Limb problems are one of the most common problems with fast-growing meat-type chickens. Various bone abnormalities, which can lead to limping, bone weakness, or even fractures, bring overall discomfort to birds and a loss of production. Genetic aspects are often associated with these side effects on bone stability and are also cited as the dominant cause. These points to a close negative relationship of genetic selection for rapid growth with traits involved in bone integrity. Due to the assumption of an additive genetic background, improvements through genetic tools can be used. Our study is focused on selected genes of important signaling pathways for bone metabolism. We tried to detect polymorphisms that would show associations with selected bone parameters in a total of 48 broilers. Those were fast-growing Ross 308 hybrids and slow-growing Hubbard M22BxJA87A hybrids. The TNFRSF11A and WISP1 genes were tested. A total of fourteen polymorphisms were found, three of them were synonymous and five in the intron. In the case of four polymorphisms found in exons of the TNFRSF11A gene (c.11G > T, c.31G > A, c.37C > G, c.514G > A), associations with the observed bone parameters (bone strength, bone dimensions and bone mass) were demonstrated. The genetic architecture of bone traits is not fully understood, therefore the present study and the knowledge gained can help to increase the potential in poultry breeding processes and thus reduce the death of individuals.

Department of Animal Morphology Physiology and Genetics Mendel University in Brno Faculty of AgriSciences Zemedelska 1 1665 613 00 Brno Czech Republic

Department of Food Technology Mendel University in Brno Faculty of AgriSciences Zemedelska 1 1665 613 00 Brno Czech Republic

See more in PubMed

Acar E.F., Sun L. A generalized Kruskal-Wallis test incorporating group uncertainty with application to genetic association studies. Biometrics. 2013;69:427–435. doi: 10.1111/biom.12006. PubMed DOI

Arnez M.F.M., Ribeiro L.S.N., Barretto G.D., Monteiro P.M., Ervolino E., Stuani M.B.S. RANK/RANKL/OPG expression in rapid maxillary expansion. Braz. Dent. J. 2017;28:296–300. doi: 10.1590/0103-6440201601116. PubMed DOI

Azizieh F.Y., Shehab D., Jarallah K.A., Gupta R., Raghupathy R. Circulatory levels of RANKL, OPG, and oxidative stress markers in postmenopausal women with normal or low bone mineral density. Biomark. Insights. 2019;14 doi: 10.1177/1177271919843825. PubMed DOI PMC

Azmal S.A., Nan J., Bhuiyan A.A., Elokil A.A., Ali M.I., Adetula A.A., Ma S., Sun Ch, Han Z., Yuan J., Li S. A genome-wide single nucleotide polymorphism scan reveals genetic markers associated with fertility rate in Chinese Jing Hong chicken. Poultry Sci. 2020;99:2873–2887. doi: 10.1016/j.psj.2019.12.068. PubMed DOI PMC

De Koning D.J., Dominguez-Gasca N., Fleming R.H., Gill A., Kurian D., Law A., McCormack A., Morrice D., Sanchez-Rodriguez E., Rodriguez-Navarro A.B., Preisinger R., Schmutz M., Šmídová V., Turner F., Wilson P.W., Zhou R., Dunn I.C. An eQTL in the cystathionine beta synthase gene is linked to osteoporosis in laying hens. Genet. Sel. Evol. 2020;52:13. doi: 10.1186/s12711-020-00532-y. PubMed DOI PMC

Dinev I., Kanakov D., Kalkanov I., Nikolov S., Denev S. Comparative pathomorphologic studies on the incidence of fractures associated with leg skeletal pathology in commercial broiler chickens. Avian Dis. 2019;63:641–650. doi: 10.1637/aviandiseases-D-19-00108. PubMed DOI

Grupioni N.V., Stafuzza N.B., Carvajal A.B., A, Ibelli A.M.G., Peixoto J.O., Ledur M.C., Munari D.P. Association of RUNX2 and TNFSF11 gene with production traits in a paternal broiler line. Genet. Mol. Res. 2017;16 doi: 10.4238/gmr16019443. PubMed DOI

Guo J., Sun C., Qu L., Shen M., Dou T., Ma M., Wang K., Yang N. Genetic architecture of bone quality variation in layer chickens revealed by a genome-wide association study. Sci. Rep. 2017;7 doi: 10.1038/srep45317. PubMed DOI PMC

Horecka E., Horecky C., Kovarikova L., Musilova A., Knoll A., Nedomova S., Pavlik A. Association between single nucleotide polymorphisms of ATP2B1 gene and bone parameters of laying hens. Avian Biol. Res. 2018;11:178–182. doi: 10.3184/175815618X15269357438898. DOI

