Dietary Indispensable Amino Acid Research Articles

Digestible indispensable amino acid score (DIAAS): 10 years on

The objective of the review is to revisit the findings of the 2011 Food and Agriculture Organization of the United Nations (FAO) Expert Consultation on Dietary Protein Quality Evaluation in Human Nutrition, and to report on progress on uptake of the findings. It is evident that since 2011 there has been a concerted research effort to enhance an understanding of the protein quality of foods. The validity of the growing pig ileal protein digestibility assay has been confirmed and numerous studies reported using the growing pig as a model to give true ileal amino acid digestibility values for foods as consumed by humans. This has allowed for the determination of digestible indispensable amino acid scores (DIAAS) for a range of foods. A new non-invasive true ileal amino acid digestibility assay in humans which can be applied in different physiological states, called the dual-isotope assay, has been developed and applied to determine the DIAAS values of foods. It is concluded that DIAAS is currently the most accurate score for routinely assessing the protein quality rating of single source proteins. In the future, the accuracy of DIAAS can be enhanced by improved information on: the ideal dietary amino acid balance including the ideal dispensable to indispensable amino acid ratio; dietary indispensable amino acid requirements; effects of processing on ileal amino acid digestibility and lysine bioavailability. There is a need to develop rapid, inexpensive in vitro digestibility assays. Conceptual issues relating DIAAS to food regulatory claims, and to holistic indices of food nutritional and health status are discussed. The first recommendation of the 2011 Consultation regarding treating each indispensable amino acid as an individual nutrient has received little attention. Consideration should be given to providing food label information on the digestible contents of specific indispensable amino acids.

62 Sulphur Amino Acids: Assessing Requirements and Utilization in Dogs and Cats

Abstract Methionine (Met), cyst(e)ine (Cys), homocysteine and taurine are the four sulfur-containing amino acids. Met can serve as a substrate for protein synthesis or to produce homocysteine, which can be remethylated to Met or irreversibly catabolized to Cys. Unlike dogs, cats are unable to further metabolize Cys to taurine, making the latter an indispensable amino acid for the cat. Met, on the other hand, is a dietary indispensable amino acid for all mammals with Cys having the ability to spare part of the Met requirement (total sulfur amino acid requirement; Met + Cys). The amount of Met that cannot be replaced by Cys is defined as the Met requirement (NRC, 2006). Empirical determinations of the Met requirement in adult dogs and cats are still limited and the recommendations proposed by the NRC (2006) are based on studies that used insensitive techniques for mature animals. When a more robust technique, the indicator amino acid oxidation, and more practical experimental diets were used to empirically determine the Met requirement, a greater estimate was found in adult cats (Pezzali et al., 2022) and adult Labrador Retrievers and Beagles, but not in miniature Dachshunds (Mansilla et al., 2020), compared with the current recommendations proposed by the NRC (2006) and AAFCO (2023). Harrison et al. (2020), however, found a decreased estimate for the Met requirement in adult Labrador Retrievers compared with Mansilla et al. (2020), likely due to differences in experimental design and diet composition. This discrepancy highlights the need to standardize methodologies and to investigate the effect of other dietary factors on the Met and total sulfur amino acid requirements owing to their complex metabolism. Indeed, the metabolism of sulfur amino acids may differ between dog breeds, resulting in differences in Met requirements and potentially making taurine a conditionally indispensable amino acid in some breeds under specific dietary conditions. While dietary Met is usually provided in excess in commercial diets, excess of Met may lead to hyperhomocysteinemia, which has been associated with detrimental health effects in other species. Due to the complex metabolism of sulfur amino acids and the complex matrix of pet foods, determining the ideal provision of sulfur amino acids in commercial diets to maximize animal health is a challenge. More long-term studies investigating the effect of different doses and ratios (Met:Cys) of sulfur amino acids on overall metabolism are necessary, in addition to requirement studies, to optimize the dietary delivery of sulfur amino acids in dogs and cats.

Population protein intakes and food sustainability indices: The metrics matter

It is common practice in food sustainability studies to calculate population-based average protein intakes and express environmental footprints on a gross protein basis. Such analyses, however, assume that the requirement for protein is also expressed as gross protein which is not the case. This ignores the critical role of dietary protein as a supply of dietary indispensable amino acids, and does not account for differences in the protein quality of foods.Previously published datasets are re-analysed here to express population protein intakes and environmental footprint data on an available protein basis, accounting for both amino acid digestibility and utilization. When protein quality is taken into account, for many countries of the world protein is found to be deficient for the average adult, and different rankings are found for various environmental footprints.

