentiated less clearly between these two entities than the macroscopic judgement by the endoscopist. The development of carcinoma must be considered even in children when the disease has lasted for seven years or longer. Ulcerative colitis must be differentiated from infectious diseases of the bowel by direct search for the relevant microorganisms and by serological testing.

Adolescent↗

Reevaluation of nitrate and nitrite levels in the human intestine.

Analyses of human fecal and ileostomy samples by a method that is insensitive and free from interferences indicate that nitrate and nitrite levels in the intestine are lower than reported previously. Fecal nitrate and nitrite concentrations ranged from 0 to 14 mumol/kg (0 to 0.9 ppm) and 5 to 19 mumol/kg (0.3 to 0.9 ppm), respectively. Ileostomy samples contained from 0 to 7 mumol/kg (0 to 0.4 ppm) and 0 to 15 mumol/kg (0 to 0.7 ppm) for nitrate and nitrite, respectively. We also showed that, when deliberately added to feces samples, nitrate and nitrite were destroyed during a two-hr incubation period in a reaction that depended on the presence of microorganisms. The results suggest that conditions in the lower gastrointestinal tract favor denitrification, not nitrification as had been proposed previously.

Adolescent↗

Modulation of chloride secretory responses and barrier function of intestinal epithelial cells by the Salmonella effector protein SigD.

The Salmonella effector protein SigD is an inositol phosphate phosphatase that inhibits phosphatidylinositol 3-kinase-dependent signaling. Because epidermal growth factor (EGF) inhibits chloride secretion via phosphatidylinositol 3-kinase, we explored whether Salmonella infection might modify the inhibitory effect of EGF. As expected, EGF inhibited chloride secretion induced by carbachol in T(84) epithelial cells. Infection with wild-type (WT) but not sigD(-) mutant S. typhimurium SL1344 decreased CCh-stimulated chloride secretion. Moreover, WT but not sigD(-) Salmonella reduced the inhibitory effect of EGF on carbachol-stimulated chloride secretion. Complementation of sigD restored the ability of mutant Salmonella to reverse the inhibitory effect of EGF. EGF-induced EGF receptor phosphorylation was similar in cells infected with either WT or mutant Salmonella, and neither WT nor sigD(-) Salmonella altered recruitment of the p85 subunit of phosphatidylinositol 3-kinase to EGF receptor, implying that SigD acts downstream of these signaling events. Furthermore, transepithelial resistance fell more rapidly in cells infected with WT vs. sigD(-) Salmonella, indicating an early role for SigD in reducing barrier function, perhaps via activation of protein kinase C. We conclude that the Salmonella bacterial effector protein SigD may play critical roles in the pathogenesis of disease caused by this microorganism.

Animals↗

Cathepsin E in follicle associated epithelium of intestine and tonsils: localization to M cells and possible role in antigen processing.

A specific rabbit anti-human serum was used selectively to localize the aspartic proteinase cathepsin E to follicle associated epithelium (FAE) of human and rat intestine, including jejunum, ileum, appendix, colon and rectum, as well as of human palatine, pharyngeal and lingual tonsils. Coexpression of class II histocompatibility antigen HLA-DR antigen has been observed in some of the cathepsin E-positive epithelial cells. In addition, cathepsin E has been detected in a few mononuclear cells of intestinal lymphoid structures and tonsils resembling interdigitating reticulum cells of lymph nodes. Another aspartic proteinase, cathepsin D, has been found to be poorly represented in FAE and intensely expressed by macrophages. Electron immunocytochemistry localized cathepsin E to endosomal vesicles and endoplasmic reticulum of M cells in rat and human ileum as well as of M-like cells in human palatine tonsil. The results suggest a possible role of endosomal cathepsin E in the processing of macromolecules and microorganisms transported by M cells and related epithelial cells to mucosal associated lymphoid tissue (MALT).

Adolescent↗

Abundant and diverse fungal microbiota in the murine intestine.

Enteric microbiota play a variety of roles in intestinal health and disease. While bacteria in the intestine have been broadly characterized, little is known about the abundance or diversity of enteric fungi. This study utilized a culture-independent method termed oligonucleotide fingerprinting of rRNA genes (OFRG) to describe the compositions of fungal and bacterial rRNA genes from small and large intestines (tissue and luminal contents) of restricted-flora and specific-pathogen-free mice. OFRG analysis identified rRNA genes from all four major fungal phyla: Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota. The largest assemblages of fungal rRNA sequences were related to the genera Acremonium, Monilinia, Fusarium, Cryptococcus/Filobasidium, Scleroderma, Catenomyces, Spizellomyces, Neocallimastix, Powellomyces, Entophlyctis, Mortierella, and Smittium and the order Mucorales. The majority of bacterial rRNA gene clones were affiliated with the taxa Bacteroidetes, Firmicutes, Acinetobacter, and Lactobacillus. Sequence-selective PCR analyses also detected several of these bacterial and fungal rRNA genes in the mouse chow. Fluorescence in situ hybridization analysis with a fungal small-subunit rRNA probe revealed morphologically diverse microorganisms resident in the mucus biofilm adjacent to the cecal and proximal colonic epithelium. Hybridizing organisms comprised about 2% of the DAPI (4',6-diamidino-2-phenylindole, dihydrochloride)-positive organisms in the mucus biofilm, but their abundance in fecal material may be much lower. These data indicate that diverse fungal taxa are present in the intestinal microbial community. Their abundance suggests that they may play significant roles in enteric microbial functions.

Animals↗

The spectrum of skin, mucosa and other extra-articular manifestations.

