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Elderly individuals experience age-related declines in digestive function, which can hinder the digestion of dietary proteins and thereby negatively impact overall health. This study provides a comprehensive understanding of the protein digestive fate of beef with Protease-DS supplementation by analyzing gastric emptying behavior and amino acid and peptide profiles during a simulated in vitro elderly digestion process. The adult model exhibited superior gastric emptying capacity, with relatively rapid and early expulsion of gastric digesta into the intestinal segment. Additionally, the underperforming elderly model showed notable improvement in the emptying process with Protease-DS supplementation, achieving a rate of 0.0110 min -1 , which was close to the adult model of 0.0125 min -1 and significantly faster than the elderly model of 0.0080 min -1 . Protein digestibility in the elderly model was significantly enhanced with Protease-DS supplementation, increasing from 55.62 % to 63.60 %, approaching that of the adult model (69.60 %). Protease-DS significantly improved the essential amino acid score (EAAS), particularly for methionine + cysteine, phenylalanine + tyrosine, lysine, and leucine. Furthermore, it enhanced the release of unique polypeptide fragments, distinct from those in both the adult and elderly models, and demonstrated significant anti-amnesic and antithrombotic bioactive functions, highlighting its potential to improve memory and reduce blood clotting. This study provides guidance for elderly consumers on enhancing dietary protein digestion through protease supplementation, presenting it as an effective strategy to address protein deficiency and improve health and well-being., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

With an ever-increasing global population and dwindling natural resources, a shift towards more sustainable food systems is required. Important aspects to aid in this transition are the reduction of food waste, and a movement towards non-animal protein sources. Brewers spent yeast (BSY) is an abundant by-product of the brewing industry, which is generally regarded as waste, despite its high nutritional value. Previous work has shown that fermentation of BSY with Lactobacillus amylovorus FST 2.11 resulted in changes in composition, functionality, and improved palatability of the processed raw material (PBSY). In this study, in vitro protein digestibility, amino acid bioaccessability, and protein quality of PBSY was explored using the static INFOGEST in vitro model. In vitro protein digestibility of PBSY (73.0 %) was almost two-fold higher than that of CBSY (40.0 %), while PBSY also displayed significantly higher in vitro bioaccessability values for all essential amino acids, except for tryptophan. Investigation of protein quality using the digestible indispensable amino acid score (DIAAS) values and the FAO recommended amino acid scoring pattern for individuals >3 years old showed that the protein quality for CBSY was low (DIAAS of 17.0 %), while PBSY was considered to be of "good" protein quality (DIAAS of 98.2 %). Investigation of the modulation potential of PBSY on the gut microbiome using an in vitro colon model system showed an increase in gut microbiome α-diversity indices and an abundance of beneficial Mediterranean diet-responsive taxa after 24 h. Overall, this study highlights the potential of BSY as raw material for the production of a high-quality food ingredient with potential prebiotic effects, aiding in the reduction food waste and supporting global food systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This study aimed to explore the effects of egg albumin protein addition (5, 15 and 20 g/100 g db) on the textural characteristics, as well as in the in vitro digestibility of protein and starch of wheat bread. Egg albumin addition resulted in smoother bread loaves as compared to traditional wheat bread. Reduced hardness and increased cohesiveness were correlated to the protein secondary structure, mainly with the content of β-sheets. The in vitro protein digestibility decreased with the albumin addition, suggesting the mediation of protein-starch interactions. The in vitro starch digestibility was also decreased, reflected in a huge decrease of the slowly digestible starch fraction, but with non-significant changes in the rapidly digestible starch fraction. The supplemented albumin can form a physical barrier around the starch granules, which hampers the access of the amylolytic enzymes to the starch chains. Overall, the results of this study suggest that the addition of egg albumin is a viable alternative for producing wheat bread with reduced glycemic index and improved nutritional properties., Competing Interests: Declarations. Competing Interests: The authors declare no competing interests., (© 2025. The Author(s).)

Advanced glycation end products (AGEs) are a heterogeneous group of compounds formed both endogenously and exogenously through reactions between reducing sugars and amino acids within the proteins. The digestive tract may also serve as a site for endogenous AGEs generation. This study examined whether additional AGEs are formed during the digestion of glycated protein diets and meal-resembling systems (dietary proteins with fructose or glyoxal). The digestion of glycated protein showed that free AGEs were gradually released, but no additional AGEs were generated. In contrast, co-digestion of dietary proteins with fructose or glyoxal resulted in the formation of additional AGEs, and the reaction substrates (fructose or glyoxal) were depleted during digestion. Additionally, the lysine released from proteins decreased, leading to a loss of nutritional value of the food during co-digestion. The formation of AGEs and the depletion of essential amino acids in the gut may have significant implications for human health., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2024. Published by Elsevier Ltd.)

This study aimed to evaluate the impact of in vitro digestive characteristics of five different protein sources (fish, crab, beef, chicken, and soy) in adults and elderly individuals. Simulated gastrointestinal tract degradation in the elderly significantly reduced the digestibility of all proteins. The Elderly model had difficulties digesting any protein that had a firm structure or was prone to aggregation. Of the five protein types, soy protein was the least easily digested. Ultraviolet spectroscopy and SDS-PAGE analysis revealed that beef and crab proteins contain myoglobin and hemocyanin, respectively, making their structure firm. Simultaneously, alkali-soluble proteins in meat were more likely to be digested by the elderly than salt-soluble proteins. The alkali-soluble protein percentage in fish protein was the highest, including the best in vitro digestion characteristics in the elderly population. This study provides novel and significant recommendations for the elderly's daily intake of various protein sources., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

This study aimed to evaluate the inclusion of insect larvae meal (Tenebrio molitor) on the apparent total digestibility of dry matter (DM), gross energy (GE), ether extract (EE), crude protein (CP), neutral and acid detergent fiber (NDF and FDA), and the apparent and standardized ileal digestibility of amino acids (Exp.1) and to evaluate the inclusion of insect meal with iron and manganese supplementation on growth performance and serum hemoglobin concentration by collecting blood (day zero), on the 14th and 32nd days of the experiment (Exp.2). The standardized ileal digestibility values for amino acids and crude protein of insect meal were similar to those found for soybean meal, with high metabolizable energy values (Exp.1), and there was no difference in performance for any of the variables analyzed (p ≥ 0.05) in any of the periods (Exp.2). For serum hemoglobin values, there was no interaction (p ≥ 0.05) between the treatments used and the collection days; however, hemoglobin values increased (p < 0.05) at each collection time. It can therefore be concluded that insect meal for pigs can be considered an alternative ingredient to soybean meal as a source of protein in piglet nutrition due to high digestibility of amino acids without deleterious effect on productive performance., (© 2025 Japanese Society of Animal Science.)

The development of a healthier and more sustainable food supply is a main concern of consumers, industry, governments, and international institutions. Foods containing high levels of rapidly digestible starches have been linked to a rise in the number of people suffering from diet-related chronic diseases. Consequently, there is interest in reducing the digestibility of starch to improve their healthiness. The ability of natural additives including proteins, dietary fibers, and polyphenols, and sustainable processing technologies such as high-intensity ultrasonic, pulsed electric field, non-thermal plasma, γ-ray irradiation that regulate reduce starch digestibility in foods are reviewed. The potential mechanisms of action, advantages, and disadvantages of each approach at inhibiting starch digestibility is highlighted. The potential for commercializing these technologies is discussed, and areas where further research are required are emphasized. Natural additives and sustainable processing operations can effectively reduce the digestibility of starch and inhibit postprandial sugar "spikes" in the bloodstream by adjusting the structural changes, which can be used to create healthier and more sustainable foods and have broad application prospects.

