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    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 PJ
    Background: 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.
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    Protein Intake and Protein Quality Patterns in New Zealand Vegan Diets: An Observational Analysis Using Dynamic Time Warping
    (MDPI (Basel, Switzerland), 2025-05-26) Soh BXP; Vignes M; Smith NW; Von Hurst PR; McNabb WC; Hayes M; Naik AS
    Background/Objectives: Inadequate intake of indispensable amino acids (IAAs) is a significant challenge in vegan diets. Since IAAs are not produced or stored over long durations in the human body, regular and balanced dietary protein consumption throughout the day is essential for metabolic function. The objective of this study is to investigate the variation in protein and IAA intake across 24 h among New Zealand vegans with time-series clustering, using Dynamic Time Warping (DTW). Methods: This data-driven approach objectively categorised vegan dietary data into distinct clusters for protein intake and protein quality analysis. Results: Total protein consumed per eating occasion (EO) was 11.1 g, with 93.5% of the cohort falling below the minimal threshold of 20 g of protein per EO. The mean protein intake for each EO in cluster 1 was 6.5 g, cluster 2 was 11.4 g and only cluster 3 was near the threshold at 19.0 g. IAA intake was highest in cluster 3, with lysine and leucine being 3× higher in cluster 3 than cluster 1. All EOs in cluster 1 were below the reference protein intake relative to body weight, closely followed by cluster 2 (91.5%), while cluster 3 comparatively had the lowest EOs under this reference (31.9%). Conclusions: DTW produced three distinct dietary patterns in the vegan cohort. Further exploration of plant protein combinations could inform recommendations to optimise protein quality in vegan diets.
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    Concentrations of Plasma Amino Acids and Neurotransmitters in Participants with Functional Gut Disorders and Healthy Controls
    (MDPI (Basel, Switzerland), 2023-02-20) James SC; Fraser K; Cooney J; Günther CS; Young W; Gearry RB; Heenan PE; Trower T; Keenan JI; Talley NJ; McNabb WC; Roy NC; Jang C
    Amino acids are important in several biochemical pathways as precursors to neurotransmitters which impact biological processes previously linked to functional gastrointestinal disorders (FGIDs). Dietary protein consumption, metabolic host processes, and the gut microbiome can influence the plasma concentration of amino acids and neurotransmitters, and their uptake by tissues. The aim of this analysis was to quantify 19 proteogenic and 4 non-proteogenic amino acids and 19 neurotransmitters (including precursors and catabolites, herein referred to as neurotransmitters) to ascertain if their circulating concentrations differed between healthy participants and those with FGIDs. Plasma proteogenic and non-proteogenic amino acids and neurotransmitters were measured using ultra-performance liquid chromatography and liquid chromatography-mass spectrometry, respectively, from 165 participants (Rome IV: irritable bowel syndrome (IBS-constipation, IBS-diarrhea), functional constipation, functional diarrhea, and healthy controls). There were significant differences (p < 0.05) in pairwise comparisons between healthy controls and specific FGID groups for branched-chain amino acids (BCAAs), ornithine, and alpha-aminobutyric acid. No other significant differences were observed for the neurotransmitters or any other amino acids analyzed. Multivariate and bivariate correlation analyses between proteogenic and non-proteogenic amino acids and neurotransmitters for constipation (constipation (IBS-C and functional constipation) and phenotypes diarrhea (IBS-D and functional diarrhea)) and healthy controls suggested that associations between BCAAs, 5-hydroxytryptophan, and kynurenine in combination with tyrosine, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxyphenylacetic acid and associations with gamma-aminobutyric acid, glutamate, asparagine, and serine are likely disrupted in FGID phenotypes. In conclusion, although correlations were evident between some proteogenic and non-proteogenic amino acids and neurotransmitters, the results showed minor concentration differences in plasma proteogenic and non-proteogenic amino acids, amino acid-derived metabolites, and neurotransmitters between FGID phenotypes and healthy controls.