203 Efficiency of SID lysine utilization and maximum SID lysine retention for gilts in early, mid and late gestation

Abstract Efficiency of amino acid (AA) use is presumed constant across gestation but may not reflect changes in metabolic demand during gestation nor consider changes in efficiency depending on level of AA intake. Two experiments were conducted to determine efficiency of SID Lys utilization in gilts during early (d 48-52), mid (d 75-79) and late gestation (d 103-107). Each experiment provided 4 isocaloric (3,335 kcal ME/kg) and isoproteic (11.75 % CP) diets containing 4 SID Lys levels (Table 1). Diets were randomly assigned to 45 gilts (PIC 1050, 158.0 ± 8.0 kg at d 39.4 ± 1 of gestation) in Exp. 1 and 27 gilts (PIC 1050, 169.0 ± 7.5 kg at d 41 ± 1 of gestation) in Exp. 2. Dietary indispensable AA were set to meet or exceed 100% of AA:Lys ratios in both experiments. The SID Lys retention was estimated from whole body nitrogen (N) retention balance studies in each period (7 d diet adaptation, 5 d total urine collection and grab fecal sampling) according to the NRC (2012) equations. The relationship between SID Lys intake and SID Lys retention was determined by nonlinear regression models using the CurveExpert Professional software. According to the Hoerl regression model: E(y)=exp(β 0+β 1X)[Xβ2] best-fitting line, maximum efficiency of SID Lys utilization (i.e. g SID Lys retention/g SID Lys intake) was 65%, 57%, and 53% in early, mid and late gestation and occurred at 6.6, 8 and 12 g of SID Lys intake/d, respectively. Maximum SID Lys retention occurred at 8.1 and 9.8 g of SID Lys intake/d for early and mid-gestation. The SID Lys retention did not reach a maximum value in late gestation. These results suggest that efficiency of SID Lys utilization is not constant across gestation and that maximal efficiency occurs at intake below current recommendations.

A review on fish immuno-nutritional response to indispensable amino acids in relation to TOR, NF-κB and Nrf2 signaling pathways: Trends and prospects.

Indispensable amino acids (IAAs) are important regulators of key metabolic pathways associated with protein synthesis, tight junction proteins, inflammatory cytokines and immune-antioxidant related signaling molecules. However, the information pertaining to the immune functions of IAAs in relation to molecular approaches for commercially important fish species are scarce and discordant. This review summarizes the dietary requirements for IAAs necessary for improved growth and immune response in variety of fish species, using molecular approaches (nutrigenomics), particularly the interrelationships between IAAs and genes. Briefly, antioxidant status of fish as well as gene transcriptions regulating antioxidant enzymes are profoundly governed by the nutritional factors including a set of IAAs, and these genes expression are often regulated by the nuclear factor erythoid 2-related factor 2 signaling pathway (Nrf2). IAAs level could also attenuate the inflammatory response in fish partly by down-regulating the expression levels of pro-inflammatory cytokines and up-regulating the anti-inflammatory cytokines. The regulation of these cytokines by IAAs could be mediated by the signaling molecules nuclear transcription factor-κB (NF-κB) and target of rapamycin (TOR). Overall, this review provides clear and recent molecular mechanisms of fish immuno-nutritional interrelation and highlights regulatory pathways underlying dietary IAAs mediated enhancement in the antioxidant, anti-inflammatory, and immune defense capacities, presenting trends and future perspectives.

Nutritional composition of farm chinchilla (Chinchilla lanigera) meat

This study examined the nutritional composition of farm chinchilla meat from Lithuania and compared with the nutrient levels found in other studies for rodents’ meat. The chinchilla meat was characterised as a good source of animal protein (19.96–21.44 g/100 g) and was found to be rich in lysine (8.67 ± 0.13 g/100 g protein), glutamic acid (15.42 ± 0.19 g/100 g protein) and sulphur-containing methionine (5.43 ± 0.09 g/100 g protein). For the first time, the score based on individual dietary indispensable amino acid digestibility was determined for chinchilla and other rodents’ meat. The DIAAS values ranged from 89% (SSA) in nutria meat to 114% (valine) in chinchilla meat. Lipids of chinchilla meat had lower SFA content, and the content of PUFA n-3 and n-6 was always higher than those reported for nutria, capybara and rabbit meat. The mineral composition of chinchilla meat is presented for the first time in this study. It was different from that of other rodents with a small amount of sodium (32.65 ± 0.78 mg/100 g muscle). Nutritional information on chinchilla meat is limited. The data obtained in this study might be useful for further discussions on farm chinchilla meat as potential meat sources.

