Metabolomic analysis reveals the molecular mechanism related to leg abnormality in broilers

Abstract

Rapid body weight gain in broilers overloads the metabolic system of the organism, resulting in leg abnormalities, which seriously affects animal welfare and industry economics. In this study, broilers with normal and deformed leg bones were examined. Serum biochemical indices showed that the serum calcium to phosphorus ratio was extremely decreased in leg deformed group. In addition, abnormal serum lipid levels suggested a disruption in lipid metabolism. Based on widely targeted metabonomic analysis of serum and cartilage tissues, a total of nine differential metabolites (DMs) significantly associated with leg abnormalities and serum calcium and phosphorus levels were screened, including carnitine C16:0, carnitine C18:1, 3-hydroxymethyl-L-tyrosine, cis-4-hydroxy-D-proline, cis-L-3-hydroxyproline, trans-4-hydroxy-L-proline, and so on. Pathway analysis revealed that fatty acid degradation and arachidonic acid metabolism were enriched. Analysis of DMs in these two pathways showed that prostaglandin D₂, prostaglandin J₂, prostaglandin A₂, 15-keto prostaglandin F_2α, and Δ12-prostaglandin J₂ significantly differed between the normal and abnormal groups. It was hypothesized that these important metabolic pathways and metabolites were involved in the metabolic regulation of leg abnormalities.

Keywords

arachidonic acid metabolism / broiler / fatty acid degradation / leg abnormality / metabolomic

References

[1] Zuidhof, M. J., Schneider, B. L., Carney, V. L., Korver, D. R., & Robinson, F. E. (2014). Growth, efficiency, and yield of commercial broilers from 1957, 1978, and 2005. Poultry Science, 93(12), 2970-2982. https://doi.org/10.3382/ps.2014-04291

[2] Weeks, C. A., Danbury, T. D., Davies, H. C., Hunt, P., & Kestin, S. C. (2000). The behaviour of broiler chickens and its modification by lameness. Applied Animal Behaviour Science, 67(1–2), 111-125. https://doi.org/10.1016/s0168-1591(99)00102-1

[3] Dixon, L. M. (2020). Slow and steady wins the race: The behaviour and welfare of commercial faster growing broiler breeds compared to a commercial slower growing breed. PLoS One, 15(4), e0231006. https://doi.org/10.1371/journal.pone.0231006

[4] van der Sluis, M., Asher, L., Rodenburg, T. B., de Haas, Y., de Klerk, B., & Ellen, E. D. (2022). Early locomotor activity in broilers and the relationship with body weight gain. Poultry Science, 101(10), 102086. https://doi.org/10.1016/j.psj.2022.102086

[5] Kestin, S. C., Gordon, S., Su, G., & Sørensen, P. (2001). Relationships in broiler chickens between lameness, liveweight, growth rate and age. The Veterinary Record, 148(7), 195-197. https://doi.org/10.1136/vr.148.7.195

[6] Santos, M. N., Widowski, T. M., Kiarie, E. G., Guerin, M. T., Edwards, A. M., & Torrey, S. (2022). In pursuit of a better broiler: Walking ability and incidence of contact dermatitis in conventional and slower growing strains of broiler chickens. Poultry Science, 101(4), 101768. https://doi.org/10.1016/j.psj.2022.101768

[7] Tahamtani, F. M., Hinrichsen, L. K., & Riber, A. B. (2018). Welfare assessment of conventional and organic broilers in Denmark, with emphasis on leg health. The Veterinary Record, 183(6), 192. https://doi.org/10.1136/vr.104817

[8] Xu, T., Yue, K., Zhang, C., Tong, X., Lin, L., Cao, Q., & Huang, S. (2022). Probiotics treatment of leg diseases in broiler chickens: A review. Probiotics Antimicrob Proteins, 14(3), 415-425. https://doi.org/10.1007/s12602-021-09869-2

[9] Wideman, R. F., Hamal, K. R., Stark, J. M., Blankenship, J., Lester, H., Mitchell, K., Lorenzoni, G., & Pevzner, I. (2012). A wire-flooring model for inducing lameness in broilers: Evaluation of probiotics as a prophylactic treatment. Poultry Science, 91(4), 870-883. https://doi.org/10.3382/ps.2011-01907

[10] Sanotra, G. S., Lund, J. D., & Vestergaard, K. S. (2002). Influence of light-dark schedules and stocking density on behaviour, risk of leg problems and occurrence of chronic fear in broilers. British Poultry Science, 43(3), 344-354. https://doi.org/10.1080/000716601201036023611

[11] Edwards, H. M. (2000). Nutrition and skeletal problems in poultry. Poultry Science, 79(7), 1018-1023. https://doi.org/10.1093/ps/79.7.1018

