Dietary Protein Reduction Research Articles

Low protein diet influences mineral absorption and utilization in medium-growing yellow-feathered broilers from 1 to 30 days of age

Reduced-protein diet can save protein ingredients and reduce nitrogen (N) losses. However, the effect of low protein diet on the mineral uptake and utilization in broilers needs to be explored. The aim of this study was to investigate the effect of low-protein diet on the growth performance, N deposition, mineral contents in serum, tissues and excreta, and the activities and gene expression of related enzymes in tissues of medium-growing yellow-feathered broilers, so as to elucidate the relationship between dietary protein level and the absorption and utilization of minerals in broilers. A total of 72 1-d-old Spotted-Brown male broilers were randomly allotted to 1 of 2 treatments with 6 replicate cages of 6 birds per cage for each treatment. The dietary crude protein (CP) levels for the two treatments were 21 % (the control treatment) and 19 % (low protein treatment), respectively. The experimental period was 30 d. The results showed that no differences (P > 0.05) were detected in average daily gain, average daily feed intake and feed: gain ratio of broilers during 1 to 30 d between the two treatments. However, low protein intake increased (P < 0.05) N retention rate, serum P, Cu and Mn, and excreta Cu, Mn and Zn, and decreased (P < 0.05) liver P and excreta P. In addition, birds fed low protein diet had higher (P < 0.05) manganese superoxide dismutase, and total superoxide dismutase activities in liver, and total antioxidant capacity and malondialdehyde content in heart, and lower (P < 0.05) copper-zinc superoxide dismutase (CuZnSOD) and succinate dehydrogenase activities in liver and CuZnSOD mRNA level in heart. In conclusion, the reduction of dietary CP content from 21 % to 19 % improved N retention, the absorption of P, Cu and Mn, as well as the antioxidant ability of liver and heart, and influenced metabolic utilization of P, Cu, Zn, Fe and Mn in medium-growing yellow-feathered broilers from 1 to 30 d of age.

Effect of Dietary Crude Protein Reduction Levels on Performance, Nutrient Digestibility, Nitrogen Utilization, Blood Parameters, Meat Quality, and Welfare Index of Broilers in Welfare-Friendly Environments.

We investigated the effects of feeding different levels of crude protein (CP) on broiler performance, blood characteristics, meat quality, nutrient and nitrogen (N) availability, stress levels, and welfare indicators in a welfare environment. A total of 625 eight-day-old male broilers were assigned to five dietary CP levels (control (CON) and 1-4% CP reductions) during the grower (8-21 days) and finisher (22-35 days) periods. The CON diet contained 20% CP in the grower phase and 19% in the finisher phase, while all birds received a 22% CP diet during the starter period (1-7 days). Intriguingly, reducing CP levels in the diet did not significantly affect growth performance. These results may be due to the adequate supplementation of essential amino acids in the diets of all treatments and the compensatory growth observed in the finisher phase. Low-CP diets reduced CP digestibility, N intake, and N excretion, while N retention efficiency improved (p < 0.001). Serum total protein (TP) levels decreased equally with decreasing CP in the diet, and the crude fat content in meat increased linearly (p < 0.01). The litter moisture content and pH decreased with lower CP levels, and the welfare indices (footpad dermatitis (FPD), serum corticosterone, etc.) improved (p < 0.01). In conclusion, an appropriate reduction in CP levels in their diet can enhance broiler welfare by reducing N excretion and improving litter quality without adversely affecting performance.

Nutrient utilisation and growth performance of broiler chickens fed standard or moderately reduced dietary protein diets with and without β-mannanase supplementation

Use of reduced protein diets in broiler chicken production provides potential benefits for performance and environmental footprint of production. The effectiveness of β-mannanase supplementation in wheat and soy based standard protein (SP) and reduced protein (RP) diets was tested for growth performance, nutrient utilisation and selected intestinal gene expression of broiler chickens. In a 2×2 factorial arrangement of treatments, two main factors included dietary protein (standard and reduced protein) and β-mannanase supplementation (with or without). All diets contained phytase and carbohydrases (xylanase and glucanase). A total of 480 Ross 308 male off-sex day-old chickens were assigned to the four experimental diets in a 35-d study. Each diet was replicated 12 times with 10 birds per replicate. Using an additional 160 birds, separate apparent metabolizable energy (AME) and nutrient digestibility assays were undertaken for the 4 experimental diets from d 21 to 24 of age. Selected gene involved in gut integrity, inflammation and immune response were quantified using quantitative PCR assays. There was no interaction between β-mannanase and dietary protein for any of the studied parameters except ileal viscosity. Enzyme had no effect on feed intake but tended to increase body weight gain (BWG) from d 0 to 35 of age (P=0.079). Birds fed RP diet consumed more feed when assessed from d 0 to 35 of age (P=0.029). At the same time, β-mannanase tended to reduce feed conversion ratio independent of dietary protein (P=0.069). β-mannanase reduced ileal viscosity of the birds fed RP diet (P<0.001). Reducing dietary protein increased nitrogen retention, nitrogen digestibility coefficient and digestibility coefficients of 11 amino acids (P<0.001). β-mannanase significantly improved digestibility coefficients of nitrogen and Arg, Gly, Thr, Lys, and Ile (P<0.05). Dietary treatments had no effect on AME or gene expression of selected tight junction proteins, interleukin-10, interleukin-1β, mucin-2 and nuclear factor-kappa B. In conclusion, supplementation of β-mannanase tended to improve feed efficiency and increased nutrient digestibility of broilers fed wheat-based diets independent of a moderate reduction in dietary protein. Complementary mode of actions of β-mannanase for intestinal health requires further investigation.

