Amino acids (AA) are essential nutrients in human milk (HM) and critical for infant growth and development. Several maternal lifestyle factors have been suggested to influence HM AA composition, with possible consequences for the breastfed infant. Whether maternal dietary protein and AA intake is associated with AA concentrations in HM is still largely unknown. Therefore, the aim of this study was to investigate the association between maternal dietary AA intake and AA concentrations in HM over the first month postpartum. Data from the observational longitudinal Amsterdam Mother’s Milk study were used, consisting of 123 lactating women in their first postpartum month. HM samples were collected three times, on day 10, 17 and 24 postpartum. Maternal dietary protein and AA intake on these collection days was assessed using three 24-h recalls. HM protein-bound and free AA (BAA and FAA, respectively) were analysed by liquid chromatography. Associations between maternal AA intake and AA concentrations in HM were assessed using linear mixed models. Maternal intake was negatively associated with milk concentrations of free arginine (–0·0003; P = 0·01) and free lysine (–0·0004; P = 0·03) and was positively associated with free glutamine (0·002; P = 0·03) and free threonine (0·0008; P = 0·03). However, these associations were attenuated after correction for multiple testing. Both the quality and quantity of dietary protein intake in lactating women do not seem to influence the amino composition of their breast milk when living in an affluent environment.

This experiment aimed to investigate the impacts of tributyrin (TB) dietary supplementation on serum biochemical indices and meat quality characteristics of longissimus thoracis et lumborum (LTL) muscle of lambs after weaning. Thirty healthy Small-Tailed Han female lambs (27.5 ± 4.1 kg; mean ± standard deviation) were randomly assigned to five treatments: basal diet (1) without TB, (2) with 0.5 g/kg TB, (3) with 1.0 g/kg TB, (4) with 2.0 g/kg TB or (5) with 4.0 g/kg TB. Each treatment consisted of six lambs, and the lambs were weaned on d 90 and were raised until d 165. Results showed that supplementing TB significantly promoted serum immunoglobulin concentrations of lambs such as immunoglobulins G, A and M. Besides, TB significantly increased muscle ether extract content, intermuscular fat length, pH value and redness but decreased lightness, drip loss and shear force. In addition, TB significantly elevated inosine-5ʹ-phosphate content and upregulated the relative expressions of genes related to lipid metabolism such as SREBP-1C, SCD, PPARγ, FAS and LPL. The mostly important, TB significantly enhanced essential amino acids (EAAs) and conjugated linoleic acids contents of the LTL muscle, despite it decreased total unsaturated fatty acids level. In conclusion, supplementing TB not only could promote the healthy status of weaned lambs via promoting serum immunity but also may improve nutritional quality of LTL muscle by improving EAA and conjugated linoleic acid contents.

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.

Protein-rich animal foods are highly digestible, high-quality sources or protein, whereas the protein quality of plant-based foods can vary considerably. Given the growing interest in alternative non-animal-based sources of protein, it is important to establish the protein digestibility of these new foods and protein concentrates which have important health implications especially for vulnerable groups who don’t consume sufficient dietary protein. The human ileostomy model is ideal for measuring protein digestibility as it enables protein digestion to be quantified independent of protein degradation in the large intestine. The aim of this study was to determine the protein digestibility and quality of a wheat-based food containing legume flours. This randomised, double-blinded, controlled cross-over intervention was conducted in 4 proctocolectomised adults with conventional and well-functioning permanent ileostomies. The study was conducted over 2 weeks and on each testing day, the participant consumed 2 test muffins (125 g each) or 2 protein-free cookies in the morning (breakfast and morning tea) followed by a standardised low-protein lunch and afternoon tea. Test muffins were made using a standard muffin recipe using wheat flour and for 2 of the test muffins 50% of the flour was substituted with soy or lupin flour. An indigestible marker, titanium dioxide was added to the muffins so that the completeness of muffin recovered in ileal digesta could be calculated. The digestible indispensable amino acid score (DIAAS) was determined by comparing concentrations of true ileal digestible indispensable amino acids to recommended amino acid requirements(1). Data was reported as mean ± SD and repeated measures ANOVA was used to compare means between treatment groups with significance reported at P < 0.05. Substituting 50% of wheat flour in muffins with soy or lupin flour doubled the protein content of muffins (soy 11.8 g/100g and lupin 10.6 g/100g) compared to muffins that only contained wheat flour (wheat 5.1 g/100g). However, substituting wheat with legume flour did not affect protein digestibility which was similar for all muffin types; wheat (76.8 ± 7.0%), soy (77.9 ± 7.4%) and lupin (81.6 ± 6.9%) (P = 0.181). The DIAAS values for all muffins were below 75% which is classified as the cut off for a good quality protein food. In conclusion, substitution of wheat-based muffins with soy and lupin flour increased the protein content of wheat-based muffins but protein digestibility and overall protein quality was similar.