Early nutritional regulation has become a research hotspot. The present study was undertaken to assess dietary early addition enzyme-treated soybean meal (ESBM) on nutrient digestibility, amino acid absorption, and intestinal development of broilers. Four hundred and fifty 1-day-old broilers were divided into three groups with 10 replicates of 15 broilers and fed with a basic diet, 2.5 or 5% ESBM during 1-10 days. Then, all groups were fed the same basic diets until 42 d. The obtained results indicated that supplementation of ESBM in early stage resulted in heavier BW and a better feed conversion ratio during the experimental periods compared with the control group. Supplementing the broiler's diet with 5% ESBM led to enhance digestibility of DM, CP and decrease abdominal fat yield (P < 0.05). Meanwhile, 5% ESBM in the early diet increased villus height (P < 0.05, d 21) and reduced the crypt depth in jejunum and ileum (P < 0.05, d 42). Dietary 5% ESBM supplementation improved enzyme activity and upregulated nutrient transporters expression in jejunum, increased plasma amino acid (AA) concentrations (P < 0.05, d 10). In conclusion, our data demonstrated that supplementation ESBM promote intestinal development, upregulate AAs and peptide transporters gene expression for physiological absorption of AAs, and improve protein digestibility caused by intestinal enzyme activity, thereby improving growth performance., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The effects of tiger nut insoluble dietary fiber (TNF) (1%-3% w/w) on techno-functional properties and in vitro protein digestibility of low-fat meat emulsions were examined. The results showed that TNF (especially 2% and 3%) could improve techno-functional performances of low-fat meat emulsions in terms of water binding capacity and textural attributes, which were even preferable to the high-fat control. The improvements were associated with the attenuated fluidity of inner water within the gels, the increased storage modulus (G'), the formation of a well-joined gel network with uniformly-dispersed fat particles and the uncoiling of α-helix and the formation of β-sheet. The weakened water mobility, the heat-triggered conformational shift from α-helix to β-sheet, as well as less and evenly distributed fat particles ulteriorly led to elevated protein digestibility in meat emulsions. Therefore, TNF shows promise as a fat substitute to develop fat-reduced emulsified pork meat products with desirable quality and digestion behaviors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Protein digestion and microbial metabolism play crucial roles in overall health. However, the mechanisms that differentiate the digestion and metabolism of dietary proteins from different sources in the organism remain poorly understood. This study investigated the digestive properties and microbial fermentation of various animal proteins (chicken, pork, beef, and casein) and plant proteins (soy bean, mung bean, kidney bean, rice, and wheat) in an in vitro simulation. The results indicated that animal-derived proteins had higher essential amino acid content (33.97-37.12 g/100 g) and digestibility levels (49.15-60.94 %), and provided more small molecule peptides upon digestion. Nevertheless, soy bean and wheat proteins also exhibited higher digestibility (54.70 % and 60.94 %), probably due to the extraction process. The fermentation results showed that distinct metabolic profiles that emerged for different protein sources. Plant-derived proteins (especially kidney bean, rice and wheat) promoted the proliferation of beneficial bacteria and microbial diversification and stimulated short-chain fatty acids (SCFA) production. Conversely, meat proteins (pork, chicken, beef) had significantly lower microbial diversity and SCFA than these plant proteins. These findings provide valuable insights into the effects of dietary protein sources on digestion and gut microbiome, and offer scientific guidance for optimizing dietary choices to improve health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The objectives of this experiment were to determine the digestible indispensable amino acid score (DIAAS) for eggs cooked in different forms and in traditional egg-bread or egg-hash brown combinations, and to test the hypothesis that DIAAS in eggs is greater than in breads or potatoes. Nine ileal cannulated gilts (average initial body weight: 51.1 ± 6.0 kg) were allotted to a 9 × 6 Youden square design with nine diets and six 7-day periods. Fried egg, boiled egg, scrambled egg, English muffin, Texas toast, and hash brown were included in the experiment. Six diets each contained one source of protein and three diets were combinations of fried eggs and English muffin, boiled eggs and Texas toast, or scrambled egg and hash brown. A nitrogen-free diet was also used and fed to all pigs in one period. The standardised ileal digestibility (SID) of crude protein (CP) and amino acids (AA) was calculated, and DIAAS was calculated for the individual ingredient and combined meals for children between 6 and 36 months and individuals older than 3 years. For both age groups, all cooked eggs had greater (P < 0.001) DIAAS compared with the other foods, and hash brown had greater (P < 0.001) DIAAS than both breads. All combined meals had DIAAS greater than 75 and there were no differences between measured and predicted DIAAS for the combined meals. In conclusion, eggs have 'excellent' protein quality for individuals older than 6 months and can compensate for the lower protein quality in plant-based foods, and DIAAS obtained from individual ingredients are additive in mixed meals., Competing Interests: The authors have no conflicts of interest., (© The Author(s) 2024.)

Background: Although not every food protein contributes sufficient quantities of all of the indispensable amino acids (AAs) required for human health, the human diet is a mixture of foods. It is therefore important to evaluate protein quality in mixed meals., Objectives: The hypothesis was that the true ileal digestible AA content of mixed meals can be predicted from the amounts of digestible AA determined for individual food ingredients, allowing further prediction of the digestible indispensable amino acid score (DIAAS) for meals., Methods: Four diets contained either pea protein isolate (PPI), whey protein concentrate (WPC), or mixtures of PPI:WPC at a ratio of 25:75 (PP25) or 75:25 (PP75). Six ileal cannulated pigs were allocated to a 4 × 6 Latin Square design. Experimental diets at 10% protein and a protein-free diet were fed for 7 d with ileal digesta collection on days 6 and 7. True ileal digestibility (TID) of AA, amount of digestible AA, and DIAAS were calculated for each diet and predicted for the 2 mixed meals., Results: Predicted and determined digestible AA contents of the mixed diets were not different. The predicted values fell within the confidence interval (95%) of the determined values. DIAAS for PPI was 61% and for WPC it was 96%. DIAAS calculated based on the predicted amounts of digestible AA for PP25 was 104% and for PP75 83%. These values were nearly identical to the determined values of 104% and 84% for PP25 and PP75, respectively., Conclusions: The amount of digestible AA and DIAAS can be accurately predicted in mixtures of PP and WPC from the AA digestibility values for the individual ingredients., Competing Interests: Conflict of interest All authors report no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Background: Protein quality, evaluated using Digestible Indispensable Amino Acid Score (DIAAS), requires ileal digestibility values of individual indispensable amino acids (IAAs) in each protein. However, true tryptophan (Trp) digestibility has rarely been quantified in humans., Objective: To measure the true Trp digestibility and DIAAS of 2 H-intrinsically labeled plant and animal protein sources in humans, using the dual isotope tracer technique., Methods: The true Trp digestibility of 2 H intrinsically labeled plant proteins such as whole mung bean (n = 6) and dehulled mung bean (n = 6), chickpea (n = 5), and yellow pea (n = 5), and protein from animal source foods such as egg white (n = 6), whole egg (n = 6), chicken meat (n = 6), and goat milk (n = 7) was determined against the known digestibility of U- 13 C spirulina whole cell protein as reference, except for goat milk protein that was measured against free crystalline 13 C-Trp as reference. Banked samples from earlier studies conducted to determine true IAA digestibility of different protein sources were used for the analysis. DIAAS was calculated for each test protein using digestibility corrected IAA scores (mg IAA/g of protein) in comparison with the IAA requirement score for adults., Results: The true Trp digestibility of whole mung bean, dehulled mung bean, chickpea, yellow pea, egg white, whole egg, chicken meat, and goat milk were 67.6 ± 3.7%, 74.5 ± 4.4%, 72.6 ± 2.3%, 72.5 ± 2.2%, 89.7 ± 2.5%, 91.4 ± 2.6%, 95.9 ± 2.2%, and 92.8 ± 2.9%, respectively. The true Trp digestibility of plant protein sources was significantly lower than that of animal protein sources (P < 0.05). Trp was not a limiting IAA in all the tested proteins., Conclusion: The true Trp digestibility determined in this study ranged from 67.6 ± 3.7% to 95.9 ± 2.2% for whole mung bean and chicken meat, respectively, and adds to the database of individual true IAA digestibility of different protein sources., Trial Registration Number: This study was registered in Clinical Trials Registry of India (CTRI) with registration numbers CTRI/2017/11/010468, CTRI/2020/04/024512, and CTRI/2018/03/012265., Competing Interests: Conflict of interest The authors report no conflicts of interest., (Copyright © 2024 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