Dietary Crude Protein Reduction with Addition of Crystalline Amino Acids in Growing Pekin Ducks Housed in Cascading Cages: Influence on Growth Performance, Carcass Traits, and Apparent Nutrient Digestibility

The intensive raising model for meat duck production has widely adopted cascading cages. However, investigations on low-protein diets for Pekin ducks under this model are scarce. Thus, a 3 × 6 factorial experiment was conducted using six dietary crude protein (CP) levels (17.61%, 16.25%, 15.34%, 14.55%, 13.38%, and 12.64%, analyzed) and three cage layers (lower, middle, and upper). The study aimed to examine the effects of dietary CP reduction with crystalline amino acid supplementation on growth performance, carcass traits, and apparent nutrient digestibility in growing Pekin ducks from 21 to 42 days of age housed at different cage layers. A total of 1620 21-day-old Pekin ducks were randomly divided into 18 groups, each with six replicates of 15 ducks per replicate. Ducks were distributed across different cage layers and were fed one of six experimental diets, respectively, each varying in dietary CP levels but maintaining similar dietary energy and amino acid profiles. The results showed that the reduction in dietary CP levels affected growth performance (p &lt; 0.05), while weight gain, feed intake, and feed/gain of Pekin ducks had no difference among 17.61%, 16.25%, and 15.34% CP groups (p &gt; 0.05). As dietary CP decreased, abdominal fat yield increased (p &lt; 0.05), but breast and leg meat yields were unaffected (p &gt; 0.05). CP intake, nitrogen intake, and nitrogen excretion decreased linearly (p &lt; 0.05) with decreasing dietary CP levels, while apparent nutrient digestibility in Pekin ducks increased proportionally (p &lt; 0.05). Additionally, body weight, weight gain, feed intake, CP intake, nitrogen intake, and nitrogen excretion of Pekin ducks decreased (p &lt; 0.05) due to different cage layers, while carcass characteristics remained unaffected by cage layer (p &gt; 0.05). In summary, it is feasible to reduce the dietary CP level with crystalline amino acid supplementation in growing Pekin ducks from 21 to 42 days of age housed in cascading cages, while the cage layers affected growth performance of ducks. Based on broken-line models, it was determined that the optimal dietary CP requirement for achieving maximum weight gain and feed efficiency in growing Pekin ducks is approximately 15%.

Effects of methionine supplementation in a reduced protein diet on growth performance, oxidative status, intestinal health, oocyst shedding, and methionine and folate metabolism in broilers under Eimeria challenge

BackgroundThis study investigated effects of different methionine (Met) supplementation levels in a reduced protein diet on growth performance, intestinal health, and different physiological parameters in broilers under Eimeria challenge. A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E. maxima, E. tenella, and E. acervulina, and randomly allocated in a 2 × 5 factorial arrangement. Birds received normal protein diets (20% crude protein, NCP) or reduced protein diets (17% crude protein, LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met.ResultsOn 6 and 9 days post inoculation (DPI), increasing Met level linearly improved the growth performance (P < 0.05). Total oocyst shedding linearly increased as Met level increased (P < 0.05). Duodenal villus height (VH):crypt depth (CD) in the LCP groups were higher on 6 DPI (P < 0.01) while lower on 9 DPI (P < 0.05) compared to the NCP groups. Jejunal CD and duodenal VH:CD changed quadratically as Met level increased (P < 0.05). On 6 DPI, liver glutathione (GSH) and glutathione disulfide (GSSG) linearly increased as Met level increased (P < 0.05). On 9 DPI, GSSG quadratically increased, whereas GSH:GSSG quadratically decreased as Met levels increased (P < 0.05). The expression of amino acid transporters linearly decreased as Met level increased (P < 0.05). The expression of zonula occludens 2 and claudin-1 linearly increased on 6 DPI whereas decreased on 9 DPI as Met level increased (P < 0.05). The expressions of cytokines were lower in the LCP groups than the NCP groups (P < 0.05). Interaction effects were found for the expression of IL-10 and TNFα on 6 DPI (P < 0.05), where it only changed quadratically in the NCP group as Met level increased. The expression of Met and folate metabolism genes were lower in the LCP groups than the NCP groups on 9 DPI (P < 0.05). The expression of these genes linearly or quadratically decreased as Met level increased (P < 0.05).ConclusionThese results revealed the regulatory roles of Met in different physiological parameters including oxidative status, intestinal health, and nutrient metabolism in birds fed reduced protein diet and challenged with Eimeria.