[12] Kapell, D. N., Hill, W. G., Neeteson, A. M., McAdam, J., Koerhuis, A. N., & Avendaño, S. (2012). Twenty-five years of selection for improved leg health in purebred broiler lines and underlying genetic parameters. Poultry Science, 91(12), 3032-3043. https://doi.org/10.3382/ps.2012-02578

[13] Judge, A., & Dodd, M. S. (2020). Metabolism. Essays in Biochemistry, 64(4), 607-647. https://doi.org/10.1042/ebc20190041

[14] Bowling, F. G., & Thomas, M. (2014). Analyzing the metabolome. Methods in Molecular Biology, 1168, 31-45. https://doi.org/10.1007/978-1-4939-0847-9_3

[15] Schrimpe-Rutledge, A. C., Codreanu, S. G., Sherrod, S. D., & McLean, J. A. (2016). Untargeted metabolomics strategies-challenges and emerging directions. Journal of the American Society for Mass Spectrometry, 27(12), 1897-1905. https://doi.org/10.1007/s13361-016-1469-y

[16] Giera, M., Yanes, O., & Siuzdak, G. (2022). Metabolite discovery: Biochemistry's scientific driver. Cell Metabolism, 34(1), 21-34. https://doi.org/10.1016/j.cmet.2021.11.005

[17] Simó, C., Ibáñez, C., Valdés, A., Cifuentes, A., & García-Cañas, V. (2014). Metabolomics of genetically modified crops. International Journal of Molecular Sciences, 15(10), 18941-18966. https://doi.org/10.3390/ijms151018941

[18] Sumner, L. W., Lei, Z., Nikolau, B. J., & Saito, K. (2015). Modern plant metabolomics: Advanced natural product gene discoveries, improved technologies, and future prospects. Natural Product Reports, 32(2), 212-229. https://doi.org/10.1039/c4np00072b

[19] Ashokan, M., Rana, E., Sneha, K., Namith, C., Naveen Kumar, G. S., Azharuddin, N., Elango, K., Jeyakumar, S., & Ramesha, K. P. (2022). Metabolomics-a powerful tool in livestock research. Animal Biotechnology, 34(7), 1-13. https://doi.org/10.1080/10495398.2022.2128814

[20] Zhou, M., Jing, J. H., Mao, R. H., Guo, J., & Wang, Z. P. (2019). [Applications of metabonomics in animal genetics and breeding]. Yi Chuan, 41(2), 111-124. https://doi.org/10.16288/j.yczz.18-226

[21] Goldansaz, S. A., Guo, A. C., Sajed, T., Steele, M. A., Plastow, G. S., & Wishart, D. S. (2017). Livestock metabolomics and the livestock metabolome: A systematic review. PLoS One, 12(5), e0177675. https://doi.org/10.1371/journal.pone.0177675

[22] Shi, S., Shen, Y., Zhang, S., Zhao, Z., Hou, Z., Zou, J., & Guo, Y. (2017). Combinatory evaluation of transcriptome and metabolome profiles of low temperature-induced resistant ascites syndrome in broiler chickens. Scientific Reports, 7(1), 2389. https://doi.org/10.1038/s41598-017-02492-8

[23] Jastrebski, S. F., Lamont, S. J., & Schmidt, C. J. (2017). Chicken hepatic response to chronic heat stress using integrated transcriptome and metabolome analysis. PLoS One, 12(7), e0181900. https://doi.org/10.1371/journal.pone.0181900

[24] Florencio-Silva, R., Sasso, G. R., Sasso-Cerri, E., Simões, M. J., & Cerri, P. S. (2015). Biology of bone tissue: Structure, function, and factors that influence bone cells. BioMed Research International, 2015, 421746. https://doi.org/10.1155/2015/421746

[25] Suzuki, A., & Iwata, J. (2021). Amino acid metabolism and autophagy in skeletal development and homeostasis. Bone, 146, 115881. https://doi.org/10.1016/j.bone.2021.115881

[26] Wawrzyniak, A., & Balawender, K. (2022). Structural and metabolic changes in bone. Animals, 12(15), 1946. https://doi.org/10.3390/ani12151946

[27] Suzuki, A., Minamide, M., Iwaya, C., Ogata, K., & Iwata, J. (2020). Role of metabolism in bone development and homeostasis. International Journal of Molecular Sciences, 21(23), 8992. https://doi.org/10.3390/ijms21238992

[28] Armas, L. A., & Recker, R. R. (2012). Pathophysiology of osteoporosis: New mechanistic insights. Endocrinology and Metabolism Clinics of North America, 41(3), 475-486. https://doi.org/10.1016/j.ecl.2012.04.006