The amount of chitin, a nitrogen-containing dietary fiber, in edible insects can mislead the exact nitrogen-to-protein conversion factor (NPF) and true protein content. We determined the amino acid score (AAS), protein digestibility-corrected AAS (PDCAAS), chitin content, and net NPF of five edible insects. Additionally, the effect of the amino acid composition of migratory locust on rat growth were investigated. The AAS of the insects were ranged from 63 to 94. The chitin contents were ranged from 1.6 g/100 g to 10.7 g/100 g. The PDCAAS, calculated by AAS and gut-intestinal digestibility, ranged from 44 to 81, which was lower than casein (97). The net NPF ranged from 4.93 to 5.76, which were lower than the conventional value. Dietary migratory locust, whose PDCAAS was the lowest, decreased growth and altered lipid metabolism. Therefore, a lower PDCAAS and overestimation of net NPF of insects can affect the true protein calculations and growth., Competing Interests: Declaration of competing interest All the authors have no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Amino acids (AA) are an expensive nutritional components of poultry diets. Distillers dried grains with solubles (DDGS) is the primary co-product produced by the dry grind bioethanol industry, although new technologies are being implemented to produce high protein distillers dried grains (HP-DDG) and corn fermented protein (CFP), but data on their nutritive value in poultry are lacking. Two experiments (EXP) were conducted to determine the energy and AA digestibility of DDGS, HP-DDG, and CFP in poults in addition to a feeding trial to evaluate increasing dietary levels of HP-DDG and CFP on growth performance and intestinal characteristics. In EXP 1, 6 different DDGS sources were evaluated using poults to determine their nitrogen-corrected apparent metabolizable energy (AME n ) concentrations, and cecectomized roosters were used to determine their standardized ileal (SID) AA digestibility (SID-AA). In EXP 2, AME n and SID-AA for HP-DDG and CFP were determined in young poults, and a feeding trial was conducted to evaluate growth performance and intestinal morphology and permeability of poults fed diets containing 7.5 and 15% HP-DDG or CFP from 1 to 32 d of age. In EXP 1, the AME n concentration among the DDGS samples ranged from 2,530 to 3,573 kcal/kg DM but was not different (P = 0.57) among the samples, with an average SID for LYS of 66.6%. In EXP 2, different (P = 0.001) AME n concentrations for HP-DDG and CFP were observed (3,114 and 3,760 kcal/kg DM, respectively), with the SID for LYS being 66.55 and 77.00% for HP-DDG and CFP, respectively. Including HP-DDG or CFP into the diet at 7.5 and 15% had no effect (P > 0.05) on growth, feed intake, or feed conversion. Neither co-product nor its inclusion rate affected intestinal morphology and permeability (P > 0.05). Overall, DDGS, HP-DDG, and CFP are excellent sources of AME n and digestible AA, with dietary inclusion rates of up to 15% of HP-DDG or CFP having no impact on growth or intestinal characteristics., Competing Interests: DISCLOSURES The authors declare that a portion of this manuscript was funded by The Anderson Inc. who donated the HP-DDG and CFP used in Experiment 2 in this manuscript and provided financial assistance to support a portion of manuscript. The financial support supported the first authors in their graduate program, farm costs associated with the trial, and assay costs. This declaration is provided in the manuscript under the disclosure section. Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA, University of Minnesota, or Iowa State University and does not imply approval to the exclusion of other products that may be suitable. The USDA is an equal opportunity provider and employer., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The present study investigated the effects of low protein diets with different starch sources and starch to protein ratio on growth, digestibility, intestinal health, caecal short chain fatty acids (SCFAs), serum cholesterol and triglycerides in broiler chickens. Eight hundred one-day-old male broiler chicks (Ross 308) were randomly allotted to one of 4 dietary treatments with 10 repeats and 20 birds in each repeat. The dietary treatments included 1) a standard protein corn-SBM based diet (SP), 2) a low protein corn-SBM based diet (LPI) without reduced starch: protein ratio, 3) a low protein corn-SBM based diet (LPII) with reduced starch: protein ratio, and 4) a low protein corn-SBM-peas based diet (LPP) and reduced starch: protein ratio. Soy hulls were added in the LPII and LPP diets to reduce starch: protein ratio. During the experiment period from 11 to 24 d, FI was not affected by the dietary treatments (P > 0.05). The BWG was significantly reduced in the LPI diet compared to the SP diet (P < 0.05). Likewise, FCR deteriorated in LPI and LPII but was better in the SP diet followed by the LPP diet (P < 0.05). The apparent total tract digestibility (ATTD) of dry matter (DM) varied significantly among the dietary treatments (P < 0.01). While ATTD of starch was similar for all the diets except the LPP diet wherein the ATTD of starch was significantly lower (P < 0.001). Ether extract digestibility was also significantly different between the SP and LPII dietary treatments (P < 0.01). The AME and AME n values were significantly lower in the LPP diet compared with other dietary treatments (P < 0.001). Nitrogen retention (%) was increased in all the LP diets compared with the SP diet (P < 0.001), but it was significantly better in both LPII and LPP diets compared to the LPI diet. The data showed that cecal SCFAs production was increased in the LPII and LPP compared to the SP and LPI diets (P < 0.001). Further, the production of acetic, butyric, and propionic acids was substantially higher in the LPP diet (P < 0.001). There was no significant difference in gene expression of Claudin-1 and ZO-1 (P > 0.05). However, MUC-2 and GLUT-1 gene expression were significantly downregulated in the LPI diet (P < 0.05). The concentration of cholesterol and triglycerides was significantly increased in the LPI diet (P < 0.001). In conclusion, the addition of peas as a slowly digestible starch source combined with soy hulls in low protein diet helped to partly recover the growth performance and improved cecal SCFAs production compared to other low protein diets with and without reduced starch: protein ratio in broiler chickens., Competing Interests: DISCLOSURES The authors are employees of Adisseo France S.A.S., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose of Review: Dietary proteins are broken down into peptides across the gastrointestinal tract, with skeletal muscle being a primary deposition site for amino acids in the form of incorporation into, for example, metabolic and structural proteins. It follows that key research questions remain as to the role of amino acid bioavailability, of which protein digestibility and splanchnic sequestration (absorption and utilization) of amino acids are determining factors, impact upon muscle protein synthesis (MPS) in clinical states., Recent Findings: Elevated splanchnic amino acid uptake has been implicated in anabolic resistance (i.e. attenuated anabolic responses to protein intake) observed in ageing, though it is unclear whether this limits MPS. The novel 'dual stable isotope tracer technique' offers a promising, minimally invasive approach to quantify the digestion of any protein source(s). Current work is focused on the validation of this technique against established methods, with scope to apply this to clinical and elderly populations to help inform mechanistic and interventional insights., Summary: Considerations should be made for all facets of protein quality; digestibility of the protein, absorption/utilization and subsequent peripheral bioavailability of amino acids, and resultant stimulation of MPS. Stable isotope tracer techniques offer a minimally invasive approach to achieve this, with wide-ranging clinical application., (Copyright © 2024 The Author(s). Published by Wolters Kluwer Health, Inc.)

Many improvements have been made to bring infant formula (IF) closer to human milk (HM) regarding its nutritional and biological properties. Nevertheless, the protein components of HM and IF are still different, which may affect their digestibility. This study aimed to evaluate and compare the protein digestibility of HM and IF using the infant INFOGEST digestion method. Pooled HM and a commercial IF were subjected to the infant INFOGEST method, which simulates the physiological digestion conditions of infants, with multiple directions, i.e. the curd state, gel images of SDS-PAGE, molecular weight distribution, free amino acid concentrations and in vitro protein digestion rate. HM underwent proteolysis before digestion and tended to have a higher protein digestion rate with finer curds during gastric digestion, than the IF. However, multifaceted analyses showed that the protein digestibility of HM and IF was not significantly different after gastrointestinal digestion. In conclusion, the infant INFOGEST method showed that the digestibility of HM and IF proteins differed to some extent before digestion and after gastric digestion, but not at the end of gastrointestinal digestion. The findings of this study will contribute to the refinement of IF with better protein digestibility in infant stomach.

Food proteins and their peptides play a significant role in the important biological processes and physiological functions of the body. The peptides show diverse biological benefits ranging from anticancer to antihypertensive, anti-obesity, and immunomodulatory, among others. In this review, an overview of food protein digestion in the gastrointestinal tract and the mechanisms involved was presented. As some proteins remain resistant and undigested, the multifarious factors (e.g. protein type and structure, microbial composition, pH levels and redox potential, host factors, etc.) affecting their colonic fermentation, the derived peptides, and amino acids that evade intestinal digestion are thus considered. The section that follows focuses on the mechanisms of the peptides with anticancer, antihypertensive, anti-obesity, and immunomodulatory effects. As further considerations were made, it is concluded that clinical studies targeting a clear understanding of the gastrointestinal stability, bioavailability, and safety of food-based peptides are still warranted., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bioactive peptides derived from native proteins modulate physiological processes in the metabolic pathways. Given that multiple protocols in the literature mimic the digestion of dietary components, gathering studies that use such models directed at protein digestion processes is critical. This systematic review aimed to gather evidence that adopted adequate experimental models to simulate human protein digestion. The databases searched were PubMed, Web of Science, ScienceDirect, Embase, Virtual Health Library, and Scopus. A total of 1985 articles were found, resulting in 20 eligible in vitro studies. The Office of Health Assessment and Translation was used to evaluate methodological quality. Seven studies used plant-based protein sources, twelve used animal protein sources, and one used both. The duration of the oral phase varied, although 60% of the studies employed a protein digestion period of 120 min. Amylase, pepsin, and pancreatin enzymes were utilized in 40% of the studies, with pH levels of 7, 3, and 7, respectively, during the oral, gastric, and intestinal phases. The INFOGEST harmonized static model was adopted by 65% of the studies; INFOGEST is the most effective model for simulating gastrointestinal protein processes in humans and can be used to answer several research questions because it describes experimental conditions close to the human physiological situation.