Responses of Pigs of Different Genotypes to a Variation in the Dietary Indispensable Amino Acid Content in Terms of Their Growth and Carcass and Meat Quality Traits.

Simple SummaryThe aim of the experiment was to study the response of pigs of different genotypes to a variation in their dietary amino acid allowance. Ninety-six crossbred barrows of two lean paternal genetic lines (Hypor Maxter and PIC 337) were housed in eight pens from body weight 60 to 145 kg and fed quasi ad libitum on feeds with either high or low ileal digestible indispensable amino acid contents. The high and low amino acid feeds were formulated to have the same standardized ileal digestible (SID) lysine, methionine, tryptophan and threonine contents per unit of crude protein, but provided 9.4 to 8.0 (assumed to be non-limiting) or 8.5 to 6.5 (close to National Research Council recommendations) g/kg of SID lysine, respectively. The low amino acid feed reduced the estimated N excretion compared with the high, without affecting growth, carcass yield, carcass anatomical composition and meat quality traits. Genetic line had trivial effects and the amino acid level × genotype interaction was never significant. We concluded that the two pig genotypes did not differ sufficiently in growth potential and leanness to respond differently.We studied the response of pigs from two crossbred genetic lines (GL) in the 60 to 145 kg body weight interval to a variation in the indispensable amino acid (AA) content of their feed. Ninety-six barrows of two paternal GLs (Hypor Maxter and PIC 337) were housed in eight pens and fed quasi ad libitum on feeds differing in their standardized ileal digestible (SID) indispensable AA contents. Pigs in four pens received feeds containing 9.4 to 8.0 g/kg of SID Lys (HAA), considered non-limiting, while the others received feeds containing 8.5 to 6.5 g/kg of SID Lys (LAA). The two feeds had identical indispensable lysine, methionine, tryptophan and threonine contents per unit of crude protein (CP). Feed intake, growth, carcass weight, and the weights of the lean and fat cuts were recorded, and samples of longissimus lumborum were analyzed. Data were analyzed using a two-way factorial mixed model. The LAA feed lowered the estimated N excretion (p < 0.001) compared with HAA, without affecting growth, carcass or meat quality traits. Genetic line had trivial effects and the AA level × genotype interaction was never significant. The two pig genotypes did not differ sufficiently in growth potential and leanness to respond differently.

Nutritional evaluation of 3 types of novel ethanol coproducts

Five 48-h precision-fed rooster assays were conducted with the objective to determine true metabolizable energy (TMEn) using conventional roosters and/or standardized amino acid digestibility using cecectomized roosters for distillers dried grains with solubles (DDGS) produced from human food waste at high solids content (FWDDGS), DDGS produced from 4 corn hybrids with increases in grain protein concentration or the concentrations of several dietary indispensable amino acids, and a coproduct that is produced by a process which separates a high protein and yeast fraction from ethanol stillage (Still Pro, Fluid Quip Process Technologies, Cedar Rapids, IA). These results from the first cecectomized rooster assay indicated that the standardized digestibility values for Lys, Met, Cys, Thr, and Val were 61, 75, 70, 70, and 72%, respectively. Using conventional roosters, the TMEn for the FWDDGS was 3,890kcal/kg DM. The DDGS produced from the high protein mutant corn hybrid had a higher protein content of 34% compared with 28% protein for DDGS from the control corn hybrid. Using Lys as an example, there was a large difference between the 2 samples; the high protein mutant DDGS contained 1.60% Lys vs 1.05% for the control DDGS. Standardized digestibility of amino acids was generally not different for the mutant DDGS and the control DDGS. Similar results were observed for Lys, Arg, and Trp for the DDGS produced from the second mutant corn hybrid. The Still Pro sample was analyzed to contain 53% protein (DM basis) with 2.22% Lys, 1.05% Met, 0.90% Cys, 2.06% Thr, and 3.08% Val. The standardized digestibility values for these amino acids were 84, 92, 87, 86, and 87%, respectively. The TMEn of the Still Pro sample was determined to be 3,372kcal/kg DM. In conclusion, the results of this study indicate that the nutritional value of the 3 types of novel ethanol coproducts is equal to or superior to the nutritional values generally reported for conventional DDGS.