Optically active cationic complexes adsorbed on montmorillonite can be used for the resolution of racemic mixtures. Montmorillonite-Cu-lysine systems were used as a solid phase in high-pressure liquid chromatography for the resolution of the optical isomers of α-amino acids. Selectivity constants > 1.5 were measured for phenylalanine and tryptophan. The selectivity constants for the amino acids containing saturated-hydrocarbon side chains were in the range of 1.25–1.44. The montmorillonite-Cu-l-lysine complex displayed a stronger affinity for the l-isomers of α-amino acids than for the d-isomers at pHs near neutrality. Inasmuch as surface-catalyzed peptide formation on clays has been proposed as a step in chemical evolution, this stronger affinity between the clay-Cu-l-amino acid complex and l-amino acids might have been significant in prebiotic evolution. The mechanism of optical resolution probably involved ligand exchange. Optimizing the choice of the optically active ligands and of the chelating cation in the chiral agent may improve the resolution of the optical isomers.

Intracellular levels of glutathione, the major mammalian antioxidant, are reported to decline with age in several species. To understand whether ageing affects circulating glutathione levels in cats, blood was sampled from two age groups, < 3 years and > 9 years. Further, to determine whether dietary supplementation with glutathione precursor glycine (GLY) affects glutathione concentrations in senior cats (> 8 years), a series of free GLY inclusion level dry diets were fed. Subsequently, a 16-week GLY feeding study was conducted in senior cats (> 7 years), measuring glutathione, and markers of oxidative stress. Whole blood and erythrocyte total, oxidised and reduced glutathione levels were significantly decreased in senior cats, compared with their younger counterparts (P ≤ 0·02). The inclusion level study identified 1·5 % free GLY for the subsequent dry diet feeding study. Significant increases in erythrocyte total and reduced glutathione were observed between senior cats fed supplemented and control diets at 4 weeks (P ≤ 0·03; maximum difference of 1·23 µM). Oxidative stress markers were also significantly different between groups at 8 (P = 0·004; difference of 0·68 nG/ml in 8-hydroxy-2'-deoxyguanosine) and 12 weeks (P ≤ 0·049; maximum difference of 0·62 nG/mG Cr in F2-isoprostane PGF2α). Senior cats have lower circulating glutathione levels compared with younger cats. Feeding senior cats a complete and balanced dry diet supplemented with 1·5 % free GLY for 12 weeks elevated initial erythrocyte glutathione and altered markers of oxidative stress. Dietary supplementation with free GLY provides a potential opportunity to restore age-associated reduction in glutathione in cats.

Intercalation of amino acids into 10.0-Å hydrated kaolinite was studied by powder X-ray diffraction (XRD), differential thermal analysis-thermal gravimetry (DTA-TG), and infrared (IR) spectroscopy. Intercalation was found to be dependent on the chain-length, pH, and the concentration of the amino acid zwitterion. Near the isoelectric point, fully intercalated phases were obtained in solutions of concentration >0.5–1 M for glycine (Gly), 2–3 M for β-alanine (β-Ala), and 12 M for both γ-aminobutyric acid (γ-Aba) and δ-aminovaleric acid (δ-Ava). ∊-aminocaproic acid (∊-Aca) with a long chain (C = 6) was only partially intercalated. Intercalated amino acid formed a mono-molecular arrangement with the alkyl chain tilting toward the layer at an angle related to H2O content. The compositions of the intercalates of the Gly and β-Ala are Al2Si2O5(OH)4·(Gly)0.67·0.24H2O and Al2Si2O5(OH)4·(β-Ala)0.63·0.25H2O, respectively, based on TG data. From IR data, Gly and β-Ala molecules are found intercalated as zwitterions and these molecules form hydrogen bonds with both the Al-OH and Si-O surfaces of kaolinite. Washing the intercalate with water produced a hydrated kaolinite, which may form a second amino-acid intercalate of high order. Thus, hydrated kaolinite intercalates or deintercalates amino acids depending on concentration and conditions.

