N-carbamylglutamate Research Articles

Dietary Supplementation with Rumen-Protected Arginine or N-Carbamylglutamate Enhances Fetal Liver Development in Nutrient-Restricted Pregnant Hu Ewes.

This study was conducted in nutrient-restricted pregnant Hu ewes to determine whether rumen-protected arginine (RP-Arg) or N-carbamylglutamate (NCG) supplementation affects fetal liver growth and development. From 35 d to 110 d of gestation, 32 Hu ewes were randomly divided into four groups: a control group (100% of the National Research Council (NRC) requirements), a nutrient-restricted group (50% of the NRC requirements), and two treatment groups (ARG and NCG, 50% of the NRC requirements, supplemented with 20 g/day RP-Arg or 5 g/day NCG, respectively). Fetal body weights, fetal liver growth performance, the capability of antioxidation, and the expression of the mRNA and proteins of apoptosis-related genes in the fetal liver were determined and analyzed at 110 d of gestation. The dry matter, water, fat, protein, and ash components of the fetal livers in the RG group were found to be lower than in the CG group, and these components were significantly higher in the NCG group than in the RG group (p < 0.05). A decrease in DNA, RNA, and protein concentrations and contents, as well as in protein/DNA ratios, was observed in the RG group in comparison to the CG group (p < 0.05). Compared with the RG group, the NCG group had higher concentrations of DNA, RNA, and protein, as well as higher protein/DNA ratios (p < 0.05). The RG group had lower concentrations of cholinesterase, nitric oxide, nitric oxide synthase, superoxide dismutase, alanine aminotransferase, and total protein than the CG group (p < 0.05). The RG group had higher levels of glutathione peroxidase, maleic dialdehyde, and aspartate aminotransferase than the CG group (p < 0.05). In the RG group, the mRNA and protein expression of p53 and Bax was significantly increased (p < 0.05) compared with the CG group, and the gene expression of FasL and Bcl-2, the ratio of Bcl-2 to Bax, and the protein expression of Bcl-2 in the RG group were lower (p < 0.05) than in the CG group. It appears that RP-Arg and NCG supplementation during pregnancy could influence fetal liver growth and development. A nutrition-based therapeutic intervention to alleviate reduced fetal growth can be developed based on this study, which has demonstrated that maternal undernutrition during pregnancy induces the maldevelopment of the fetal liver.

IMPACTS OF CONCENTRATE FEEDING LEVEL WITH OR WITHOUT N-CARBAMYLGLUTAMATE ON PRODUCTIVE PERFORMANCE OF GOATS

This study was conducted to evaluate the effect of feeding three levels of concentrate 2%, 3%, and 4% with or without additives of N-carbamylglutamate (NCG) on the digestibility, feed efficiency, and daily gain of Shami goats. Twenty-four male goats were used aged 8-10 months with an initial weight 32.687 ± 1.52kg divided into six treatments in a 2×3 factorial experiment/ Completely Randomized Design. Individual feeding was used for 70 days preceded by 14 days as an adaptation period. The results of increasing concentrate showed a non-significant increase in nutrients intake with superiority of NCG treatments and a significant increase (P&lt;0.01) of ether extract intake. Non-significant increase in digestibility with increasing feed intake. Significant increases (P&lt;0.01) in total and daily gain of 3% concentrate with or without NCG and non-significant increases for final weight. The better feed efficiency with 3% concentrate then 2% and 4% respectively. In conclusion: A level of feeding 3% had a positive effect on animal performance and the best for fattening.

N-Carbamylglutamate in ovo feeding improves carcass yield, muscle fiber development, and meat quality in broiler chickens.

Insufficient endogenous nutrients in the broiler embryo can lead to muscle gluconeogenesis, which ultimately affects the post-hatching performance of chicks. This study investigated the effects of in ovo feeding (IOF) of N-carbamylglutamate (NCG) on the growth hormones, carcass yield, and meat quality in broilers. Fertile eggs from a 30-week-old Ross 308 breeder flock were divided into three treatment groups: NC (non-injection), SC (100 μL saline-injection), and NCG (2 mg NCG injection). Each group had six replicates, with 70 eggs per replicate during incubation. Injections were administered on the 17.5th day of embryonic development. After hatching, 270 chicks were selected for 42-day rearing for further sampling. Chicks in the NCG group had significantly higher body weight (BW) and average daily gain (ADG) at the growing phase, increased growth and testosterone hormone in both feeding phases (21 and 42 days), and improved average daily gain (ADG) and food conversion ratio (FCR) in both grower and entire feeding phases (P < 0.05). Triiodothyronine (T3) and tetraiodothyronine (T4) levels, carcass yield, dressing, drum weight, breast muscle weight, drumstick weights, thighs, pectoralis major, and their part percentage of carcass were improved in the NCG group (P < 0.05), these effects were varied along feeding phases. Moreover, IOF of the NCG also improved pectoralis breast muscle color values at 24 h post mortem (P < 0.05). These results suggest that NCG injection at the late embryonic age of broiler enhances growth performance and meat quality throughout the lifespan and this can probably be attributed to an increase in thyroid and testosterone hormones, indicating potential involvement in metabolic and nutrient partitioning pathway regulation. © 2024 Society of Chemical Industry.