Indispensable Amino Acid–Deficient Diets Induce Seizures in Ketogenic Diet–Fed Rodents, Demonstrating a Role for Amino Acid Balance in Dietary Treatments for Epilepsy

Low protein amounts are used in ketogenic diets (KDs), where an essential (indispensable) amino acid (IAA) can become limiting. Because the chemically sensitive, seizurogenic, anterior piriform cortex (APC) is excited by IAA limitation, an imbalanced KD could exacerbate seizure activity. We questioned whether dietary IAA depletion worsens seizure activity in rodents fed KDs. In a series of 6 trials, male rats or gerbils of both sexes (6-8/group) were given either control diets (CDs) appropriate for each trial, a KD, or a threonine-devoid (ThrDev) diet for≥7 d, and tested for seizures using various stimuli. Microchip analysis of rat APCs was also used to determine if changes in transcripts for structures relevant to seizurogenesis are affected by a ThrDev diet. Glutamate release was measured in microdialysis samples from APCs during the first meal after 7 d on a CD or a ThrDev diet. Adult rats showed increased susceptibility to seizures in both chemical (58%) and electroshock (doubled) testing after 7 d on a ThrDev diet compared with CD (each trial, P≤ 0.05). Seizure-prone Mongolian gerbils had fewer seizures after receiving a KD, but exacerbated seizures (68%) after 1 meal of KD minus Thr (KD-T compared with CD, P<0.05). In kindled rats fed KD-T, both counts (19%) and severities (77%) of seizures were significantly elevated (KD-T compared with CD, P<0.05). Gene transcript changes were consistent with enhanced seizure susceptibility (7-21 net-fold increases, P=0.045-0.001) and glutamate release into the APC was increased acutely (4-fold at 20 min, 2.6-fold at 60 min, P<0.05) after 7 d on a ThrDev diet. Seizure severity in rats and gerbils was reduced after KDs and exacerbated by ThrDev, both in KD- and CD-fed animals, consistent with the mechanistic studies. We suggest that a complete protein profile in KDs may improve IAA balance in the APC, thereby lowering the risk of seizures.

Effect of phytase on amino acid digestibility in pig: A meta-analysis

Phytic acid represents about 60% of the phosphorus in plant-based ingredients used in pig feed. It is known to bind to proteins and amino acids and thereby interfere with digestion. The effectiveness of microbial phytase to improve the digestibility of amino acid in pig feed appears inconsistent. The objective of this study was to estimate the effect of microbial phytase on the digestibility of dietary indispensable amino acids, as a function of dietary crude protein content, amino acid content, calcium, phytic acid, neutral detergent fibre and acid detergent fibre, using a meta-analysis tool. A database derived from 34 papers published between 1994 and 2015 and describing 138 experimental treatments was compiled. Adding microbial phytase to pig feed appears to improve the digestibility of amino acids. Digestibility increased linearly for Arg (R2 = 0.99), His 36 (R2 = 0.98), Ile (R2 = 0.97), Leu (R2 = 0.97), Met (R2 = 0.99), Thr (R2 = 0.97), Phe (R2 = 0.98) 37 and Val (R2 = 0.85), and was quadratic for Lys (R2 = 0.99), and Tyr (R2 = 0.99). Fibre and phytase do not interact. The current study showed that microbial phytase supplementation affected positively the apparent ileal digestibility of amino acids and this effect was not modified by other dietary components such as amino acids, calcium, phytic acid and fibre. The current models allow quantifying the effect of microbial phytase on amino acids which is important to use phytase feed enzyme accurately in diet formulation.