[29] Zheng, J., Liu, D., Zhang, G., Ding, J., He, Z., Li, Q., Liu, R., Wen, J., & Zhao, G. (2023). Improve the accuracy of chicken leg disease assessment by combination of gait score and X-ray scan. The Journal of Applied Poultry Research, 32(2), 100339. https://doi.org/10.1016/j.japr.2023.100339

[30] Kanehisa, M., & Goto, S. (2000). KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Research, 28(1), 27-30. https://doi.org/10.1093/nar/28.1.27

[31] Proszkowiec-Weglarz, M., & Angel, R. (2013). Calcium and phosphorus metabolism in broilers: Effect of homeostatic mechanism on calcium and phosphorus digestibility1 1Presented as a part of the informal nutrition symposium “metabolic responses to nutrition and modifiers” at the poultry science association's annual meeting in Athens, Georgia, July 9, 2012. The Journal of Applied Poultry Research, 22(3), 609-627. https://doi.org/10.3382/japr.2012-00743

[32] Wei, H., Bi, Y., Wang, Y., Zhao, Q., Zhang, R., Li, J., & Bao, J. (2023). Serum bone remodeling parameters and transcriptome profiling reveal abnormal bone metabolism associated with keel bone fractures in laying hens. Poultry Science, 102(4), 102438. https://doi.org/10.1016/j.psj.2022.102438

[33] Civitelli, R., & Ziambaras, K. (2011). Calcium and phosphate homeostasis: Concerted interplay of new regulators. Journal of Endocrinological Investigation, 34(7 Suppl), 3-7.

[34] Taylor, J. G., & Bushinsky, D. A. (2009). Calcium and phosphorus homeostasis. Blood Purification, 27(4), 387-394. https://doi.org/10.1159/000209740

[35] Suki, W. N., & Moore, L. W. (2016). Phosphorus regulation in chronic kidney disease. Methodist DeBakey Cardiovascular Journal, 12(4 Suppl), 6-9. https://doi.org/10.14797/mdcj-12-4s1-6

[36] Huang, S. C., Zhang, L. H., Zhang, J. L., Rehman, M. U., Tong, X. L., Qiu, G., Jiang, X., Iqbal, M., Shahzad, M., Shen, Y. q., & Li, J. k. (2018). Role and regulation of growth plate vascularization during coupling with osteogenesis in tibial dyschondroplasia of chickens. Scientific Reports, 8(1), 3680. https://doi.org/10.1038/s41598-018-22109-y

[37] During, A. (2020). Osteoporosis: A role for lipids. Biochimie, 178, 49-55. https://doi.org/10.1016/j.biochi.2020.08.004

[38] Erken, H. Y., Ofluoglu, O., Aktas, M., Topal, C., & Yildiz, M. (2012). Effect of pentoxifylline on histopathological changes in steroid-induced osteonecrosis of femoral head: Experimental study in chicken. International Orthopaedics, 36(7), 1523-1528. https://doi.org/10.1007/s00264-012-1497-6

[39] Wang, B., Wang, H., Li, Y., & Song, L. (2022). Lipid metabolism within the bone micro-environment is closely associated with bone metabolism in physiological and pathophysiological stages. Lipids in Health and Disease, 21(1), 5. https://doi.org/10.1186/s12944-021-01615-5

[40] Tintut, Y., Morony, S., & Demer, L. L. (2004). Hyperlipidemia promotes osteoclastic potential of bone marrow cells ex vivo. Arteriosclerosis, Thrombosis, and Vascular Biology, 24(2), e6-e10. https://doi.org/10.1161/01.ATV.0000112023.62695.7f

[41] Baggio, B. (2002). Fatty acids, calcium and bone metabolism. Journal of Nephrology, 15(6), 601-604.

[42] Longo, N., Frigeni, M., & Pasquali, M. (2016). Carnitine transport and fatty acid oxidation. Biochimica et Biophysica Acta, 1863(10), 2422-2435. https://doi.org/10.1016/j.bbamcr.2016.01.023

[43] Lee, N. K., Sowa, H., Hinoi, E., Ferron, M., Ahn, J. D., Confavreux, C., Dacquin, R., Mee, P. J., McKee, M. D., Jung, D. Y., Zhang, Z., Kim, J. K., Mauvais-Jarvis, F., Ducy, P., & Karsenty, G. (2007). Endocrine regulation of energy metabolism by the skeleton. Cell, 130(3), 456-469. https://doi.org/10.1016/j.cell.2007.05.047

[44] Lee, N. K., Choi, Y. G., Baik, J. Y., Han, S. Y., Jeong, D. W., Bae, Y. S., Kim, N., & Lee, S. Y. (2005). A crucial role for reactive oxygen species in RANKL-induced osteoclast differentiation. Blood, 106(3), 852-859. https://doi.org/10.1182/blood-2004-09-3662

