Viabilitas dan Fragmentasi DNA Spermatozoa pada Semen Beku Ayam Kampung Menggunakan Pengencer Ringer Laktat Kuning Telur yang Ditambahkan Glisin dan Fruktosa
Article Sidebar
Fruktosa,
Glisin,
Kriopreservasi semen ayam,
Viabilitas
Main Article Content
Abstract
Penelitian ini bertujuan untuk mengetahui pengaruh penambahan glisin dan fruktosa pada ringer laktat kuning telur terhadap viabilitas dan fragmentasi DNA spermatozoa beku ayam kampung. Penelitian ini menggunakan rancangan acak lengkap dengan 4 perlakuan: P0 (ringer laktat + kuning telur), P1 (ringer laktat + kuning telur + glisin 60 mM), P2 (ringer laktat + kuning telur + fruktosa 80 mM), dan P3 (ringer laktat + kuning telur + glisin 60 mM + fruktosa 80 mM), masing-masing dengan 6 ulangan. Semen diekuilibrasi 2 jam pada suhu 5 °C, prefreezing dengan ketinggian 3 cm pada uap nitrogen selama 10 menit, penyimpanan 24 jam dan thawing pada suhu 37 °C selama 30 detik. Evaluasi dilakukan terhadap viabilitas spermatozoa menggunakan pewarnaan eosin nigrosin serta integritas DNA dengan pewarnaan toluidine blue. Hasil penelitian menunjukkan bahwa perlakuan berpengaruh (P<0,05) terhadap viabilitas post thawing namun tidak berpengaruh (P>0,05) terhadap fragmentasi DNA post thawing. Penambahan glisin mampu mempertahankan viabilitas spermatozoa selama pembekuan, sedangkan penambahan fruktosa tidak memberikan pengaruh. Viabilitas spermatozoa post thawing dapat dipertahankan hingga 46.57-77,12%, sedangkan kerusakan DNA yang terjadi setelah pembekuan mengalami peningkatan sebesar 6,51-9,61% Penggunaan glisin 60 mM memberikan hasil terbaik dalam mempertahankan viabilitas spermatozoa ayam kampung selama pembekuan.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Abou El-Ela, A., Kandeel, A., Hassan, E., Nasr, M., & Esam, Y. (2022). Utility of magnetic activated cell sorting (MACS) in assisted reproduction. Al-Azhar International Medical Journal, 3(11), 22-29.
Abruzzese, G. A., Sanchez‐Rodriguez, A., & Roldan, E. R. (2024). Sperm metabolism. Molecular Reproduction and Development, 91(10), e23772.
Agarwal, A., Gupta, S., & Sharma, R. (2016). Eosin-nigrosin staining procedure. In: Agarwal, A., Gupta, S., & Sharma, R., editors. Andrological Evaluation of Male Infertility: A Laboratory Guide. Switzerland: Springer.
Blesbois, E., Grasseau, I. & Seigneurin, F., (2005). Membrane fluidity and the ability of domestic bird spermatozoa to survive cryopreservation. Reproduction, 129(3), 371-378.
Bustani, G. S., & Baiee, F. H. (2021). Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Veterinary World, 14(5), 1220.
Darni, D., Saili, T., & Rahadi, S. (2021). Kualitas spermatozoa ayam kampung dengan penambahan vitamin E dalam pakan. Jurnal Ilmiah Peternakan Halu Oleo, 3(1), 19-22.
Davoodian, N., Kadivar, A., Ahmadi, E., & Mohebbi, A. (2017). Effects of two amino acids on motion parameters and enzymatic antioxidant activity of freeze-thawed stallion spermatozoa. Journal of Equine Veterinary Science, 59, 49-56.
Di Santo, M., Tarozzi, N., Nadalini, M., & Borini, A. (2012). Human sperm cryopreservation: update on techniques, effect on DNA integrity, and implications for ART. Advances in Urology, 2012(1), 854837.
Duplaix, M., & Sexton, T. J. (1983). Effects of prefreeze treatments on the fertilizing capacity of unfrozen and frozen chicken semen: extender characteristics and dilution method. Poultry Science, 62(11), 2255-2260.
