Self-reinoculation with fecal flora changes microbiota density and composition leading to an altered bile-acid profile in the mouse small intestine

Background: The upper gastrointestinal tract plays a prominent role in human physiology as the primary site for enzymatic digestion and nutrient absorption, immune sampling, and drug uptake. Alterations to the small intestine microbiome have been implicated in various human diseases, such as non-alcoholic steatohepatitis and inflammatory bowel conditions. Yet, the physiological and functional roles of the small intestine microbiota in humans remain poorly characterized because of the complexities associated with its sampling. Rodent models are used extensively in microbiome research and enable the spatial, temporal, compositional, and functional interrogation of the gastrointestinal microbiota and its effects on the host physiology and disease phenotype. Classical, culture-based studies have documented that fecal microbial self-reinoculation (via coprophagy) affects the composition and abundance of microbes in the murine proximal gastrointestinal tract. This pervasive self-reinoculation behavior could be a particularly relevant study factor when investigating small intestine microbiota. Modern microbiome studies either do not take self-reinoculation into account, or assume that approaches such as single housing mice or housing on wire mesh floors eliminate it. These assumptions have not been rigorously tested with modern tools. Here, we used quantitative 16S rRNA gene amplicon sequencing, quantitative microbial functional gene content inference, and metabolomic analyses of bile acids to evaluate the effects of self-reinoculation on microbial loads, composition, and function in the murine upper gastrointestinal tract. Results: In coprophagic mice, continuous self-exposure to the fecal flora had substantial quantitative and qualitative effects on the upper gastrointestinal microbiome. These differences in microbial abundance and community composition were associated with an altered profile of the small intestine bile acid pool, and, importantly, could not be inferred from analyzing large intestine or stool samples. Overall, the patterns observed in the small intestine of non-coprophagic mice (reduced total microbial load, low abundance of anaerobic microbiota, and bile acids predominantly in the conjugated form) resemble those typically seen in the human small intestine. Conclusions: Future studies need to take self-reinoculation into account when using mouse models to evaluate gastrointestinal microbial colonization and function in relation to xenobiotic transformation and pharmacokinetics or in the context of physiological states and diseases linked to small intestine microbiome and to small intestine dysbiosis

Intestinal transplantation in composite visceral grafts or alone

Under FK 506-based immunosuppression, the entire cadaver small bowel except for a few proximal and distal centimeters was translated to 17 randomly matched patients, of whom two had antigraft cytotoxic antibodies (positive cross-match). Eight patients received the intestine only, eight had intestine in continuity with the liver, and one received a full multivisceral graft that included the liver, stomach, and pancreas. One liver-intestine recipient died after an intestinal anastomotic leak, sepsis, and graft- versus-host disease. The other 16 patients are alive after 1 to 23 months, in one case after chronic rejection, graft removal, and retransplantation. Twelve of the patients have been liberated from total parenteral nutrition, including all whose transplantation was 2 months or longer ago. The grafts have supported good nutrition, and in children, have allowed growth and weight gain. Management of these patients has been difficult and often complicated, but the end result has been satisfactory in most cases, justifying further clinical trials. The convalescence of the eight patients receiving intestine only has been faster and more trouble free than after liver-intestine or multivisceral transplantation, with no greater difficulty in the control of rejection

Regulation of bicarbonate secretion in marine fish intestine by the calcium-sensing receptor

In marine fish, high epithelial intestinal HCO3- secretion generates luminal carbonate precipitates of divalent cations that play a key role in water and ion homeostasis. The present study was designed to expose the putative role for calcium and the calcium-sensing receptor (CaSR) in the regulation of HCO3- secretion in the intestine of the sea bream (Sparus aurata L.). Effects on the expression of the CaSR in the intestine were evaluated by qPCR and an increase was observed in the anterior intestine in fed fish compared with unfed fish and with different regions of intestine. CaSR expression reflected intestinal fluid calcium concentration. In addition, anterior intestine tissue was mounted in Ussing chambers to test the putative regulation of HCO3- secretion in vitro using the anterior intestine. HCO3- secretion was sensitive to varying calcium levels in luminal saline and to calcimimetic compounds known to activate/block the CaSR i.e., R 568 and NPS-2143. Subsequent experiments were performed in intestinal sacs to measure water absorption and the sensitivity of water absorption to varying luminal levels of calcium and calcimimetics were exposed as well. It appears, that CaSR mediates HCO3- secretion and water absorption in marine fish as shown by responsiveness to calcium levels and calcimimetic compounds.European Social Funds through the Portuguese National Science Foundation (FCT) [SFRH/BD/113363/2015, PTDC/MAR-BIO/3034/2014]; Portuguese Foundation for Science and Technology (FCT) [UID/Multi/04326/2013]; Ministry of Science and Higher Educatio

Effect of ischemia on the canine large bowel: A comparison with the small intestine