The aim of this study was to explore the effect of replacing protein pellets with soybean grain in high-concentrate diets with or without the addition of silage, on the intake, digestibility, and rumen and blood parameters of feedlot cattle in tropical regions. Four cannulated, crossbred steers were used, 4.5 ± 0.5 years old, with an average weight of 685.55 ± 111.78 kg. The steers were distributed in a 4 × 4 Latin square, in a 2 × 2 factorial scheme (two sources of protein: protein pellets or whole soybean grain, with or without added dietary bulk). There was no effect (P ≥ 0.109) from the interaction between the source of protein and the addition of silage to the diet on dry matter (DM) and nutrient intake, or the digestibility (P ≥ 0.625) of DM or crude protein (CP). However, both factors affected (P ≤ 0.052) the intake of DM, neutral detergent fiber (NDF), and non-fiber carbohydrates (NFC), as well as the independent digestibility (P ≤ 0.099) of fat, NFC, total carbohydrates (TC), and total cholesterol concentration. There was an effect (P ≤ 0.053) from the interaction between the source of protein and the addition of silage to the diet on the digestibility of NDF and total digestible nutrients (TDN), as well as on the glycose concentration (P = 0.003). Blood parameters (i.e. protein, albumin, creatinine, triglycerides, aspartate aminotransferase (AST), and alanine aminotransferase (ALT)) were not affected (P ≥ 0.139) by the source of protein, the addition of silage, or their interaction. Lastly, including 150 g/kg silage DM in a high-grain diet, and using soybean grain as a source of protein in substitution of protein pellet could be a suitable nutritional strategy to ensure adequate DM and nutrient intake and digestibility, with no detrimental effects on rumen and blood parameters of feedlot cattle in the tropics., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The study aims to assess the impact of oven-drying and decontamination on crude protein concentration and in vitro crude protein digestibility of yellow mealworms. Two kilograms of 12-wk-old mealworm larvae were subjected to freezing prior to the drying process. Approximately 1.5 kg of mealworm larvae were divided into 3 groups and exposed to oven-drying at temperatures of 50 °C for 36 h, 60 °C, and 70 °C for 24 h each. At intervals of 2 h, sets of 3 replicates were withdrawn to record water loss. Consistent weight stabilization was observed at 36 h for 50 °C (T50), 18 h for 60 °C (T60), and 14 h for 70 °C (T70). The remaining 0.5 kg of mealworm larvae was divided and dried under treatments T50, T60, and T70. Each treatment was then split into 2 portions, with one portion subjected to 90 °C for 15 min (denoted as T50-90, T60-90, T70-90) to eliminate microbial contamination. The 6 treatments were then used to determine concentrations of dry matter, crude ash, crude protein, pre-caecal protein digestibility, and dry matter residues after neutral detergent fiber, acid detergent fiber, and acid detergent lignin treatments. No interaction was observed between drying and decontamination treatments (P > 0.17). Pre-caecal crude protein digestibility increased with decreasing temperature (T50: 58% crude protein; T60: 51% crude protein; T70: 50% crude protein). Therefore, lower temperatures for longer times preserve crude protein digestibility. These findings are crucial for understanding how drying temperature and time impact protein bioavailability., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Our objectives were to determine the effects of reducing dietary CP concentration on nutrient digestibility, rumen function, N balance, and serum AA concentration for dairy cows in late lactation. At the initiation of the experimental period, we stratified Holstein cows (n = 128; mean ± SD 224 ± 54 DIM) by parity and days pregnant (86 ± 25 d) and assigned them to 1 of 16 pens. For 3 wk, all cows received a covariate diet containing 16.9% CP (DM basis). For the subsequent 12 wk, we assigned pens to 1 of 4 treatments containing 16.2%, 14.4%, 13.4%, or 11.9% CP (DM basis) in a randomized complete block design. Diets were fed as a TMR once daily. To reduce dietary CP, we replaced soybean meal with soybean hulls in the concentrate mix (DM basis). Diet evaluations suggested that several EAA, especially His, limited productivity as dietary CP declined. Digestibility of DM and CP decreased linearly with dietary CP reduction. Digestibility of NDF and potentially digestible NDF tended to respond in a quadratic pattern with the greatest digestibility at intermediate treatments. The reduction in dietary CP did not affect ruminal pH, but ruminal ammonia-N and branched-chain VFA concentrations declined linearly. The concentration of milk urea-N and plasma urea-N, secretion of milk N, and excretions of fecal N, urinary N, urinary urea-N, and unaccounted N decreased linearly with the reduction in dietary CP concentration. Urinary N expressed as a percentage of N intake was unaffected by dietary CP. Serum concentrations of total essential AA and NEAA were unaffected by dietary CP concentration. However, the ratio of essential to NEAA decreased with decreasing dietary CP. Serum 3-methylhistidine concentration increased linearly with decreasing dietary CP concentration, indicating greater skeletal muscle breakdown. Although our trial confirmed that reducing dietary CP decreased absolute excretion of urinary N, diet evaluations suggested that milk protein production decreased as certain essential AA became increasingly limited. Thus, reduced-CP diets have the potential to lessen reactive-N outputs of late-lactation cows, but more research is needed to design diets that minimize deleterious effects on productivity., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

To avoid a high body protein mobilization in modern lean sows during lactation, an adequate dietary amino acid (AA) supply and an efficient AA utilization are crucial. This study evaluated the effects of dietary CP and in vitro protein digestion kinetics on changes in sow body condition, litter weight gain, milk composition, blood metabolites, protein utilization efficiency and subsequent reproductive performance. We hypothesized that a slower digestion of dietary protein would improve AA availability and utilization. In total, 110 multiparous sows were fed one of four lactation diets in a 2 × 2 factorial design, with two CP concentrations: 140 g/kg vs 180 g/kg, and two protein digestion kinetics, expressed as a percentage of slow protein (in vitro degradation between 30 and 240 min): 8 vs 16% of total protein. Feeding sows the high CP diets reduced sow weight loss (Δ = 7.6 kg, P < 0.01), estimated body fat loss (Δ = 2.6 kg, P = 0.02), and estimated body protein loss (Δ = 1.0 kg, P = 0.08), but only at a high percentage of slow protein. A higher percentage of slow protein increased litter weight gain throughout lactation (Δ = 2.6 kg, P = 0.04) regardless of CP concentrations, whereas a higher CP only increased litter weight gain during week 3 of lactation (Δ = 1.2 kg, P = 0.01). On Day 15 postfarrowing, serial blood samples were taken from a subsample of sows fed with the high CP diets. In these sows, a high percentage of slow protein resulted in higher plasma AA concentrations at 150 and 180 min after feeding (Δ = 0.89, P = 0.02, Δ = 0.78, P = 0.03, mmol/L, respectively) and lower increases in urea at 90 and 120 min after feeding (Δ = 0.67, P = 0.04, Δ = 0.70, P = 0.03, mmol/L, respectively). The higher dietary CP concentration increased total nitrogen loss to the environment (Δ = 604 g, P < 0.01) with a reduction of protein efficiency (Δ = 14.8%, P < 0.01). In the next farrowing, a higher percentage of slow protein increased subsequent liveborn litter size (Δ = 0.7, P < 0.05). In conclusion, feeding sows with a high dietary CP concentration alleviated maternal weight loss during lactation when the dietary protein digestion rate was slower, but lowered protein efficiency. A slower protein digestion improved litter weight gain, possibly by reducing AA oxidation and improving plasma AA availability, thus, improving protein efficiency., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Reducing dietary CP is a well-established means to improve N use efficiency. Yet, few studies have considered if transient restrictions in dietary CP could reduce the environmental footprint of late-lactation cows. We hypothesized that the effects of CP feeding pattern on digestibility and environmental outputs would be amplified at lower dietary CP. We tested CP levels below and near predicted requirements (low protein [LP], 13.8%; high protein [HP], 15.5%) offered in 2 feeding patterns: where diets alternated ±1.8 percentage units CP every 2 d (oscillating [OF]) or remained static. Our study used a 2 × 2 factorial design with 16 mid- to late-lactation Holsteins (mean = 128, SD = 12 DIM), divided into rumen-cannulated (n = 8) and noncannulated subsets (n = 8). For each 28-d experimental period, we recorded feed intake and milk production and took samples of orts (1×/d) and milk (2×/d) for 4 d. For the cannulated subset, we measured and sampled from the total mass of feces and urine production and collected plasma 2×/d across 4 d. For the noncannulated subset, we sampled carbon dioxide and methane emissions 3×/d for 4 d. For each subset, we fit linear mixed models with fixed effects for CP level, CP feeding pattern, the interaction of CP level and CP feeding pattern, period, and a random effect for cow. For plasma and urinary urea-N, we conducted time series analysis. Contrary to our hypothesis, we found no evidence that dietary CP level and CP feeding pattern interacted to influence N balance, nutrient digestibility, or gas emissions. Results showed HP resulted in similar milk N but increased manure N, reducing N use efficiency (milk true protein N/intake N) relative to LP. For OF, urea-N in urine and plasma peaked 46 to 52 h after the first higher-CP phase feeding. Nutrient digestibility and gas emissions were similar across treatments, except CO 2 production was greater for OF-HP. In summary, measured variables were minimally affected by dietary CP alternating ±1.8 percentage units every 48 h, even when average dietary CP was fed below predicted requirements (LP). Although our findings suggest that mid- to late-lactation cows are resilient to oscillation in dietary CP, oscillating CP neither reduced the environmental footprint by improving nutrient use efficiencies nor reduced the potential for direct and indirect greenhouse gas emissions., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Low crude protein (CP) diets can reduce nitrogen (N) excretion and costs by increasing N utilization efficiency. Exogenous proteases may further improve protein digestibility in low CP diets. This study first evaluated in vitro the efficacy of a multiprotease on amino acid (AA) release from feedstuffs and broiler feed. Later, a broiler study evaluated the effect of feeding corn-soybean meal diets containing 3 CP levels (17, 19, and 21% CP) with supplementation on top of 0 or 2,400 U/kg multiprotease on chicken growth performance, total tract CP, and ileal AA digestibilities, and energy utilization. Ross 708 male chickens were placed in 42 cages and assigned to 6 treatments resulting from a 3 × 2 factorial arrangement. Three isocaloric basal diets were formulated to reduce CP, but all diets maintained digestible Lys:CP in 5.47% and the same ideal protein profile. Data were analyzed in a completely randomized design. On average, the multiprotease increased (P < 0.05) in vitro free AA release by 27.81% in most feedstuffs evaluated compared to the control. For broiler feed, 1,200 U/g multiprotease addition improved (P < 0.001) in vitro free AA release by 18.90%. This multiprotease showed interaction effects (P < 0.05) on chicken FCR, energy, and CP digestibility. As expected, BW at 24 d, BW gain, and FCR (8-24 d) worsened (P < 0.001) as dietary CP reduced from 21 to 17%, and multiprotease addition did not improve (P > 0.05) these parameters. BW gain decreased by 12.9% when N intake was reduced from 49.32 to 38.49 g/bird. Multiprotease supplementation improved (P < 0.01) AMEn by 71 kcal/kg, CP digestibility from 59.45 to 63.51%, ileal AA digestibility, and DM digestibility from 67.08 to 73.49%, but only in the 21% CP diet. No differences in ileal AA digestibility due to CP level (P > 0.05) were detected, except for Cys digestibility (P < 0.01). In conclusion, low CP diets reduced growth performance and improved N utilization but negatively affected energy utilization efficiency. Exogenous multiprotease supplementation improved AME, AMEn, protein, ileal AA, and DM digestibility in the 21% CP diet without significantly affecting growth performance., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

