Sperm Longevity and Motility in Ringer’s Lactate Solution with Addition of Egg Yolk among Five Phenotypes of Kokok Balenggek Chicken


Ananda Ananda(1*), Jaswandi Jaswandi(2), Rusfidra Rusfidra(3), Harif Gusdinal(4)

(1) Department of Livestock Production Technology, Faculty of Animal Science, Universitas Andalas, Padang, 25163
(2) Department of Livestock Production Technology, Faculty of Animal Science, Universitas Andalas, Padang, 25163
(3) Department of Livestock Production Technology, Faculty of Animal Science, Universitas Andalas, Padang, 25163
(4) Postgraduated Program, Faculty of Animal Science, Universitas Andalas, Padang, 25163
(*) Corresponding Author


The application of cryopreservation to preserve germplasm in such specific breed requires preliminary studies, primarily related to the resistance of spermatozoa to low temperatures (4 – 5 °C) as measured by their motility and longevity. In this study, semen taken from five phenotypes of Kokok Balengeek Chicken (KBC) (Biriang, Jalak, Kinantan, Kuriak, and Taduang) was used to evaluate the effect of Ringer’s Lactate-egg yolk diluent on longevity and motility of spermatozoa. The treatments consisted of Ringer’s Lactate (RL) solution added with egg yolk at a concentration of 1% (RLKT1), 3% (RLKT3), and 5% (RLKT5). Evaluation of fresh semen showed that the spermatozoa of Jalak had the highest motility, namely 75.63 ± 0.5% (P<0.05). Post-dilution longevity and motility observations were carried out at 0, 24, 48, and 72 h, significantly decreasing each time (P<0.05). The lowest range of reduction was found in Jalak spermatozoa diluted with RL with longevity of 7.75 ± 0.70 days. Overall, the RL diluent showed the highest motility after 24 h, namely 41.13 ± 2.27%. Adding egg yolks to Ringer's Lactate solution could not maintain the motility of KBC spermatozoa when stored at 4-5°C for 48-72 h.


Kokok Balenggek chicken; semen; egg yolk; ringer’s lactate solution

Full Text:

PDF. Ananda


Al-Ahmad, M. Z. A., G. Chatagnon, L. Amirat-Briand, M. Moussa, D. Tainturier, M. Anton, and F. Fieni. 2008. Use of glutamine and low density lipoproteins isolated from egg yolk to improve buck semen freezing. Reproduction in Domestic Animals 4: 429–436. https://doi.org/10.1111/j.14390531. 2007.00930.x

Almahdi, A. B., Y.S. Ondho, and Sutopo. 2014. Comparative studies of semen quality on different breed of chicken in Poultry Breeding Center Temanggung-Central Java. International Refereed Journal of Engineering and Science (IRJES) 3: 94–103.

Amirat, L., D. Tainturier, L. Jeanneau, C. Thorin, O. Gérard, J. L. Courtens, and M. Anton. 2004. Bull semen in vitro fertility after cryopreservation using egg yolk LDL: A comparison with Optidyl®, a commercial egg yolk extender. Theriogenology 61: 895–907. https://doi.org/10.1016/S0093691X (03)00259-0

Andaruisworo, S. and E. Yuniati. 2021. Teknologi IB pada ayam Hutan Hijau jantan dengan ayam Kampung betina dalam upaya pelestarian plasma nuftah ayam Bekisar. Jurnal Ahli Muda Indonesia 2: 57–62. https://doi.org/10.46510/jami.v2i1.59

Arifiantini, R. I. 2012. Teknik Koleksi dan Evaluasi Semen pada Hewan. Penerbit IPB Press, Bogor.

Barbas, J. P. and R. D. Mascarenhas. 2009. Cryopreservation of domestic animal sperm cells. Cell and Tissue Banking 10: 49–62. https://doi.org/10.1007/s10561-008-9081-4

Bebas, W. and D. N. D. I. Laksmi. 2013. Konsentrasi spermatozoa dan motilitas spermatozoa ayam Hutan Hijau (Gallus varius). Buletin Veteriner Udayana 5: 57–62.