Echeverría, C. E., Oyarzún, V. I., López‐Cortés, A., Cancino, J., Sotomayor, P. C., Goncalves, M. D., & Godoy, A. S. (2024). Biological role of fructose in the male reproductive system: Potential implications for prostate cancer. The Prostate, 84(1), 8-24.
El-Sheshtawy, R. I., El-Sisy, G. A., & El-Nattat, W. S. (2008). Use of selected amino acids to improve buffalo bull semen cryopreservation. Global Veterinaria, 2, 146-150.
Giambelluca, M. S., & Gende, O. A. (2009). Effect of glycine on the release of reactive oxygen species in human neutrophils. International Immunopharmacology, 9(1), 32-37.
Gliozzi, T. M., Zaniboni, L., & Cerolini, S. (2011). DNA fragmentation in chicken spermatozoa during cryopreservation. Theriogenology, 75(9), 1613-1622.
Hai, E., Li, B., Zhang, J., & Zhang, J. (2024). Sperm freezing damage: the role of regulated cell death. Cell Death Discovery, 10(1), 239.
Haryuni, N., Lestariningsih, L., & Khopsoh, B. (2022). Perbaikan kualitas semen ayam kampung melalui peningkatan energi metabolisme pakan. Journal of Science Nusantara, 2(3), 123-129.
Isnaeni, M., Faidiban, O. R., & Tethool, A. N. (2019). Konsentrasi dan motilitas spermatozoa ayam kampung (Gallus domesticus) dalam pengencer ringer laktat yang diberi tambahan minyak buah merah (Pandanus conoideus Lam). Jurnal Ilmu Peternakan dan Veteriner Tropis, 9(2), 44-49.
Junaedi, J., Isnaini, N., Natsir, M. H., & Susilawati, T. (2024a). Supplementation of glycine and glucose into egg yolk lactated Ringer diluent on the quality of local chicken semen stored at 5 °C for 120 Hours. Jurnal Medik Veteriner, 7(1).
Junaedi, J., Isnaini, N., Natsir, M. H., & Susilawati, T. (2024b). The quality of cryopreserved semen of local chickens treated with Ringer’s lactate-egg yolk extender supplemented with glycine and glucose. Tropical Animal Science Journal, 47(2), 161.
Junaedi, J., Sumantri, C., & Gunawan, A. (2016). Penggunaan dimethyl sulfoxide sebagai krioprotektan dalam pembekuan semen ayam kampung. Jurnal Veteriner, 17(2), 300-308.
Khaeruddin, A. N., Ardi, N., Fattah, A. H., & Armayanti, A. K. (2020a). Penentuan konsentrasi susu skim terbaik dalam pengencer semen ayam kampung berbahan dasar Ringer laktat. Jurnal Veteriner, 21(2), 300-308.
Khaeruddin, K., Arismunandar, A., & Nurda, N. (2020b). Karakteristik semen ayam kampung yang diberi minyak hati ikan kod sebagai feed suplement. Musamus Journal of Livestock Science, 3(1), 15-24.
Khaeruddin, K., Ciptadi, G., Yusuf, M., & Wahjuningsih, S. (2024). Effect of addition of glucose or fructose in three types of crystalloid fluid on spermatozoa quality of Gaga' chicken from South Sulawesi, Indonesia. Pakistan Journal of Zoology, 56(5), 2441-2448.
Khaeruddin, K., Hermawansyah, H., Syamsuryadi, B., Armayanti, A. K., Fattah, A. H., Mangalisu, A., Kurniawan, M. E., Yustisia, D., Alamsyah, R., & Pardi, P. (2026). Practical use of amino acids in diluent to sustain native chicken sperm quality during 5 °C storage. Journal of Animal Health and Production, 14(2), 670-677.
Khaeruddin, K., Junaedi, J., & Hastuti, H. (2020c). Cryopreservation of Indonesian native chicken semen by using dimethyl sulfoxide and various level of ethylene glycol as cryoprotectants. Biodiversitas, 21(12), 5718-5722.