To improve animal performance and modify growth by increasing lean tissue accretion, beef cattle production has relied on use of growth promoting technologies such as beta-adrenergic agonists. These synthetic catecholamines, combined with the variable inclusion of rumen degradable (RDP) and undegradable protein (RUP), improve feed efficiency and rate of gain in finishing beef cattle. However, research regarding the impact of beta-adrenergic agonists, protein level, and source on the ruminal microbiome is limited. The objective of this study was to determine the effect of different protein concentrations and beta-adrenergic agonist (ractopamine hydrochloride; RAC) on ruminal bacterial communities in finishing beef heifers. Heifers (n = 140) were ranked according to body weight and assigned to pens in a generalized complete block design with a 3 × 2 factorial arrangement of treatments of 6 different treatment combinations, containing 3 protein treatments (Control: 13.9% CP, 8.9% RDP, and 5.0% RUP; High RDP: 20.9% CP, 14.4% RDP, 6.5% RUP; or High RUP: 20.9% CP, 9.7% RDP, 11.2% RUP) and 2 RAC treatments (0 and 400 mg/day). Rumen samples were collected via orogastric tubing 7 days before harvest. DNA from rumen samples were sequenced to identify bacteria based on the V1-V3 hypervariable regions of the 16S rRNA gene. Reads from treatments were analyzed using the packages 'phyloseq' and 'dada2' within the R environment. Beta diversity was analyzed based on Bray-Curtis distances and was significantly different among protein and RAC treatments (P < 0.05). Alpha diversity metrics, such as Chao1 and Shannon diversity indices, were not significantly different (P > 0.05). Bacterial differences among treatments after analyses using PROC MIXED in SAS 9 were identified for the main effects of protein concentration (P < 0.05), rather than their interaction. These results suggest possible effects on microbial communities with different concentrations of protein but limited impact with RAC. However, both may potentially act synergistically to improve performance in finishing beef cattle., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

This study was performed to investigate the nitrogen (N) and carbohydrate digestive traits of grazing heifers. The experiment was carried out at the Federal University of Lavras. The treatments were a Marandu palisadegrass ( Urochloa brizantha [Syn, Brachiaria brizantha ] Stapf. A. Rich. cv. Marandu) monoculture fertilised with 150 kg N/[ha ∙ year] (FP) or Marandu palisadegrass mixed pasture with forage peanut (MP). The pastures were grazed by six rumen-cannulated zebu heifers. A double cross-over design was used in four periods. Nutritive value, intake and apparent digestibility of forage, ruminal traits and kinetics and N balance were evaluated. Apparent total-tract digestibility of dry matter (DM) and neutral detergent fibre (NDF) were greater for FP than for MP. There was no effect in apparent total-tract digestibility of N. The estimated intestinal digestibility of nutrients was greater on MP than FP. Even though N intake and faecal N output were greater on MP than FP, there was no effect in urine N output. The N balance tended to be greater on MP than FP. The forage peanut, which contains condensed tannins, decreased ruminal fibre degradation, apparent digestibility and ruminal protein degradation, increased N flow from the rumen. Inclusion of forage peanut in the mixed pasture decreased the ruminal fibre degradability but increased N retention by the animals.

Duckweed has attracted increasing attention as a high-quality and sustainable novel plant-based protein source. However, little research has been conducted in dogs and cats. We evaluated the effects of inclusion of duckweed protein (Lemna; MCSelect; Parabel; Vero Beach, FL) primarily in replacement of pea protein in dog diets at 0%, 5%, and 10% and cat diets at %, 10%, and 15% on stool quality, nutrient digestibility, and palatability. We hypothesized that duckweed protein would be a viable protein source in both dog and cat diets by showing no detriment to nutritional outcomes. All feeding tests were conducted at an independent research facility (Susquehanna, PA). A standard 2-bowl palatability test over a 2-d period was conducted with adult animals (n = 30 each) to determine intake ratio between test diets (duckweed-containing diets) and control diets (0% duckweed protein). Apparent total tract nutrient digestibility was conducted with 18 adult dogs and 21 adult cats (n = 6 to 7 per diet) with 5 d of diet acclimation followed by 5 d of total fecal collection. Stool quality was evaluated on a 1 to 5 scale where 1 = non-formed or diarrhea and 5 = hard, formed. Palatability data were analyzed using paired t-test (daily consumption) and chi-square test (first choice). All other data were analyzed by ANOVA and contrast (SAS version 9.4). For cats, 10% duckweed had greater (P < 0.05) palatability than control, while no difference was observed between 15% duckweed protein and control. For dogs, 5% and 10% duckweed protein had (P < 0.05) lower palatability, demonstrating a preference to control. Both cats and dogs fed duckweed diets had acceptable stool quality (Mean = 3.4 and 3.3, respectively). No detriments in nutrient digestibility were observed in dogs fed 5% and 10% duckweed protein; however, cats fed 10% and 15% duckweed protein had (P < 0.05) lower dry matter, protein, and energy digestibility vs. control. In conclusion, the data collected indicate that duckweed can be a viable replacement for other plant-based proteins in dog diets at inclusion levels up to 10%; more development is needed for duckweed protein inclusion into cat diets., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The study determined the effects of replacing different levels of soybean meal (SBM) with rocket seed cake (RSC) in the diets of growing lambs on feed utilization and growth performance. Twenty-eight male lambs (180 ± 5 d old) were divided into four groups in a complete randomized design with repeated measures for 105 d. Soybean meal was replaced with RSC at 0% (RSC0), 25% (RSC25), 50% (RSC50), and 75% (RSC75). The RSC75 group had the lowest final weight, total weight gain, and daily weight gain. The RSC25 increased (P < 0.001) the intakes of DM, starch value (SV), total digestible nutrients (TDN), digestible energy (DE), and digestible crude protein (DCP) compared to the other diets, while the RSC75 decreased these values. Moreover, the RSC25 decreased (P < 0.05) feed conversion of DM compared to other diets. Treatments did not affect nutrient digestibility or diet's nutritive values expressed as true SV, TDN, DCP, and DE. The RSC linearly increased albumin and urea and lowered the high-density lipoprotein concentrations in lamb's blood. The inclusion of RSC in the diet increased economic efficiency, with the highest relative percentages of net revenue with the RSC25. Overall, RSC can replace SBM at 25% in the diet of growing lambs., (© 2024 The Author(s). Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.)