Effect of dietary protein reduction on growth performance and water quality of the blue streak hap Labidochromis caeruleus (Fryer, 1956) reared in a biofloc system

The 8-week study evaluated the effect of dietary protein reduction on growth performance and water quality of blue streak hap (Labidochromis caeruleus) using biofloc technology (BFT). Fish averaging 0.67 ± 0.13 g in weight were distributed into 15 polyester tanks, each with a water volume of 60 L within 100 L tanks, accommodating 15 fish per tank. Four isolipidic and isoenergetic diets were formulated with gradually decreasing protein levels (40%, 35%, 30%, and 25%). Four biofloc groups (40P + BF, 35P + BF, 30P + BF, and 25P + BF) and control (C) (40P + without BF) were fed twice daily (09:00 and 17:00) at a rate of 5% of their body weight. Molasses was added to the experimental tanks on a daily basis as an organic carbon source. This ensured that the biofloc tanks had a balanced carbon/nitrogen (C/N) ratio and facilitated control of total ammonia nitrogen (TAN), nitrite-nitrogen (NO2-N), and nitrate-nitrogen (NO3-N) levels. The nutritional composition of the bioflocs obtained from the experimental groups revealed that the crude protein and crude lipid contents were 37.00–38.14% and 1.45–1.52%, respectively (P > 0.05). The best specific growth rate (SGR) (1.68%) and feed conversation ratio (FCR) (2.21) were determined in the 35P group. Based on the overall evaluation of the study’s data, it can be concluded that the dietary protein for the blue streak hap (L. caeruleus) can be reduced from 40 to 35% in a biofloc system without any negative effects on health. Improving water quality and providing additional food to fish through zero water exchange and organic carbon addition (BFT) can be considered a sustainable aquaculture technique that can be used in ornamental fish farming.

Effects of Methionine Supplementation Levels in Normal or Reduced Protein Diets on the Body Composition and Femur Bone Characteristics of Broilers Challenged with Coccidia.

This study investigated the effects of dietary methionine (Met) levels on the bone quality of broilers challenged with coccidia. A total of 600 fourteen-day-old male Cobb500 broilers were gavaged with mixed Eimeria spp. and randomly allocated into 10 treatment groups by a 2 × 5 factorial arrangement. Birds received normal protein diets (NCP) or reduced-protein diets (LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. Data were analyzed via two-way ANOVA and orthogonal polynomial contrast. At 9 days post-inoculation (DPI), whole body bone mineral density (BMD) and bone mineral content (BMC) linearly decreased as Met levels increased (p < 0.05). For the femoral metaphysis bone quality at 9 DPI, BMD linearly decreased, and porosity linearly increased as Met levels increased (p < 0.05) in the cortical bone. The increased Met levels linearly improved trabecular bone quality in LCP groups (p < 0.05) while not in NCP groups. For the femoral diaphysis cortical bone at 6 DPI, LCP groups had higher BMD and BMC than NCP groups (p < 0.05). Bone volume linearly increased as Met levels increased in LCP groups (p < 0.05) while not in NCP groups. In summary, the results suggested that increased Met levels decreased the cortical bone quality. However, in the context of reduced-protein diets, the increased Met levels improved trabecular bone quality.

Ammonium phosphate is a potential nitrogen supplement in nitrogen-limiting diets for growing pigs

Reduced protein diets may be limiting in dietary nitrogen (N). The objective of this study was to determine the impact of non-protein nitrogen (NPN) inclusion and lysine content on growth performance of growing pigs. Pen-housed growing pigs were randomly assigned to one of six dietary treatments to test the impact of NPN inclusion and dietary lysine content for 28 days. NPN inclusion increased feed efficiency and lean depth ( P &lt; 0.05). Final body weight increased with greater lysine ( P &lt; 0.05) but was not impacted by NPN ( P &gt; 0.05). Ammonium phosphate may be an appropriate N source in N-limiting diets for pigs.

Effects of Amino Acid Supplementation to a Low-Protein Diet on the Growth Performance and Protein Metabolism-related Factors in Broiler Chicks.

