# N-Carbamylglutamate Promotes Growth and Immunity in Danzhou Chickens via Gut Microbiota-Metabolite Interactions Involving Sphingolipid and mTOR Pathways

**Authors:** Hongzhi Wu, Dexin Zhao, Xilong Yu, Haoliang Chai, Hanyang Liu, Weiqi Peng, Liangmei Xu, Guanyu Hou

PMC · DOI: 10.3390/microorganisms14020370 · 2026-02-04

## TL;DR

N-Carbamylglutamate improves growth and immunity in chickens by affecting gut microbes and key metabolic pathways.

## Contribution

This study reveals how N-Carbamylglutamate enhances chicken growth and immunity through gut microbiota and metabolite interactions.

## Key findings

- NCG at 400 mg/kg improved growth and immune function in Danzhou chickens.
- NCG modulated sphingolipid and mTOR pathways, affecting metabolites like L-arginine and ceramide.
- NCG altered gut microbiota composition, particularly Bacteroidota and Firmicutes.

## Abstract

This study evaluated the effects of dietary supplementation with N-Carbamylglutamate (NCG) on growth performance, immune function, intestinal metabolites, and microbiota in Danzhou chickens. In a 35-day feeding trial, a total of 480 one-day-old female chicks were randomly assigned to a control group (basal diet) and three experimental groups supplemented with 400, 800, or 1200 mg/kg NCG, with 120 chicks in each group (n = 120). The results demonstrated that NCG, particularly at 400 mg/kg, significantly improved growth parameters, including average daily gain and feed conversion ratio, while enhancing immune function by increasing serum levels of immunoglobulins (IgA, IgY) and malate dehydrogenase (p < 0.05). Metabolomic analysis revealed that NCG modulated key pathways such as sphingolipid metabolism and mTOR signaling pathway, leading to significant changes in metabolites including L-arginine, ceramide, and docosahexaenoic acid (p < 0.05). 16S rDNA sequencing indicated that NCG induced structural shifts in the gut microbiota, primarily affecting Bacteroidota and Firmicutes, with several bacterial genera showing strong correlations with the observed metabolic changes (p < 0.05). Mechanistically, NCG promotes growth by facilitating arginine synthesis via the urea cycle and activating the mTOR signaling pathway, while its regulation of sphingolipid metabolism enhances immunomodulatory capacity. In conclusion, NCG enhances feed efficiency and immune competence by orchestrating the gut microbiota-metabolite network, demonstrating its potential for poultry production.

## Linked entities

- **Chemicals:** N-Carbamylglutamate (PubChem CID 121396), L-arginine (PubChem CID 232), ceramide (PubChem CID 139583739), docosahexaenoic acid (PubChem CID 445580)

