Journal Articles
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Item True Ileal Amino Acid Digestibility of Human Foods Classified According to Food Type as Determined in the Growing Pig(Elsevier Inc on behalf of the American Society for Nutrition, 2025-10-18) Hodgkinson SM; Stroebinger N; Stein HH; Fanelli NS; de Vries S; van der Wielen N; Hendriks WH; Moughan PJBackground: A Food and Agriculture Organization Expert Consultation recommended the use of digestible indispensable amino acid score (DIAAS) to evaluate protein quality of foods for humans. Calculation of DIAAS requires true ileal digestibility (TID) of amino acid (AA) values but currently insufficient data are available. Objectives: This study aims to generate in pigs TID of AA for a wide range of foods commonly consumed by humans and determine the range of differences in TID of AA among food types. Methods: A standardized protocol was followed to determine TID of AA in 97 foods across 3 laboratories. Female pigs (25–100 kg during study, n ≥ 6) received foods for 7 d following a Youden Square design with ileal digesta collected via T-cannula on days 6–7. Endogenous AA losses were determined by feeding a protein-free diet. Foods, diets, and digesta were analyzed for nitrogen, AA, reactive lysine, titanium and dry matter. Foods were categorized into food types with the degree of variation within each food type evaluated using descriptive statistics. Results: The TID (mean of AA) ranged from 0.247 (apples) to 0.988 (beef tenderloin). The median TID of AA was high (mean of AA > 0.90) for foods categorized as dairy products, eggs, fish and seafood, isolates and concentrates, meat, nuts, plant-based burgers, soy products and wheat products. Food categories with median TID < 0.80 were baked products, fruit, pulses and seeds, and wheat bran cereal, yeast, and zein. Food categories with low variations between foods were fish and seafood (1% units), dairy products (3% units), and eggs (5% units), whereas categories with the greatest variation were grains (18% units), vegetables (16% units), seeds (14% units), and fruit (12% units). There was considerable variation in TID for individual AA both within and among foods. Conclusions: The database with TID of AA of 97 foods generated by 3 laboratories using a standardized methodology can be utilized for protein quality evaluation.Item Exploring in vitro production of colonic microbial metabolites from diverse protein sources using human ileal digesta(Elsevier Ltd, 2025-12-15) van der Wielen N; Zhang H; Schouten PJC; Meulenbroeks E; Stroebinger N; Hodgkinson SM; Mensink M; Hendriks W; Capuano EWe explored the relationship between protein fermentation metabolites and ileal digesta composition, using ileal digesta from ileostomates, who ingested nine different protein sources, incubated in the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®). NH3, short-chain fatty acids, branched-chain fatty acids (BCFA), H2S, tryptophan derivatives, and biogenic amines were measured in proximal and distal colon vessels. The relative changes in most metabolites were positively correlated with their amino acid precursors in ileal digesta. In both colon vessels, the relative change of NH3 was a good predictor for the production of other metabolites. Indole was strongly associated with oxindole, 5-HT, and tryptamine and the sum of Trp metabolites in the distal colon. Per gram ingested protein, zein and whey may produce the highest levels of NH3 and BCFA in the proximal colon and BCFA in the distal colon, whereas whey and pigeon peas may result in the highest levels of H2S.Item Protein extraction from Buckwheat, Chondrus crispus, and Spelt and assessment of nutritional benefits and limitations in vitro(Springer Nature Limited, 2025-09-24) Cain E; Hodgkinson SM; McNabb W; Brodkorb A; Giblin L; Hayes MPlant protein consumption has increased globally but concerns exist regarding their ability to provide sufficient amino acids to consumers. Extraction methods that can separate protein from anti-nutritional factors have potential to increase the nutritional value of this biomass. Few studies concerning analysis of the amino acid content of plant protein extracts exist. In this work, three different protein extraction methods were used to generate protein extracts from Buckwheat, Spelt and the red seaweed Chondrus crispus. Methods used include ultra-sonication in water combined with ammonium sulphate-induced protein precipitation; an enzymatic extraction method using the enzymes Alcalase and Viscozyme, and an iso-electric precipitation extraction method using alkaline protein solubilization followed by acidic protein precipitation. Proteins extracted using the enzymatic method contained the highest proportion of essential amino acids (EAA) in viable quantities, and the method holds promise for use in the generation of alternative marine and cereal protein extracts for human consumption.Item Gastric protein digestion of cow, goat, and sheep milk is not reflected in the amino acid appearance in the blood of suckling piglets(Elsevier Inc on behalf of the American Dairy Science Association, 2025-06) Roy D; Montoya CA; Stroebinger N; Hodgkinson SM; Ye A; McNabb WC; Moughan PJ; Singh HStructural changes in