Bilgili, S. F., K. J. Sexton, and J. A. Renden. 1987. Fluorometry of poultry semen: influence of dilution and storage on chicken spermatozoal viability and fertility. Poultry Science 66: 2032–2035. https://doi.org/10.3382/ps. 0662032

Bootwalla, S. M. and R. D. Miles. 1992. Development of diluents for domestic fowl semen. World’s Poultry Science Journal 48: 121–128. https://doi.org/10.1079/ WPS19920012

Bozkurt, T., G. Türk, and S. Gür. 2007. The time-dependent motility and longevity of stallion spermatozoa diluted in different spermatozoal concentrations and extenders during cool-storage. Revue de Medecine Veterinaire 158: 67–72.

Bustani, G. S. and F. H. Baiee. 2021. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Veterinary World 14: 1220–1233. https://doi.org/10.14202/ vetworld.2021.1220-1233

Danang, D. R., N. Isnaini, and P. Trisunuwati. 2012. Pengaruh lama simpan semen terhadap kualitas spermatozoa ayam kampung dalam pengencer ringer’s pada suhu 4°C. Jurnal Ternak Tropika 13: 47–57.

Donoghue, A. M. and G. J. Wishart. 2000. Storage of poultry semen. Animal Reproduction Science 62: 213–232. https://doi.org/10. 1016/S0378-4320(00)00160-3

Fumihito, A., T. Miyake, M. Takada, R. Shingu, T. Endo, T. Gojobori, N. Kondo, and S. Ohno. 1996. Monophyletic origin and unique dispersal patterns of domestic fowls. Proc. Natl. Acad. Sci. USA 93: 6792–6795. https://doi.org/10.1073/pnas.93.13.6792

Gee, G. F., H. Bertschinger, A. M. Donoghue, J. Blanco, and J. Soley. 2004. Reproduction in nondomestic birds: Physiology, semen collection, artificial insemination and cryopreservation. Avian and Poultry Biology Reviews 15: 47–101. https://doi.org/10. 3184/147020604783637435

Hammerstedt, R. H. and J. K. Graham. 1992. Cryopreservation of Poultry Sperm: The Enigma of Glycerol. Cryobiology 29: 26–38. https://doi.org/10.1016/0011-2240(92)90004-L

Hardiyanti, and M. E. Kurniawan. 2020. Pengaruh penambahan sari wortel (Daucus carota L) pada pengencer terhadap kualitas spermatozoa ayam kampung selama penyimpanan. Musamus Journal of Livestock Sci. 3: 12–20.

Helfenstein, F., M. Podevin, and H. Richner. 2010. Sperm morphology, swimming velocity, and longevity in the house sparrow Passer domesticus. Behavioral Ecology and Sociobiology 64: 557–565. https://doi.org/ 10.1007/s00265-009-0871-x

Hidayat, T. 2001. Pengaruh Bahan Pengencer Laktat Ringer Kuning Telur dan Lama Penyimpanan Semen Terhadap Fertilitas Telur Ayam Buras. Institut Pertanian Bogor, Bogor.

Hill, G. E., W. R. Hood, Z. Ge, R. Grinter, C. Greening, J. D. Johnson, N. R. Park, H. A. Taylor, V. A. Andreasen, M. J. Powers, N. M. Justyn, H. A. Parry, A. N. Kavazis, and Y. Zhang. 2019. Plumage redness signals mitochondrial function in the house finch. Proceedings of the Royal Society B: Biological Science 286: 1–9. https://doi. org/10.1098/rspb.2019.1354

Indrawati, D., W. Bebas, and I. Trilaksana. 2013. Motilitas dan daya hidup spermatozoa ayam kampung dengan penambahan astaxanthin pada suhu 3-5°C. Indonesia Medicus Veterinus 2: 445–452.