A low-protein (LP) diet may alleviate the environmental impact of chicken meat production by reducing nitrogen excretion and ammonia emissions. Thus, this study investigated the effect of a 15% reduced protein diet with or without amino acid (AA) supplementation on the growth performance of broiler chicks from 10 to 35 days of age and the underlying mechanism for loss of skeletal muscle mass. Thirty-six male broiler chicks were allocated to three experimental groups based on body weight: control, LP, and essential AA-supplemented LP (LP+AA). The body weight gain, feed conversion ratio, and weight of breast muscles and legs significantly decreased only in the LP group at the end of the feeding period. Plasma uric acid levels were significantly lower in the LP+AA group than those of the other groups. In the LP group, mRNA levels of microtubule-associated protein 1 light chain 3 isoform B were significantly higher in the pectoralis major, whereas those of atrogin-1, muscle RING-finger protein-1, and myoblast determination protein 1 were significantly higher in the biceps femoris compared to those in the control group. There were no significant differences in insulin-like growth factor 1 mRNA levels in the liver or skeletal muscle between groups. These findings suggested that supplementation with essential AAs ameliorated the impaired effects of an LP diet on growth performance in broiler chicks, and that the transcriptional changes in proteolytic genes in skeletal muscles might be related to the impaired effects of the LP diet.

The Order of Limiting Amino Acids in a Wheat–Sorghum-Based Reduced-Protein Diet for Laying Hens

Understanding the order of limiting amino acids (AA) in reduced-protein (RP) diets for laying hens will facilitate precise feed formulation and ensure that AA requirements are met cost-effectively. The order of the first three limiting AAs—lysine (Lys), methionine (Met), and threonine (Thr)—has been well established in RP laying hen diets. This study aimed to determine the priority order of eight additional limiting AAs (critically important AAs) when formulating wheat–sorghum-based RP diets for laying hens: tryptophan (Trp), valine (Val), isoleucine (Ile), arginine (Arg), leucine (Leu), histidine (His), phenylalanine (Phe), and glycineequivalent (Gly). A total of 330 Hy-Line Brown laying hens were randomly assigned to 11 dietary treatments (30 replicates of individual birds per treatment) from 20 to 39 weeks of age (WOA). Treatments were a standard-protein (17.24% CP) diet as the control (SP); a reduced-protein (15.00% CP) diet with sufficient levels of Lys, Met, and Thr but insufficient levels of the eight experimental essential AA (RP); a reduced-protein diet with sufficient levels of all essential AAs (RP-EAA); and eight subsequent dietary treatments of the RP-EAA diet with one of the experimental essential AAs removed: Trp (RP-EAA-Trp), Val (RP-EAA-Val), Ile (RP-EAA-Ile), Arg (RP-EAA-Arg), Leu (RP-EAA-Leu), His (RP-EAA-His), Phe (RP-EAA-Phe), and Gly (RP-EAA-Gly). Eggs were collected and weighed daily, and feed intake and feed conversion ratio (FCR) were calculated weekly. External and internal egg quality was measured at 29 and 39 WOA. Nutrient digestibility, serum uric acid concentration, caecal microbiota composition, and tibia parameters were measured at 40 WOA. Overall, hens fed the RP-EAA-Val, RP-EAA-Ile, and RP diets presented significantly lower egg mass compared to hens fed the SP, RP-EAA-His, and RP-EAA-Gly diets (p &lt; 0.001). Hens fed the RP diet and RP-EAA-Val diet had a higher FCR compared to those offered the RP-EAA, RP-EAA-Leu, RP-EAA-Phe, and RP-EAA-Gly diets (p = 0.046). Lower protein intake and excretion were observed in hens offered the RP diets compared to hens fed the SP diet (p = 0.001 and 0.018, respectively). Based on the egg mass, Ile may be considered the fourth and Val the fifth limiting AA, after Lys, Met, and Thr, in laying hens fed wheat–sorghum-based RP diets during peak lay. However, if ranked based on FCR, Val may be considered the fourth limiting AA, followed by Trp, Ile, Arg, and His as the co-fifth limiting AAs. Leu, Phe, and Gly may be considered as non-essential AAs for laying hens fed RP diets.

Growth, not digestibility, in chickens receiving reduced-protein diets is independent of non-specific amino-nitrogen sources when the essential-to-total-nitrogen ratio is constant and lower than 50%