The Effect of N-Carbamylglutamate Supplementation during the Last Third of Gestation on the Growth and Development of Fetuses Born to Nutrient-Restricted Twin-Bearing Ewes.

N-carbamylglutamate (NCG) is postulated to improve fetal growth in nutrient-restricted gestations when supplemented from day 35 to 110 of gestation, but the effects of supplementation from 100 days of gestation to birth have not been evaluated. The aim of this study was to evaluate the effect of oral NCG supplementation from 100 days of gestation (dga) to term in naturally nutrient-restricted grazing twin-bearing ewes, on the maternal body weight (BW), body condition score (BCS), placental morphology, fetal body and organ weights and blood biochemistry and antioxidant status in the ewe and fetuses. Eighteen twin-bearing ewes maintained under grazing management were randomly allocated to either a treatment group (NCG; n = 10), orally dosed once daily with 60 mg/kg of NCG from day 100 until 140 dga, or an unsupplemented control group (CON; n = 8). At 140 dga, blood gases, redox status, maternal and fetal plasma and fetal biometrics were obtained after caesarian section. The serum concentration of NCG was increased 15-fold in the NCG ewes compared to the CON. No major effects on dam or fetal body weight nor on blood biochemistry or antioxidant parameters were observed. These results indicate that NCG supplementation in mid-to-late gestation to grazing ewes was unable to rescue the negative production effects of severe natural nutritional restriction on both the dam and fetuses.

Dietary N-carbamylglutamate (NCG) supplementation improves the survival, digestive and immunological parameters by promoting ammonia excretion in Chinese mitten crab (Eriocheir sinensis), under ammonia nitrogen stress

Ammonia nitrogen is one of the most common environmental stress factors in aquaculture. The aim of this experiment was to evaluate the effects of ammonia nitrogen stress on ammonia metabolism, digestion and immune function as well as the protective effects of dietary N-carbamylglutamate (NCG) in the Chinese mitten crab (Eriocheir sinensis). Two feeds were used, namely a control feed (regular commercial feed) and an NCG feed (NCG (302.96 ± 4.07) mg/kg added). Subsequently, animals fed by these two feeds were exposed to 30 mg/L or 60 mg/L ammonia nitrogen stress (named A30 or A60) for 21 days, respectively. The rate of hemolymph ammonia excretion was significantly increased in NCG group compared to C (control) group (P < 0.05). Hemolymph ammonia levels were significantly lower in NCG A30 (NCG feed and A30) and NCG A60 (NCG feed and A60) groups compared to A30 (control feed and A30) and A60 (control feed and A60) groups (P < 0.05). Glutamate dehydrogenase and glutamine synthetase activities in the gills and hepatopancreas of NCG A60 group were significantly increased compared with the A60 group, which promoted glutamine synthesis (P < 0.05). The cumulative mortality rate of Eriocheir sinensis in NCG A30 and NCG A60 groups were effectively reduced (P < 0.05). The arginine levels of hepatopancreas were significantly increased in NCG A60 group (P < 0.05). Under ammonia nitrogen stress, NCG significantly increased the activities of trypsin, pepsin, lipase and α-Amylase in NCG feed group, and promoted feeding (P < 0.05). In hemolymph, hepatopancreas and intestine, NCG significantly increased nitric oxide synthase activity, lysozyme and phenol oxidase activities compared to the ammonia nitrogen stress group (P < 0.05). The results indicated that, NCG added in feed could decelerate the toxic effects of ammonia on Eriocheir sinensis by promoting ammonia excretion and accelerating glutamine and endogenous arginine synthesis by metabolism. Moreover, the activity of digestive enzymes and immune-related enzymes is improved.

Effects of dietary N-carbamylglutamate supplementation on the modulation of microbiota and Th17/Treg balance-related immune signaling after lipopolysaccharide challenge.

