The The Efectiveness of Jamblang (Syzygium cumini) Leaves Extract Inclusion Skim Milk-Egg Yolk Extender on Motility and Viability of Aceh Cattle Spermatozoa during Pre-Freezing and Post-Thawing

  • Cut Intan Novita Department of Animal Science, Faculty of Agriculture, Universitas Syiah Kuala, Darussalam, Banda Aceh 23111, Indonesia, and Research Centre of Aceh Cattle and Local Animal, USK, Indonesia
  • Faqihuddin Nasution Department of Animal Science, Faculty of Agriculture, Universitas Syiah Kuala, and Research Centre of Aceh Cattle and Local Animal, Indonesia
  • Eka Meutia Sari Department of Animal Science, Faculty of Agriculture, Universitas Syiah Kuala, and Research Centre of Aceh Cattle and Local Animal, Indonesia
Keywords: Spermatozoa of Aceh Cattle, Jamblang (Syzygium cumini) leaves extract, motility, viability, pre-freezing, post-thawing


The process of semen freezing causes an increase in free radicals concentration which can damage spermatozoa. The addition of natural ingredients in semen diluent is expected to solve this challenges. One of the natural ingredients that can be used is jamblang (Syzygium cumini) leaves. The objective of the current study was to investigate the quality of spermatozoa in Aceh cattle which was added with jamblang leaves extract in skim milk- egg yolk extender during pre-freezing and post-thawing. This study applied Completely Randomized Design (CRD) with 5 treatments and 5 replications. The treatments consisted of J0 = skim milk-egg yolk; J1 = skim milk-egg yolk + jamblang leaves extract 0.2%; J2 = skim milk-egg yolk + jamblang leaves extract 0.4%; J3 = skim milk-egg yolk + jamblang leaves extract 0.6%; and J4 = skim milk-egg yolk + jamblang leaves extract 0.8%. The parameters observed in this study were the percentage of motility and viability of frozen semen of Aceh cattle. The data obtained were analyzed using Analysis of Variance (ANOVA) and if differences were found, then it would be continued with Duncan's Multiple Distance test. The results showed that the addition of jamblang leaves extract in egg yolk skim milk significantly affected the percentage of motility during pre-freezing and post-thawing, significantly affected spermatozoa viability during pre-freezing and significantly affected the spermatozoa viability during post-thawing. J3 treatment (jamblang leaves extract 0.6 gram/100 ml) it should be higher than the other treatment, where the percentage of motility at pre-freezing and post-thawing were 55.48% and 52.71%, respectively, and the percentage of viability during pre-freezing and post-thawing were 56.59% and 53.94%, respectively. It was concluded that the addition of jamblang leaves extract in the skim milk-egg yolk extender affected the percentage of spermatozoa motility and viability of Aceh cattle during pre-freezing and post-thawing.