Mucosal injury caused by ischemia and reperfusion has been well documented with the small intestine, but little is known about the colon. In the present study, the effect of warm and cold ischemia on the canine colon was studied and compared to that on the small intestine. After in situ flushing, the small intestine and the colon from six beagle dogs were removed and stored for 0.5, 1.5, and 3 hr at 37°C (warm ischemia) or for 1, 6, 12, 24, 36, and 48 hr at 4°C (cold ischemia). Electrophysiology, permeability, biochemistry, and histopathology of the specimens at each ischemic period and after reperfusion in the Ussing chamber were determined. Warm and cold ischemia induced duration-dependent suppression of electrophysiology in both organs, but the colonic mucosa retained higher activity of absorptive enterocytes and cryptic cells than the small intestine. Only the colon showed increased permeability of FITC-conjugated Dextran from the mucosal surface to the submucosal layer after prolonged ischemia. Changes in adenine nucleotides and purine catabolites were not markedly different between the organs. Histopathologic abnormalities during ischemia and after reperfusion were more serious with the small intestine than with the colon. Compared to warm ischemia, hypothermia lessened or delayed these morphofunctional derangements in both organs, which became universally worsened after reperfusion. Colonic mucosa receives morphofunctional derangements from ischemia and reperfusion, but the severity of the damage was much less severe in the colon than in the small intestine

Factors influencing the development and carbohydrate metabolism of Echinococcus granulosus in dogs

Echinococcus granulosus adult worms, 35 days postinfection, were measured for dispersion in the intestines of 10 dogs, a range of morphological characters, and the excreted end products of carbohydrate catabolism following 4 hr incubation in vitro. Most worms were found in the proximal sections of the small intestine, but the pattern of dispersion differed between dogs. Worm development varied both between dogs and between different regions of the small intestine of individual dogs. Overall there was a high level of variability with no simple patterns. Worm metabolism was related to worm development and, also independently, to local population density within the intestine. Larger, more mature worms produced less lactate and, at higher densities. worms tended to produce more acetate and succinate (pathways with a higher energy yield than lactate) and less ethanol. Thus, both more developed worms and high population density are associated with a shift from cytosolic to mitochondrial metabolism. The variation between worm populations along the small intestine along with the observed variation between worm populations from sibling dogs infected with genetically identical parasites suggests that the local host environment has a significant effect on parasite development

To respond or not to respond - a personal perspective of intestinal tolerance

For many years, the intestine was one of the poor relations of the immunology world, being a realm inhabited mostly by specialists and those interested in unusual phenomena. However, this has changed dramatically in recent years with the realization of how important the microbiota is in shaping immune function throughout the body, and almost every major immunology institution now includes the intestine as an area of interest. One of the most important aspects of the intestinal immune system is how it discriminates carefully between harmless and harmful antigens, in particular, its ability to generate active tolerance to materials such as commensal bacteria and food proteins. This phenomenon has been recognized for more than 100 years, and it is essential for preventing inflammatory disease in the intestine, but its basis remains enigmatic. Here, I discuss the progress that has been made in understanding oral tolerance during my 40 years in the field and highlight the topics that will be the focus of future research

Mathematical Modeling of Transport and Degradation of Feedstuffs in the Small Intestine

We describe a mathematical modeling of the digestion in the small intestine. The main interest of our work is to consider, at the same time, different aspects of the digestion i.e. the transport of the bolus all along the intestine, feedstuffs degradation according to the enzymes and local physical conditions, and nutrients absorption. A system of coupled ordinary differential equations is used to model these phenomena. The major unknowns of this system are the position of the bolus and its composition. This system of equations is solved numerically. We present different numerical computations for the degradation, absorption and transport of the bolus with acceptable accuracy with experimental data. The main feature and interest of this model are its generality. Even if we are at an early stage of development, our approach can be adapted to treat any kind of feedstuffs in any non-ruminant animal to predict the composition and velocity of bolus in the small intestine

Clinical intestinal transplantation: New perspectives and immunologic considerations

Background: Although tacrolimus-based immunosuppression has made intestinal transplantation feasible, the risk of the requisite chronic high- dose treatment has inhibited the widespread use of these procedures. We have examined our 1990-1997 experience to determine whether immunomodulatory strategies to improve outlook could be added to drug treatment. Study Design: Ninety-eight consecutive patients (59 children, 39 adults) with a panoply of indications received 104 allografts under tacrolimus-based immunosuppression: intestine only (n = 37); liver and intestine (n = 50); or multivisceral (n = 17). Of the last 42 patients, 20 received unmodified adjunct donor bone marrow cells; the other 22 were contemporaneous control patients. Results: With a mean followup of 32 ± 26 months (range, 1-86 months), 12 recipients (3 intestine only, 9 composite grafts) are alive with good nutrition beyond the 5-year milestone. Forty-seven (48%) of the total group survive bearing grafts that provide full (91%) or partial (9%) nutrition. Actuarial patient survival at 1 and 5 years (72% and 48%, respectively) was similar with isolated intestinal and composite graft recipients, but the loss rate of grafts from rejection was highest with intestine alone. The best results were in patients between 2 and 18 years of age (68% at 5 years). Adjunct bone marrow did not significantly affect the incidence of graft rejection, B-cell lymphoma, or the rate or severity of graft-versus-host disease. Conclusions: These results demonstrate that longterm rehabilitation similar to that with the other kinds of organ allografts is achievable with all three kinds of intestinal transplant procedures, that the morbidity and mortality is still too high for their widespread application, and that the liver is significantly but marginally protective of concomitantly engrafted intestine. Although none of the endpoints were markedly altered by donor leukocyte augmentation (and chimerism) with bone marrow, establishment of the safety of this adjunct procedure opens the way to further immune modulation strategies that can be added to the augmentation protocol