This study evaluated the importance of a correction for amino acids (AA) released into the hindgut on a measure of AA absorption kinetics and tested whether AA absorption kinetics are related to the extent of AA absorption using the growing pig as a model for humans. Thirty-six nine-week-old pigs (22·3 kg) received a diet containing whey protein as the sole protein source for 8 d. Pigs received their last meal containing the indigestible marker titanium dioxide before being euthanised at 1, 2, 3, 4, 6 and 12 h post-feeding. The entire content of each gastrointestinal tract (GIT) region was collected to determine AA released into the hindgut, and the kinetics and extent of AA absorption (uncorrected and corrected for AA entering the hindgut). Amounts of AA released into the hindgut increased over time (e.g. 33 and 180 mg of Glu for 4 and 6 h post-feeding). The corrected apparent amount of each AA absorbed from the GIT lumen after 4 h post-feeding was generally lower (P ≤ 0·05) than the uncorrected counterpart. Differences in both the kinetics and extent of AA absorption were observed across AA. For example, the time to reach half of the apparent AA absorption (T50) was 1·5 and 3·4 h for Met and Arg, respectively, whereas their extent of apparent absorption was 93 and 73 %. Negative correlations between parameters related to kinetics and the extent of apparent absorption were observed (e.g. for T50 r = −0·81; P < 0·001). The kinetics of AA absorption is related to the extent of AA absorption.

This review explores the evolution of dietary protein intake requirements and recommendations, with a focus on skeletal muscle remodelling to support healthy ageing based on presentations at the 2023 Nutrition Society summer conference. In this review, we describe the role of dietary protein for metabolic health and ageing muscle, explain the origins of protein and amino acid (AA) requirements and discuss current recommendations for dietary protein intake, which currently sits at about 0⋅8 g/kg/d. We also critique existing (e.g. nitrogen balance) and contemporary (e.g. indicator AA oxidation) methods to determine protein/AA intake requirements and suggest that existing methods may underestimate requirements, with more contemporary assessments indicating protein recommendations may need to be increased to >1⋅0 g/kg/d. One example of evolution in dietary protein guidance is the transition from protein requirements to recommendations. Hence, we discuss the refinement of protein/AA requirements for skeletal muscle maintenance with advanced age beyond simply the dose (e.g. source, type, quality, timing, pattern, nutrient co-ingestion) and explore the efficacy and sustainability of alternative protein sources beyond animal-based proteins to facilitate skeletal muscle remodelling in older age. We conclude that, whilst a growing body of research has demonstrated that animal-free protein sources can effectively stimulate and support muscle remodelling in a manner that is comparable to animal-based proteins, food systems need to sustainably provide a diversity of both plant and animal source foods, not least for their protein content but other vital nutrients. Finally, we propose some priority research directions for the field of protein nutrition and healthy ageing.

The transition towards more plant-based diets may pose risks for bone health such as low vitamin D and Ca intakes. Findings for the contribution of animal and plant proteins and their amino acids (AA) to bone health are contradictory. This 6-week clinical trial aimed to investigate whether partial replacement of red and processed meat (RPM) with non-soya legumes affects AA intakes and bone turnover and mineral metabolism in 102 healthy 20–65-year-old men. Participants were randomly assigned to diet groups controlled for RPM and legume intake (designed total protein intake (TPI) 18 E%): the meat group consumed 760 g RPM per week (25 % TPI) and the legume group consumed non-soya legume-based products (20 % TPI) and 200 g RPM per week, the upper limit of the Planetary Health Diet (5 % TPI). No differences in bone (bone-specific alkaline phosphatase; tartrate-resistant acid phosphatase 5b) or mineral metabolism (25-hydroxyvitamin D; parathyroid hormone; fibroblast growth factor 23; phosphate and Ca) markers or Ca and vitamin D intakes were observed between the groups (P > 0·05). Methionine and histidine intakes were higher in the meat group (P ≤ 0·042), whereas the legume group had higher intakes of arginine, asparagine and phenylalanine (P ≤ 0·013). Mean essential AA intakes in both groups met the requirements. Increasing the proportion of non-soya legumes by reducing the amount of RPM in the diet for 6 weeks did not compromise bone turnover and provided on average adequate amounts of AA in healthy men, indicating that this ecologically sustainable dietary change is safe and relatively easy to implement.

