In-ovo mineral supplementation: a comprehensive analysis of broiler bone morphometry and structural implications

Main Article Content

Imran Ahmed
Nasir Rajput
Imdad Hussain Laghari
Nasir Mukhtar
Zahid Iqbal Rajput


This study aimed to investigate the impact of in-ovo feeding of macro (Ca, P, and Mg) and micro (Cu, Mn, and Zn) minerals on various bone parameters, gait score, black bone syndrome, lameness, and curled toe paralysis in broiler chickens. The experiment was conducted using a completely randomized design with seven treatment groups, including a control group. Results revealed that in-ovo administration of Ca significantly influenced (P<0.05) tibia, femur, keel bone, and metatarsus morphometry. Femur bone, keel bone and metatarsus bone ash contents were higher (P<0.05) in in-ovo Calcium administered birds, however, tibia ash was same across the groups. The Ca, P, Mg, Cu, Mn and Zn mineralization was higher (P<0.05) in respective in-ovo administered group. Moreover, in ovo Ca feeding positively influenced pododermatitis score, gait score, black bone syndrome score, lameness score, and curled toe paralysis score. In conclusion, in-ovo mineral supplementation, particularly with Ca, effectively enhanced broiler bone morphometry, bone ash and bone mineralization with positive impact on bone health and gait.

Article Details

How to Cite
Imran Ahmed, Rajput, N., Imdad Hussain Laghari, Nasir Mukhtar, & Zahid Iqbal Rajput. (2024). In-ovo mineral supplementation: a comprehensive analysis of broiler bone morphometry and structural implications. Pakistan Journal of Multidisciplinary Research, 4(2), 165-175. Retrieved from


Angel, R., 2007. Metabolic disorders: limitations to growth of and mineral deposition into the broiler skeleton after hatch and potential implications for leg problems. Journal of Applied Poultry Research 16(1), 138-149.

Biesek, J., Banaszak, M., K?dzio?ka, K., Wla?lak, S., Adamski, M., 2022. Growth of broiler chickens, and physical features of the digestive system, and leg bones after aluminosilicates used. Scientific Reports 12(1), 20425.

Ciosek, ?., Kot, K., Kosik-Bogacka, D., ?anocha-Arendarczyk, N., Rotter, I., 2021. The effects of calcium, magnesium, phosphorus, fluoride, and lead on bone tissue. Biomolecules 11(4), 506.

Oliveira, T., Bertechini, A., Bricka, R., Hester, P., Kim, E., Gerard, P., Peebles, E., 2015. Effects of in ovo injection of organic trace minerals and post-hatch holding time on broiler performance and bone characteristics. Poultry Science 94(11), 2677-2685.

Abbasi, T., Shakeri, M., Zaghari, M., & Kohram, H. (2017). Growth performance parameters, bone calcification and immune response of in ovo injection of 25-hydroxycholecalciferol and vitamin K3 in male ross 308 broilers. Theriogenology, 90, 260-265.

Yair, R., Uni, Z., 2011. Content and uptake of minerals in the yolk of broiler embryos during incubation and effect of nutrient enrichment. Poultry Science 90(7), 1523-1531.

Bello, A., Bricka, R. M., Gerard, P. D., & Peebles, E. D. (2014). Effects of commercial in ovo injection of 25-hydroxycholecalciferol on broiler bone development and mineralization on days 0 and 21 posthatch. Poultry science, 93(5), 1053-1058.

Das, R., Mishra, P., Jha, R., 2021. In ovo feeding as a tool for improving performance and gut health of poultry: a review. Frontiers in Veterinary Science 8, 754246.

de Carvalho, B.R., Hélvio da Cruz Ferreira, J., da Silva Viana, G., Alves, W.J., Muniz, J.C.L., Rostagno, H.S., Pettigrew, J.E., Hannas, M.I., 2021. In-feed organic and inorganic manganese supplementation on broiler performance and physiological responses. Animal Bioscience 34(11), 1811.

Fleming, R.H., 2008. Nutritional factors affecting poultry bone health: Symposium on ‘Diet and bone health’. Proceedings of the Nutrition Society 67(2), 177-183.