An investigation into the relationship between small intestinal fluid secretion and systemic arterial blood pressure in the anesthetized rat

In the absence of an ability to absorb fluid by cellular uptake mechanisms, fluid movement in vivo from the perfused rat intestine is absorptive when the diastolic blood pressure is normal or very low but is secretory when blood pressure falls below normal. This pattern of fluid movement is consistent with changes in capillary pressure within the villus. Whether flow moves into or out of the intestine is determined by changes in the Starling forces across intestinal capillaries. These observations indicate that secretion caused by some bacterial enterotoxins may act solely on the vasculature of the small intestine. This contradicts a major current theory of secretion that requires the source of the fluid to be from the epithelial cell. The significance of this work is that the intestinal arterioles rather than the epithelial cells may determine secretion. If substantiated, this may allow the development of the effective anti-secretory drugs that have not been forthcoming with development strategies based on the enterocyte model of deranged intestinal secretion

Feed interventions and skatole deposition

Skatole produced in the large intestine of the pig and the testicular steroid androstenone are the main substances contributing to boar tainted meat from entire male pigs. Boar taint decreases the quality of the meat and is not accepted by consumers. Until now boar taint has been avoided by castrating male pigs. Surgical castration reduces lean meat percentage, growth rate and feed efficiency, and it causes pain to the animal. This constitutes a problem in relation to productivity and welfare. Different attempts on avoiding surgical castration were either not fully effective, not accepted by the market, or they have a long time horizon for implementation. However, when focusing on the effect of feed interventions on boar taint, previous studies have showed a reducing effect through reduced skatole production in the large intestine after a one week application period. Skatole is produced from the microbial fermentation of L-tryptophan in the large intestine. In the literature it is well documented that skatole production in the large intestine is positively correlated with skatole deposition in adipose tissue. Moreover skatole production can be decreased by adding non-digestible and easy fermentable carbohydrates to the feed. However, little is known about skatole producing bacteria from the large intestine of pigs, and how these bacteria can be affected. This thesis reviews skatole production and metabolism in the pig, and how skatole production can be reduced by affecting the microbial production of skatole in the large intestine. A skatole producing bacterium SK9 K4 was isolated from the gastrointestinal tract of pigs. No such bacterium has previously been described. The bacterium was characterised by 16S RNA sequencing, gram stain, analysis of DNA G-C content, cellular fatty acids composition and DNA hybridisation with closely related bacteria. The fermentation of different carbohydrate sources, the growth pattern, the production of organic acids and the skatole production were studied in vitro. The production of skatole in the large intestine was correlated with skatole deposition in adipose tissue. Skatole production could be reduced when adding a minimum of 20 % raw potato starch or 9 % inulin to the feed. The problem concerning deposition of skatole in adipose tissue seems to be solved through the introduction of feed interventions. However high concentrations of androstenone deposited in adipose tissue remains a challenge. Thus, the feed interventions were not fully effective against boar taint. SK9 K4 was described as cells being strictly anaerobic, occurred singly or in pairs and were gram positive. It was identical with an Olsenella sp. strain isolated from the rumen, an uncultured Olsenella sp. clone isolated from sludge and an uncultured bacterium colon isolated from the oral cavity. Moreover SK9 K4 was closely related to Olsenella uli, Olsenella profusa, Olsenella umbonata and Atopobium parvulum. SK9 K4 and O. uli produced skatole from idole-3-acetic acid but not from L-tryptophan. The major fermentation products were lactic acid together with acetic acid and formic acid. SK9 K4 was not able to ferment raw potato starch, inulin and raw corn starch. Thus, when feeding resistant starch or inulin, the growth of skatole producing bacteria might be reduced followed by a reduced ability to produce skatole. The characterisation of a skatole producing bacterium isolated from the gastrointestinal tract of pigs gives the opportunity to further study the bacterium in vivo. Studies should be conducted to investigate the effect of a control diet compared to a diet added a non-digestible and easy fermentable carbohydrate on the growth of SK9 K4 in the large intestine of the pig