This study aimed to evaluate the effects of N-carbamylglutamate (NCG) on piglets' growth performance and immune response, and to unravel the mechanisms of such effects. In a 2 × 2 factorial design including diet (with or without NCG) and immunological challenge (saline or lipopolysaccharide (LPS)), 24 piglets were randomly distributed into four groups. After being fed a basic diet or a NCG-supplemented diet for 21 days, piglets were administered LPS or saline intraperitoneally. The results showed that NCG increased the average daily gain and average daily feed intake, and the feed conversion ratio of piglets, and alleviated the adverse effects of LPS stimulation on intestinal morphology. At the phylum level, NCG reversed the increase in the abundance of Firmicutes and the reduction in that of Actinomycete caused by LPS stimulation. At the genus level, NCG increased the abundance of Lactobacillus, Blautia, norank_Butyricicoccaceae, Subdoligranulum, and Ruminococcus_gauvreauii_group, and LPS decreased the abundance of Escherichia-Shigella and Ruminococcus_gauvreauii_group. The short-chain fatty acid content was increased by NCG, but LPS reduced its content. N-Carbamylglutamate also inhibited significantly the LPS-induced increase in the relative expression of signal transducer and activator of transcription (STAT) 3, related orphan receptor (RAR) c, and pro-inflammatory cytokines, and the decrease in the relative expression of STAT5, forkhead box P3, IL-10 and transforming growth factor beta 1 mRNA. A significant correlation was found between intestinal microbiota and inflammatory cytokines and short-chain fatty acids. N-Carbamylglutamate can improve piglets' growth performance. It can also attenuate LPS-induced intestinal inflammation by modulating microbiota and Th17/Treg balance-related immune signaling pathways. © 2023 Society of Chemical Industry.

PSI-21 The Role of N-Carbamylglutamate Supplementation During Last Third of Gestation in Nutrient Restricted Twin-Bearing Ewes

Abstract Nutrient restriction in sheep during pregnancy normally occurs in Patagonia with negative effects on ewe body condition (BCS) and reproductive performance. N-carbamylglutamate (NCG), an analogue of N-acetylglutamate, is postulated to improve urea recycling, nitrogen utilization and antioxidant capacity in ruminants. The objective of this study was to evaluate the effect of oral NCG supplementation in naturally nutrient restricted grazing twin-bearing ewes, on maternal body weight (BW) and body condition score (BCS), serum biochemical indexes and antioxidant capacity in mid-late gestation. Ten twin-bearing ewes were orally dosed once-daily with 60 mg/kg BW NCG in a water carrier from 100 to 140 days of gestation (dg) and were compared with unsupplemented controls (CON; n = 8). Maternal BW, and BCS were recorded every 10 days, from 100 to 140 dg. Blood samples were collected at the same time points to evaluate plasma metabolites (NEFA, urea, total protein, albumin, globulin, Ca, P, Mg, Cholesterol, ASAT, CK, GGT, GLDH, total bilirubin and triglycerides), antioxidant status [malondialdehyde (MDA)], and total antioxidant capacity (TAC). Ewes were managed as a single mob in a paddock with natural pasture (3.3% CP, 1.9 Mcal/kg ME), supplemented with alfalfa hay (600 g· ewe-1· /day-1; 13% CP and ME 2.2 Mcal/kg ME) from 130 dg, with a stocking rate of 0.9 ewes per hectare and a dry matter availability of 525 kg per hectare, representative of Patagonian prairie conditions. The effect of treatment on ewe BW, BCS and plasma metabolites were analyzed using REML analysis in R using lme4 R package with time (dg) and treatment (and their interaction) as fixed effects and animal as the random effect. Ewe BW increased (P &amp;lt; 0.0001) and BCS decreased (P = 0.005) with advancing gestation, but there was no effect of treatment. There was a tendency for a treatment effect (P = 0.07) on plasma urea with ~10% less concentrations in NCG than CON ewes at 130 dg (P = 0.08) and 140 dg (P = 0.04). There was a treatment by time interaction for total bilirubin (P = 0.005), phosphorus (P = 0.02), and a similar trend for albumin (P = 0.09). Ewes supplemented with NCG had~10% lower concentrations of bilirubin at 100 (P = 0.01) and 120 dg (P = 0.04), less concentrations of phosphorus (P = 0.003) and albumin (P = 0.02) at 140 dg compared with CON ewes. No other effects of treatment on blood metabolites or antioxidant status or capacity were observed. Collectively these results suggest that supplementation of nutritionally restricted ewes with 60 mg/kg BW from 100 to 140 dg did not influence BW or LWT but may have some effects on liver function, protein metabolism, urea recycling, feed intake and/or feed efficiency which warrant further investigation.*Funded by FONDECYT Project N°1221042; AgResearch Strategic Science Investment Fund.

Datamining approaches for examining the low prevalence of N-acetylglutamate synthase deficiency and understanding transcriptional regulation of urea cycle genes.