The Ussing chamber is a tool for analyzing drug absorption. We investigated whether the Ussing chamber can be used to analyze the process from digestion to absorption of protein in the gastrointestinal tract. Mixtures containing infant formula, whole cow's milk, processed soy milk, enteral nutrition, or human breast milk, were placed in the apical membrane side equipped with Caco-2 cells. After the addition of first pepsin then pancreatin, samples from the apical and basal membranes were collected. Infant formula showed the highest digestibility and absorption rate. This may be attributed to the presence of whey protein, which is rapidly digested and absorbed. The digestion and absorption of human breast milk showed different results in each donor, suggesting that digestion and absorption may vary among individuals. We concluded that the Ussing chamber can continuously analyze the process from digestion to absorption of proteins in the gastrointestinal tract.

Microalgae are booming as a sustainable protein source for human nutrition and animal feed. Nevertheless, certain strains were reported to have robust cell walls limiting protein digestibility. There are several disruption approaches to break down the cell integrity and increase digestive enzyme accessibility. This review's intent is to discuss the digestibility of microalgae proteins in intact cells and after their disruption. In intact single cells, the extent of protein digestibility is chiefly related to cell wall structural properties (differing among strains) as well as digestion method and when added to food or feed protein digestibility changes depending on the matrix's composition. The degree of effectiveness of the disruption method varies among studies, and it is complicated to compare them due to variabilities in digestibility models, strains, disruption method/conditions and their consequent impact on the microalgae cell structure. More exhaustive studies are still required to fill knowledge gaps on the structure of microalgal cell walls and to find efficient and cost-effective disruption technologies to increase proteins availability without hindering their quality.

The rates of dietary protein digestion and absorption can be significantly increased or decreased by food processing treatments such as heating, gelling and enzymatic hydrolysis, with subsequent metabolic impacts, e.g. on muscle synthesis and glucose homeostasis.This review examines in vivo evidence that industrial and domestic food processing modify the kinetics of amino acid release and absorption following a protein-rich meal. It focuses on studies that used compositionally-matched test meals processed in different ways.Food processing at extremely high temperature at alkaline pH and/or in the presence of reducing sugars can modify amino acid sidechains, leading to loss of bioavailability. Some protein-rich food ingredients are deliberately aggregated, gelled or hydrolysed during manufacture. Hydrolysis accelerates protein digestion/absorption and increases splanchnic utilisation. Aggregation and gelation may slow or accelerate proteolysis in the gut, depending on the aggregate/gel microstructure.Milk, beef and eggs are heat processed prior to consumption to eliminate pathogens and improve palatability. The temperature and time of heating affect protein digestion and absorption rates, and effects are sometimes non-linear. In light of a dietary transition away from animal proteins, more research is needed on how food processing affects digestion and absorption of non-animal proteins.Food processing modifies the microstructure of protein-rich foods, and thereby alters protein digestion and absorption kinetics in the stomach and small intestine. Exploiting this principle to optimise metabolic outcomes requires more human clinical trials in which amino acid absorption rates are measured and food microstructure is explicitly considered, measured and manipulated.

Estimating protein fractions and their degradation rate are vital to ensure optimum protein supply and degradation in the digestive system of ruminants. This study investigated the possibility of using the ANKOM gas production system and preserved rumen fluid to estimate the protein fractions and in vitro degradability of protein-rich feeds. Three in vitro methods: (1) gas production method (2) Cornell Net Carbohydrate and Protein System (CNCPS), and (3) the unavailable nitrogen assay of Ross (uN Ross ) were used to quantify protein fractions of four feeds (lupin meal, vetch grain, Desmanthus hay, and soybean meal). Rumen fluid mixed with 5% dimethyl sulfoxide and frozen at -20 °C was also compared against fresh rumen fluid in the gas production and uN Ross methods. All three methods ranked the feeds identically in the proportions of available (degradable or 'a + b') protein fractions as vetch grain, soybean meal, lupin meal, and Desmanthus hay in decreasing order. The use of fresh rumen fluid produced greater available protein fractions than preserved rumen fluid in all feeds. However, there was no difference between total gas production from lupin meal and vetch grain fermented for 16 h in either rumen fluid source. The in vitro degradable CP (IVDP) was higher for vetch grain (46 and 70%) at the 4th and 8th hours of incubation than other feeds, whereas soybean meal (85%) exceeded the other feeds after the 16th hour of incubation (P < 0.001). The greatest ammonia-N concentration was from soybean meal (1.27 mg/g) and lupin meal (0.87 mg/g) fermented for four hours using fresh rumen fluid. The proportion of fraction 'b' for soybean (82.1% CP) and lupin meals (39.4% CP) from the CNCPS method were not different (P = 0.001) from the fraction 'b' estimation of the gas production method for the same feeds (r = 0.99). Regardless of the methods, a greater water-soluble protein fraction was found from vetch grain (39.6-46.6% CP), and the proportion of fraction 'c' or unavailable protein in Desmanthus hay (39.1-41.5% CP) exceeded other substrates (P < 0.001). The strong positive correlation between fractions across different methods and identical ranking of feeds suggests the possibility of using ANKOM gas production apparatus for protein fractionation., (Copyright © 2023. Published by Elsevier B.V.)

The future of precision nutrition requires treating amino acids as essential nutrients. Currently, recognition of essential amino acid requirements is embedded within a generalized measure of protein quality known as the PDCAAS (Protein Digestibility-Corrected Amino Acid Score). Calculating the PDCAAS includes the FAO/WHO/UNU amino acid score, which is based on the limiting amino acid in a food, that is, the single amino acid with the lowest concentration compared to the reference standard. That "limiting" amino acid score is then multiplied by a bioavailability factor to obtain the PDCAAS, which ranks proteins from 0.0 (poor quality) to 1.0 (high quality). However, the PDCAAS has multiple limitations: it only allows for direct protein quality comparison between 2 proteins, and it is not scalable, transparent, or additive. We therefore propose that shifting the protein quality evaluation paradigm from the current generalized perspective to a precision nutrition focus treating amino acids as unique, metabolically active nutrients will be valuable for multiple areas of science and public health. We report the development and validation of the Essential Amino Acid 9 (EAA-9) score, an innovative, nutrient-based protein quality scoring framework. EAA-9 scores can be used to ensure that dietary recommendations for each essential amino acid are met. The EAA-9 scoring framework also offers the advantages of being additive and, perhaps most importantly, allows for personalization of essential amino acid needs based on age or metabolic conditions. Comparisons of the EAA-9 score with PDCAAS demonstrated the validity of the EAA-9 framework, and practical applications demonstrated that the EAA-9 framework is a powerful tool for precision nutrition applications., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Amino acid (AA) digestibility in humans has been determined conventionally based on oro-ileal AA disappearance. With this approach, it is necessary to account for undigested AAs of body origin (endogenous AAs) found in the ileal digesta. Determination of the endogenous AAs under physiological conditions is not straightforward, and the use of isotopes (labeled foods or body tissues) has been pivotal to advancing our understanding. The application of isotopes for determining gut endogenous AAs and AA digestibility is discussed as well as the types of digestibility coefficient generated (apparent, true, real) dependent upon methodology. Recently a new dual isotope-based method for determining ileal AA digestibility in humans has been developed that obviates the collection of ileal digesta. The dual isotope method, which awaits full validation, offers considerable promise for making noninvasive measures of AA digestibility in humans of different ages and physiological states., (Copyright © 2023 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

The 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, R 2  = 0.89, P < 0.0001) with a mean difference of 0.1 %., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The digestible indispensable amino acid score uses ileal digestibility of each indispensable amino acid (IAA) of a dietary protein to calculate its protein quality. However, true ileal digestibility, which is the exclusive sum of digestion and absorption of a dietary protein up to the terminal ileum, is difficult to measure in humans. It is traditionally measured using invasive oro-ileal balance methods but can be confounded by endogenous secreted protein in the intestinal lumen, although the use of intrinsically labeled protein corrects for this. A recent, minimally invasive dual isotope tracer technique is now available to measure true IAA digestibility of dietary protein sources. This method involves simultaneous ingestion of 2 intrinsically but differently (stable) isotopically labeled proteins, a ( 2 H or 15 N-labeled) test protein and ( 13 C-labeled) reference protein whose true IAA digestibility is known. Using a plateau-feeding protocol, the true IAA digestibility is determined by comparing the steady state ratio of blood to meal test protein IAA enrichment to the similar reference protein IAA ratio. The use of intrinsically labeled protein also distinguishes between IAA of endogenous and dietary origin. The collection of blood samples makes this method minimally invasive. As the α- 15 N and α- 2 H atoms of AAs of the intrinsically labeled protein are prone to label loss because of transamination, underestimation of digestibility, appropriate correction factors need to be employed when using 15 N or 2 H labeled test protein. The true IAA digestibility values of highly digestible animal protein by the dual isotope tracer technique are comparable to that measured by direct oro-ileal balance measurements, but no data are yet available for proteins with lower digestibility. A major advantage is that the minimally invasive method allows for true IAA digestibility measurement in humans across different age groups and physiological conditions., (Copyright © 2022 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

