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Attempts were made to find and characterize an antibacterial activity (ABA) factor in porcine pancreatic juice (PJ). Its isolation requires several steps. Since ABA factor was found to be heat resistant, the first step was heating for 30 min at 65°C. Afterwards column chromatography, ethanol precipitation and polyacrylamide gel electrophoresis were involved. Finally, we obtained a pancreatic juice fraction with antibacterial activity against Escherichia coli strain AB1157. In the presence of this fraction the number of living bacterial cells in overnight culture decreased about 10,000 fold and a spot-test gave clearly positive results. The results of analysis suggest that the antibacterial factor is a polypeptide active in a pH range 8.0 - 8.5, that migrates in polyacrylamide gel electrophoresis as a band under 14,000 Da. Mass spectroscopy analysis of active fraction showed high concentration of porcine pancreatic spasmolytic polypeptide (PSP). In conclusion, a polypeptide controlling bacterial homeostasis has been found in the porcine pancreatic juice.
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Histamine H3 receptors modulate reactive hyperemia in rat gut

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Reactive hyperemia (RH) is an abrupt blood flow increase following release from mechanical occlusion of an artery, with restoration of intra-arterial pressure. The mechanism of this postocclusion increase in blood flow in the gut is multifactorial. Relaxation of intestinal resistance vessels, observed during RH, may involve myogenic, metabolic, hormonal and neurogenic factors. Evidence exists that histamine is an important endogenous mediator of various functions of the gut, including blood flow. The vascular effects of histamine in the intestinal circulation are due its agonistic action on histamine H1, H2 and H3 receptors. In the present study the hypothesis was tested that peripheral histamine H3 receptors are involved in the mediation of RH in the intestinal circulation. In anesthetized rats, anterior mesenteric artery blood flow (MBF) was determined with ultrasonic Doppler flowmeter, and arterial pressure (AP) was determined with a transducer. The increase in the volume of blood accumulating during RH (RH-volume), the peak increase of arterial blood flow (RH-peak response) and the duration of the hyperemia (RH-duration) were used to quantify RH after occluding the anterior mesenteric artery for 30, 60 and 120 s. Hyperemia parameters were determined before and after administration of the selective histamine H3 receptor antagonist clobenpropit. Pretreatment with clobenpropit was without any effect on control MBF and AP but significantly reduced most of RH responses. These findings support the hypothesis that histamine H3 receptors do not play any role in the control of intestinal vasculature at basal conditions but these receptors participate in the intestinal hyperemic reaction in response to complete temporal intestinal ischemia.
The effects of genetically modified (GM) soybean meal (SBM) and maize on the diversity and activity of microbiota inhabiting terminal gut segments in broiler chickens Were studied. Eight diets were prepared, based on conventional or GM SBM combined with maize cvs Clarica or PR39 F58, or their isogenic MON 810 counterparts cvs Bacilla or PR39 F56. Diets were fed from age 1 to 28 days to 144 Ross broilers, allocated to eight groups of 18 birds each. The microbiota was analysed by terminal restriction fragment length polymorphism (T-RFLP) analysis and its activity was measured. In the ileum and caecum of all groups, members representing the orders Clostridiales, Lactobacillales and Selenomonadales were present, accompanied by Bifidobacteriales in the caecum. The diversity of the order Lactobacillales in the ileum and caecum of birds fed GM maize was reduced, while that of Lactobacillales in the ileum and Bifidobacteriales in the caecum of birds fed GM SBM was higher compared with conventional maize and SBM. The use of GM and conventional maize and SBM did not affect the activity of microbiota measured as bacterial enzyme activity and the concentration of short-chain fatty acids in the ileal and caecal digesta. The GM maize did not change resistance of E.