Ammonia, which is toxic to the brain, is converted into non-toxic urea, through a pathway of six enzymatically catalyzed steps known as the urea cycle. In this pathway, N-acetylglutamate synthase (NAGS, EC 2.3.1.1) catalyzes the formation of N-acetylglutamate (NAG) from glutamate and acetyl coenzyme A. NAGS deficiency (NAGSD) is the rarest of the urea cycle disorders, yet is unique in that ureagenesis can be restored with the drug N-carbamylglutamate (NCG). We investigated whether the rarity of NAGSD could be due to low sequence variation in the NAGS genomic region, high NAGS tolerance for amino acid replacements, and alternative sources of NAG and NCG in the body. We also evaluated whether the small genomic footprint of the NAGS catalytic domain might play a role. The small number of patients diagnosed with NAGSD could result from the absence of specific disease biomarkers and/or short NAGS catalytic domain. We screened for sequence variants in NAGS regulatory regions in patients suspected of having NAGSD and found a novel NAGS regulatory element in the first intron of the NAGS gene. We applied the same datamining approach to identify regulatory elements in the remaining urea cycle genes. In addition to the known promoters and enhancers of each gene, we identified several novel regulatory elements in their upstream regions and first introns. The identification of cis-regulatory elements of urea cycle genes and their associated transcription factors holds promise for uncovering shared mechanisms governing urea cycle gene expression and potentially leading to new treatments for urea cycle disorders.

N-carbamylglutamate supplementation induces functional egg production in layers by modulating liver transcriptome profiles

Eggs rich in polyunsaturated fatty acids (PUFA), known as functional eggs, are animal products deemed beneficial to human health and possess high economic value. The production of functional eggs involves supplementing exogenous additives with the ability to regulate lipid metabolism. As N-Carbamylglutamate (NCG) serves as an endogenous arginine synthesizer, and arginine acts as the substrate for the formation of nitric oxide (NO), the biological function of NCG is partially mediated by NO. NO is a key regulatory molecule in lipid metabolism, suggesting that NCG may also have the ability to modulate lipid metabolism. In order to assess the capacity of NCG in regulating liver lipid metabolism and its potential application in producing functional eggs, we conducted a study to investigate the effects of dietary supplementation of NCG on production performance, serum, and liver NO levels, yolk fatty acid composition, and the liver transcriptome of layers. In this study, we utilized 30 layers of the Jinghong No.1 breed, all aged 45 wk. All the birds were randomly divided into 2 groups. Each group had 5 replicates, and each replicate had 3 birds. We provided them with different diets: one group received the basic diet, and the other group's diet was supplemented with 0.08% NCG. The experiment lasted for 14 wk. The results did not reveal any positive impact of NCG on production performance. However, NCG supplementation elevated NO levels in serum and liver, along with an increase in yolk PUFA, ω-3, and ω-6 fatty acids. Liver transcriptome analysis identified 124 upregulated differentially expressed genes (DEGs) and 43 downregulated DEGs due to NCG supplementation. Functional annotation using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database highlighted 3 upregulated DEGs (CPT1A, MOGAT1, and CHKA) and 2 downregulated DEGs (FASN and ETNPPL) associated with lipid metabolism. Pathway enrichment analysis revealed that CPT1A was enriched in the AMPK signaling pathway and the PPAR signaling pathway, while FASN was enriched in the AMPK signaling pathway. Thus, CPT1A and FASN are potential functional genes related to lipid metabolism facilitated by NCG supplementation. In summary, our study suggests that NCG supplementation modulates liver lipid metabolism, leading to the production of functional eggs in layers.

Effect of N-Carbamylglutamate Supplementation on Growth Performance, Jejunal Morphology, Amino Acid Transporters, and Antioxidant Ability of Weaned Pigs.

Weaning is an important period that affects the performance of piglets. However, the regulation of dietary amino acid levels is considered to be an effective way to alleviate the weaning stress of piglets. N-carbamylglutamate (NCG) plays an important role in improving the growth performance and antioxidant capacity of animals. A total of 36 weaned piglets were randomly assigned to two treatment groups, a control group (CON) and a 500 mg/kg NCG group (NCG), and the experiment lasted for 28 days. The results show that the NCG treatment group showed an increased 0-28 days average weight gain and average daily feed intake, and also increased contents of GLU and HDL, and lower SUN in serum, and an upregulation of the expression of the amino acid transporters SNAT2, EAAC1, SLC3A1, and SLC3A2 mRNA in the jejunum (p < 0.05), as well as an increased villus length and VH:CD ratio, and claudin-1, occludin, and ZO-1 mRNA expression in the jejunum (p < 0.05). The NCG treatment group showed an increased content of GSH-Px in serum and T-AOC and SOD in the jejunum, and a lower content of MDA (p < 0.05); and the upregulation of the mRNA expression related to antioxidant enzymes (CAT, SOD1, Gpx4, GCLC, GCLM and Nrf2, AhR, CYP1A1) in the jejunal mucosa (p < 0.05). In addition, compared with the control group, the NCG treatment group saw an upregulation in the mRNA expression of IL-10 and a decrease in the expression of IL-1β and IL-4 in the jejunal mucosa (p < 0.05). In summary, the results of this study suggest that NCG improved growth performance and jejunal morphology, improved the jejunal transport of amino acids related to the ornithine cycle, and improved the antioxidant capacity in weaned pigs.