Rumen-protected methionine during the peripartal period in dairy cows and its effects on abundance of major species of ruminal bacteria

BackgroundExtensive degradation of amino acids in the rumen via microbial deamination decreases the post-ruminal availability of dietary indispensable amino acids. Together with the normal decrease in voluntary dry matter intake (DMI) around parturition in dairy cows, microbial metabolism contributes to a markedly negative balance of indispensable amino acids, including methionine which may be the first-limiting for milk production. The main objective of the current study was to profile changes in major bacterial species with key functions in cellulose and hemicellulose digestion, xylan breakdown, proteolytic action, propionic acid production, lactate utilization and ruminal biohydrogenation in cows supplemented with rumen-protected Methionine (SM; Smartamine M, Adisseo NA, Alpharetta, GA, USA) from −23 through 30 d relative to parturition. Because ~90% of the methionine in SM bypasses the rumen, ~10% of the methionine is released into the rumen and can be utilized by microbes.ResultsAs expected, there was an increase in overall DMI after parturition (Day, P < 0.05) during which cows consumed on average 19.6 kg/d versus 13.9 kg/d in the prepartum period. The postpartum diet contained greater concentrations of lipid and highly-fermentable carbohydrate from corn grain, which likely explains the increases in the relative abundance of Anaerovibrio lipolytica, Megasphaera elsdenii, Prevotella bryantii, Selenomonas ruminantium, Streptococcus bovis, and Succinimonas amylolytica. Despite similar DMI prepartum, cows fed SM had greater (Treatment × Day, P < 0.05) abundance prepartum of Fibrobacter succinogenes, Succinimonas amylolytica, and Succinivibrio dextrinosolvens. However, the greater DMI in cows fed SM after parturition (19.6 kg/d versus 13.9 kg/d) was associated with lower abundance of Fibrobacter succinogenes (2.13 × 10−3 versus 2.25 × 10−4) and Selenomonas ruminantium (2.98 × 10−1 versus 4.10 × 10−1). A lower abundance (Day, P < 0.05) was detected on d 20 compared with d −10 for Fibrobacter succinogenes and Succinivibrio dextrinosolvens. The relative abundance of Butyrivibrio proteoclasticus and Eubacterium ruminantium was stable across treatment and time.ConclusionsIn diets with proper balance of rumen-degradable protein and fermentable carbohydrate, the small fraction of Methionine released from the rumen-protected supplement did not seem to compromise growth of major bacterial species in the rumen. In fact, it had a positive effect on 3 major species prepartum when DMI was similar between groups. Because the actual requirements of Methionine (and Lysine, for example) by the cow during the transition period are unknown, it appears warranted to study the rumen microbiome as it relates to supply of rumen-protected amino acids.

Influence of mild feed restriction and mild reduction in dietary amino acid content on feeding behaviour of group-housed growing pigs

This study investigates changes in the feeding behaviour of pigs as a result of a restriction in their feed allowance and a reduction in dietary indispensable amino acid (AA) content. Ninety-six Topig Talent×PIC barrows were housed in 8 pens and individually fed either ad libitum (AL) or a restricted diet (RF) from 47 to 145kg body weight (BW). The amount of feed given to RF pigs was close to their expected voluntary intake, but it was limited to proportions of 0.33, 0.66 and 1.00 of the estimated daily amount of feed in 3 time intervals, 00:01 to 8:00, 8:01 to 16:00 and 16:01 to 24:00h, respectively. From 86kg BW, the pigs in 4 of the pens were fed diets with conventional standardized ileal digestible AA content (CAA), while the pigs in the other pens received diets (LAA) in which the proportions of dietary indispensable AA were lowered with respect to CAA by 0.91 from 86 to 118kg BW and by 0.82 from 118 to 145kg BW. Automated feeders monitored individual feeding behaviour. Data were analysed by pig and feeding phase with a 2×2 factorial design. Over the whole experimental period, feed restriction resulted in a decrease in daily feed intake (7%, P <0.001), the number of visits (27%, P <0.001) and the time spent feeding (14%, P <0.001), but an increase in feed consumption per visit (20%, P=0.001) and feeding rate (10%, P=0.032). The reduction in AA increased daily feed intake (7%, P=0.031), tended to increase feeding rate (14%, P=0.07) and interacted with feeding regime with respect to the number and duration of feeding visits. During growing and finishing, we observed high, negative, non-linear relationships between feed consumption per visit and visit frequency (R2=0.989 to 0.876), between visit duration and visit frequency (R2=0.648 to 0.695), and between feeding rate and time spent feeding in a day (R2=0.802 to 0.707), and positive linear relationships between visit duration and feed consumption per visit (R2=0.614 to 0.570). The individual feeding rate during growing was positively correlated with that during finishing (R2=0.458). We conclude that pigs try to adapt their feeding pattern to compensate for a reduction in feed allowance or nutrient restriction by, for example, increasing their feeding rate, which may reflect increased feeding motivation.