milk during gastric digestion are a key driving factor for the rate of digestion of nutrients in the gastrointestinal tract. Thus, the influence of gastric coagulation behavior on the kinetics of protein digestion of raw cow, goat, and sheep whole milk in the stomach was investigated using the 3-wk-old suckled male piglet as an animal model for human infants. Piglets received a single meal of fresh raw milk normalized for protein content, and were slaughtered at 0, 30, 90, 150, or 210 min postprandially. Gastric chyme and cardiac blood samples were collected. Gastric pepsin activity, rate of protein hydrolysis, and gastric emptying of AA were determined along with how these changes influence the appearance of AA in the plasma. The disappearance rates of individual proteins (especially β-LG and αS-CN), total digested proteins entering the small intestine, as well as the gastric emptying of some AA (proline, leucine) were (or tended to be) greater for goat and sheep milk than for cow milk. Differences in plasma concentrations for some AA (e.g., leucine) were observed across milk types, but they did not directly reflect changes in gastric protein digestion and the gastric emptying of AA. In conclusion, a combination of protein (and AA) composition, susceptibility of specific proteins to hydrolysis, and the nature of the curd structure formed influenced the digestion behavior of milk proteins in the stomach and their subsequent release into the small intestine.Item Editorial: Dietary protein for human health.(Frontiers Media S.A., 2025-01-15) Moughan PJ; Hendriks WH; Hodgkinson SM; Chungchunlam SMS; Lim WXJ; Mensink M; Stroebinger N; van der Wielen N; Pivovarova-Ramich OItem The kinetics of amino acid disappearance in the small intestine is related to the extent of amino acids absorbed in growing pigs(Cambridge University Press on behalf of The Nutrition Society, 2024-03-14) Montoya CA; van Bemmel M; Kreutz K; Hodgkinson SM; Stroebinger N; Moughan PJThis study evaluated the importance of a correction for amino acids (AA) released into the hindgut on a measure of AA absorption kinetics and tested whether AA absorption kinetics are related to the extent of AA absorption using the growing pig as a model for humans. Thirty-six nine-week-old pigs (22·3 kg) received a diet containing whey protein as the sole protein source for 8 d. Pigs received their last meal containing the indigestible marker titanium dioxide before being euthanised at 1, 2, 3, 4, 6 and 12 h post-feeding. The entire content of each gastrointestinal tract (GIT) region was collected to determine AA released into the hindgut, and the kinetics and extent of AA absorption (uncorrected and corrected for AA entering the hindgut). Amounts of AA released into the hindgut increased over time (e.g. 33 and 180 mg of Glu for 4 and 6 h post-feeding). The corrected apparent amount of each AA absorbed from the GIT lumen after 4 h post-feeding was generally lower (P ≤ 0·05) than the uncorrected counterpart. Differences in both the kinetics and extent of AA absorption were observed across AA. For example, the time to reach half of the apparent AA absorption (T50) was 1·5 and 3·4 h for Met and Arg, respectively, whereas their extent of apparent absorption was 93 and 73 %. Negative correlations between parameters related to kinetics and the extent of apparent absorption were observed (e.g. for T50 r = -0·81; P < 0·001). The kinetics of AA absorption is related to the extent of AA absorption.Item Protein and amino acid digestibility: definitions and conventional oro-ileal determination in humans(Frontiers Media S.A., 2024-06-20) Hodgkinson SM; Sergi DWhen assessing protein quality, a correction needs to be made to take into consideration the availability of the amino acids. This correction is based on the digestibility of the amino acids. It is recommended to use ileal (end of small intestine) digestibility as opposed to faecal digestibility. A correction needs to be made for endogenous (gut sourced as opposed to diet sourced) amino acids to give true digestibility as opposed to apparent digestibility. Also, this correction should be made by correcting the amino acid composition for individual amino acid digestibilities as opposed to correcting all amino acids for nitrogen digestibility. Determination of true ileal amino acid digestibility requires the collection of ileal digesta. In the human there are two methods that can be used; naso-ileal intubation and using the ileostomy model. Both are discussed in detail and it is concluded that both are appropriate methods to collect ileal digesta.Item In Vitro Ileal Fermentation is Affected More by the Fiber Source Fermented than the Ileal Microbial Composition in Growing Pigs(Elsevier Inc. on behalf of American Society for Nutrition, 2023-05) Hoogeveen AM; Moughan PJ; Hodgkinson SM; Stroebinger N; Yu W; Rettedal EA; McNabb WC; Montoya CABACKGROUND: The fermentation of undigested material in the ileum is quantitatively important. However, the respective contributions of the microbial composition and the substrate to ileal fermentation are unclear. OBJECTIVE: This aim was to investigate the contribution of microbial composition and fiber source to in vitro ileal fermentation outcomes. METHODS: Thirteen