Hul L.M., Ibelli A.M.G., Savoldi I.R., Mercelino D.E.P., Fernandes L.T., Peixoto J.O., Cantão M.E., Higa R.H., Giachetto P.F., Coutinho L.L., Ledur M.C. Differentially expressed genes in the femur cartilage transcriptome clarify the understanding of femoral head separation in chickens. Sci. Rep. 2021;11 doi: 10.1038/s41598-021-97306-3. PubMed DOI PMC

Huybrechts Y., Mortier G., Boudin E., Van Hul W. Wnt signaling and bone: lessons from skeletal dysplasias and disorders. Front. Endocrinol. 2020;11:165. doi: 10.3389/fendo.2020.00165. PubMed DOI PMC

Infante M., Fabi A., Cognetti F., Gorini S., Caprio M., Fabbri A. RANKL/RANK/OPG system beyond bone remodeling: involvement in breast cancer and clinical perspectives. J. Exp. Clin. Cancer Res. 2019;38:12. doi: 10.1186/s13046-018-1001-2. PubMed DOI PMC

Jansen S., Baulain U., Habig Ch, Ramzan F., Schauer J., Schmitt A.O., Scholz A.M., Sharifi A.R., Weigend A., Weigend S. Identification and functional annotation of genes related to bone stability in laying hens using random forests. Genes. 2021;12:702. doi: 10.3390/genes12050702. PubMed DOI PMC

Jiang B., Wang M., Tang Z., Du X., Feng S., Ma G., Ye D., Cheng H., Wang H., Liu X. Genome-wide association study of bone mineral density trait among three pig breeds. Animal. 2020;14:2443–2451. doi: 10.1017/S1751731120001111. PubMed DOI

Johnnson M., Wall H., Pinto F.A.L., Fleming R.H., McCormack H.A., Benavides-Reyes C., Dominguez-Gasca N., Sanchez-Rodriguez E., Dunn I.C., Rodriguez-Navarro A.B., Kindmark A., De Koning D.J. Genetics of tibia bone properties of cross-bred commercial laying hens in different housing systems. G3-Genes Genom. Genet. 2023;13 PubMed PMC

Koromani F., Trajanoska K., Rivadeneira F., Oei L. Recent advances in the genetics of fractures in osteoporosis. Front. Endocrinol. 2019;10:337. doi: 10.3389/fendo.2019.00337. PubMed DOI PMC

Kryeziu A.R. Proceedings of the Days of Veterinary Medicine 2012: 3rd International Scientific Meeting. University Ss. Cyril and Methodius in Skopje, Republic of Macedonia; 2012. Housing factors effecting on meat type poultry bone strength. 2-4 September.

Li Y.D., Liu X., Li Z.W., Wang W.J., Li Y.M., Cao Z.P., Luan P., Xiao F., Gao H.H., Guo H.S., Wang N., Li H., Wang S.Z. A combination of genome-wide association study and selection signature analysis dissects the genetic architecture underlying bone traits in chickens. Animal. 2021;15 doi: 10.1016/j.animal.2021.100322. PubMed DOI

Liu K., Wang K., Wang L., Zhou Z. Changes of lipid and bone metabolism in broilers with spontaneous femoral head necrosis. Poultry Sci. 2021;100 doi: 10.1016/j.psj.2020.10.062. PubMed DOI PMC

Liu W., Zhang X. Receptor activator of nuclear factor-κB ligand (RANKL)/RANK/osteoprotegerin system in bone and other tissues. Mol. Med. Rep. 2015;11:3212–3218. doi: 10.3892/mmr.2015.3152. review) PubMed DOI

Maeda K., Kobayashi Y., Koide M., Uehara S., Okamoto M., Ishihara A., Kayama T., Saito M., Marumo K. The regulation of bone metabolism and disorders by Wnt signaling. Int. J. Mol. Sci. 2019;20:5525. doi: 10.3390/ijms20225525. PubMed DOI PMC

Miao J., Wang X., Bao J., Jin S., Chang T., Xia J., Yang L., Zhu B., Xu L., Zhang L., Gao X., Chen Y., Li J., Gao H. Multimarker and rare variants genomewide association studies for bone weight in Simmental cattle. J. Anim. Breed. Genet. 2018;135:159–169. doi: 10.1111/jbg.12326. PubMed DOI

Nan J.H., Yin L.L., Tang Z.S., Xiang T., Ma G.J., Li X.Y., Liu X., Zhao S.H., Liu X.D. Identification of novel variants and candidate genes associated with porcine bone mineral density using genome-wide association study. J. Anim. Sci. 2020;98 doi: 10.1093/jas/skaa052. skaa052. PubMed DOI PMC

Niu Q., Zhang T., Xu L., Wang T., Wang Z., Zhu B., Gao X., Chen Y., Zhang L., Gao H., Li J., Xu L. Identification of candidate variants associated with bone weight using whole genome sequence in beef cattle. Front. Genet. 2021;12 doi: 10.3389/fgene.2021.750746. PubMed DOI PMC