Low birth weight (LBW) neonates show impaired growth compared with normal birth weight (NBW) neonates. Glutamine (Gln) supplementation benefits growth of weaning piglets, while the effect on neonates is not sufficiently clear. We examined the effect of neonatal Gln supplementation on piglet growth, milk intake and metabolic parameters. Sow-reared pairs of newborn LBW (0·8–1·2 kg) and NBW (1·4–1·8 kg) male piglets received Gln (1 g/kg body mass (BM)/d; Gln-LBW, Gln-NBW; n 24/group) or isonitrogenous alanine (1·22 g/kg BM/d; Ala-LBW; Ala-NBW; n 24/group) supplementation at 1–5 or 1–12 d of age (daily in three equal portions at 07:00, 12:00 and 17:00 by syringe feeding). We measured piglet BM, milk intake (1, 11–12 d), plasma metabolite, insulin, amino acid (AA) and liver TAG concentrations (5, 12 d). The Gln-LBW group had higher BM (+7·5%, 10 d, P = 0·066; 11–12 d, P < 0·05) and milk intake (+14·7%, P = 0·015) than Ala-LBW. At 5 d, Ala-LBW group had higher plasma TAG (+34·7%, P < 0·1) and lower carnosine (–22·5%, P < 0·05) than Ala-NBW and Gln-LBW, and higher liver TAG (+66·9%, P = 0·029) than Ala-NBW. At 12 d, plasma urea was higher (+37·5%, P < 0·05) with Gln than Ala supplementation. Several proteinogenic AA in plasma were lower (P < 0·05) in Ala-NBW v. Gln-NBW. Plasma arginine was higher (P < 0·05) in Gln-NBW v Ala-NBW piglets (5, 12 d). Supplemental Gln moderately improved growth and milk intake and affected lipid metabolism in LBW piglets and AA metabolism in NBW piglets, suggesting effects on intestinal and liver function.

Adequate protein intake is essential for the maintenance of whole-body protein mass. Different methodological approaches are used to substantiate the evidence for the current protein recommendations, and it is continuously debated whether older adults require more protein to counteract the age-dependent loss of muscle mass, sarcopenia. Thus, the purpose of this critical narrative review is to outline and discuss differences in the approaches and methodologies assessing the protein requirements and, hence, resulting in controversies in current protein recommendations for healthy older adults. Through a literature search, this narrative review first summarises the historical development of the Food and Agriculture Organization/World Health Organization/United Nations University setting of protein requirements and recommendations for healthy older adults. Hereafter, we describe the various types of studies (epidemiological studies and protein turnover kinetic measurements) and applied methodological approaches founding the basis and the different recommendations with focus on healthy older adults. Finally, we discuss important factors to be considered in future studies to obtain evidence for international agreement on protein requirements and recommendations for healthy older adults. We conclude by proposing future directions to determine ‘true’ protein requirements and recommendations for healthy older adults.

This study was designed to investigate the effects of dietary starch structure on muscle protein synthesis and gastrointestinal amino acid (AA) transport and metabolism of goats. Twenty-seven Xiangdong black female goats (average body weight = 9·00 ± 1·12 kg) were randomly assigned to three treatments, i.e., fed a T1 (normal maize 100 %, high amylose maize 0 %), T2 (normal maize 50 %, high amylose maize 50 %) and T3 (normal maize 0 %, high amylose maize 100 %) diet for 35 d. All AA in the ileal mucosa were decreased linearly as amylose:amylopectin increased in diets (P < 0·05). The plasma valine (linear, P = 0·03), leucine (linear, P = 0·04) and total AA content (linear, P = 0·03) increased linearly with the increase in the ratio of amylose in the diet. The relative mRNA levels of solute carrier family 38 member 1 (linear, P = 0·01), solute carrier family 3 member 2 (linear, P = 0·02) and solute carrier family 38 member 9 (linear, P = 0·02) in the ileum increased linearly with the increase in the ratio of amylose in the diet. With the increase in the ratio of amylose:amylopectin in the diet, the mRNA levels of acetyl-CoA dehydrogenase B (linear, P = 0·04), branched-chain amino acid transferase 1 (linear, P = 0·02) and branched-chain α-keto acid dehydrogenase complex B (linear, P = 0·01) in the ileum decreased linearly. Our results revealed that the protein abundances of phosphorylated mammalian target of rapamycin (p-mTOR) (P < 0·001), phosphorylated 4E-binding protein 1 (P < 0·001) and phosphorylated ribosomal protein S6 kinases 1 (P < 0·001) of T2 and T3 were significantly higher than that of T1. In general, a diet with a high amylose ratio could reduce the consumption of AA in the intestine, allowing more AA to enter the blood to maintain higher muscle protein synthesis through the mTOR pathway.