Heffernan, S.M., Horner, K., De Vito, G., Conway, G.E., 2019. The role of mineral and trace element supplementation in exercise and athletic performance: a systematic review. Nutrients 11(3), 696.

Matuszewski, A., ?ukasiewicz, M., Niemiec, J., Kamaszewski, M., Jaworski, S., Domino, M., Jasi?ski, T., Chwalibog, A., Sawosz, E., 2021. Calcium Carbonate nanoparticles—Toxicity and effect of in ovo inoculation on chicken embryo development, broiler performance and bone status. Animals 11(4), 932.

Nakhon, S., Numthuam, S., Charoensook, R., Tartrakoon, W., Incharoen, P., Incharoen, T., 2019. Growth performance, meat quality, and bone-breaking strength in broilers fed dietary rice hull silicon. Animal Nutrition 5(2), 152-155.

Nouri, S., Ghalehkandi, J.G., Hassanpour, S., Aghdam-Shahryar, H., 2018. Effect of in ovo feeding of folic acid on subsequent growth performance and blood constituents levels in broilers. International Journal of Peptide Research and Therapeutics 24, 463-470.

Oikeh, I., Sakkas, P., Blake, D.P., Kyriazakis, I., 2019. Interactions between dietary calcium and phosphorus level, and vitamin D source on bone mineralization, performance, and intestinal morphology of coccidia-infected broilers. Poultry science 98(11), 5679-5690.

Oliveira, T., Bertechini, A., Bricka, R., Hester, P., Kim, E., Gerard, P., Peebles, E., 2015. Effects of in ovo injection of organic trace minerals and post-hatch holding time on broiler performance and bone characteristics. Poultry Science 94(11), 2677-2685.

Rath, N.C., Durairaj, V., 2022. Avian bone physiology and poultry bone disorders, Sturkie's Avian Physiology. Elsevier, pp. 549-563.

Retes, P., Clemente, A., Neves, D., Espósito, M., Makiyama, L., Alvarenga, R., Pereira, L., Zangeronimo, M., 2018. In ovo feeding of carbohydrates for broilers—A systematic review. Journal of animal physiology and animal nutrition 102(2), 361-369.

Riber, A.B., Casey-Trott, T.M., Herskin, M.S., 2018. The influence of keel bone damage on welfare of laying hens. Frontiers in veterinary science 5, 6.

Saeed, A.A., Sandhu, M.A., Khilji, M.S., Yousaf, M.S., Rehman, H.U., Tanvir, Z.I., Ahmad, T., 2017. Effects of dietary chromium supplementation on muscle and bone mineral interaction in broiler chicken. Journal of Trace Elements in Medicine and Biology 42, 25-29.

Salary, J., Hemati Matin, H., Ghafari, K., Hajati, H., 2017. Effect of in ovo injection of calcium carbonate nanoparticles on bone post hatched characteristics and broiler chicken performance. Iranian Journal of Applied Animal Science 7(4), 663-667.

Tang, S., Dong, Z., Ke, X., Luo, J., Li, J., 2021. Advances in biomineralization-inspired materials for hard tissue repair. International journal of oral science 13(1), 42.

Trautvetter, U., Neef, N., Leiterer, M., Kiehntopf, M., Kratzsch, J., Jahreis, G., 2014. Effect of calcium phosphate and vitamin D 3 supplementation on bone remodelling and metabolism of calcium, phosphorus, magnesium and iron. Nutrition journal 13, 1-9.

Waldenstedt, L., 2006. Nutritional factors of importance for optimal leg health in broilers: A review. Animal feed science and technology 126(3-4), 291-307.

Webster, A., 2004. Welfare implications of avian osteoporosis. Poultry science 83(2), 184-192.

Yair, R., Shahar, R., Uni, Z., 2013. Prenatal nutritional manipulation by in ovo enrichment influences bone structure, composition, and mechanical properties. Journal of animal science 91(6), 2784-2793.

Yair, R., Shahar, R., Uni, Z., 2015. In ovo feeding with minerals and vitamin D3 improves bone properties in hatchlings and mature broilers. Poultry science 94(11), 2695-2707.

Zhu, Y., Li, S., Sun, Q., Yang, X., 2019. Effect of in ovo feeding of vitamin C on antioxidation and immune function of broiler chickens. Animal 13(9), 1927-1933.