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    The impact of heat treatment of bovine milk on gastric emptying and nutrient appearance in peripheral circulation in healthy females: a randomized controlled trial comparing pasteurized and ultra-high temperature milk
    (Elsevier Inc on behalf of the American Society for Nutrition, 2024-05-01) Milan AM; Barnett MPG; McNabb WC; Roy NC; Coutinho S; Hoad CL; Marciani L; Nivins S; Sharif H; Calder S; Du P; Gharibans AA; O'Grady G; Fraser K; Bernstein D; Rosanowski SM; Sharma P; Shrestha A; Mithen RF
    BACKGROUND: Heat treatments of dairy, including pasteurization and ultra-high temperature (UHT) processing, alter milk macromolecular structures, and ultimately affect digestion. In vitro, animal, and human studies show faster nutrient release or circulating appearance after consuming UHT milk (UHT-M) compared with pasteurized milk (PAST-M), with a faster gastric emptying (GE) rate proposed as a possible mechanism. OBJECTIVES: To investigate the impact of milk heat treatment on GE as a mechanism of faster nutrient appearance in blood. We hypothesized that GE and circulating nutrient delivery following consumption would be faster for UHT-M than PAST-M. METHODS: In this double-blind randomized controlled cross-over trial, healthy female (n = 20; 27.3 ± 1.4 y, mean ± SD) habitual dairy consumers, consumed 500 mL of either homogenized bovine UHT-M or PAST-M (1340 compared with 1320 kJ). Gastric content volume (GCV) emptying half-time (T50) was assessed over 3 h by magnetic resonance imaging subjective digestive symptoms, plasma amino acid, lipid and B vitamin concentrations, and gastric myoelectrical activity were measured over 5 h. RESULTS: Although GCV T50 did not differ (102 ± 7 min compared with 89 ± 8 min, mean ± SEM, UHT-M and PAST-M, respectively; P = 0.051), GCV time to emptying 25% of the volume was 31% longer following UHT-M compared with PAST-M (42 ± 2 compared with 32 ± 4 min, P = 0.004). Although GCV remained larger for a longer duration following UHT-M (treatment × time interaction, P = 0.002), plasma essential amino acid AUC was greater following UHT-M than PAST-M (55,324 ± 3809 compared with 36,598 ± 5673 μmol·min·L-1, P = 0.006). Heat treatment did not impact gastric myoelectrical activity, plasma appetite hormone markers or subjective appetite scores. CONCLUSIONS: Contrary to expectations, GE was slower with UHT-M, yet, as anticipated, aminoacidemia was greater. The larger GCV following UHT-M suggests that gastric volume may poorly predict circulating nutrient appearance from complex food matrices. Dairy heat treatment may be an effective tool to modify nutrient release by impacting digestion kinetics. CLINICAL TRIAL REGISTRY: www.anzctr.org.au (ACTRN12620000172909).
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    Protein and amino acid digestibility: definitions and conventional oro-ileal determination in humans
    (Frontiers Media S.A., 2024-06-20) Hodgkinson SM; Sergi D
    When 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.
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    Dynamic Gastrointestinal Digestion of Bovine, Caprine and Ovine Milk Reconstituted from Commercial Whole Milk Powders
    (MDPI AG, 2024-05-02) Li S; Ye A; Cui J; Zhang Y; Ware L; Miller JC; Abbotts-Holmes H; Roy NC; Singh H; McNabb W; Polidori P
    The global dairy market has been increasingly diversified with more dairy product offerings of milk products from different animal species. Meanwhile, milk powders remain the main exported dairy product format due to their ease of transportation. In this work, we studied the structural changes, protein hydrolysis and nutrient delivery during dynamic gastric digestion and small intestinal digestion of cow, goat and sheep milk reconstituted from commercial whole milk powders. The results show that the reconstituted milks digest similarly to processed fresh milk. The digestion behaviors of the three reconstituted ruminant milks are broadly similar (gastric coagulation, kinetics of gastric emptying of protein and fat and the high digestibility in the small intestine) with some differences, which are likely contributed by the processing history of the milk powders. The delivery of individual amino acids to the small intestine differed between the early and late stages of gastric digestion, which were primarily affected by the abundance of amino acids in caseins and whey proteins but also by the difference between milk types associated with their gastric coagulation behaviors. This work showed that powdered milk is similar to fresh processed milk in digestion behavior, and the inherent differences between ruminant milks can be modified by processing treatments.