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Luminal sensing in the gut: an overview

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The wall of the gut responds to an impressive array of signals originating in the lumen, including nutrient and non-nutrient chemicals, mechanical factors, and micro-organisms. The idea that the gut wall exhibits luminal chemo-sensitivity is implied in the original discovery of secretin by Bayliss and Starling, and has become an integral part of models of neurohumoral control of gastrointestinal function. Entero-endocrine cells are specialised for luminal nutrient sensing but sub-epithelial nerve fibres may also respond to luminal chemicals that freely diffuse across the epithelium eg short chain fatty acids. The molecular recognition mechanisms include G-protein coupled receptors (GPCRs) eg the extracellular Ca2+ sensing receptor which also responds to aromatic amino acids. There are also GPCRs sensing fatty acids, as well as bitter or noxious compounds. In addition, though, gating of ion channels including events secondary to energy availability eg ATP, may be involved in sensing some luminal chemicals. There is likely to be integration of luminal signals at several levels including at the level of entero-endocrine cells and at sub-epithelial nerve fibers. For example, the intestinal hormone CCK acts on primary afferent nerve fibers of the vagal trunk. The same fibers also express leptin receptors that are thought to respond to leptin released from gastric chief cells, orexin receptors (activation of which inhibits CCK) and possibly ghrelin receptors. Multiple signalling mechanisms allow specific responses to be matched to meals of differing content.
Cholecystokinin (CCK) is a major peptide hormone in the gut and a major peptide transmitter in the brain. Its synthesis requires endoproteolytic cleavage of proCCK at several mono- and dibasic sites by prohormone convertases (PCs). Of these, PC1 and PC2 are expressed in cerebral neurons and intestinal endocrine cells. Characteristically, however, the processing of proCCK varies markedly between the brain and the gut. In neurons, CCK-8 is always the predominating form, whereas the endocrine gut cells (I-cells) contain a mixture of small and larger CCK-peptides of which CCK-33 or CCK-22 often predominate. The role of PC1 and PC2 in the processing of proCCK have now been examined by measuring the concentrations of prohormone, processing intermediates and amidated end-products in jejunal and cerebral extracts of PC1 and PC2 deficient mice and corresponding wild type controls. The PC1 null mice revealed a pattern opposite to that of the PC2 null mice, in whom only the cerebral processing of proCCK was affected. Thus PC1 knockouts reveal a severe block in the processing of intestinal proCCK. Accordingly, the intestinal concentration of proCCK was many fold increased, and also the concentrations of different processing intermediates were raised; but the concentration of bioactive, a-amidated and O-sulfated CCK was reduced to a few percent of normal. The only bioactive CCK peptide in the gut of PC1 deficient mice was CCK-22, and it was present only in trace amounts. The cerebral processing of proCCK was, however, not at all affected by the lack of PC1 - in sharp contrast to the effect of PC2. The results show that the tissue-specific processing of proCCK to a large extent can be explained by prohormone convertases, as PC1 plays a decisive role in the maturation of hormonal CCK in the gut, whereas PC2 governs the processing of brain proCCK.