Plant-derived proteins have been suggested to have less anabolic properties when compared with animal-derived proteins. Whether blends of plant- and animal-derived proteins can compensate for their lesser anabolic potential has not been assessed. The present study compares post-prandial muscle protein synthesis rates following the ingestion of milk protein with wheat protein or a blend of wheat plus milk protein in healthy, young males. In a randomised, double-blind, parallel-group design, 36 males (23 (sd 3) years) received a primed continuous L-[ring-13C6]-phenylalanine infusion after which they ingested 30 g milk protein (MILK), 30 g wheat protein (WHEAT) or a 30 g blend combining 15 g wheat plus 15 g milk protein (WHEAT+MILK). Blood and muscle biopsies were collected frequently for 5 h to assess post-prandial plasma amino acid profiles and subsequent myofibrillar protein synthesis rates. Ingestion of protein increased myofibrillar protein synthesis rates in all treatments (P < 0·001). Post-prandial myofibrillar protein synthesis rates did not differ between MILK v. WHEAT (0·053 (sd 0·013) v. 0·056 (sd 0·012) %·h−1, respectively; t test P = 0·56) or between MILK v. WHEAT+MILK (0·053 (sd 0·013) v. 0·059 (sd 0·025) %·h−1, respectively; t test P = 0·46). In conclusion, ingestion of 30 g milk protein, 30 g wheat protein or a blend of 15 g wheat plus 15 g milk protein increases muscle protein synthesis rates in young males. Furthermore, muscle protein synthesis rates following the ingestion of 30 g milk protein do not differ from rates observed after ingesting 30 g wheat protein or a blend with 15 g milk plus 15 g wheat protein in healthy, young males.

To evaluate the responses of some important agronomic traits of 14 bread wheat cultivars, a split plot experiment was conducted based on a randomized complete block design under non-stress, moderate and severe salinity stress conditions. In this study, Backcross and Roshan were identified as the most salinity-tolerant cultivars, while Mihan and Shirudi were considered as the most salinity-sensitive cultivars. The proline content, as well as Δ1-pyrroline-5-carboxylate synthase (P5CS) and vacuolar-type H+-ATPase subunit E (W36) gene expression levels, were examined in these cultivars under normal, moderate and high salinity stress conditions. The proline content and P5CS gene expression level increased with a rise in NaCl concentration. Further, a direct relationship was observed between the proline content and P5CS gene expression in all samples. Our results showed that W36 gene expression in Backcross and Roshan cultivars, as the most resistant ones, strongly increased with elevation of the NaCl concentrations. On the other hand, in the sensitive cultivars such as Mihan and Shirudi, small changes were observed in the gene expression levels with rising salinity levels. Additionally, Backcross and Roshan cultivars had the highest proline content as well as P5CS and W36 genes expression levels under moderate and high salinity stress levels.

Gallic acid (GA) is widely used as a dietary supplement due to several health-promoting effects, although its effects on intestinal-epithelial-cell integrity and transport remain mostly unknown. The present study aims to clarify the effects of GA on tight junctions and intestinal nutrient uptake through in vitro and ex vivo models. Both intestinal porcine enterocyte cell line-J2 cells and porcine middle-jejunum segments were treated with 5 (T5), 25 (T25) and 50 (T50) µm GA and mounted in Ussing chambers to determine transepithelial resistance (TEER), claudin-1 (CLDN1), occludin (OCLN), zonula occludens-1 (ZO-1) protein (in tissues and cells) and mRNA (in cells) expression. In addition, uptake of l-glutamate (l-Glut), l-arginine (l-Arg), l-lysine (l-Lys) and l-methionine (l-Meth) together with cationic-amino-acid transporter-1 (CAT-1) and excitatory-amino-acid transporter-3 (EAAT3) expression was evaluated. No apoptosis was observed in GA-treated cells, but TEER and CLDN1 protein abundance was lower with T50 compared with untreated cells. l-Arg and l-Lys uptake was greater with T5 than with T25 and T50. Ex vivo, T50 decreased the TEER values and the protein levels of CLDN1, OCLN and ZO-1, whereas T5 and T25 only decreased CLDN1 protein expression compared with untreated tissues. Moreover, T25 increased l-Glut and l-Arg uptake, the latter confirmed by an increased protein expression of CAT-1. GA influences intestinal uptake of the tested cationic amino acids at low concentrations and decreases the intestinal-cell barrier function at high concentrations. Similarities were observed between in vitro and ex vivo, but different treatment times and structures must be considered.

