Paul J. Moughan

Regulation of Tight Junction Permeability by Intestinal Bacteria and Dietary Components

The human intestinal epithelium is formed by a single layer of epithelial cells that separates the intestinal lumen from the underlying lamina propria. The space between these cells is sealed by tight junctions (TJ), which regulate the permeability of the intestinal barrier. TJ are complex protein structures comprised of transmembrane proteins, which interact with the actin cytoskeleton via plaque proteins. Signaling pathways involved in the assembly, disassembly, and maintenance of TJ are controlled by a number of signaling molecules, such as protein kinase C, mitogen-activated protein kinases, myosin light chain kinase, and Rho GTPases. The intestinal barrier is a complex environment exposed to many dietary components and many commensal bacteria. Studies have shown that the intestinal bacteria target various intracellular pathways, change the expression and distribution of TJ proteins, and thereby regulate intestinal barrier function. The presence of some commensal and probiotic strains leads to an increase in TJ proteins at the cell boundaries and in some cases prevents or reverses the adverse effects of pathogens. Various dietary components are also known to regulate epithelial permeability by modifying expression and localization of TJ proteins.

The effect of microbial phytase on ileal phosphorus and amino acid digestibility in the broiler chicken

1. The study aimed to assess the effect of a commercially available microbial phytase on phytate phosphorus and total phosphorus content at the terminal ileum as well as true ileal amino acid digestibility. 2. Five diets, each containing a different plant-based feedstuff, were supplemented with microbial phytase and fed, along with a non-supplemented corresponding diet, to 28-d-old broiler chickens, Chromic oxide was used as an indigestible marker. Ileal contents were collected and analysed, along with the diets, for total phosphorus, phytate phosphorus and amino acids. 3. Endogenous phosphorus determined at the terminal ileum was 272 ± 108 mg/kg food dry matter (mean ± SE). Endogenous ileal amino acid flows ranged from 58 ± 10 mg/kg food dry matter for methionine to 568 ± 47 mg/kg food dry matter for glutamic acid. 4. Supplementation with microbial phytase resulted in a significantly greater phytate P disappearance from the terminal ileum for rice bran (17% units), but not for soyabean meal, maize, wheat or rapeseed meal. Similarly total phosphorus digestibility was significantly ( P < 0.05) higher when microbial phytase was added to the rice-bran-based diet but not for any of the other feedstuffs. 5. Amino acid digestibility was significantly greater in the presence of microbial phytase for all the amino acids examined in wheat, for several of the amino acids each in maize and rapeseed meal and for one amino acid in rice bran and soyabean meal. The average increase in amino acid digestibility for those amino acids affected, was 13, 6, 10, 7 and 12% units for wheat, maize, rapeseed meal, rice bran and soyabean meal, respectively. 6. It appears that microbial phytase improves phosphorus digestibility and amino acid digestibility for certain plant-based feedstuffs.

Comparison of the ileal and faecal digestibility of dietary amino acids in adult humans and evaluation of the pig as a model animal for digestion studies in man.

The aim of the study was to determine if there is a difference between ileal and faecal assays for determining amino acid and N digestibilities in adult human subjects. Comparison of true ileal amino acid and N digestibilities was also made between adult human subjects and growing pigs to establish the usefulness of the pig as a model animal. Five subjects with established ileostomies and six subjects with intact large bowels consumed a constant diet consisting of meat, vegetables, fruit, bread and dairy products for 7 d with collection of ileostomy contents or faeces respectively over the last 4 d. The study was repeated using 25 kg body weight ileostomized and intact pigs. Apparent amino acid and N digestibility coefficients were determined. For human subjects the faecal digestibility values were significantly higher (P < 0.05) than the ileal values for Arg, Asp, Gly, Phe, Pro, Ser, Thr and Trp. The faecal digestibility of Met was significantly lower than the ileal value. Determination of DNA, diaminopimelic acid (DAPA) and the digestibilities of pectin, hemicellulose and cellulose in human subjects indicated that some microbial colonization had occurred at the terminal ileum after formation of an ileostomy; however, this was not as extensive as in the large intestine. True ileum amino acid and N digestibilities were calculated after correcting for the endogenous contribution of amino acids at the terminal ileum determined using a protein-free diet. There were no significant differences between adult human subjects and pigs for true ileal dietary amino acid digestibility except for Thr, Phe, Cys and Met. There were no significant differences between adult humans and pigs for the ileal digestibility of dry matter and the faecal digestibility of gross energy.