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    Effects of a novel consensus bacterial 6-phytase variant on the apparent ileal digestibility of amino acids, total tract phosphorus retention, and tibia ash in young broilers
    (Oxford University Press, 2022-02-01) Dersjant-Li Y; Abdollahi MR; Bello A; Waller K; Marchal L; Ravindran V
    The effect of a novel consensus bacterial 6-phytase variant (PhyG) on apparent ileal digestibility (AID) of amino acids (AA) and phosphorus (P) utilization in young broilers when added to diets with high phytate-P (PP) content without added inorganic phosphate (Pi) and deficient in digestible (dig) AA and metabolizable energy (ME) was investigated. A total of 256 Ross 308 male broilers were assigned to 4 treatments (8 birds/cage, 8 cages/treatment) in a completely randomized design. Treatments comprised a positive control (PC, 2,975 kcal/kg ME, 3.7 g/kg dig P, 2.83 g/kg PP, 8.4 g/kg Ca, 10.6 g/kg dig lysine), a negative control (NC) without added Pi (ME −68 kcal/kg, crude protein −10 g/kg, dig AA −0.1 to −0.4 g/kg, Ca −2.0 g/kg, dig P −2.2 g/kg, Na −0.4 g/kg vs. PC), and NC plus 500 or 1,000 FTU/kg of PhyG. Test diets were corn/soy/rapeseed-meal/rice-bran-based and fed from 5 to 15 d of age. Ileal digesta and tibias were collected on day 15. Excreta was collected during days 12 to 15 to determine P retention. The NC (vs. PC) reduced (P < 0.05) P retention (−10.4% units), tibia ash (−14.3% units), weight gain (−109 g), feed intake (−82 g) and increased FCR (from 1.199 to 1.504), confirming that the NC was extremely deficient in nutrients and energy. Phytase addition to the NC linearly (P < 0.001) improved performance, but did not fully recover it to the level of the PC due to the severe nutrients/energy reduction in NC. Phytase linearly increased P retention (P < 0.001), tibia ash (P < 0.001), AID of dry matter (P < 0.05), nitrogen (P < 0.01), gross energy (P < 0.05), and all 17 individual AA (P < 0.01). At 1,000 FTU/kg, phytase increased (P < 0.05) P retention vs. PC and NC (+14.5 and +24.9% units, respectively) and increased tibia ash vs. NC (+13.8% units), equivalent to PC. The NC decreased AID of Cys, Gly, Thr, and Met vs. PC (P < 0.05). At 1,000 FTU/kg, phytase increased AID of all 17 AA vs. NC (P < 0.01), equivalent to PC. At 1,000 FTU/kg, AID AA responses (above NC) ranged from +4.5% (Met) to +15.0% (Cys), being maximal for essential Thr (+10.4%) and Val (+8.2%) and non-essential Cys (+15.0%) and Gly (+10.4%). The results highlight the efficacy of PhyG at a dose level of 500 to 1,000 FTU/kg in young broilers for improving the ileal digestibility of nitrogen, AA, and energy alongside P retention and tibia ash. The performance data emphasize the need to consider digestible nutrient intake as a response variable in exogenous enzyme studies.
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    Modeling improvements in ileal digestible amino acids by a novel consensus bacterial 6-phytase variant in broilers
    (Elsevier, 2022-03) Dersjant-Li Y; Bello A; Stormink T; Abdollahi MR; Ravindran V; Babatunde OO; Adeola O; Toghyani M; Liu SY; Selle PH; Marchal L
    Data from 13 datasets from 4 trials on the effect of a novel consensus bacterial 6-phytase variant (PhyG) on the apparent ileal digestibility (AID) of amino acids (AA) in broilers were used to model AID AA responses. The datasets were obtained from 3 trial locations (New Zealand, Australia and United States) and collectively incorporated variations in diet composition (feedstuff composition, phytate-P (PP) level, limestone solubility), feed form (mash or pellet), bird genetics (strain), and age at sampling (11–35 d of age). In total, 384 observations were analyzed. First, the relationships between AID of AA (as coefficients) and increasing phytase dose level from 0 to 4,000 FTU/kg were evaluated across all datasets using exponential curve fitting. Second, the percentage unit change in AID of AA at each phytase dose level from baseline (basal diet [BD] without phytase) was calculated separately for each dataset and the data then modeled together using exponential curve fitting. The model-predicted mean coefficient of AID of total AA in basal diets was 0.76 (range 0.56 [Cys] to 0.83 [Glu]), which was increased by PhyG to 0.80 and 0.81 at 2,000 and 4,000 FTU/kg, respectively. Exponential increases in the percentage unit improvement in AID of 18 individual and of total AA with increasing phytase dose level were evident (P < 0.05). Improvements (vs. BD) at 2,000 FTU/kg and 4,000 FTU/kg, respectively, were greatest for Cys (+9.2 and +11.0% units), Met (after deduction of synthetic Met, +8.4 and +9.0% units), and Thr (after deduction of synthetic Thr, +6.2 and +7.3% units). The data demonstrated consistent improvements in the AID of AA by the phytase. The modeling results generated from data gathered from birds sampled at different ages and from different dietary settings with correction of synthetic AA for Lys, Met, Thr, and Trp, enabled a more accurate prediction of the digestible AA contribution from the diet by this novel phytase. This will allow diet-specific AA matrix recommendations to be made in commercial feed formulations.