ABSTRACT 1. A 21 d experiment was conducted to investigate whether growth performance and coefficients of amino acids digestibility (cAID) in broilers receiving reduced-protein diets supplemented with different non-essential amino acids (NEAA) were dependent on supplemented NEAA in diets with the same essential-to-total N (eN-to-tN) ratio kept at <50%. 2. The experiment used 240 male broiler chicks, allocated to eight treatments with six replicate pens per treatment, and five chicks per replicate. The diets were either adequate in protein diet (PC), reduced protein (NC) diet or the NC diet supplemented with Gly, Gln, Ser, Ala, Gly + Ser or Ala + Ser. Digesta from the distal half of the ileum were collected on d 21. Tissue samples were collected for analysis for gene expression of protein synthesis and degradation (pectoralis major and liver) and peptide and AA transporters (jejunum). 3. The treatments had no effects on growth performance. Generally, cAID was greater (P < 0.05) in NC compared to the PC diet. Individual supplementation of the NC diet with Gly, Gln, Ser, Ala or Ala+Ser increased (P < 0.01) cAID of Cys compared to the PC diet. There were no treatment effects on mRNA levels for the AA or peptide transporters in the jejunum. Supplementation of the NC diet with Gln, Ser, Ala, or Gly + Ser produced an upward expression (P < 0.05) of S6 kinase in the liver compared to PC and NC. In addition, there was greater (P < 0.05) expression of TRIM36 in the pectoralis major of broiler chickens receiving the NC diet supplemented with Gly. 4. When reduced-protein diets have an eN-to-tN ratio of <50% and the ratio is kept constant in all the diets, growth performance response was independent of the source of non-specific amino-N, but the treatments may influence ileal digestibility of individual AA.

Beta-hydroxy-β-methyl butyrate-supplemented diet for broiler chickens is more conducive to dietary protein reduction than a leucine-supplemented diet until 21 days old.

Dietary l-leucine or its metabolite 𝛽-hydroxy-𝛽-methylbutyrate (HMB) has a crucial role in the muscle protein metabolism of broilers during the first few-week growing period. The present study aimed to evaluate the effects of l-leucine (LLPD) or HMB (HLPD) supplementation in a low-protein diet (20%, LPD) until 21 days old on performance, carcass weight, muscle yield and meat quality, as well as intestinal morphometry, in broiler chickens. From days 1-42, LPD decreased body weight gain and feed intake (FI) and increased feed conversion ratio compared to a standard protein diet (22%, SPD). The LLPD and HLPD did not affect FI, but the LLPD decreased the body weight gain and increased the feed conversion ratio compared to the SPD. The LPD group had lower body and muscle weights than other groups. Compared to LPD, HLPD increased dressing percentage. The LPD decreased the serum insulin-like growth factor-1 content compared to the SPD and LLPD. The duodenal villus height of the LPD and LLPD broilers was smaller than those of the SPD and HLPD birds. The HLPD broilers had lower duodenal villus width than the SPD birds. The duodenal crypt depth and ileal mucosal thickness were higher in the HLPD group than in other groups. The HLPD and LLPD enhanced the ileal villus height compared to the SPD. The LLPD and HLPD treatments did not affect meat quality traits compared to the SPD treatment. Dietary HMB could be a conducive approach to reducing dietary protein for broilers until 21 days old. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effects of reduced-protein diets with protease supplementation on growth, carcass yield, intestinal morphology, organ development, nutrient digestibility, and blood biochemical of broiler chickens.

This study was conducted to evaluate growth performance, carcass yield, intestinal morphology, organ development, nutrient digestibility, and blood biochemical parameters of broiler fed 1% reduced-protein diets with/without protease supplementation. A total of 1,120 one-day-old male broiler chickens with average initial body weight (BW), 46.45 ± 0.49 g, were divided into five groups with seven replications and 32 birds per replication. The treatment varied according to the protein and protease enzyme levels: positive control (PC), negative control (NC, PC with reduction of 1% protein), PC supplemented with 50 g/t protease (PC + 50), NC supplemented with 50 g/t protease (NC + 50), and NC supplemented with 100 g/t protease (NC + 100). The results showed that there was no significant effect of 1% reduced-protein diets, with or without protease on feed intake, final BW, average daily gain, feed conversion ratio, and nutrient digestibility. The changes in dietary protein level and supplementation of protease did not affect carcass yield, but significantly affected abdominal fat content, PC + 50 group had significantly lower abdominal fat content than NC-based diet including NC, NC + 50, NC + 100. Reduced-protein with protease supplementation strongly affected organ weight, especially on day 21: the pancreas was heavier in PC and NC + 50 group than other groups, spleen was heaver in NC group than in NC + 100 group, thymus was heavier in NC + 50 group than in PC, NC and NC + 100 group, small intestine was heavier in NC + 50 and NC + 100 group than in PC group, and large intestine was also heavier in NC + 50 group than in NC group. Villus height sampled at 35-d was significantly increased with protease supplement, and which was significantly higher in NC + 100 group than NC group. Regarding on blood metabolites, only urea and uric acid were affected by the reduction of dietary protein, broiler fed PC diet had higher urea and uric acid content than fed NC diet. In conclusion, supplementation of 50 g/t protease in 1% reduced-protein diets does not negatively affect on growth, nutrient digestibility, carcass yield, organ development, and blood metabolites. Moreover, supplementation of protease in low-protein diet could effectively promote organ development and benefit intestine morphology.