N-carbamylglutamate supplementation regulates hindgut microbiota composition and short-chain fatty acid contents in Charollais and Small Tail Han crossbred sheep.

The objective of this study was to investigate the effects of N-carbamylglutamate (NCG) supplementation on the growth performance, hindgut microbiota composition, and short-chain fatty acid (SCFA) contents in Charollais and Small Tail Han crossbred sheep. A total of 16 female crossbred mutton sheep (Charollais × Small Tail Han), aged 4 months, with an initial body weight of 30.03 ± 0.08 kg, were utilized in a 60 days experiment. The sheep were divided into two groups based on their initial body weight. Each group consisted of 8 replicates, with each individual sheep considered as a replicate. The dietary treatments comprised a basal diet supplemented with either 0.00% or 0.12% NCG. Our findings indicate that NCG supplementation did not have a significant effect on the growth performance of mutton sheep. However, it did lead to changes in hindgut SCFA contents. Specifically, NCG supplementation increased the content of propanoic acid while decreasing acetic acid and hexanoic acid in the hindgut. Through microbiota analysis using the 16S rRNA technique, we identified Lachnospiraceae_NK3A20_group and Parasutterella as biomarkers for the hindgut microbiota in mutton sheep fed a diet containing NCG. Further analysis of the microbiota composition revealed that NCG supplementation significantly increased the abundance of Lachnospiraceae_NK3A20_group and Parasutterella, while decreasing unclassified_f_Lachnospiraceae and Lachnoclostridium. Correlation analysis between hindgut SCFA contents and microbiota composition revealed that the abundance of Lachnoclostridium was positively correlated with the contents of acetic acid and hexanoic acid, but negatively correlated with propanoic acid. Additionally, the abundance of Lachnospiraceae_NK3A20_group and Parasutterella was positively correlated with the content of propanoic acid, while being negatively correlated with acetic acid and hexanoic acid. Based on these findings, we conclude that dietary supplementation of 0.12% NCG can modulate hindgut SCFA contents in mutton sheep by regulating the composition of the hindgut microbiota.

Dietary rumen-protected L-arginine or N-carbamylglutamate enhances placental amino acid transport and suppresses angiogenesis and steroid anabolism in underfed pregnant ewes

This study aimed to investigate the effects of dietary supplementation of underfed Hu ewes from d 35 to 110 of gestation with either rumen-protected L-arginine (RP-Arg) or N-carbamylglutamate (NCG) on placental amino acid (AA) transport, angiogenic gene expression, and steroid anabolism. On d 35 of gestation, 32 Hu ewes carrying twin fetuses were randomly divided into four treatment groups, each consisting of eight ewes, and were fed the following diets: A diet providing 100% of NRC's nutrient requirements for pregnant ewes (CON); A diet providing 50% of NRC's nutrient requirements for pregnant ewes (RES); RES diet plus 5 g/d NCG (RES + NCG); or RES diet plus 20 g/d RP-Arg (RES + ARG). On the d 110 of pregnancy, blood samples were taken from the mother, and samples were collected from type A cotyledons (COT; the fetal portions of the placenta). The levels of 17β-estradiol and progesterone in the maternal serum and both the capillary area density (CAD) and capillary surface density (CSD) in type A COT were decreased in response to Arg or NCG supplementation when compared to the RES group. The concentrations of arginine, leucine, putrescine and spermidine in type A COT were higher (P < 0.05) in the RES + ARG or RES + NCG group than in the RES group. The mRNA expression levels of inducible nitric oxide synthase (iNOS) and solute carrier family 15, member 1 (SLC15A1) were increased (P < 0.05) while those of progesterone receptor (PGR) and fibroblast growth factor 2 (FGF2) were decreased in type A COT by supplementation with either NCG or RP-Arg compared to the RES group. The results suggest that providing underfed pregnant ewes from d 35 to 110 of gestation with a diet supplemented with NCG or RP-Arg improves placental AA transport, and reduces the expression of angiogenic growth factor genes and steroid anabolism, leading to better fetal development.

Dietary N-carbamylglutamate supplementation improves ammonia tolerance of juvenile yellow catfish Pelteobagrus fulvidraco.