Jaswandi, Ananda, Rusfidra, K. Subekti, H. Gusdinal, R. S. Wahyuni, and F. A. Caniago. 2023. Fertility rate, fertility period, and DOC sex ratio of Kokok Balenggek chicken after artificial insemination. Advances in Animal and Veterinary Sciences 11: 795–801. https://doi.org/10. 17582/journal.aavs/2023/11.5.795.801

Junaedi, R. I. Arifiantini, C. Sumantri, and A. Gunawan. 2016. Penggunaan dimethyl sulfoxide sebagai krioprotektan dalam pembekuan semen ayam kampung. Jurnal Veteriner 17: 300–308. https://doi.org/10. 19087/jveteriner.2016.17.2.300

Junaedi and Husnaeni. 2019. Kaji banding kualitas semen segar empat genetik ayam lokal Indonesia. Jurnal Veteriner 20: 397–402. https://doi.org/10.19087/jveteriner.2019.20.3.397

Koch, R. E., K. J. Mcgraw, and G. E. Hill. 2016. Effects of diet on plumage coloration and carotenoid deposition in red and yellow domestic canaries (Serinus canaria). Wilson Journal of Ornithology 128: 328–333. https://doi.org/10.1676/wils-128-02-328-333.1

Kusuma, P. W., W. Bebas, and M. K. Budiasa. 2018. Motilitas dan daya hidup spermatozoa ayam Pelung dalam pengencer kuning telur fosfat yang disimpan pada suhu 29°C. Indonesia Medicus Veterinus 7: 115–122. https://doi.org/10.19087/imv.2018.7.2.115

Lake, P. E. 1960. Studies on the dilution and storage of fowl semen. Journal of Reproduction and Fertility 1: 30–35. https://doi.org/10.1530/jrf.0.0010030

Lake, P. E. 1983. Factors affecting the fertility level in poultry, with special reference to artificial insemination. World’s Poultry Science Journal 39: 106–117. https://doi.org/10. 1079/WPS19830011

Lake, P. E. and O. Ravie. 1981. An attempt to improve the fertility of stored fowl semen with certain additives in a basic diluent. Reproduction, Nutrition, Development 21: 1077–1084. https://doi.org/10.1051/r nd:19810806

Long, J. A. 2006. Avian semen cryopreservation: What are the biological challenges? Poultry Science 85: 232–236. https://doi.org/10. 1093/ps/85.2.232

Łukaszewicz, E., M. Chrzanowska, A. Jerysz, and B. Chełmońska. 2004. Attempts on freezing the Greylag (Anser anser L.) gander semen. Animal Reproduction Science 80: 163–173. https://doi.org/10.1016/S0378-4320(03) 00119-2

Magfira, R. I. Arifiantini, N. W. K. Karja, and S. Darwati. 2017. Efektivitas low density lipoprotein dan kuning telur ayam dan puyuh pada pengawetan semen ayam Merawang. Jurnal Veteriner 18: 345. https://doi.org/10.19087/jveteriner.2017.18.3.345

Manjunath, P. and I. Thérien. 2002. Role of seminal plasma phospholipid-binding proteins in sperm membrane lipid modification that occurs during capacitation. Journal of Reproductive Immunology 53: 109–119. https://doi.org/10.1016/S0165-0378(01)00098-5

Moussa, M., V. Martinet, A. Trimeche, D. Tainturier, and M. Anton.2002. Low density lipoproteins extracted from hen egg yolk by an easy method: Cryoprotective effect on frozen-thawed bull semen. Theriogenology, 57: 1695–1706. https://doi.org/10.1016/ S0093-691X(02)00682-9

Mphaphathi, M. L., M. M. Seshoka, D. Luseba, B. Sutherland, and T. Nedambale. 2016. The characterisation and cryopreservation of Venda chicken semen. Asian Pacific Journal of Reproduction 5: 132–139. https://doi.org/10.1016/j.apjr.2016.01.009

Muryanto and D. Pramono. 2014. Sukses Budi Daya Ayam Kampung. B. Sudaryanto & Y. Winarti (Eds.), LOKA AKSARA.

Rochmi, S. E. and M. S. Sofyan. 2019. A diluent containing coconut water, fructose, and chicken egg yolk increases rooster sperm quality at 5°C. Veterinary World 12: 1116–1120. https://doi.org/10.14202/vetworld. 2019.1116-1120

Rusfidra. 2004. Karakteristik Sifat-sifat Fenotipik Sebagai Strategi Awal Konservasi Ayam Kokok Balenggek di Sumatera Barat. Institut Pertanian Bogor.

Rusfidra, Y. Y. Tumatra, M. H. Abbas, Y. Heriyandi, and F. Arlina. 012. Identifikasi marka bioakustik suara kokok ayam Kokok Balenggek di Kandang Penangkaran “Agutalok”, Kabupaten Solok. Jurnal Peternakan Indonesia 14: 303–307.

