Integrating blood biochemistry and plasma metabolomics to define age-related metabolic biomarkers in Alope village chickens

Vet World Vol.19 February-2026 Article - 12

Research Article

Veterinary World, 19(2): 619-630

https://doi.org/10.14202/vetworld.2026.619-630

Andi Mujnisa¹, Athhar Manabi Diansyah², Muhammad Ridwan Bahar³, Andi Muhammad Fuad Al Kautsar Walinono³, Aji Praba Baskara⁴, Aeni Nurlatifah⁴, Herdis Herdis⁵, Fitra Aji Pamungkas⁵, and Syahruddin Said⁶

1. Department of Animal Nutrition, Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi, Indonesia.

2. Department of Animal Production, Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi, Indonesia.

3. Faculty of Vocational Study, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea Makassar, South Sulawesi, Indonesia.

4. Department of Nutrition and Feed Technology, Faculty of Animal Science, Gadjah Mada University, Depok, Sleman Regency, Special Region of Yogyakarta, Indonesia.

5. Research Center for Animal Husbandry, National Research and Innovation Agency, Raya Jakarta, Bogor, West Java 16915, Indonesia.

6. Research Center for Applied Zoology, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta, Bogor, West Java 16915, Indonesia.

Background and Aim: The Alope village chicken is an indigenous Indonesian line developed to combine local adaptability with improved growth potential. However, age-specific physiological and metabolic reference data remain limited, constraining evidence-based feeding and health management. This study aimed to integrate blood biochemical profiling and plasma metabolomics to characterize age-related metabolic changes, establish baseline reference values, and identify putative age-discriminant biomarkers in Alope village chickens.

Materials and Methods: A cross-sectional design was applied using 30 clinically healthy female Alope village chickens (Gallus gallus domesticus), stratified into three age groups: G1 (8 weeks), G2 (12 weeks), and G3 (16 weeks) (n = 10 per group). Blood samples were collected for biochemical analysis of glucose, total protein, urea, and triglycerides using enzymatic colorimetric assays. Plasma metabolomic profiling was conducted using untargeted gas chromatography–mass spectrometry. Multivariate analyses, including principal component analysis and partial least squares–discriminant analysis, were used to explore age-associated metabolic patterns. Discriminant metabolites were identified based on variable importance in projection values and descriptive receiver operating characteristic analysis.

Results: All biochemical parameters increased significantly with age (p < 0.05), remaining within physiological reference ranges, indicating normal metabolic development. Multivariate metabolomic analysis revealed clear age-dependent separation, with distinct metabolic signatures characterizing each growth stage. Exploratory pathway mapping indicated age-associated involvement of amino acid metabolism (glycine, serine, and threonine), lipid metabolism (glycerolipid and glycerophospholipid turnover), purine metabolism, the citrate cycle, and nodes linked to the pentose phosphate pathway. Seven metabolites demonstrated strong discriminatory performance (variable importance in projection ≈ 1.17–1.35; area under the curve ≥ 0.985) and were identified as putative age-related biomarkers, reflecting coordinated shifts in one-carbon/redox balance, membrane remodeling, and nucleotide and energy metabolism.

Conclusion: Age was the primary determinant of metabolic organization in Alope village chickens from 8 to 16 weeks. The integration of blood biochemistry and plasma metabolomics revealed coordinated physiological transitions from early growth toward enhanced energy handling and metabolic stabilization. The identified candidate metabolites provide an exploratory framework for defining physiological age and support the potential application of age-informed metabolic indicators for precision nutrition and health monitoring in indigenous village chicken production systems.

Keywords: age-related biomarkers, Alope village chicken, blood biochemistry, Gallus gallus domesticus, GC–MS metabolomics, growth physiology, plasma metabolomics, precision nutrition.

How to cite this article: Mujnisa A., Diansyah A.M., Bahar M.R., Walinono A.M.F.A.K., Baskara A.P., Nuralitah A., et al. Integrating blood biochemistry and plasma metabolomics to define age-related metabolic biomarkers in Alope village chickens. Vet World. 2026;19(2):619-630.

Received: 21-10-2025 Accepted: 05-01-2026 Published online: 17-02-2026

Corresponding author: Andi Mujnisa E-mail: mujnisa@unhas.ac.id

DOI: 10.14202/vetworld.2026.619-630

Copyright: Mujnisa, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.