Introduction: Ammonia has been of concern for its high toxicity to animals. N-carbamylglutamate (NCG) can reduce blood ammonia levels in mammals, but studies on ammonia tolerance in fish are insufficient. Methods: Juvenile yellow catfish were fed two levels of NCG (0.00% and 0.05%) for 84days under three ammonia levels (0.00, 0.08, and 0.16mg/L NH3). Results and Discussion: The results showed that survival rate (SUR), final body weight (FBW), weight gain (WG), and serum total protein (TP), triglycerides (TG), glucose (Glu), ornithine (Orn), citrulline (Cit) contents, and liver superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), arginase (ARG), ornithine transcarbamylase (OTC) activities decreased with the increase of ammonia levels, on the contrary, feed conversion ratio (FCR), hepatosomatic index (HSI), and serum ammonia, urea, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamine (Gln), arginine (Arg) contents, and liver malondialdehyde (MDA), tumor necrosis factor (TNF), interleukin (IL) 1, IL 8 contents, and mRNA expressions of cu/zn sod, cat, gpx, gr, tnf ɑ, il 1, and il 8 were significantly increased. Dietary 0.05% NCG supplementation had higher SUR, FBW, WG, feed intake (FI), whole-body protein, and serum TP, total cholesterol (TC), Glu, citrulline (Cit) contents, and liver SOD, GPx, argininosuccinate synthetase (ASS), argininosuccinate lyase (ASL), inducible nitric oxide synthase (iNOS) activities compared to 0.00% NCG group, but had lower serum ammonia, urea, ALT, AST, Gln, Arg contents, and liver MDA, TNF, IL 1, IL 8 contents, and neuronal nitric oxide synthase activity. At the end of bacterial challenge, cumulative mortality (CM) increased with ammonia levels increased, but serum antibody titer (AT), lysozyme (LYZ) activity, 50% hemolytic complement, immunoglobulin (Ig) contents, respiratory burst (RB), phagocytic indices decreased with ammonia levels increased. CM in 0.05% NCG group was lower than that in 0.00% NCG group, but serum AT, LYZ activity, Ig content, RB in 0.05% NCG group were significantly higher. The correlation analysis found that iNOS was positively correlated with ASS activity. This study indicates that dietary NCG supplementation can improve the ammonia tolerance of yellow catfish, and ASS may also be the target of NCG to activate the urea cycle.

Effect of N- carbamylglutamate and feeding level on amino acids and biochemical parameters in goats

The effect of feed additive N- carbamylglutamate (NCG) with different feeding levels on some biochemical parameters and amino acids in the serum of goats was investigated. Thirty male goats aged 10-11 months were distributed into three levels of concentrate 70%, 50%, and 30% with or without N- carbamylglutamate (NCG) in a 2×3 factorial experiment/ Completely Randomized Design (CRD). After 77 days of individual feeding, jugular blood was sampled before morning feeding. The results showed that additive NCG with high concentrate (70%) led to a decrease (P&lt;0.01) in total blood protein, triglycerides, and globulins, and a decrease (P&lt;0.05) for creatinine, uric acid, and VLDL. Blood magnesium increased (P&lt;0.01) with NCG of low concentrate (30%) treatment, while phosphor increased with NCG of 50% concentrate treatment. Calcium increased with all NCG treatments. The results of adding N- carbamylglutamate with high roughage led to a negative effect on blood amino acids. In conclusion, increasing roughage feeding without NCG is the best for blood homeostasis, and don't use N- carbamylglutamate without non-protein nitrogen additives.&#x0D; Keywords: Amino acids, blood traits, N-carbamylglutamate, feeding level.

Production performance, egg quality, and uterine gene expression for layers as affected by N-Carbamylglutamate supplementation.

Supplementation of exogenous additives is a strategy to improve laying performance of layers by regulating uterine function. N-Carbamylglutamate (NCG) as an activator for endogenous arginine synthesis has the potential to regulate the laying performance of layers, but its effects have not been fully understood. This study investigated the effects of dietary supplementation of NCG on production performance, egg quality, and uterine gene expression in layers. A total of 360 45-week-old layers with a genetic line of Jinghong No. 1 were used in this study. The experimental period was 14 weeks. All birds were divided into 4 treatments with 6 replicates per treatment and 15 birds per replicate. Dietary treatments were based on a basal diet and supplemented with 0, 0.08, 0.12, or 0.16% NCG to form C, N1, N2, and N3 groups. We found that layers in group N1 had higher egg production rate than those in group C. Egg weight was significantly reduced, while eggshell thickness was significantly improved, by treatment. However, the albumen height and Haugh unit were the lowest in group N3. Based on the above results, groups C and N1 were selected for further transcriptomics analysis of uterine tissue by RNA-seq. More than 7.4 Gb clean reads and 19,882 tentative genes were obtained using the Gallus gallus genome as a reference. Transcriptomics analysis in uterus tissue revealed that 95 differentially expressed genes (DEGs) were upregulated and 127 DEGs were downregulated. Functional annotation and pathway enrichment analysis showed that DEGs in uterine tissue were mainly enriched in glutathione metabolism, cholesterol metabolism, and glycerolipid metabolism, etc. Vitamin A metabolism-related gene, RBP1, nutrient transport-related gene, ALB, protein synthesis-related gene, METTL21C, and calcium transport-related gene, RYR2, CACNB2, RAMP3, and STAC, were significantly regulated by 0.08% NCG supplementation. Therefore, we concluded that NCG supplementation at a dose of 0.08% improved production performance and egg quality of layers by regulating uterus function.