Amino Acid Absorption in the Large Intestine of Humans and Porcine Models

Dietary protein quality has been recognized as a critical issue by international authorities because it can affect important functions of the body. To predict protein quality, the FAO introduced the Digestible Indispensable Amino Acid Score. This score depends on ileal amino acid (AA) digestibility; therefore, the assumption is made that AAs are not absorbed in nutritionally relevant amounts from the large intestine. This article reviews the evidence for this assumption by considering the role of the mammalian large intestine in dietary protein and AA digestion and absorption, with particular reference to adult humans. Although most dietary AAs and peptides are absorbed in the small intestine, substantial amounts can enter the large intestine. Nitrogen is absorbed in the large intestine, and a series of animal experiments indicate a potential small degree of AA absorption. In humans, colonocytes have the capacity for AA absorption because AA transporters are present in the large intestine. The absorption of nutritionally relevant amounts of dietary indispensable AAs and peptides in the human large intestine has not been convincingly demonstrated, however.

Dietary indispensable amino acids profile affects protein utilization and growth of Senegalese sole larvae

In diet formulation for fish, it is critical to assure that all the indispensable amino acids (IAA) are available in the right quantities and ratios. This will allow minimizing dietary AA imbalances that will result in unavoidable AA losses for energy dissipation rather than for protein synthesis and growth. The supplementation with crystalline amino acids (CAA) is a possible solution to correct the dietary amino acid (AA) profile that has shown positive results for larvae of some fish species. This study tested the effect of supplementing a practical microdiet with encapsulated CAA as to balance the dietary IAA profile and to improve the capacity of Senegalese sole larvae to utilize AA and maximize growth potential. Larvae were reared at 19°C under a co-feeding regime from mouth opening. Two microdiets were formulated and processed as to have as much as possible the same ingredients and proximate composition. The control diet (CTRL) formulation was based on commonly used protein sources. A balanced diet (BAL) was formulated as to meet the ideal IAA profile defined for Senegalese sole: the dietary AA profile was corrected by replacing 4% of encapsulated protein hydrolysate by CAA. The in vivo method of controlled tube-feeding was used to assess the effect on the larvae capacity to utilize protein, during key developmental stages. Growth was monitored until 51 DAH. The supplementation of microdiets with CAA in order to balance the dietary AA had a positive short-term effect on the Senegalese sole larvae capacity to retain protein. However, that did not translate into increased growth. On the contrary, larvae fed a more imbalanced (CTRL group) diet attained a better performance. Further studies are needed to ascertain whether this was due to an effect on the voluntary feed intake as a compensatory response to the dietary IAA imbalance in the CTRL diet or due to the higher content of tryptophan in the BAL diet.

The supplementation of a microdiet with crystalline indispensable amino-acids affects muscle growth and the expression pattern of related genes in Senegalese sole (Solea senegalensis) larvae