The development and maintenance of body composition and functions require an adequate protein intake with a continuous supply of amino acids (AA) to tissues. Body pool and AA cellular concentrations are tightly controlled and maintained through AA supply (dietary intake, recycled from proteolysis and de novo synthesis), AA disposal (protein synthesis and other AA-derived molecules) and AA losses (deamination and oxidation). Different molecular regulatory pathways are involved in the control of AA sufficiency including the mechanistic target of rapamycin complex 1, the general control non-derepressible 2/activating transcription factor 4 system or the fibroblast growth factor 21. There is a tight control of protein intake, and human subjects and animals appear capable of detecting and adapting food and protein intake and metabolism in face of foods or diets with different protein contents. A severely protein deficient diet induces lean body mass losses and ingestion of sufficient dietary energy and protein is a prerequisite for body protein synthesis and maintenance of muscle, bone and other lean tissues and functions. Maintaining adequate protein intake with age may help preserve muscle mass and strength but there is an ongoing debate as to the optimal protein intake in older adults. The protein synthesis response to protein intake can also be enhanced by prior completion of resistance exercise but this effect could be somewhat reduced in older compared to young individuals and gain in muscle mass and function due to exercise require regular training over an extended period.

A 6-week growth trial was conducted to evaluate the influences of dietary valine (Val) levels on growth, protein utilisation, immunity, antioxidant status and gut micromorphology of juvenile hybrid groupers. Seven isoenergetic, isoproteic and isolipidic diets were formulated to contain graded Val levels (1·21, 1·32, 1·45, 1·58, 1·69, 1·82 and 1·94 %, DM basis). Each experimental diet was hand-fed to triplicate groups of twelve hybrid grouper juveniles. Results showed that weight gain percentage (WG%), protein productive value, protein efficiency ratio, and feed efficiency were increased as dietary Val level increased, reaching a peak value at 1·58 % dietary Val. The quadratic regression analysis of WG% against dietary Val levels indicated that the optimum dietary Val requirement for hybrid groupers was estimated to be 1·56 %. Gut micromorphology and expression of growth hormone in pituitary, insulin-like growth factor 1, target of rapamycin and S6 kinase 1 in liver were significantly affected by dietary Val levels. In serum, fish fed 1·58 % dietary Val had higher superoxide dismutase, catalase, lysozyme activities and IgM concentrations than fish fed other dietary Val levels. Fish fed 1·58 % dietary Val had higher expression of NF-E2-related factor 2 in head kidney than fish fed other dietary Val levels. Generally, the optimum dietary Val requirement for maximal growth of hybrid groupers was estimated to be 1·56 % of DM, corresponding to 3·16 % of dietary protein, and dietary Val levels affected growth, protein utilisation, immunity and antioxidant status in hybrid groupers.

The objective of this study was to assess the nutritional quality of pea protein isolate in rats and to evaluate the impact of methionine (Met) supplementation. Several protein diets were studied: pea protein, casein, gluten, pea protein–gluten combination and pea protein supplemented with Met. Study 1: Young male Wistar rats (n 8/group) were fed the test diets ad libitum for 28 d. The protein efficiency ratio (PER) was measured. Study 2: Adult male Wistar rats (n 9/group) were fed the test diets for 10 d. A protein-free diet group was used to determine endogenous losses of N. The rats were placed in metabolism cages for 3 d to assess N balance, true faecal N digestibility and to calculate the Protein Digestible-Corrected Amino Acid Score (PDCAAS). They were then given a calibrated meal and euthanised 6 h later for collection of digestive contents. The true caecal amino acid (AA) digestibility was determined, and the Digestible Indispensable Amino Acid Score (DIAAS) was calculated. Met supplementation increased the PER of pea protein (2·52 v. 1·14, P < 0·001) up to the PER of casein (2·55). Mean true caecal AA digestibility was 94 % for pea protein. The DIAAS was 0·88 for pea protein and 1·10 with Met supplementation, 1·29 for casein and 0·25 for gluten. Pea protein was highly digestible in rats under our experimental conditions, and Met supplementation enabled generation of a mixture that had a protein quality that was not different from that of casein.