Responses of Micropterus salmoides under Ammonia Stress and the Effects of a Potential Ammonia Antidote.

Ammonia is a common environmental limiting factor in aquaculture. To investigate the effects of ammonia stress and explore the protective effect of N-carbamylglutamate (NCG) on Micropterus salmoides (M. salmoides), tissue sections and parameters related to oxidative stress and the inflammatory response in M. salmoides were carried out during the ammonia stress test and feeding test. The results demonstrated that the LC50 for 24 h, 48 h, 72 h, and 96 h under ammonia stress in M. salmoides were 25.78 mg/L, 24.40 mg/L, 21.90 mg/L, and 19.61 mg/L, respectively. Under ammonia stress, the structures of the tissues were damaged, and the GSH content decreased, while the MDA content increased with the increase in stress time and ammonia concentration. The NO content fluctuated significantly after the ammonia nitrogen stress. In the 15-day feeding test, with the increased NCG addition amount and feeding time, the GSH content increased while the MDA and NO contents decreased gradually in the NCG addition groups (NL group: 150 mg/kg; NM group: 450 mg/kg; NH group: 750 mg/kg) when compared with their control group (CK group: 0 mg/kg). In the ammonia toxicology test after feeding, the damage to each tissue was alleviated in the NL, NM, and NH groups, and the contents of GSH, MDA, and NO in most tissues of the NH group were significantly different from those in the CK group. The results suggested that ammonia stress caused tissue damage in M. salmoides, provoking oxidative stress and inflammatory response. The addition of NCG to the feed enhances the anti-ammonia ability of M. salmoides. Moreover, the gill and liver might be the target organs of ammonia toxicity, and the brain and kidney might be the primary sites where NCG exerts its effects. Our findings could help us to find feasible ways to solve the existing problem of environmental stress in M. salmoides culture.

Effect of N- carbamylglutamate on amino acids and blood biochemistry in young male goats

To study the effect of feed additive N- carbamylglutamate (NCG) with different levels of feeding on amino acids profile and some analytes in serum of goats, thirty male goats aged 7-8 months were distributed into three levels of concentrate 2%, 3%, 4% with or without NCG in a 2×3 factorial experiment/ Completely Randomized Design (CRD). After 72 days of individual feeding, the blood from jugular vein was obtained before feeding in the morning. Results showed that additive NCG with 3% concentrate led to an increase (P&lt;0.01) in total protein and albumin (P&lt;0.05), globulins increased (P&lt;0.01) with increasing concentrate without NCG. Creatinine decreased with increasing concentrate without NCG. Additive NCG led to a linear decrease in uric acid with increasing feed intake (P&lt;0.05) and a linear increase without NCG. Blood magnesium, zinc, and copper increased with NCG. No difference in calcium between treatments. The level of feeding without NCG didn't affect fat parameters, while LDL, HDL, and cholesterol increased with NCG and 4% concentrate. Additive N- carbamylglutamate affected negatively blood amino acids, especially with 4% concentrate. In conclusion, 3% concentrate was the best for blood homeostasis with or without NCG, and preferred to use NCG with urea.

Dietary N-carbamylglutamate and L-arginine supplementation improves redox status and suppresses apoptosis in the colon of intrauterine growth-retarded suckling lambs.

Previous studies have revealed that dietary N-carbamylglutamate (NCG) or L-arginine (Arg) improves small intestinal integrity and immune function in suckling Hu lambs that have experienced intrauterine growth retardation (IUGR). Whether these nutrients alter redox status and apoptosis in the colon of IUGR lambs is still unknown. This study, therefore, aimed at investigating whether dietary supplementation of Arg or NCG alters colonic redox status, apoptosis and endoplasmic reticulum (ER) stress and the underlying mechanism of these alterations in IUGR suckling Hu lambs. Forty-eight 7-d old Hu lambs, including 12 with normal birth weight (4.25±0.14kg) and 36 with IUGR (3.01±0.12kg), were assigned to 4 treatment groups (n=12 each; 6 males and 6 females) for 3 weeks. The treatment groups were control (CON), IUGR, IUGR+Arg and IUGR+NCG. Relative to IUGR lambs, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) content, as well as proliferation index, were higher (P<0.05) whereas reactive oxygen species (ROS), malondialdehyde (MDA) levels and apoptotic cell numbers were lower (P<0.05) in colonic tissue for both IUGR+Arg and NCG lambs. Both mRNA and protein levels of C/EBP homologous protein 10 (CHOP10), B-cell lymphoma/leukaemia 2 (Bcl-2) -associated X protein (Bax), apoptosis antigen 1 (Fas), activating transcription factor 6 (ATF6), caspase 3, and glucose-regulated protein 78 (GRP78) were lower (P<0.05) while glutathione peroxidase 1 (GPx1), Bcl-2 and catalase (CAT) levels were higher (P<0.05) in colonic tissue for IUGR+Arg and IUGR+NCG lambs compared with IUGR lambs. Based on our results, dietary NCG or Arg supplementation can improve colonic redox status and suppress apoptosis via death receptor-dependent, mitochondrial and ER stress pathways in IUGR suckling lambs.