The role of cell surface architecture of lactobacilli in host-microbe interactions in the gastrointestinal tract.

Lactobacillus species can exert health promoting effects in the gastrointestinal tract (GIT) through many mechanisms, which include pathogen inhibition, maintenance of microbial balance, immunomodulation, and enhancement of the epithelial barrier function. Different species of the genus Lactobacillus can evoke different responses in the host, and not all strains of the same species can be considered beneficial. Strain variations may be related to diversity of the cell surface architecture of lactobacilli and the bacterias ability to express certain surface components or secrete specific compounds in response to the host environment. Lactobacilli are known to modify their surface structures in response to stress factors such as bile and low pH, and these adaptations may help their survival in the face of harsh environmental conditions encountered in the GIT. In recent years, multiple cell surface-associated molecules have been implicated in the adherence of lactobacilli to the GIT lining, immunomodulation, and protective effects on intestinal epithelial barrier function. Identification of the relevant bacterial ligands and their host receptors is imperative for a better understanding of the mechanisms through which lactobacilli exert their beneficial effects on human health.

Ileal amino acid digestibility assay for the growing meat chicken--comparison of ileal and excreta amino acid digestibility in the chicken

1. The apparent and true amino acid digestibilities in sorghum, wheat, soyabean meal, meat-and-bone meal, fish meal and blood meal for growing meat chickens were determined using an assay based on the collection of digesta from the terminal ileum and comparison was made with digestibility values determined using an excreta-based assay. 2. Five-week-old meat chickens were given maize-soyabean meal basal diet or mixtures of the basal diet and test diets containing the 6 ingredients as the sole source of dietary protein (50:50 on weight basis). Apparent amino acid digestibility values of assay diets at ileal and excreta levels were calculated using chromic oxide as the indigestible marker. True digestibility values were calculated using endogenous outputs determined by feeding a protein-free diet. Amino acid digestibilities of the ingredients were calculated by difference. 3. The site of measurement had no influence on endogenous amino acid output, the exceptions being aspartic acid and glutamic acid. The output of these two amino acids was higher in the excreta. 4. Significant differences were found between ileal and excreta-based digestibility of certain amino acids in some ingredients, with excreta values being usually higher than the ileal values, indicating a net catabolism of amino acids in the large intestine. The degree of net amino acid disappearance was found to be variable among amino acids and ingredients. In general, threonine had the lowest digestibility at the ileal level and, compared with other amino acids, the highest degredation during passage through the hindgut. 5. The results showed that digestibility determination based on excreta collection will overestimate the uptake for some amino acids in some feeds. The degree of overestimation was often considerable, ranging from 8.9% (apparent digestibility of threonine in soyabean meal) to 56% (apparent digestibility of aspartic acid in wheat). It is concluded that digestibility values measured at the terminal ileum provide a more reliable measure of amino acid availability than those measured in the excreta.

Long-Chain Polyunsaturated Fatty Acids and the Regulation of Bone Metabolism

The role of prostaglandin E2 (PGE2) in the regulation of bone remodeling is well established. There is increasing evidence that various long-chain polyunsaturated fatty acids (LCPUFAs), as well as nonprostanoid LCPUFA metabolites, also have critical roles in regulating bone metabolism and may have therapeutic potential in the management of postmenopausal osteoporosis. Although only the 18-carbon precursors for the n-3 and n-6 LCPUFAs are deemed “dietary essential,” the ability of the body to convert these precursor fatty acids into the more highly unsaturated 20- and 22-carbon LCPUFAs decreases with aging, menopause, and various lifestyle factors (e.g., smoking). Increasing dietary LCPUFA intake increases tissue and blood LCPUFA concentrations, as well as the concentrations of their metabolites. Modification of dietary LCPUFA content, particularly increasing the intake of n-3 LCPUFAs, has been shown to minimize the decline in bone mass caused by menopause in women and ovariectomy in animal models. This review summarizes findings from both in vivo and in vitro studies and outlines the effects of LCPUFAs and their metabolites on calcium balance, osteoblastogenesis, osteoclastogenesis, and osteoblast and osteoclast function.