The full expression of growth potential in fish larvae largely depends on an efficient protein utilization, which requires that all the indispensable amino acids (IAAs) are provided at an optimum ratio. The effect of supplementing a practical microdiet with encapsulated crystalline-AA to correct possible IAA deficiencies was evaluated in Senegalese sole larvae. Two isonitrogenous and isoenergetic microdiets were formulated and processed to have approximately the same ingredients and proximate composition. The control diet (CTRL) was based on protein sources commonly used in the aquafeed industry. In the supplemented diet (SUP) 8% of an encapsulated fish protein hydrolysate was replaced by crystalline-AA in order to increase the dietary IAA levels. The microdiets were delivered from mouth-opening upon a co-feeding regime until 51days after hatching (DAH). The larvae capacity to utilize protein was evaluated using an in vivo method of controlled tube-feeding during relevant stages throughout development: pre-metamorphosis (13DAH); metamorphosis climax (19DAH) and metamorphosis completion (25DAH). Somatic growth was monitored during the whole trial. A possible effect on the regulation of muscle growth was evaluated through muscle cellularity and the expression of related genes (myf5, myod2, myogenin, mrf4, myhc and mstn1) at metamorphosis climax (19DAH) and at a juvenile stage (51DAH). The SUP diet had a negative impact on larvae somatic growth after the metamorphosis, even though it had no effect on the development of Senegalese sole larvae capacity to retain protein. Instead, changes in somatic growth may reflect alterations on muscle growth regulation, since muscle cellularity suggested delayed muscle development in the SUP group at 51DAH. Transcript levels of key genes regulating myogenesis changed between groups, during the metamorphosis climax and at the 51DAH. The group fed the SUP diet had lower dnmt3b mRNA levels compared to the CTRL group. Further studies are needed to ascertain whether this would possibly lead to an overall DNA hypomethylation in skeletal muscle. Statement of relevanceIn farmed fish species, there has been a great effort over the years to provide the best conditions for successful development of embryos and small larvae, as early environmental conditions can strongly affect muscle growth during early stages and influence the subsequent growth potential at later life stages. In spite of increased efforts to understand the regulation of myogenesis by intrinsic factors like genotype and extrinsic factors such as photoperiod and temperature, studies evaluating the impact of nutritional factors on fish larvae muscle development are still very scarce.This work will raise interest to the discussion on whether a nutritional cue during an early developmental stage can impact on the regulation of muscle growth and on further growth potential in a metamorphosing farmed fish species, such as Senegalese sole. Supplementing microdiets with crystalline-AA in order to correct dietary IAA did not improve Senegalese sole larvae somatic growth and led to changes on the regulation of muscle growth associated with changes in expression patterns of muscle growth markers during the trial (secondary MRFs myogenin and mrf4, myhc and mstn1). Dietary IAA affected the expression of a DNA methyltransferase essential for de novo methylation, dnmt3b, suggesting that an epigenetic effect at the transcriptional regulation level may explain differences found in somatic growth as a response to a nutritional cue.This work may contribute to lay down basis for future studies on nutritional programming of muscle growth in fish larvae of important farmed species.

Comparative ileal amino acid digestibility and growth performance in growing pigs fed different level of canola meal.

The digestibility of different vegetable protein sources were investigated and the effects of supplementing canola meal (CM) as partial inclusions were studied in growing pigs, to determine the performance parameters and its economic importance. In Exp. 1, four pigs (average initial BW = 15.4 ± 0.35 kg, 5 weeks of age) fitted with simple T-cannula at terminal ileum, were fed four diets following repeated 4 × 4 Latin square design having adoption period of 7 days. Diet 1 was Nitrogen free diet containing corn starch. Diets 2, 3, and 4 were the basal diet supplemented with soybean meal (SBM), rapeseed meal (RSM), and domestic CM respectively. The AID of crude protein was decrease in RSM in comparison to SBM supplementation. The AID of Dietary indispensable amino acids (DIAA) such as Lys, Meth, Pha, and dispensable amino acid Ala, Pro, Asp were decreased (P < 0.05) in RSM supplemented diets. The SID of DIAA does not differ but the SID of Asp was higher (P < 0.05) in RSM and CM diets while SID of Pro was lower (P < 0.05) in RSM in comparison to SBM supplemented diets. In Exp. 2, 192 growing pigs (average initial BW 24.76 ± 2.55 kg) were randomly allotted to four dietary treatments with increasing levels of CM i.e. 0, 3.75, 7.50, and 11.25 % respectively. Diets were fed in meal form for 35 days. Increasing CM levels in diets had no effects (P > 0.05) on growth performance and apparent total tract digestibility (ATTD) of nutrients and energy. Total weight gain, total feed intake, and feed cost per kg weight gain were not affected by increasing levels of CM in diets but total feed cost (TFC) per pigs was linearly reduced (26.463 to 25.674; P < 0.05). Broadly, the AID, and SID of amino acid was reduced in RSM but was not effected in CM in comparison to SBM supplemented pigs. Moreover, increasing levels of CM in pigs diet had no effect on the ATTD and performance but TFC per pig was reduced. Thus CM inclusion of up to 11.25 % in diets can be used for reducing the production cost in growing pigs without any negative effect.

Current issues in determining dietary protein and amino-acid requirements

Pregnancy and the first two years of life are periods of rapid growth and yet the knowledge of requirements for protein and dietary indispensable amino acids is very limited. The development of carbon oxidation methods opens the way to studies that should fill these important gaps in knowledge.

No protein intake compensation for insufficient indispensable amino acid intake with a low-protein diet for 12days.