Rafter P., Gormley I.C., Purfield D., Parnell A.C., Naderi S., Berry D.P. Genome-wide association analyses of carcass traits using copy number variants and raw intensity values of single nucleotide polymorphisms in cattle. BMC Genom. 2021;22:757. doi: 10.1186/s12864-021-08075-2. PubMed DOI PMC

Rath N.C., Huff G.R., Huff W.E., Balog J.M. Factors regulating bone maturity and strength in poultry. Poultry Sci. 2000;79:1024–1032. doi: 10.1093/ps/79.7.1024. PubMed DOI

Raymond B., Johansson A.M., Mccormack H.A., Fleming R.H., Schmutz M., Dunn I.Ch, De Koning D.J. Genome-wide association study for bone strength in laying hens. J. Anim. Sci. 2018;96:2525–2535. doi: 10.1093/jas/sky157. PubMed DOI PMC

Regard J.B., Zhong Z., Williams B.O., Yang Y. Wnt signaling in bone development and disease: making stronger bone with Wnts. Cold Spring Harbor Perspect. Biol. 2012;4 https://10.1101/cshperspect.a007997 PubMed DOI PMC

Song Y., Du Z., Yang Q., Ren M., Wang Q., Wang A., Chen G., Zhao H., Yu T., Zhang G. Association of genes variants in RANKL/RANK/OPG signaling pathway with the development of osteonecrosis of the femoral head in Chinese population. Int. J. Med. Sci. 2017;14:690–697. doi: 10.7150/ijms.19124. PubMed DOI PMC

Steinerova M., Horecky C., Knoll A., Nedomova S., Pavlik A. Proceedings of the Mendelnet 2019: 26th International PhD Students Conference. Mendel University; 2019. The search for single nucleotide polymorphisms in genes encoding non-collagenous proteins in bone tissue of laying hens; pp. 490–493. (In Brno, Czech Republic).

Steinerova M., Horecky C., Horecka E., Knoll A., Nedomova S., Pavlik A. Variability of selected genes in relation to the parameters of bones in laying hens: a pilot study. J. Microbiol. Biotechnol. Food Sci. 2019;9:449–452. doi: 10.15414/jmbfs.2019.9.special.449-452. DOI

Steinerova M., Horecky C., Knoll A., Nedomova S., Pavlik A. Proceedings of the Mendelnet 2020: 27th International PhD Students Conference. Mendel University in Brno, Czech Republic; 2020. Study of selected signaling pathways genes that play an important role in bone metabolism in laying hens; pp. 424–429. 10-11 November.

Steinerova M., Horecky C., Horecka E., Knoll A., Nedomova S., Slama P., Pavlik A. Study of selected genes of Wnt signaling pathway in relation to the parameters in the bone tissue of the laying hens. Saudi J. Biol. Sci. 2021;29:2526–2531. https://10.1016/j.sjbs.2021.12.024 PubMed DOI PMC

Ventura M.V.A., da Silva R.M. Bone problems caused by the deficiency of calcium and phosphorus in the feeding of broilers. J. Sci. Tech. Res. 2019;16:12223–12226. doi: 10.26717/BJSTR.2019.16.002886. DOI

Wang Q.F., Bi H.S., Qin Z.L., Wang P., Nie F.F., Zhang G.W. Associations of LRP5 gene with bone mineral density, bone turnover markers, and fractures in the elderly with osteoporosis. Front. Endocrinol. 2020;11 doi: 10.3389/fendo.2020.571549. PubMed DOI PMC

Wang X., Salimi S., Deng Z., Perry J., Ryan K.A., Li Z., Liu D., Streeten E., Shuldiner A.R., Fu M. Evaluation of WISP1 as a candidate gene for bone mineral density in the Old Order Amish. Sci. Rep. 2018;8:7141. doi: 10.1038/s41598-018-25272-4. PubMed DOI PMC

Won S., Jung J., Park E., Kim H. Identification of genes related to intramuscular fat content of pigs using genome-wide association study. Asian-Australas. J. Anim. Sci. 2017;31:157–162. doi: 10.5713/ajas.17.0218. PubMed DOI PMC

Xia Y. Correlation and association analyses in microbiome study integrating multiomics in health and disease. Prog. Mol. Biol. Transl. Sci. 2020;171:309–491. doi: 10.1016/bs.pmbts.2020.04.003. PubMed DOI

Zhu X., Bai W., Zheng H. Twelve years of GWAS discoveries for osteoporosis and related traits: advances, challenges and applications. Bone Res. 2021;9:23. doi: 10.1038/s41413-021-00143-3. PubMed DOI PMC