## Full-text entities

- **Genes:** ASL1 (argininosuccinate lyase 1) [NCBI Gene 396498] {aka ASL, CRYD1, D-CRY}, ME1 (malic enzyme 1) [NCBI Gene 374189] {aka NADP-ME}, SMPD2 (sphingomyelin phosphodiesterase 2) [NCBI Gene 770663] {aka ISC1, nSMase}, ASS1 (argininosuccinate synthase 1) [NCBI Gene 417185], LITAF (lipopolysaccharide induced TNF factor) [NCBI Gene 374125] {aka TNF-alpha}, TIPRL (TOR signaling pathway regulator) [NCBI Gene 418374], GAPDH (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 374193] {aka GAPD, KNC-NDS6}, MTOR (mechanistic target of rapamycin) [NCBI Gene 419455] {aka FRAP1}, LGR5 (leucine rich repeat containing G protein-coupled receptor 5) [NCBI Gene 427867] {aka GPR49}, IGF1 (insulin like growth factor 1) [NCBI Gene 418090] {aka IGF-1, IGF-I}, EIF4EBP1 (eukaryotic translation initiation factor 4E binding protein 1) [NCBI Gene 426773], MDH2 (malate dehydrogenase 2) [NCBI Gene 417517], ACTB (actin, beta) [NCBI Gene 396526] {aka Bact, actin}, CAT (catalase) [NCBI Gene 423600], CRPL1 (C-reactive protein like 1) [NCBI Gene 429786] {aka APCS, CRP}, CPS1 (carbamoyl-phosphate synthase 1) [NCBI Gene 428994], IL6 (interleukin 6) [NCBI Gene 395337] {aka CHIL-6, IL-6, interleukin-6}, IL10 (interleukin 10) [NCBI Gene 428264] {aka IL-10, interleukin-10}, MDH1 (malate dehydrogenase 1) [NCBI Gene 421281], JCHAIN (joining chain of multimeric IgA and IgM) [NCBI Gene 374117] {aka IGJ}
- **Diseases:** lipid metabolic disorders (MESH:D052439), infectious bronchitis (MESH:D001991), necrosis (MESH:D009336), Gumboro disease (MESH:D004194), injury to (MESH:D014947), inflammation (MESH:D007249), gut dysbiosis (MESH:D064806), weight gain (MESH:D015430), Newcastle disease (MESH:D009521), Marek's disease (MESH:D008380)
- **Chemicals:** polyunsaturated fatty acid (MESH:D005231), hematoxylin (MESH:D006416), 1-(4-Hydroxyphenyl)-1-decene-3,5-dione (-), ornithine (MESH:D009952), H&amp;E (MESH:D006371), sphingomyelin (MESH:D013109), propionate (MESH:D011422), creatine (MESH:D003401), carbohydrates (MESH:D002241), Arginine (MESH:D001120), fatty acid (MESH:D005227), butyrate (MESH:D002087), urea (MESH:D014508), amino acids (MESH:D000596), DHA (MESH:D004281), lipid (MESH:D008055), paraformaldehyde (MESH:C003043), N-Carbamylglutamate (MESH:C006895), agarose (MESH:D012685), polyamines (MESH:D011073), lignin (MESH:D008031), carbon dioxide (MESH:D002245), short-chain fatty acids (MESH:D005232), sphingosine (MESH:D013110), eosin (MESH:D004801), acetate (MESH:D000085), Sphingolipid (MESH:D013107), carbamoyl phosphate (MESH:D002221), hydrogen (MESH:D006859), formate (MESH:C030544), ammonia (MESH:D000641), ceramide-1-phosphate (MESH:C065576), cellobiose (MESH:D002475), paraffin (MESH:D010232), Succinate (MESH:D019802), methanol (MESH:D000432), sodium chloride (MESH:D012965), acetonitrile (MESH:C032159), monosaccharides (MESH:D009005), Sphinganine (MESH:C005682), tricarboxylic acid (MESH:D014233), lactate (MESH:D019344), polysaccharides (MESH:D011134), nitrogen (MESH:D009584), Trizol (MESH:C411644), sphingosine-1-phosphate (MESH:C060506), eicosapentaenoic acid (MESH:D015118), phospholipids (MESH:D010743), water (MESH:D014867), luminal (MESH:D010634), nitric ox-ide (MESH:D009569), ceramide (MESH:D002518), citrulline (MESH:D002956), N-Acetylglutamate (MESH:C016195), HCl (MESH:D006851)
- **Species:** Jeotgalicoccus (genus) [taxon 227979], Homo sapiens (human, species) [taxon 9606], Bacteroides (genus) [taxon 816], Cutibacterium (genus) [taxon 1912216], Bos taurus (bovine, species) [taxon 9913], Pseudomonas (RNA similarity group I, genus) [taxon 286], Photobacterium frigidiphilum (species) [taxon 264736], Bacillota (clostridial firmicutes, phylum) [taxon 1239], Gallus gallus (bantam, species) [taxon 9031], Mailhella (genus) [taxon 1981028], Glycine max (soybean, species) [taxon 3847], Photobacterium sp. (species) [taxon 660], Faecalibacterium (genus) [taxon 216851], Lactobacillus (genus) [taxon 1578], Phocaeicola (genus) [taxon 909656], Brachybacterium (genus) [taxon 43668], Victivallis (genus) [taxon 172900], Sus scrofa (pig, species) [taxon 9823]

## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12943504/full.md

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Source: https://tomesphere.com/paper/PMC12943504