Santiago-Moreno, J., B. Bernal, S. Pérez-Cerezales, C. Castaño, A. Toledano-Díaz, M. C. Esteso, A. Gutiérrez-Adán, A. López-Sebastián, M. G. Gil, H. Woelders, and E. Blesbois. 2019. Seminal plasma amino acid profile in different breeds of chicken: Role of seminal plasma on sperm cryoresistance. PLoS ONE 14: 1–19. https://doi.org/10. 1371/journal.pone.0209910

Sexton, T. J. 1978. A new poultry semen extender 3. Effect of storage conditions on the fertilizing capacity of chicken semen stored at 5 degrees C. Poultry Science 57: 285–289. https://doi.org/10.3382/ps.0570285

Siudzińska, A. and E. Łukaszewicz. 2008. Effect of semen extenders and storage time on sperm morphology of four chicken breeds. Journal of Applied Poultry Research 17: 101–108. https://doi.org/10.3382/japr.2007-00048

Solihati, N., R. Idi, R. Setiawan, I. Asmara, and B. Sujana. 2006. Pengaruh lama penyimpanan semen cair ayam buras pada suhu 5°c terhadap periode fertil dan fertilitas sperma. JIT 6: 11–17. http://zsp.com.pk/pdf46/ Contents 46 (1) 2014.pdf

Suprijatna, E., U. Atmomarsono, and R. Kartasudjana. 2005. Ilmu Dasar Ternak Unggas (Cetakan Ke). Penebar Swadaya.

Tarvis, K. M. 2013. New Methods for Cryopreserving Rooster Spermatozoa. Colorado State University.

Telnoni, S. P., R. I. Arifiantin, T. L. Yusuf, and S. Darwati. 2017. SK Kedu semen cryopreservation in Beltsville Poultry semen extender and lactated ringer’s-egg yolk extender using dimethyl sulfoxide. Asian Journal of Poultry Science 11: 14–19. https://doi.org/10.3923/ ajpsaj.2017.14.19

Triques, G. E., A. B. De Cristo, M. Canevese, P. F. Dos Santos Marques, A. M. Burin, and J. I. M. Fernandes. 2019. Effect of antioxidant supplementation in diets of roosters during the post-peak phase on reproduction and production characteristics of offspring. Ciencia Animal Brasileira 20: 1–13. https://doi.org/10.1590/1089-6891v20e-43072

Vera-Munoz, O., L. Amirat-Briand, D. Bencharif, M. Anton, S. Desherces, E. Shmitt, C. Thorin, and D. Tainturier. 2011. Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4°C. Asian Journal of Andrology 13: 281–286. https://doi.org/10.1038/aja. 2010.84

Watson, P. F. 2000. The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science 60: 481–492. https://doi.org/10. 1016/S0378-4320(00)00099-3

Wicaksono, A. and R. I. Arifiantini. 2009. Uji Banding empat bahan pengencer untuk preservasi semen anjing retriever. JITV 14: 50–57.

Widiastuti, W. A., W. Bebas, and I. G. N. B. Trilaksana. 2018. Penggunaan berbagai kuning telur sebagai bahan pengencer terhadap motilitas dan daya hidup spermatozoa ayam Pelung. Indonesia Medicus Veterinus 7: 251. https://doi.org/ 10.19087/imv.2018. 7.3.251

Wishart, G. J. and H. J. Staines. 1999. Measuring sperm: Egg interaction to assess breeding efficiency in chickens and turkeys. Poultry Science 78: 428–436. https://doi.org/10. 1093/ps/78.3.428

Zong, Y., Y. Li, Y. Sun, G. M. K. Mehaisen, T. Ma, and J. Chen. 2023. Chicken sperm cryopreservation: review of techniques, freezing damage, and freezability mechanisms. Agriculture (Switzerland) 13: 1–16. https://doi.org/10.3390/agriculture 13020445

DOI: https://doi.org/10.21059/buletinpeternak.v47i3.83647

Article Metrics

Abstract views : 802 | views : 577


  • There are currently no refbacks.

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Buletin Peternakan (Bulletin of Animal Science) Indexed by:

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.