Effects of L-arginine, guanidinoacetic acid and L-citrulline supplementation in reduced-protein diets on bone morphology and mineralization of laying hens.

The alterations in feed ingredients and the nutrient matrix to produce reduced-protein diets may affect bone morphology and mineralization in laying hens. This study was implemented to determine the effects of L-arginine (Arg), guanidinoacetic acid (GAA), and L-citrulline (Cit) supplementation to Arg-deficient reduced-protein diets on bone morphology, strength, and mineralization status of laying hens. Individually housed Hy-Line Brown laying hens were evenly distributed to five dietary treatments with 25 replicates per treatment from 20 to 40wk of age. Treatments consisted of a standard protein diet (17% crude protein, SP), a reduced-protein diet deficient in Arg (13% crude protein, RP), and RP supplemented with Arg (0.35% Arg, RP-Arg), GAA (0.46% GAA equivalent to 0.35% Arg, RP-GAA), or Cit (0.35% Cit equivalent to 0.35% Arg, RP-Cit) to reach the Arg level of SP diets. Birds fed the SP diet had similar bone weight, ash, length, width, Seedor index, breaking strength, and serum mineral concentration, but higher toe B level (P<0.001) compared to those fed the RP diet at wk 40. Birds fed the SP diet consumed more but also excreted more K and B compared to those fed the RP diet (P<0.01). Birds fed the SP diet had lower Cu digestibility (P=0.01) and higher B retention (P<0.01) compared to those offered the RP diet. Supplementation of Arg, GAA, and Cit to the RP diet increased relative femur weight and length (P<0.001). Citrulline supplementation also increased relative tibia and femur ash, and Zn digestibility (P<0.05). Supplementation of GAA to the RP diet decreased serum Ca, P, and Mg levels, decreased tibia Fe and Mg levels and toe Mg level, but increased Al, Fe, Zn, and Mn digestibility (P<0.05). The current findings demonstrated the capacity of laying hens to adapt to low mineral intake by increasing mineral utilization. Overall, bone morphology and breaking strength, and serum mineral level in laying hens were not influenced by dietary CP levels. Dietary Arg, GAA, or Cit supplementation were effective in improving bone morphology and mineralization in laying hens fed Arg-deficient RP diets.

Social interactions, precursors of damaging behaviours, object manipulation, straw rooting, and activity: a detailed data set in undocked pigs under protein restriction

Damaging behaviours, such as tail biting, are common problems in pig production, compromising animal welfare and causing economic losses. Detailed studies are impeded by laborious direct observations. Tail biting is a broader phenomenon that begins long before lesions manifest, and behavioural problems caused by various stressors present themselves weeks before they escalate to damaging behaviour. Therefore, we collected detailed data on behaviours considered precursors to tail biting, such as oral and nasal manipulation of conspecifics. Dietary protein reduction is a promising way to reduce nitrogen emissions in pig manure, but its implications for animal welfare are not yet clear. The CP content in the diet was reduced to 80% of the recommendations. Pigs differ in their ability to utilise dietary proteins; therefore, there might be individual differences in how they cope with the protein reduction. Here, we present detailed data of focal observations of 95 pigs at an experimental farm with undocked tails. Pigs were observed directly by the same person for five minutes on four different days. All actions directed towards objects in the pen, interactions with and confrontations among pen mates, straw rooting behaviour and general activity were recorded. After the behavioural observations, wounds on different body parts and the cleanliness were noted by the same person observing the pigs. The protein efficiency of 94 pigs was obtained. The data set comprises six tables. The first table contains information on the animals, including the identities of their parents, farrowing group, sex, and protein efficiency. The other data tables contain four 5-min observations of each pig on 10 object-manipulation behaviours; 150 interaction behaviours, including reactions; 14 confrontation behaviours and their outcomes and reactions; 10 mounting behaviours, including reactions; two rooting behaviours; seven basic behaviours; and an index of general activity. The observations took place under comparatively good housing conditions. Pigs were given fresh straw daily, ad libitum access to feed, floor space above the legal requirements, and daily cleaning of pens, and they were closely monitored for signs of damaging behaviour; all of these are favourable conditions as they limit stress and the risk of damaging behaviour. These data can be used to further explore the relationships of specific behaviours and phenomena and their association with protein efficiency. The ethogram can be used as a template for further observations. Practitioners could use the data to support pigs’ need for occupation, such as by providing sufficient straw.