Dietary L-Arginine or N-Carbamylglutamate Alleviates Colonic Barrier Injury, Oxidative Stress, and Inflammation by Modulation of Intestinal Microbiota in Intrauterine Growth-Retarded Suckling Lambs.

Our previous studies have revealed that dietary N-carbamylglutamate (NCG) and L-arginine (Arg) supplementation improves redox status and suppresses apoptosis in the colon of suckling Hu lambs with intrauterine growth retardation (IUGR). However, no studies have reported the function of Arg or NCG in the colonic microbial communities, barrier function, and inflammation in IUGR-suckling lambs. This work aimed to further investigate how dietary Arg or NCG influences the microbiota, barrier function, and inflammation in the colon of IUGR lambs. Forty-eight newborn Hu lambs of 7 d old were assigned to four treatment groups (n = 12 per group; six male, six female) as follows: CON (normal birth weight, 4.25 ± 0.14 kg), IUGR (3.01 ± 0.12 kg), IUGR + Arg (2.99 ± 0.13 kg), and IUGR + NCG (3.03 ± 0.11 kg). A total of 1% Arg or 0.1% NCG was supplemented in a basal diet of milk replacer, respectively. Lambs were fed the milk replacer for 21 d until 28 d after birth. Compared to the non-supplemented IUGR lambs, the transepithelial electrical resistance (TER) was higher, while fluorescein isothiocyanate dextran 4 kDa (FD4) was lower in the colon of the NCG- or Arg-supplemented IUGR lambs (p &lt; 0.05). The IUGR lambs exhibited higher (p &lt; 0.05) colonic interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, reactive oxygen species (ROS), and malondialdehyde (MDA) levels than the CON lambs; the detrimental effects of IUGR on colonic proinflammatory cytokine concentrations and redox status were counteracted by dietary Arg or NCG supplementation. Both IUGR + Arg and IUGR + NCG lambs exhibited an elevated protein and mRNA expression of Occludin, Claudin-1, and zonula occludens-1 (ZO-1) compared to the IUGR lambs (p &lt; 0.05). Additionally, the lipopolysaccharide (LPS) concentration was decreased while the levels of acetate, butyrate, and propionate were increased in IUGR + Arg and IUGR + NCG lambs compared to the IUGR lambs (p &lt; 0.05). The relative abundance of Clostridium, Lactobacillus, and Streptococcus was lower in the colonic mucosa of the IUGR lambs than in the CON lambs (p &lt; 0.05) but was restored upon the dietary supplementation of Arg or NCG to the IUGR lambs (p &lt; 0.05). Both Arg and NCG can alleviate colonic barrier injury, oxidative stress (OS), and inflammation by the modulation of colonic microbiota in IUGR-suckling lambs. This work contributes to improving knowledge about the crosstalk among gut microbiota, immunity, OS, and barrier function and emphasizes the potential of Arg or NCG in health enhancement as feed additives in the early life nutrition of ruminants.

Docosapentaenoic Acid (DPA, 22:5n-3) Alleviates Ulcerative Colitis via Modification of Gut Microbiota and Their Metabolism.

N-3 polyunsaturated fatty acids (n-3PUFA) are regarded as viable alternatives to aid the treatment of ulcerative colitis (UC). Most research focuses on eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA); little information is available about the effect of docosapentaenoic acid (DPA) on the gut microbiota and their metabolism in UC mice. In this study, the changes in gut microbiota and their metabolism in UC mice were studied through the 16S rRNA sequencing method and untargeted metabolomics. Moreover, the differential bacterial genus and differential metabolites in responding to DPA supplementation were screened through permutation test after orthogonal partial least squares discriminant analysis (OPLS-DA). The results indicated that DPA supplementation increased the diversity and altered the composition of the gut microbiota in UC mice; Akkermansia, Alistipes, Butyricicoccus, and Lactobacillus were selected as the differential bacterial genus. Supplementation of DPA also altered the fecal metabolite profile in the UC mice. Moreover, butyrate, N-carbamylglutamate (NCG), and histamine were screened as the differential metabolites. In conclusion, the regulation effect of DPA on the gut microbiota and their metabolism might be involved in the intervention mechanism of DPA in UC. More research needs to be carried out to elucidate the mechanism systematically.