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    Metabolizable energy and standardized ileal amino acid digestibility of full-fat soybeans for broilers are influenced by wet-heating, expansion temperature, and autoclaving time
    (Elsevier Inc. on behalf of Poultry Science Association Inc, 2022-09) Abdollahi MR; Wiltafsky-Martin M; Zaefarian F; Ravindran V
    The influence of wet-heating (WH) and expansion temperature (ET), and autoclaving time (AT) on the nitrogen-corrected apparent metabolizable energy (AMEn) and standardized ileal digestibility (SID) of AA in full-fat soybeans (FFSB) for broilers was examined in 2 experiments. The AMEn and SID AA of FFSB were determined by the difference and direct methods, respectively. In Experiment 1, raw FFSB (K0) were either treated by WH at 80°C for 1 min and expanded at 115°C (K1-115) or 125°C (K1-125), WH at 100°C for 6 min and expanded at 115°C (K2-115) or 125°C (K2-125), or WH at 100°C for 16 min and expanded at 115°C (K3-115) or 125°C (K3-125). Wet-heating and ET significantly (P < 0.001) increased the AMEn in FFSB. Among heat-treated FFSB, K1-115 and K1-125 resulted in the lowest and highest AMEn values, respectively, with all samples wet-heated at 100°C being intermediate. The K3-125 had AMEn values similar (P > 0.05) to K1-125. Among heat-treated FFSB, the highest average SID AA was recorded for K3-125. In Experiment 2, K3-125 from experiment 1 was divided into 9 batches and autoclaved at 110°C for 15 (Z1), 30 (Z2), 45 (Z3), 60 (Z4), 120 (Z5), 180 (Z6), 240 (Z7), 300 (Z8), and 360 (Z9) min. A quadratic (P < 0.01) pattern was observed for the effects of AT on AMEn. The AMEn was unaffected until 300 min AT and then declined at 360 min. The AT quadratically (P < 0.001) affected the average SID AA where the SID increased from K3-125 to Z1, plateaued to Z5, and then declined to Z9. In conclusion, the results demonstrated that WH at 100°C for 16 min followed by expansion at 125°C as the most optimal wet-heating and expansion processing, associated with the highest SID AA. Autoclaving at 110°C for 30 min enhanced energy utilization and AA digestibility in FFSB, suggesting that further advantages may be achieved by short-time autoclaving of previously wet-heated and expanded FFSB samples.
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    Basal ileal endogenous amino acid flow in broiler chickens as influenced by age
    (Elsevier Inc, 2021-08) Barua M; Abdollahi MR; Zaefarian F; Wester TJ; Girish CK; Chrystal PV; Ravindran V
    The current study was carried out to measure the basal ileal endogenous amino acid (EAA) flow in male broilers (Ross 308) at different ages (d 7, 14, 21, 28, 35, and 42), following the feeding of a nitrogen-free diet. Titanium dioxide (5 g/kg) was included as an indigestible marker. The nitrogen-free diet was offered for four days prior to ileal digesta collection to 6 replicate cages housing 14 (d 3–7), 12 (d 10–14), 10 (d 17–21), 8 (d 24–28), 8 (d 31–35), and 6 (d 38–42) birds per cage. The basal EAA flow was calculated as g/kg DM intake. The amino acid (AA) profile of endogenous protein, expressed as g/100 g protein, was also calculated. The basal endogenous flow of nitrogen and all individual and total AA decreased quadratically (P < 0.05 to 0.001), with flows being higher on d 7, then decreasing on d 14, plateauing until d 35 and decreasing further on d 42. The concentrations of Trp, Cys, and Gly in the endogenous protein increased linearly (P < 0.01 to 0.001) with advancing age, whereas a linear decrease (P < 0.001) was noted for Lys. A quadratic influence (P < 0.05 to 0.001) was observed for the concentrations of Ile, Leu, Met, Val, and Asp. These changes in the endogenous protein profile may be attributed to variations in the contribution of endogenous sources with age but delineating the exact contribution of different sources is complicated. Overall, the current findings suggest that the basal ileal EAA flow is influenced by broiler age and age-specific EAA flows may need to be considered to standardize the AA digestibility.