Negative consequences of reduced protein diets supplemented with synthetic amino acids for performance, intestinal barrier function, and caecal microbiota composition of broiler chickens

The consequences of feeding broiler chickens with reduced protein (RP) diets for gut health and barrier function are not well understood. This study was performed to elucidate the effect of reducing dietary protein and source of protein on gut health and performance parameters. Four experimental diets included 2 control diets with standard protein levels either containing meat and bone meal (CMBM) or an all-vegetable diet (CVEG), a medium RP diet (17.5% in growers and 16.5% in finisher), and a severe RP diet (15.6% in grower and 14.6% in finisher). Off-sex Ross 308 birds were assigned to each of the 4 diets and performance measurements were taken from d 7 to 42 post-hatch. Each diet was replicated 8 times (10 birds per replicate). A challenge study was conducted on additional 96 broilers (24 birds per diet) from d 13 to 21. Half of the birds in each dietary treatment were challenged by dexamethasone (DEX) to induce a leaky gut. Feeding birds with RP diets decreased weight gain (P < 0.0001) and increased feed conversion ratio (P < 0.0001) from d 7 to 42 compared with control diets. There was no difference between CVEG and CMBM control diets for any parameter. The diet containing 15.6% protein increased (P < 0.05) intestinal permeability independent of the DEX challenge. Gene expression of claudin-3 was downregulated (P < 0.05) in birds fed 15.6% protein. There was a significant interaction between diet and DEX (P < 0.05) and both RP diets (17.5% and 15.6%) downregulated claudin-2 expression in DEX-challenged birds. The overall composition of the caecal microbiota was affected in birds fed 15.6% protein having a significantly lower richness of microbiota in both sham and DEX-injected birds. Proteobacteria was the main phylum driving the differences in birds fed 15.6% protein. At the family level, Bifidobacteriaceae, Unclassified Bifidobacteriales, Enterococcaceae, Enterobacteriaceae, and Lachnospiraceae were the main taxa in birds fed 15.6% protein. Despite supplementation of synthetic amino acids, severe reduction of dietary protein compromised performance and intestinal health parameters in broilers, evidenced by differential mRNA expression of tight junction proteins, higher permeability, and changes in caecal microbiota composition.

Dietary sulfur amino acid restriction to prevent Alzheimer’s Disease

AbstractBackgroundHigh serum homocysteine is a risk factor for Alzheimer’s Disease (AD). It has been reported to influence the accumulation of its characteristic lesions ‐ senile plaques (SP) and neurofibrillary tangles (NFT) ‐ containing amyloid β protein (Abeta) and microtubule‐associated protein Tau (MAPT), respectively. Reduced food or calorie intake or branched‐chain amino acids are reported to benefit AD model mice. Restriction of Methionine (Met), an essential amino acid, contributes to increased longevity without calorie restriction.MethodWe treated APP/PS1 transgenic mice with a 75% Met restricted (MR) diet and compared it to an isocaloric isonitrogenous complete diet (CD) for up to 12 months. We subsequently evaluated amyloid loads by ELISA and other APP metabolites by western blotting as described previously (Barnwell et al., 2014, Teich et al., 2018). We collected fecal matter from the colon, extracted DNA, and evaluated the gut microbiome.ResultOur data show that MR treatment substantially lowered brain soluble Abeta at 3 and 6 months. Abeta levels are excessively and presumably saturated in older mice, accounting for the lack of statistically significant differences between the different diets evaluated after twelve months. Nevertheless, older mice showed substantial improvement in behavior in a novel object recognition task after MR treatment. MR treatment is known to be senolytic, and therefore, we are currently examining related processes, such as preservation of lysosomal capacity changes in metabolome, mRNA, and miRNA profiles. Future work involves evaluating methionine and branch‐chain amino acid restriction in AD animal models, such as in human tau‐overexpressing Tg mice, and evaluating miRNAs.MR diets significantly alter gut microbiota. As expected, the bacteria that metabolize Met and Cys were decreased. Specifically, several major species of bacteria were reduced, and the overall diversity of microbiota was increased. Senolysis and improvements in AD pathology may also be mediated by reduced inflammation due to the increased diversification of the microbiome. Thus, such diets may play an important role in strategies to prevent AD, retinal degeneration, and other age‐associated disorders.ConclusionSulfur amino acid restriction reduces amyloid production and preserves cognitive function in mice. Reduced protein diets may be useful in preventing proteinopathies that lead to neurodegeneration.

A Pilot Controlled Feeding Trial Modifying Protein Intake in Healthy Subjects to Assess Adherence and the Metabolome

Dietary protein has been shown to impact physiology and pathophysiology, including inflammation and cancer, effects believed to occur through host and microbe-mediated mechanisms. However, the majority of studies investigating this concept have been conducted in animal models, with less information on the optimal approach, tolerability and biologic effects of modifying protein intake in humans. The current study presents a longitudinal controlled feeding trial carried out in healthy humans to acutely modulate protein intake using individualized diets. Adherence to study diets was monitored through subject-reported electronic picture-based assessments and global metabolomic analysis was performed on serum and stool, following each diet stage. Subjects exhibited strong adherence to study diets, with macronutrient intake meeting study goals during each stage. Metabolomic analysis revealed shifts in both serum and feces in association with modifying protein intake, including reciprocal changes in the abundance of amino acids and amino-acid related compounds, when comparing high to reduced protein stages. Additional fecal metabolite changes consisted of reduced microbial fermentation products following the reduced protein diet stage. Collectively, this study provides a robust method to precisely modify and monitor protein intake in humans, as well as assess corresponding metabolomic alterations.