Various studies with growing ruminants report increases in nitrogen use efficiency (NUE) when feeding oscillating (OS) dietary CP, whereas limited research with lactating dairy cows demonstrates a lack of improvement in NUE when feeding OS diets. We hypothesised that a total mixed ration (TMR) delivering OS CP (48-h phases of 134 and 171 g CP/kg DM, respectively) compared to a static CP TMR (ST; 152 g CP/kg DM) would result in similar or increased urinary purine derivative excretion (as a marker of microbial protein synthesis (MPS)) and greater urinary nitrogen excretion in lactating dairy cows. Responses in intake, production, apparent total tract digestibility (ATTD), nutrient balance, and estimated MPS were evaluated using faecal and urine collection in 12 multiparous cows (172 ± 39 d in milk) in a randomised complete block design, where total urinary output was estimated indirectly. All measurements were taken during d 8 (at 1700) to d 16 (at 1700) of the 16-d study that followed a 28-d period in which cows already received their respective treatments. Dry matter intake, yields of milk, protein, fat, lactose, and fat- and protein-corrected milk were similar for ST and OS. Milk composition, BW, and body condition score also did not differ between treatments, except for a tendency for increased milk urea concentration with OS (13.7 vs 12.4 mg/dL). Feed efficiency, NUE and ATTD of organic matter, NDF, CP and gross energy did not differ, but ATTD of crude fat (658 vs 627 g/kg) and starch (980 vs 975 g/kg) increased, and ATTD of DM (702 vs 691 g/kg) tended to increase with OS. Milk energy as a proportion of digested energy tended to decrease with OS (34.6 vs 37.1%), but other energy metabolism variables were not affected by treatment. Estimated urinary nitrogen excretion increased (165 vs 144 g/d), estimated urinary nitrogen as a proportion of nitrogen intake tended to increase (25.3 vs 22.7%), and milk nitrogen as a proportion of digested nitrogen decreased (47.3 vs 51.8%) in response to OS. Estimated urinary excretion of creatinine (184 vs 165 mmol/d), uric acid (29 vs 20 mmol/d) and urea (3.1 vs 2.5 mol/d) increased, but other nitrogen metabolism parameters were not affected by OS. Overall, oscillating dietary CP content did not affect lactational performance, milk NUE, or estimated MPS. However, ATTD of some nutrients increased, postabsorptive energy use for milk synthesis tended to decrease, and estimated urinary nitrogen losses increased with OS., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

The recent Food and Agricultural Organization/World Health Organization/United Nations University expert consultations on protein requirements and quality have emphasized the need for the new Digestible Indispensable Amino Acid Score (DIAAS), as a measure of protein quality. This requires human measurements of the true ileal digestibility of individual indispensable amino acids (IAAs) until the end of the small intestine. Digestibility is measured using standard oro-ileal balance methods, which can only be achieved by an invasive naso-ileal intubation in healthy participants or fistulation at the terminal ileum. Significant efforts have been made over the last 2 decades to develop noninvasive or minimally invasive methods to measure IAA digestibility in humans. The application of intrinsically labeled (with stable isotopes like 13C, 15N, and 2H) dietary proteins has helped in circumventing the invasive oro-ileal balance techniques and allowed the differentiation between endogenous and exogenous protein. The noninvasive indicator amino acid oxidation (IAAO) technique, which is routinely employed to measure IAA requirements, has been modified to estimate metabolic availability (a sum of digestibility and utilization) of IAA in foods, but provides an estimate for a single IAA at a time and is burdensome for participants. The recently developed minimally invasive dual isotope tracer method measures small intestinal digestibility of multiple amino acids at once and is suitable for use in vulnerable groups and disease conditions. However, it remains to be validated against standard oro-ileal balance techniques. This review critically evaluates and compares the currently available stable isotope-based protein quality evaluation methods with a focus on the digestibility and metabolic availability measurements in humans. In view of building a reliable DIAAS database of various protein sources and subsequently supporting protein content claims in food labeling, a re-evaluation and harmonization of the available methods are necessary., (© The Author(s) 2021. Published by Oxford University Press.)

Precision dietary interventions (e.g., altering proportions of dietary protein fractions) has significant implications for the efficiency of nutrient use in ruminants, as well as lowering their environmental footprint, specifically nitrogen (N) emissions. Soluble protein (SP) is defined as the protein fraction that is rapidly degraded in the rumen (e.g., non-protein N and true protein), and our previous study found that regulating SP levels could improve N efficiency in Hu sheep. Thus, the present study was conducted to explore in vitro how protein fractions with different SP levels modulate the rumen microbial community and its association with N metabolism. Four dietary treatments with different SP proportions and similar crude protein (CP) content (~14%) were formulated (% of CP): 20 (S20), 30 (S30), 40 (S40) and 50 (S50). Results showed that NH3-N content increased with increasing SP levels at 4, 12 and 24 h; TVFA, acetate, propionate and valerate were higher in S30 and S40 (p < 0.05) and had quadratic effects (p < 0.05). Moreover, dry matter digestibility (DMD) and N digestibility (ND) were all decreased with S20 and S50 (p < 0.05). The S30 and S40 treatments increased the abundance of Bacteroidetes and Prevotella (Prevotella&#95;ruminicola) but decreased the abundance of Firmicutes and Proteobacteria (p < 0.05). Bacterial pathways related to amino acid and fatty acid metabolism also were enriched with S30 and S40. The abundance of Entodinium was increased with S30 and S40 and had a positive correlation with Prevotella, and these two genera also played an important role in N metabolism and VFA synthesis of this study. In conclusion, bacterial and protozoal communities were altered by the level of SP (% of CP), with higher SP levels (~50% of CP) increasing the microbial diversity but being detrimental to rumen N metabolism.

For design of healthy and sustainable diets and food systems, it is important to consider not only the quantity but also the quality of nutrients. This is particularly important for proteins, given the large variability in amino acid composition and digestibility between dietary proteins. This article reviews measurements and metrics in relation to protein quality, but also their application. Protein quality methods based on concentrations and digestibility of individual amino acids are preferred, because they do not only allow ranking of proteins, but also assessment of complementarity of protein sources, although this should be considered only at a meal level and not a diet level. Measurements based on ileal digestibility are preferred over those on faecal digestibility to overcome the risk of overestimation of protein quality. Integration of protein quality on a dietary level should also be done based on measurements on an individual amino acid basis. Effects of processing, which is applied to all foods, should be considered as it can also affect protein quality through effects on digestibility and amino acid modification. Overall, protein quality data are crucial for integration into healthy and sustainable diets, but care is needed in data selection, interpretation and integration.

Infant formulas, designed to provide similar nutritional composition and performance to human milk, are recommended when breastfeeding is not enough to provide for the nutritional needs of children under 12 months of age. In this context, the present study aimed to assess the protein quality and essential amino acid content of both starting (phase 1) and follow-up (phase 2) formulas from different manufacturers. The chemical amino acid score and protein digestibility corrected by the amino acid score were calculated. The determined protein contents in most formulas were above the maximum limit recommended by FAO and WHO guidelines and at odds with the protein contents declared in the label. All infant formulas contained lactoferrin (0.06 to 0.44 g·100 g -1 ) and α-lactalbumin (0.02 to 1.34 g·100 g -1 ) below recommended concentrations, whereas ĸ-casein (8.28 to 12.91 g·100 g -1 ), α-casein (0.70 to 2.28 g·100 g -1 ) and β-lactoglobulin (1.32 to 4.19 g·100 g -1 ) were detected above recommended concentrations. Essential amino acid quantification indicated that threonine, leucine and phenylalanine were the most abundant amino acids found in the investigated infant formulas. In conclusion, infant formulas are still unconforming to nutritional breast milk quality and must be improved in order to follow current global health authority guidelines.

The current study aimed to investigate the damage of long-term high concentrate diet feeding pattern on Yak jejunal structure, physiological function and protein composition during cold season. Twelve Datong male Yak (Bos grunniens) with the same age from cold season were randomly selected and slaughtered to determine Yak jejunal digestive enzyme activity, morphology and protein composition on different feeding patterns in Tibetan Plateau. The results showed that Yak jejunum digestive enzyme activity and morphology of grazing reared group were better than those in the intensively reared group. A total of 96 differentially expressed proteins were identified by label-free Mass Spectrometry (MS), which could be concluded to two predominant themes: protein structure and inflammatory response. Nine differentially expressed proteins were correlated in Yak jejunum damage in different feeding patterns. According to this research, we found that feeding pattern resulted the differences in Yak jejunum physiological function, morphology and protein composition. This fact was confirmed long-term high dietary concentrate feeding could damage the jejunum epithelial morphology and function.