Dietary Influences on Endogenous Ileal Protein and Amino Acid Loss in the Pig—A Review

Abstract The basis for using determined values of endogenous ileal protein and amino acid losses in the protein evaluation of pig feeds is reviewed. The amounts of endogenous protein and amino acids in ileal digesta may be determined by numerous methods. These include N-free diets, regression analysis, enzyme hydrolysed casein (EHC) coupled with ultra-filtration, modifying lysine to homoarginine, isotope dilution techniques and calculations based on the difference between the in vitro and in vivo digestibility of protein and amino acids. Endogenous ileal protein loss seems to be influenced primarily by dietary dry matter intake and secondarily by the composition of the diet and can be divided into a basal and an extra loss. The basal loss after ingestion of experimental N-free diets is 10-15 kg−1 DM intake, but, under more physiologically normal conditions when protein-containing diets are ingested, the basal loss seems to be about 20 g kg−1 DM intake. When vegetable feedstuffs are ingested there is often...

Description and validation of a model simulating growth in the pig (20–90 kg liveweight)

Abstract A deterministic computer model is described which simulates the growth of pigs (20 – 90 kg liveweight). From information on the sex, genotype, liveweight, and body lipid content at the start of growth, diet, and feeding scale, the model can be used to predict average daily liveweight gain, mean food conversion ratio, carcass weight at slaughter, and backfat thickness. A comparison of the actual and predicted growth performance of entire male and female pigs given a balanced grower diet at a daily rate of either 9 or 11% of metabolic body weight (kg 0.75) over the liveweight range 20 – 80 kg, demonstrated that the model can be used to give realistic predictions of pig growth performance.

Amino acid availability: aspects of chemical analysis and bioassay methodology

It is important to be able to characterise foods and feedstuffs according to their available amino acid contents. This involves being able to determine amino acids chemically and the conduct of bioassays to determine amino acid digestibility and availability. The chemical analysis of amino acids is not straightforward and meticulousness is required to achieve consistent results. In particular and for accuracy, the effect of hydrolysis time needs to be accounted for. Some amino acids (for example, lysine) can undergo chemical modification during the processing and storage of foods, which interferes with amino acid analysis. Furthermore, the modified amino acids may also interfere with the determination of digestibility. A new approach to the determination of available lysine using a modified in vivo digestibility assay is discussed. Research is required into other amino acids susceptible to structural damage. There is recent compelling scientific evidence that bacterial activity in the small intestine of animals and man leads to the synthesis and uptake of dietary essential amino acids. This has implications for the accuracy of the ileal-based amino acid digestibility assay and further research is required to determine the extent of this synthesis, the source of nitrogenous material used for the synthesis and the degree of synthesis net of amino acid catabolism. Although there may be potential shortcomings in digestibility assays based on the determination of amino acids remaining undigested at the terminal ileum, there is abundant evidence in simple-stomached animals and growing evidence in human subjects that faecal-based amino acid digestibility coefficients are misleading. Hindgut microbial metabolism significantly alters the undigested dietary amino acid profile. The ileal amino acid digestibility bioassay is expected to be more accurate than its faecal-based counterpart, but correction of the ileal amino acid flow for amino acids of endogenous origin is necessary. Approaches to correcting for the endogenous component are discussed.

The effect of dietary peptide concentration on endogenous ileal amino acid loss in the growing pig

The aim of the present study was to determine whether dietary peptide concentration had an effect on endogenous ileal amino acid flow in the growing pig. Eight 33 kg live weight entire male pigs had post-valve T-caecum (PVTC) cannulas surgically implanted for the collection of ileal digesta. The pigs were fed twice daily at 100 g/kg metabolic body weight per d and were given diets containing enzyme-hydrolysed casein (EHC) at 0, 50, 100 and 200 g/kg in a Latin-square design. A basal casein-based diet was fed to the pigs for 6 d periods between receiving the experimental diets. The pigs received the experimental diets for 8 d periods, with continuous collection of digesta for 24 h on each of the fifth and eighth days. The endogenous ileal amino acid flows were determined with reference to recovery of the marker, Cr, directly for pigs receiving the protein-free diet or after centrifugation and ultrafiltration (10,000 Da molecular mass cut-off) for pigs on the EHC-based diets. Mean endogenous ileal N flows were 1753, 1948, 2851 and 5743 micrograms/g DM intake when the pigs received diets containing 0, 50, 100 and 200 g EHC/kg respectively. There was a significant (P < 0.05) effect of dietary peptide concentration on the endogenous ileal flows of N and all of the amino acids, with an increase in endogenous ileal amino flow with increasing dietary EHC concentration.