BackgroundProtein quality evaluation aims to determine the capacity of food sources and diets to meet protein and indispensable amino acid (IAA) requirements. This study determined whether nitrogen balance was affected and whether dietary IAA were adequately obtained from the ad libitum consumption of diets at three levels of protein from different primary sources for 12 days.MethodsTwo 12-day randomized crossover design trials were conducted in healthy subjects [n = 70/67 (M/F); age: 19-70 y; BMI: 18.2-38.7 kg/m2]. The relative dietary protein content was lower than [5% of energy (En%)], similar to (15En%), and higher than (30En%) customary diets. These diets had a limited variety of protein sources, containing wheat protein as a single protein source (5En%-protein diet) or 5En% from wheat protein with 10En% (15En%-protein diets) or 25En% (30En%-protein diets) added from whey with α-lactalbumin, soy or beef protein.ResultsThere was a dose-dependent increase in nitrogen excretion with increasing dietary protein content, irrespective of the protein sources (P = 0.001). Nitrogen balance was maintained on the 5En%-protein diet, and was positive on the 15En%- and 30En%-protein diets (P < 0.001) over 12 days. Protein intake from the 5En%-protein diet did not reach the amount necessary to meet the calculated minimal IAA requirements, but IAA were sufficiently obtained from the 15En%- and 30En%-protein diets. In the 15En%- and 30En%-protein conditions, a higher protein intake from the soy-containing diets than from the whey with α-lactalbumin or beef containing diets was needed to meet the minimal IAA requirements.ConclusionProtein intake did not compensate for an insufficient indispensable amino acid intake with a low-protein diet for 12 days.Trial registrationThese trials were registered at clinicaltrials.gov as NCT01320189 and NCT01646749.

Criteria and markers for protein quality assessment – a review

Dietary proteins are found in animal products, plant products and single-cell organisms. Proteins are present in variable proportions in these different food sources and the different proteins also differ in their amino acid composition, dietary indispensable amino acid content and physico-chemical properties. Different criteria can be used to define dietary protein requirements and different markers can be used to assess nutritional protein quality according to the criteria used for protein requirement estimation. The current approach to determining protein requirements is related to nitrogen balance and the dietary indispensable amino acid score approach relates protein quality to the capacity of protein to allow reaching nitrogen balance by providing nitrogen and indispensable amino acids. A second approach considers more directly protein nitrogen utilization by the body and includes measurement of protein digestibility and of the efficiency of dietary nitrogen retention at maintenance or for protein deposition at the whole body level or in more specific body areas. Another approach is related to protein turnover and protein synthesis in relation to maintenance and/or efficiency for deposition or development (growth) at the whole body level or for different and more specific target tissues such as muscle or bone. Lastly, protein quality can also be evaluated from different markers used as risk factors for metabolic dysfunction and disorders related for instance to insulin resistance, diabetes and obesity or cardio-vascular disease. The accuracy and relevance of these different approaches is discussed regarding the capacity of the different protein sources (i.e. animal as meat, milk or eggs, legume as soya or pea, or cereal as wheat or rice) to satisfy protein requirements according to these different criteria and markers.

Amino acid requirements in children and the elderly population

The factorial approach is used to measure the dietary indispensable amino acid (IAA) requirements in children, although recent measurements based on the indicator amino acid oxidation (IAAO) method have begun to generate more direct evidence. Difficulties with the factorial method are that it depends on accurate estimates of the maintenance protein requirement, as well as of protein deposition during growth. Also, a value for the efficiency of utilizing dietary protein for deposition has to be selected, based on published Nitrogen (N) balance studies. In the recent 2007 WHO/FAO/UNU report, the amino acid requirement pattern for infants was taken to be similar to the amino acid composition of breast milk. For pre-school and older children, the factorial method gave values for the scoring pattern of protein that were fairly close to the earlier 1985 WHO/FAO/UNU report for children, since growth progressively became a smaller component of the factorial calculation as age progressed. However, given that there are several assumptions in the derivation of factorial estimates, direct experimental measurements of the amino acid requirement are desirable. The IAAO method, as it is non-invasive, as made it possible to measure the IAA requirements in children. Over the last decade, some of the IAA requirements have been determined by using the IAAO method in healthy school age children; however, the data on IAA requirements in developing country populations are still being conducted. In the elderly, there are not enough data to make a separate recommendation for IAA requirements from that of adults.