The seronegative spondyloarthropathies appear to be the genetically predisposed host's clinical expression to acute, subacute or chronic reaction to the invasion by environmental microorganisms. In the ensuing days or weeks, depending on the infectious load, clinical manifestations may occur ranging from constitutional complaints such as fever, to a variety of symptoms and/or signs related to the portal of entry-intestinal, genitourinary or respiratory. Within weeks or months, the initial or other target organs, such as the mucocutaneous, ocular and cardiovascular systems, may develop an acute reaction of greater or lesser specificity regarding the triggering agent (oral ulcers, circinate balanitis, erythema nodosum, acute anterior uveitis, pericarditis, heart blocks). Lastly, many years later, a minority of patients, probably those with a large genetic component, exhibit a spectrum of clinical manifestations related to those organs, with a chronic or recurrent course. Acute clinical manifestations--reactive arthritis--are prominent in the initial phase of the clinical spectrum, while chronic manifestations--ankylosing spondylitis--are seen at the other end of the spectrum.

Arthritis, Reactive↗

Detection of Lawsonia intracellularis in wild boar and fallow deer bred in one game enclosure in the Czech Republic.

The prevalence of Lawsonia intracellularis between wild boar (Sus scrofa) and fallow deer (Dama dama) bred in one game reserve was investigated using the nested PCR method. In the study, 88 clinically healthy wild boars of different age categories and two fallow deer bagged in the game reserve were examined. Lawsonia intracellularis was demonstrated in the mucous membrane of the intestine of eight (9.1%) wild boars and one fallow deer. Of the nine wild boar whose tissues of corresponding lymph nodes were examined in addition to the mucous membrane of the ileum, one tested positive for the microorganism. A relationship between the occurrence of L. intracellularis and age of wild boar was demonstrated. Because wild boar and fallow deer are bred together in one game reserve, the possibility of inter-species transfer of L. intracellularis should be borne in mind.

Age Factors↗

[The taxonomic position, ecological aspects and physiological-biochemical characteristics of microorganisms having industrial importance].

The paper presents fundamental results of the investigations obtained in the Department of Physiology of Industrial Microorganisms at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine for the recent 15-20 years, as well as the Department developments directed to creation of biotechnological preparations, products and processes, most of which have been put into practice of the national economy and health protection. Data have been presented concerning the study of systematics, biology and ecology of nocardioform and coryneform bacteria, lactic acid bacteria and yeasts. When studying physiological-biochemical peculiarities and biological effect of the studied microorganisms, considerable attention has been paid to growth physiology of lactic acid bacteria, yeasts, lectin-forming bacteria, to processes of shaping of fungal mycelium under the conditions of batch cultivation as well as to microorganisms which take part in sewage treatment for heavy metals and oil products. The work also deals with the problems of creation and application in the economy of preparations intended for treatment and prophylaxis of gastro-intestinal diseases of cattle, poultry and swine, for siloing fodder, of new lactic acid preparations of functional direction, technological grounds of sewage treatment for oil products and six-valent toxic chrome.

Bacteria↗

[Are Crohn disease and ulcerative colitis the results of an immunoregulatory disturbance?].

The increasing knowledge regarding the regulation of immunological processes has stimulated different approaches to the analysis of chronic inflammatory bowel diseases. The well documented independence of the mucosa-associated immune system favours data obtained analysing immunological reactions within the mucosa. Thus lymphocytes and antibodies isolated from the mucosa are consequently used instead of those obtained from peripheral blood. Despite of an initiation by exogenic agents such as allergenic food constituents for microorganisms there is a growing body of evidence pointing to an immunoregulatory disturbance as underlying pathogenic mechanism for these diseases. This hypothesis is discussed regarding recent observations of a contrasuppressive circuit of regulatory lymphocytes in the murine intestine.

Animals↗

In vitro activity of gemifloxacin (SB-265805) compared with 14 other antimicrobials against intestinal pathogens.

We studied the in vitro activity of gemifloxacin (SB-265805) and 14 comparator antimicrobials against 288 recent isolates of enteropathogenic bacteria (106 Salmonella: spp., 32 Hafnia alvei, 22 Yersinia enterocolitica, 21 Shigella: spp., 16 Aeromonas: spp. and 91 Campylobacter jejuni). Gemifloxacin, the other fluoroquinolones and cefotaxime were very active against all microorganisms tested except for C. jejuni. Seventy-seven per cent of isolates of C. jejuni were inhibited by erythromycin < or =0.5 mg/L. Only one strain of C. jejuni was highly resistant to this antimicrobial agent. Of the compounds tested, gentamicin was the most active in vitro. The in vitro activity of the other antibiotics tested was variable. A quinolone could be a good choice for treating gastrointestinal infections when antimicrobial therapy is indicated. For C. jejuni, another antibiotic such as erythromycin should be considered.

Acute Disease↗

Lectin binding defines and differentiates M-cells in mouse small intestine and caecum.

M-cell surface glycoconjugate expression was investigated by applying a panel of lectins to whole fixed mouse Peyer's and caecal patches. While the majority of lectins failed to identify mouse M-cells, the lectin Euonymus europaeus differentially stained the surface of M-cells in both mouse Peyer's and caecal patches, and the lectins Ulex europaeus II and Bandeiraea simplicifolia I isolectin B4 identified M-cells in the Peyer's and caecal patch follicle associated epithelium, respectively. These three mouse M-cell markers failed to identify rat and rabbit Peyer's patch M-cells, although both Euonymus europaeus and Ulex europaeus II differentially stained M-cells in the periphery of rabbit caecal patch domes. These site and species related variations in M-cell surface glycoconjugate expression may reflect the local microorganism populations and will have important implications if orally delivered vaccines and drugs are to be targeted to M-cells via their surface glycoconjugates.

Animals↗