Khaeruddin, K. (2020). Pembekuan spermatozoa ayam kampung dengan suplementasi bovine serum albumin dan putih telur dalam pengencer ringer laktat kuning telur. Ternak Tropika Journal of Tropical Animal Production, 21(2), 111-222.
Khaeruddin, K., Sumantri, C., Darwati, S., & Arifiantini, R. I. (2015). Penggunaan minyak zaitun ekstra virgin ke dalam bahan pengencer semen terhadap kualitas spermatozoa ayam lokal. Jurnal Ilmu Produksi dan Teknologi Hasil Peternakan, 3(1), 46-51.
Khaeruddin, K., Syamsuryadi, B., Fattah, A. H., & Hermawansyah, H. (2025). Teknologi Reproduksi Ternak Jantan. Jambi: PT. Sonpedia Publishing Indonesia.
Khairuddin, K., Kurniawan, M. E., & Soman, S. (2019). Cryopreservation of Kampung rooster semen using egg yolk diluent from four types of poultry with different concentrations. Jurnal Kedokteran Hewan, 13(3), 60-65.
Koohestanidehaghi, Y., Torkamanpari, M., Shirmohamadi, Z., Lorian, K., & Vatankhah, M. (2021). The effect of cysteine and glutamine on human sperm functional parameters during vitrification. Andrologia, 53(1), e13870.
Kusumawati, E. D, Krisnaningsih, A. T. N, & Walangara, A. U. K. (2020). Kualitas spermatozoa ayam kampung dan ayam Arab dengan lama simpan yang berbeda pada suhu ruang. Jurnal Sains Peternakan , 8 (1), 41-56.
Lake, P. E. (1957). Fowl semen as collected by the massage method. The Journal of Agricultural Science, 49(1), 120-126.
Long, J. A., (2014). Applied andrology in chickens and turkeys. In: Chenoweth, P. J., & Lorton, S. P. (Eds). Animal Andrology. Theories and Applications. Boston MA, USA: CABI.
Magfira, M., Karja, N. W. K., Arifiantini, R. I. & Sartika, T. (2023). Effect of carbohydrate type and phenotype on the quality of post-thawing frozen semen of KUB chicken. Jurnal Ilmu Ternak dan Veteriner, 28(1), 56-78.
Moussa, M., Martinet, V., Trimeche, A., Tainturier, D., & Anton, M. (2002). Low density lipoproteins extracted from hen egg yolk by an easy method: cryoprotective effect on frozen–thawed bull semen. Theriogenology, 57(6), 1695-1706.
Mussa, N. J., Rtchamak, R., Ratsiri, T., Vongpralub, T., Boonkum, W., Semaming, Y. & Chankitisakul, V. (2021). Lipid profile of sperm cells in thai native and commercial roosters and its impact cryopreserved semen quality. Tropical Animal Health and Production, 53,1-9.
Nasr-Esfahani, M. & Tavalaee, M. (2021). Sperm Chromatin Structure: Toluidine Blue Staining. In: A. Agarwal, R. Henkel, and A. Majzoub (Eds.) Manual of Sperm Function Testing in Human Assisted Reproduction. England: Cambridge University Press.
Nazif, M. S., Rehman, Z. U., Khan, H., Khan, F. A., Hussain, T., Ahmad, A., Farmanullah, F., Husnain, A., Muhammad, S., Murtaza, G. & Gang, L. (2022). Glycine improved cryopreserved spermatozoa quality in Achai bull. BioMed Research International, 2022(1), 8282387.
Ni, W., Xiao, S., Qiu, X., Jin, J., Pan, C., Li, Y., Fei, Q., Yang, X., Zhang, L., & Huang, X. (2014). Effect of Sperm DNA fragmentation on clinical outcome of frozen-thawed embryo transfer and on blastocyst formation. PLoS ONE, 9, e94956.
Pimprasert, M., Kheawkanha, T., Boonkum, W., & Chankitisakul, V. (2023). Influence of semen collection frequency and seasonal variations on fresh and frozen semen quality in Thai native roosters. Animals, 13(4), 573.