The effects of arginine and branched-chain amino acid supplementation to reduced-protein diet on intestinal health, cecal short-chain fatty acid profiles, and immune response in broiler chickens challenged with Eimeria spp.

We investigated the effects of supplementing arginine (Arg) and branched-chain amino acids (BCAA) in broilers fed reduced-protein diets and challenged with Eimeria spp. All birds were fed the same starter diet meeting Cobb 500 nutrient specifications from d 1 to 9. Four grower diets: positive control (PC) with 20.0% crude protein (CP); reduced-protein negative control (NC) with 17.5% CP; or NC supplemented with Arg or BCAA at 50% above recommendations (ARG or BCAA) were fed to the birds from d 9 to 28. Birds were allocated in a 2×4 factorial arrangement (4 diets, each with or without challenge), with 8 replicates per treatment. On d 14, the challenge groups were orally gavaged with mixed Eimeria spp. Intestinal permeability was higher (P < 0.05) in NC than PC, whereas the permeability of ARG and BCAA groups did not differ significantly from PC. On d 28, a significant interaction (P < 0.01) was observed in CD8+: CD4+ ratios in cecal tonsils (CT), Eimeria challenge increased the ratios in all groups except for the ARG group. On d 21, a significant interaction was found for CD4+CD25+ percentages in CT (P < 0.01) that Eimeria challenge increased the percentages only in PC and NC groups. On d 21 and 28, significant interactions (P < 0.01) were found for macrophage nitric oxide (NO) production. In nonchallenged birds, NO was higher in the ARG group than other groups, but in challenged birds, NO was higher in both ARG and BCAA groups. On d 21, a significant interaction was found for bile anticoccidial IgA concentrations (P < 0.05) that Eimeria challenge increased IgA only in NC and ARG groups. The results suggest that a reduced-protein diet exacerbates the impact of the Eimeria challenge on intestinal integrity, but this could be mitigated by Arg and BCAA supplementations. Arginine and BCAA supplementations in reduced-protein diets could be beneficial for broilers against Eimeria infection by enhancing the immune responses. The beneficial effects of Arg supplementation tended to be more pronounced compared to BCAA supplementation.

Dietary protein reduction with stepwise addition of crystalline amino acids and the effect of considering a minimum glycine-serine content in broiler diets

Reducing the dependency on soybean meal (SBM) is necessary to improve the sustainability of the poultry industry. Moreover, the recommendations for minimum contents of dietary Gly+Ser require further research. Two parallel studies were executed to determine the effects of replacing SBM with crystalline amino acids (CAA) to meet the amino acid requirements and to determine whether a minimum content of Gly+Ser is necessary for broiler diets. In study 1, 1,860 one-day-old male chicks were fed a common starter phase diet (22.8% CP). During the grower-1, grower-2, and finisher phases, the control CP content was reduced (up to 2.1%) with the stepwise addition of CAA (treatments 1-5). Within each feeding phase, AME, standardized ileal digestible Lys, and the minimum Met, Thr, Val, Gly+Ser, Ile, Arg, and Trp to Lys ratios were similar. In study 2, 1,488 male chickens were used in a 2×2 factorial design with the Gly+Ser content and feed ingredients used as main factors. Performance was monitored during 41 d in both studies. Reduction in CP content linearly increased (P < 0.05) BW, ADG, and ADFI in the grower-1, grower-2, and finisher phases. When the overall FCR was adjusted considering the BW differences (FCRadj); FCRadj linearly decreased with the weighted average CP (WACP) content (P < 0.001). In the lowest CP treatment, estimated dietary N utilization efficiency was improved by 10%, and the overall N excretion was reduced by 16% compared with the control (P < 0.001). The overall SBM and soybean oil intakes were linearly reduced relative to WACP (-12.0 and -20.2% in control vs. treatment 5, respectively; P < 0.001). Formulating with a minimum Gly+Ser content in the starter phase improved the FCR (P < 0.05) only in the corn-SBM-based diet. In grower-1, increasing Gly+Ser content improved the FCR independent of the feed ingredients used (P < 0.05). Crystalline amino acids can be used to partially replace intact protein reducing the dependency on SBM. Young birds may not synthesize enough Gly endogenously and a minimum content should be provided in the early phases.