[45] Geng, Q., Gao, H., Yang, R., Guo, K., & Miao, D. (2019). Pyrroloquinoline quinone prevents estrogen deficiency-induced osteoporosis by inhibiting oxidative stress and osteocyte senescence. International Journal of Biological Sciences, 15(1), 58-68. https://doi.org/10.7150/ijbs.25783

[46] Huang, S., Mao, J., Wei, B., & Pei, G. (2015). The anti-spasticity drug baclofen alleviates collagen-induced arthritis and regulates dendritic cells. Journal of Cellular Physiology, 230(7), 1438-1447. https://doi.org/10.1002/jcp.24884

[47] Bröer, S. (2022). Amino acid transporters as modulators of glucose homeostasis. Trends in Endocrinology and Metabolism, 33(2), 120-135. https://doi.org/10.1016/j.tem.2021.11.004

[48] Yang, J., Zhang, X., Wang, W., & Liu, J. (2010). Insulin stimulates osteoblast proliferation and differentiation through ERK and PI3K in MG-63 cells. Cell Biochemistry and Function, 28(4), 334-341. https://doi.org/10.1002/cbf.1668

[49] Williams, G. R., & Bassett, J. H. D. (2018). Thyroid diseases and bone health. Journal of Endocrinological Investigation, 41(1), 99-109. https://doi.org/10.1007/s40618-017-0753-4

[50] König, S., Marco, H., & Gäde, G. (2018). D-Proline: Comment to "An overview on D-amino acids”. Amino Acids, 50(2), 359-361. https://doi.org/10.1007/s00726-017-2511-5

[51] Zhang, Z., Liu, P., Su, W., Zhang, H., Xu, W., & Chu, X. (2021). Metabolic engineering strategy for synthetizing trans-4-hydroxy-L-proline in microorganisms. Microbial Cell Factories, 20(1), 87. https://doi.org/10.1186/s12934-021-01579-2

[52] Martyniak, K., Wei, F., Ballesteros, A., Meckmongkol, T., Calder, A., Gilbertson, T., Orlovskaya, N., & Coathup, M. J. (2021). Do polyunsaturated fatty acids protect against bone loss in our aging and osteoporotic population? Bone, 143, 115736. https://doi.org/10.1016/j.bone.2020.115736

[53] Ricciotti, E., & FitzGerald, G. A. (2011). Prostaglandins and inflammation. Arteriosclerosis, Thrombosis, and Vascular Biology, 31(5), 986-1000. https://doi.org/10.1161/atvbaha.110.207449

[54] Pilbeam, C. (2020). Prostaglandins and bone. Handbook of Experimental Pharmacology, 262, 157-175. https://doi.org/10.1007/164_2019_332

[55] Agas, D., Marchetti, L., Hurley, M. M., & Sabbieti, M. G. (2013). Prostaglandin F2α: A bone remodeling mediator. Journal of Cellular Physiology, 228(1), 25-29. https://doi.org/10.1002/jcp.24117

[56] Xiong, Z., Luo, P., Zhou, J., & Tan, M. (2019). 15-Deoxy-Δ(12,14)-prostaglandin J(2) as a potential regulator of bone metabolism via PPARγ-dependent and independent pathways: A review. Drug Design, Development and Therapy, 13, 1879-1888. https://doi.org/10.2147/dddt.S206695

[57] Fujimori, K. (2022). Prostaglandin D(2) and F(2α) as regulators of adipogenesis and obesity. Biological & Pharmaceutical Bulletin, 45(8), 985-991. https://doi.org/10.1248/bpb.b22-00210

[58] Damrongsri, D., Geva, S., Salvi, G. E., Cooper, L. F., Limwongse, V., & Offenbacher, S. (2006). Effects of Delta12-prostaglandin J2 on bone regeneration and growth factor expression in rats. Clinical Oral Implants Research, 17(1), 48-57. https://doi.org/10.1111/j.1600-0501.2005.01181.x

[59] Cho, K. I., Sakuma, I., Sohn, I. S., Jo, S. H., & Koh, K. K. (2018). Inflammatory and metabolic mechanisms underlying the calcific aortic valve disease. Atherosclerosis, 277, 60-65. https://doi.org/10.1016/j.atherosclerosis.2018.08.029

[60] Demer, L., & Tintut, Y. (2011). The roles of lipid oxidation products and receptor activator of nuclear factor-κB signaling in atherosclerotic calcification. Circulation Research, 108(12), 1482-1493. https://doi.org/10.1161/circresaha.110.234245

RIGHTS & PERMISSIONS

2024 The Author(s). Animal Research and One Health published by John Wiley & Sons Australia, Ltd on behalf of Institute of Animal Science, Chinese Academy of Agricultural Sciences.