ileal cannulated female pigs (Landrace/Large White; 9-wk-old; 30.5 kg body weight) were given diets containing black beans, wheat bread, chickpeas, peanuts, pigeon peas, sorghum, or wheat bran as the sole protein source for 7 d (100 g protein/kg dry matter diet). On day 7, ileal digesta were collected and stored at -80°C for microbial analysis and in vitro fermentation. For each diet, a pooled ileal inoculum was prepared to ferment different fiber sources (cellulose, pectin, arabinogalactan, inulin, fructooligosaccharides, and resistant starch) for 2 h at 37°C. Organic matter fermentability and organic acid production were determined following in vitro fermentation. Data were analyzed using a 2-way ANOVA (inoculum × fiber). RESULTS: Forty-five percent of the identified genera in the digesta differed across diets. For instance, the number of Lactococcus was 115-fold greater (P ≤ 0.05) in the digesta of pigs fed the pigeon pea diet than for pigs fed the wheat bran diet. For both in vitro organic matter fermentability and organic acid production, there were significant (P ≤ 0.05) interactions between the inoculum and the fiber source. For instance, pectin and resistant starch resulted in 1.6- to 31-fold more (P ≤ 0.05) lactic acid production when fermented by the pigeon pea inoculum than other inocula. For specific fiber sources, statistically significant correlations were found between the number of bacteria from certain members of the ileal microbial community and fermentation outcomes. CONCLUSIONS: Both the fiber source fermented and the ileal microbial composition of the growing pig affected in vitro fermentation; however, the effect of the fiber source was predominant.Curr Dev Nutr 2023;x:xx.Item In vitro digestibility of dietary proteins and in vitro DIAAS analytical workflow based on the INFOGEST static protocol and its validation with in vivo data(Elsevier Ltd, 2023-03-15) Sousa R; Recio I; Heimo D; Dubois S; Moughan PJ; Hodgkinson SM; Portmann R; Egger LThe FAO recommends the digestible indispensable amino acid score (DIAAS) to determine protein quality in foods, preferably tested in vivo. Here, the INFOGESTin vitrodigestion protocol was applied and supplemented with an analytical workflow allowing the assessment of protein digestibility and DIAAS calculation. The protocol was applied to selected samples WPI, zein, collagen, black beans, pigeon peas, All-Bran®, and peanuts. The total protein digestibility, digestibility of individual amino acids (AA), and DIAAS values were established and compared with in vivo data for the same substrates. Total protein digestibility (total Nitrogen, r = 0.7, P < 0.05; primary amines (OPA), r = 0.6, P < 0.02; total AA, r = 0.6, P < 0.02) and digestibility of individual AA (r = 0.6, P < 0.0001) were in good agreement, between in vitro and in vivo, with a mean difference of 1.2 %. In vitro DIAAS was highly correlated with DIAAS obtained from in vivo true ileal digestibility values (r = 0.96, R2 = 0.89, P < 0.0001) with a mean difference of 0.1 %.Item Comparison of True Ileal Amino Acid Digestibility between Adult Humans and Growing Pigs(Oxford University Press on behalf of the American Society for Nutrition, 2022-07) Hodgkinson SM; Stroebinger N; van der Wielen N; Mensink M; Montoya C; Hendriks WH; de Vries S; Stein HH; Moughan PJBACKGROUND: It is not feasible to determine the true ileal amino acid (AA) digestibility of protein sources in humans on a routine basis, and the growing pig has been recommended as an animal model for this purpose but requires further validation. OBJECTIVES: To determine and compare true ileal AA digestibility between adult human ileostomates and growing cannulated pigs for a range of food proteins. METHODS: Seven protein sources (black beans, bread, collagen, pigeon peas, wheat bran, whey protein isolate, and zein) that spanned the range of digestibilities typically seen in foods were evaluated. Six female growing pigs received each of the protein sources, as well as a protein-free diet, and digesta were collected via ileal T-cannula. Adult human ileostomates consumed the same protein sources (5-8 ileostomates, depending on the protein source), as well as a protein-free diet, and digesta were collected. Titanium dioxide and celite were included in the diets as indigestible markers. True ileal AA digestibility coefficients were determined. RESULTS: There was a significant effect of protein source (P ≤ 0.001) for all AAs. The effect of species was not significant (P > 0.05) except for total lysine (but not for available lysine). When analyzed within diets, the statistically significant species effect for true lysine digestibility was found for black beans only. Pig and human digestibility values were generally highly and significantly (P ≤ 0.05) correlated. A linear regression equation derived for true ileal AA digestibility (given as coefficients) determined in the human and pig for the overall mean of all AAs was (y = human, x = pig) y = 1.00x - 0.010, with the slope not statistically significant (P > 0.05) from unity and the intercept not different (P > 0.05) from zero. CONCLUSIONS: True ileal AA digestibility values determined in the growing pig can be directly used for predicting digestibility in adult humans.