Purpose: The purpose of the study was to determine if an actinidin protease aids gastric digestion and the protein anabolic response to dietary protein., Methods: Hayward green kiwifruit (containing an actinidin protease) and Hort 16A gold kiwifruit (devoid of actinidin protease) were given in conjunction with a beef meal to healthy older subjects. Twelve healthy older males (N = 6) and females (N = 6) were studied with a randomized, double-blinded, crossover design to assess muscle and whole-body protein metabolism before and after ingestion of kiwifruit and 100 g of ground beef. Subjects consumed 2 of each variety of kiwifruit daily for 14 d prior to each metabolic study, and again during each study with beef intake., Results: Hayward green kiwifruit consumption with beef resulted in a more rapid increase in peripheral plasma essential amino acid concentrations. There were significant time by kiwifruit intake interactions for plasma concentrations of EAAs, branched chain amino acids (BCAAs), and leucine (P < 0.01). However, there was no difference in the total amount of EAAs absorbed. As a result, there were no differences between kiwifruit in any of the measured parameters of protein kinetics., Conclusion: Consumption of Hayward green kiwifruit, with a beef meal facilitates protein digestion and absorption of the constituent amino acids as compared to Hort 16A gold kiwifruit., Clinical Trial: NCT04356573, April 21, 2020 "retrospectively registered".

A good transition from a mainly liquid diet to a solid diet of concentrates and forages is important considering the small size and development of the calf's rumen. However, the optimal amount of hay or other high-fiber ingredients in the diet of recently weaned calves is not well defined. The objective of this trial was to determine the effects of feeding 0, 5, or 10% chopped grass hay (6.5% crude protein, 64.6% neutral detergent fiber) with a textured, high-starch starter (20.5% crude protein, 38.4% starch, 14.1% neutral detergent fiber; protein pellet, whole corn, and oats) on performance and digestion in Holstein steer calves between 2 and 4 mo of age. Forty-eight calves (initial body weight = 90.7 ± 2.15 kg) were housed in group pens (4 per pen) and fed diets and water for ad libitum intake. Feed offered and refused was measured daily. Calf body weight, hip width, and body condition score (1-5 scale) were measured initially and at 28 and 56 d. Fecal samples were collected from the pen floor with care not to sample bedding material and composited by pen during d 5-9, 26-30, and 47-51 to estimate apparent total-tract digestibility using acid-insoluble ash. Data were analyzed as a completely randomized design with repeated measures and pen as the experimental unit. As grass hay increased, dry matter intake (kg/d) decreased linearly but tended to change quadratically. Dry matter intake as a % of body weight changed quadratically with increasing hay, increasing from 0 to 5%, and decreasing from 5 to 10% hay. Average daily gain (1.15, 1.12, and 0.95 kg/d), feed efficiency (0.336, 0.319, and 0.309 kg of average daily gain/kg of dry matter intake), and hip width change (4.6, 4.7, and 4.1) decreased linearly with 0, 5, and 10% hay, respectively. Overall estimates of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, and crude protein digestibility changed quadratically with hay, increasing from 0 to 5%, and decreasing from 5 to 10% hay, whereas digestibility of starch, sugar, and fat decreased linearly with increasing hay. Digestibility of dry matter, organic matter, neutral detergent fiber, and acid detergent fiber increased from 2 to 3 mo of age, and decreased from 3 to 4 mo of age. Digestibility of starch decreased linearly and digestibility of fat and crude protein increased linearly with age. Digestive capacity did not appear mature by 4 mo of age. Feeding 5% chopped hay supported optimal digestion and growth in calves 2 to 4 mo of age, which agrees with previously published research., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Purpose of Review: The current review provides an update on the recent research developments regarding amino acid bioavailability in conditions of both good health and gut disorders., Recent Findings: Determination of amino acid bioavailability is complex and invasive. Minimally invasive methods using stable isotopes have been developed for humans. Data were collected in different models - humans, pigs and rats with various procedures - leading to interstudy variability. They mainly focused on either plant protein or the effect of food processing on animal protein. Plant protein in their original food matrix (legumes, grains, nuts) are generally less digestible (about 80%) than animal protein (meat, egg, milk; about 93%). Food processing has a limited impact on animal protein but its effect might be higher on plant protein. Few studies have documented the effect of gut disorders on protein digestibility, except in gastric bypass where paradoxical effects were reported. Data are needed to identify the amplitude of protein malabsorption in diseases such as inflammatory bowel disease or environmental enteric dysfunction., Summary: The past 5 years have seen a renewed interest in amino acid bioavailability in view of assessing protein quality to support current shifts in protein sourcing. Methodological developments have been performed and several studies have reported values in various models. The question of protein digestibility in gut disorders remains poorly addressed.

The aim of present study was to determine the influence of the feed form (FF) on standardized ileal digestibility (SID) of nitrogen (N) and amino acids (AA) in 3 protein sources (PS) for broiler chickens. Six diets were tested in a 3 × 2 factorial arrangement of treatments involving 3 PS (meat and bone meal [MBM], soybean meal [SBM], and canola meal [CM]) in mash and pelleted forms. The basal endogenous N and AA losses were determined by offering a N-free diet in the mash form. From day 1 to 18, the birds were offered a broiler starter diet. The diets and the N-free diet were randomly assigned to 6 replicate cages (8 birds per cage) and fed from day 19 to 23. The ileal digesta were collected on day 23. The SID of N was higher (P < 0.05) in SBM followed by MBM and CM. The average SID of AA in SBM and MBM were similar (P > 0.05), and greater (P < 0.05) than that in CM. The FF had no influence (P > 0.05) on the SID of indispensable AA, the only exception being His, which was reduced (P < 0.05) by pelleting. Pelleting, however, resulted in reduction (P < 0.001) in the SID of all dispensable AA and average of AA. The AA most affected by pelleting was Cys, with a 15.4% decrease in the SID. The standardized ileal digestible contents of protein and the average of indispensable AA and dispensable AA were higher in MBM than in SBM, with CM being the lowest. Pelleting decreased (P < 0.05) the digestible protein and total digestible AA contents. These findings reveal that the FF has a substantial impact on AA digestibility estimates of feed ingredients and it must be considered in AA digestibility assays of ingredients with high protein and AA contents., (Copyright © 2020. Published by Elsevier Inc.)

This study was conducted to study the effect of protease as well as three dietary crude protein levels on growth and nutrient digestibility in broilers. The broiler chickens (n = 540) were assigned to six dietary treatments with six replicates. Three dietary crude protein (CP) levels (17, 19, and 21%) along with two levels of protease (0 and 30,000 IU/kg) were investigated in 3 × 2 factorial arrangement during the starter phase. Body weight was higher (P < 0.05) in group fed with CP-21; however, feed intake was significantly (P < 0.05) lower in CP-19 and CP-21. Resultantly, FCR was significantly (P < 0.05) improved in birds fed with CP-21. Similarly, body weight, feed intake, and FCR were significantly (P < 0.05) improved in enzyme-treated birds compared to the untreated birds. For practical purpose, birds fed CP-19 with protease enzyme resulted in improved FCR compared to birds fed with CP-21. The results revealed that CP and apparent metabolizable energy (AME) were significantly (P < 0.05) higher in birds fed with CP-21 along with supplementation of enzyme while abdominal fat was significantly (P < 0.05) lower in the same group compared to birds fed with CP-17. These results indicated that a diet having 21% CP supplemented with 30,000 IU/kg protease enzyme was superior in improving the performance and digestibility of nutrients in broiler during the starter phase.

The aim of the present study was to determine the apparent digestibility coefficients (ADCs) of gross energy, dry matter, crude protein, amino acids, and different carbohydrate fractions of spineless cactus meals (SCMs) for Nile tilapia juveniles, fed with extruded diets. Fish were fed a reference diet or one of five test diets, each of which included 70% reference diet and 30% corresponding SCM. Fecal collection was performed in modified Guelph method. The ADCs for dry matter, gross energy, and soluble carbohydrates of the "redonda" spineless cactus meal (RSCM) were higher (P < 0.05) than those of either the "IPA 20" spineless cactus meal (ISCM) or "gigante" spineless cactus meal (GSCM), and the ADCs for crude protein of the RSCM and "Orelha de elefante africana" spineless cactus meal (OEASCM) were higher than those of the other cactus meals (P < 0.05). In addition, the ADCs for dry matter, crude protein, and gross energy of the SCMs were similar to wheat middlings. The digestibility of the less soluble fraction of the evaluated carbohydrates, the non-fibrous carbohydrates, varied between 58.32 and 73.04%, indicating efficiency in the digestion and absorption process. The data obtained in this article can be the basis for growth experiments, with the substitution of traditional ingredients for SCMs. The determination of zootechnical, physiological, metabolic, and health variables can indicate the potential for using SCMs in diets for Nile tilapia.