Background. Microbial and fungal cellulases are known to hydrolyse cellulose, which is ingested as plant material by herbivorous/omnivorous fishes. Microbial enzymes have enormous advantage of being produced in large quantities by established fermentation techniques. The present investigation aims to optimize the environmental and nutritional parameters for fermentation to enhance cellulase production by two bacterial strains isolated from fish gastrointestinal tracts. Materials and Methods. Two bacterial strains, Bacillus subtilis CY5 and Bacillus circulans TP3, isolated from the gastrointestinal tracts of common carp, Cyprinus carpio L., and Mozambique tilapia, Oreochromis mossambicus (Peters, 1852), respectively were identified as potent cellulase producers. Both strains were cultured in tryptone soya broth for 24 h at 32 ± 2°C, when average viable count of 9.75 • 107 cells • mL-1 culture broth was obtained. This was used as the inoculum for the production medium. The fermentation medium was seeded with 1.0%, 2.0%, 3.0%, 4.0%, and 5.0% inoculum (tryptone soya broth) and incubated in static culture at 40°C to standardize the inoculum size for fermentation. The effect of different production parameters, such as fermentation condition, moisture, pH, temperature, inoculum size, and nitrogen sources on cellulase production by the isolated bacterial strains were studied. Results. Cellulase yield was highest (26 U in B. subtilis and 20.2 U in B. circulans) in solid-state fermentation (SSF). Enzyme production in both the isolates increased in an optimum pH range of 7.0 to 7.5. Minimum cellulase production was observed at 45°C, while maximum production was obtained at 40°C. To standardize the fermentation period for cellulase production, production rate was measured at 12-h intervals up to 120 h. Enzyme production increased for 96 h of fermentation in both strains, and decreased thereafter. The enzyme production increased with increased inoculum size up to 3.0 percentage points. Asparagine as the nitrogen source was most effective in B. subtilis CY5, while beef extract proved useful in optimizing enzyme production by B. circulans TP3. Conclusion. The results of this study will help to standardize the requirements for optimum production of cellulase by cellulase-producing fish gut bacteria and might contribute towards better fish feed formulation incorporating plant ingredients, especially in the larval stages when the enzyme system is not efficient.
Background. The information on gut microflora in fish is scanty and there is a paucity of knowledge regarding microbial enzyme activity in fish gastrointestinal tracts. Although some information is available on the enzyme-producing bacteria in fish digestive tracts, almost nothing is known about their distribution in different regions of the gut. In the present study, an attempt has been made to investigate the distribution of enzyme-producing microflora in the foregut and hindgut regions of seven culturable freshwater teleosts. Materials and Methods. Isolation and enumeration of aerobic bacterial flora in the foregut and hindgut regions of the gastrointestinal tracts of seven freshwater teleosts of different feeding habits, namely rohu, Labeo rohita; catla, Catla catla; mrigal, Cirrhinus mrigala; bata, Labeo bata; orange-fin labeo, Labeo calbasu; Nile tilapia, Oreochromis niloticus; and climbing perch, Anabas testudineus, have been carried out. Microbial culture of the gut mucosa on selected nutrient media, following the enrichment culture technique, was done for bacterial isolation. Bacterial isolates were qualitatively screened on the basis of their extracellular enzyme-producing ability. The selected strains were further quantitatively assayed for amylase, cellulase and protease activities. Results. In general, bacterial population was lower in the foregut region of all the seven species of fish examined. Amylolytic strains were present in higher densities in the foregut region of orange-fin labeo and bata (12.20 × 103 CFU·g-1 gut tissue and 11.50 × 103 CFU·g-1 gut tissue, respectively) in comparison to the hindgut region. The cellulolytic population exhibited maximum densities in the hindgut region of bata (7.20 × 103 CFU·g-1 gut tissue) followed by the foregut region of the same fish (5.50 × 103 CFU·g-1 gut tissue). Amylolytic and cellulolytic bacterial flora was not detected in both the fore and hindgut regions of climbing perch. Proteolytic bacterial flora was found in all the species of fish studied and the maximum count was observed in the hindgut region of bata (13.40 × 103 CFU·g-1 gut tissue), orange-fin labeo (9.00 × 103 CFU·g-1 gut tissue), Nile tilapia (8.30 × 103 CFU·g-1 gut tissue) and climbing perch (7.20 × 103 CFU·g-1 gut tissue). Minimum count of proteolytic bacterial flora was observed in the foregut region of all the fishes studied. Peak amylase and cellulase activities were exhibited by bacterial strains isolated from the foregut of orange-fin labeo (266.43 ± 0.15 U) and the hindgut of bata (64.01 ± 0.42 U), respectively. Maximum protease activity was exhibited by a strain isolated from the hindgut region of orange-fin labeo (44.33 ± 0.09 U), followed by the strains isolated from the hindgut regions of climbing perch (32.87 ± 0.12 U), bata (29.71 ± 0.11 U), and Nile tilapia (29.46 ± 0.11 U). Conclusion. The results of the present study indicate that there is a distinct microbial source of digestive enzymes apart from the endogenous sources in fish digestive tracts. The enzyme-producing bacteria isolated from the digestive tracts can be beneficially used as a probiotic while formulating aquafeeds, especially in the larval stages. However, further investigations are required to determine if the addition of such isolates to fish feeds do, in fact, provide some kind of benefit to the fish involved before advocating their use.