Robbaani, M., Khaeruddin, K., Rachmawati, A., Nursita, I. W., & Wahjuningsih, S. (2025). The use of ethylene glycol cryoprotectant with varying concentrations on the integrity of the plasma membrane and DNA damage in Gaga chickens semen. The 11th International Conference of Innovation in Animal Science (ICIAS 2024). pp. 95-103, Atlantis Press.
Rui, B. R., Angrimani, D. S., Losano, J. D. A., de Cássia Bicudo, L., Nichi, M., & Pereira, R. J. (2017). Validation of simple and cost-effective stains to assess acrosomal status, DNA damage and mitochondrial activity in rooster spermatozoa. Animal Reproduction Science, 187, 133-140.
Ruiz, E. D., Bermejo, J. V. D., González, F. J. N, Jurado, J. M. L., & Ariza, A. G. (2025). Packaging with lower sperm concentrations improves post-thawing quality parameters of rooster semen. Italian Journal of Animal Science, 24(1), 89-100.
Salam, L.O.M.A., Zulkaranin, L.O. M. I., Ramadhani, R., Junaedi, J., & Khaeruddin, K. (2025). Motility and viability of kampung chicken sperm using diluents containing glycine and glucose. Tarjih Tropical Livestock Journal, 5(2), 166-175.
Sangeeta, S., Arangasamy, A., Kulkarni, S., & Selvaraju, S. (2015). Role of amino acids as additives on sperm motility, plasma membrane integrity and lipid peroxidation levels at pre-freeze and post-thawed ram semen. Animal Reproduction Science, 161, 82-88.
Sharma, R., & Agarwal, A. (2021). Sperm vitality: Eosin-nigrosin dye exclusion. In: Agarwal, A., Henkel, R., & Majzoub, A. (Eds), Manual of Sperm Function Testing in Human Assisted Reproduction. England: Cambridge University Press.
Sun, Y., Xue, F., Li, Y., Fu, L., Bai, H., Ma, H., Xu, S., & Chen, J. (2019). Differences in semen quality, testicular histomorphology, fertility, reproductive hormone levels, and expression of candidate genes according to sperm motility in Beijing-You chickens. Poultry Science, 98(9), 4182-4189.
Wahjuningsih, S., Arif, A. A., Khaerudin, P. H., & Putri, A. R. I. (2024). The effects of equilibration time and post-thawing temperatures in cryopreservation of Gaga chicken semen. Advanced in Animal and Veterinary Sciences, 12(5), 807-814.
Wahjuningsih, S., Khaeruddin, K., Rachmawati, A., Tribudi, Y. A., Sari, A. P. Z. N. L., Wahju Nursita, I., & Iustitia Sitaresmi, P. (2026). Pioneering comparative analysis of intracellular cryoprotectants on Gaga chicken sperm quality during cryopreservation. Journal of Applied Animal Research, 54(1), 2628620.
Wang, Q., Gu, X., Chen, Y., Yu, M., Peng, L., Zhong, S., Wang, X., & Lv, J. (2023). The effect of sperm DNA fragmentation on in vitro fertilization outcomes of unexplained infertility. Clinics, 78, 100261.
Widyasanti, N. W. H., Suriansyah, S., Yaddi, Y., Archadiya, M., Khaeruddin, K., Yusuf, R., Magfira, M., Aldiyanti, A., Yunitasari, F., Sulfitrana, A., Yuska, D.A.T & Pancar, F. M. (2025). Reproduksi Ternak Unggas. Padang: Azzia Karya Bersama.
Wijaya, G. H., Fatmarischa, N., Falah, F. D. S., Salsabila, N., Yusuf, R., Toelle, N. N., Khaeruddin, K., Adam, K. A., Herlambang, N., Rizkuna, A., & Djunaidi, I. H. (2026). Ilmu dan Teknologi Ternak Unggas. Padang: Azzia Karya Bersama.
Zong, Y., Li, Y., Sun, Y., Mehaisen, G. M., Ma, T., & Chen, J. (2023). Chicken sperm cryopreservation: review of techniques, freezing damage, and freezability mechanisms. Agriculture, 13(2), 445.