In order to examine dynamics of the growth of anaerobic flora in large intestines of newly hatched chicks three experimental groups were formed. The first group of the birds received no feed for 60 h, the second one had a free access to a standard feed, and the chicks of the third group, besides the feeding, were individually provided per os with 1 dose of a preparation containing the normal gut flora (NGF). lt was demonstrated that feeding and treatment with the NGF preparation had a positive effect on the colonization and growth of the anaerobie flora in large intestines. The highest concentration of anaerobes (10⁸ CFU/g of the intestinal content) after 12, 36 and 60 h was found in large intestines of the birds receiving feed and NGF preparation. Treatment of the newly hatched chicks with a determined and specific intestinal flora acts as a physical and physiological barrier limiting the colonizing action of undesired and pathogenic bacteria.
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Surveillance of the gastrointestinal mucosa by sensory neurons

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A dense network of extrinsic and intrinsic sensory neurons supplies the gastrointestinal tract. Intrinsic sensory neurons provide the enteric nervous system with the kind of information that this brain of the gut requires for its autonomic control of digestion, whereas extrinsic afferents notify the brain about processes that are relevant to energy and fluid homeostasis and the sensation of discomfort and pain. The sensory repertoire of afferent neurons is extended by their responsiveness to mediators released from enteroendocrine and immune cells, which act like “taste buds” of the gut and serve as interface between the gastrointestinal lumen and the sensory nerve terminals in the lamina propria of the mucosa. Functional bowel disorders such as non-ulcer dyspepsia and irritable bowel syndrome are characterized by abdominal discomfort or pain in the absence of an identifiable organic cause. It is hypothesized with good reason that infection, inflammation or trauma causes sensory pathways to undergo profound phenotypic and functional alterations that outlast the acute insult. The pertinent changes involve an exaggerated sensitivity of the peripheral afferent nerve fibres as well as a distorted processing and representation of the incoming information in the brain. This concept identifies a number of receptors and ion channels that are selectively expressed by primary afferent neurons as important molecular targets at which to aim novel therapies for functional bowel disorders.
This manuscript reviews the current views on morphology and function of the distinct subpopulations of interstitial cells of Cajal (ICC) in the digestive tract and their interrelationships with surrounding cells. Three different functions have been postulated so far, i.e. a pacemaker role, a mediator in enteric excitatory and inhibitory neurotransmission and a mechanosensor. Attention will also be paid to the interstitial cells of Cajal and their possible involvement in pathophysiological conditions. Finally, perspectives for interstitial cells of Cajal as targets for therapeutic intervention will be discussed.
We studied enzyme polymorphisms in a striped field mouse Apodemus agrarius (Pallas, 1771) population from NE Poland and the relationships between hetero­zygosity and length and mass of the digestive tract organs, and the mucosal surface area of the small intestine. _Most of 38 loci studied were found monomorphic (pro­portion of polymorphic loci P = 0.053, observed average heterozygosity H0 = 0.021). Heterozygotes were found for Acy, Pgm-1, Mdh-2, Est-D, Pgi, Sdh and Trf. Hetero­zygous males had longer large intestines and ceca and smaller liver wet mass than their homozygous conterparts. However, in females there were no significant inter­actions between heterozygosity and gut parameters. We suggest that the low level of polymorphism, the particular set of the loci examined and sex have an effect on detection of differences between heterozygous and homozygous individuals.
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