Aboagla, E. M. E., & Terada, T. (2004). Effects of egg yolk during the freezing step of cryopreservation on the viability of goat spermatozoa. Theriogenology, 62(6), 1160-1172.
Affandhy, L., Adinata, Y., & Aryogi. (2013). Evaluasi Kualitas Semen Pejantan Sapi PO Kebumen Di Kabupaten Kebumen Provinsi Jawa Tengah. Jurnal Sains Dasar, 2(2).
Alcay, S. E., Gokce, M. B., Toker, N. T., Onder, B., Ustuner, E., Uzabaci, Z. Gul, & Cavus, S. (2016). Freeze-dried egg yolk based extenders containing various antioxidants improve post-thawing quality and incubation resilience of goat spermatozoa. Cryobiology, 72(3), 269-273.
Amirat, L., D. Tainturier, L. Jeanneau, C. Thorin, O., Gerard, J. L., Courtens, M., & Anton. (2004). Bull semen in vitro fertility after cryopreservation using egg yolk LDL: a comparison with optidyl, a commercial egg yolk extender. Theryogenology, 61, 895-907.
Arifiantini, R. I., Yusuf, T. L., & Yanti, D. (2005). Kaji Banding Kualitas semen beku sapi Friesian Holstein emnggunakan pengencer dari berbagai Balai Inseminasi Buatan di Indonesia. Animal Production, 7(3), 168-176.
Asadpour, R., Jafari, R., & Tayefi-Nasrabadi, H. (2012). The effect of antioxidant supplementation in semen extenders on semen quality and lipid peroxidation of chilled bull spermatozoa. Iranian Journal of Veterinary Research, Shiraz University, 13(3), Ser. No. 40, 246-249.
Basavaraja, N., & Hedge, S. N. (2004). Cryopreservation of the endangered mahseer (Tor khudree) spermatozoa: I. Effect of extender composition, cryoprotectants, dilution ratio, and storage period on post-thaw viability. Cryobiology, 49(2), 149-156.
Baumber J., Ball, B. A., & Linfor, J. (2005). Assesment of the cryopreservation of equine spermatozoa in the presence of enziyme scavengers and antioxidants. Am J Vet Red, 66, 772-779.
Baust, J. G., Gao, D., & Baust, J. M. (2009). Cryopreservation: An emerging paradigm change. Review Series. Organogenesis, 5(3), 90-96.
Boujenane, I., & Boussaq, K. (2013). Environmental effects and repeatability estimates for sperm production and semen quality of Holstein bulls. Arch. Anim. Breed., 56, 971-979.
Brito, L. F. C., Silva, A. E. D. F., Rodrigues, L. H., Vieira, F. V., Deragon, L. A. G., & Kastelic, J. P. (2002). Effects of environmental factors, age and genotype on sperm production and semen quality in Bos indicus and Bos taurus AI bulls in Brazil. Animal Reproduction Science, 70(3-4), 181-190.
Bucak, M. N., Atessahin, A., & Yuce, A. (2008). Effect of anti-oxidants and oxidative stress parameters on ram semen after the freeze-thawing process. Small Ruminant Research, 75(2-3), 128-134.
Buffone, M. G., Calamera, J. C., Brugo-Olmedo, S., De Vincentiis, S., Calamera, M. M., Storey, B. T., Doncel, G. F., & Alvarez, J. G. (2012). Superoxide dismutase content in sperm correlates with motility recovery after thawing of cryopreserved human spermatozoa. Fertility and Sterility, 97(2), 293-298.
Chatterjee, S., & Gagnon, C. (2001). Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Moleculer Reproduction and Development, 59(4), 451-458.
Dewi, A. S., Ondho, Y. S., & Kurnianto, E. (2012). Kualitas semen berdasarkaan umur pada sapi jantan jawa. Indian Vet J., 63, 909-912.
Ezzati, M., Shanehbandi, D., Hamdi, K., Rahbar, S., & Pashaiasl, M. (2020). Influence of cryopreservation on structure and function of mammalian spermatozoa: an overview. Cell and Tissue Banking, 21, 1-15.
Feradis. (2009). Peranan antioksidan dalam pembekuan semen. Jurnal Peternakan, 6(2), 63-70.
Ferramosca, A., & V. Zara. (2014). Bioenergetics of mammalian sperm capacitation. Review Article. Biomed Research International, 2014, 1-8.
Fraser, L., & Strzezek, J. (2005). Effects of freezing-Thawing on DNA integrity of boar spermatozoa assessed by neutral comet assay. Reproduction in Domestic Animals, 40(6), 530-536.
Fuerst-Waltl, B., Schwarzenbacher, H., Perner, C., & Solkner, J. (2006). Effects of age and environmental factors on semen production and semen quality of Austrian Simmental bulls. Animal Reproduction Science, 95(1-2), 27-37.
Garcia J. C., Dominguez, J. C., Pena, F. J., Alegre, B., Gonzalez, R., Castro, M. J., Habing, G. G., & Kirkwood, R. N. (2010). Thawing boar semen in the presence of seminal plasma: Effects on Sperm quality and fertility. Animal Reproduction Science, 119, 160-165.
Gillan, L., Maxwell, W. M. C., & Evans, G. (2004). Preservation and evaluation of semen for artificial insemination. Reproduction, Fertility and Development, 16(4), 447-454.
Gowri, S. S., & Vasantha, K. (2010). Phytochemical screening and antibacterial activity of Syzygium cumini (L.) (Myrtaceae) Leaves Extracts. PhamTech Res., 2(2), 1569-1573.
Hafez, E. S. E. (2004). Reproduction in farm animals. 7th edition. Lea and Febiger. Philadelphia.
Hanifi, H., Ihsan, M. N., & Susilawati, T. (2016). Pengaruh Lama Ekuilibrasi pada Proses Pembekuan terhadap Kualitas Semen Sapi Wagyu menggunakan Pengencer Andromed®. J. Ternak Tropika, 17(1), 31-41.
Holden, S. A., Fernandez-Fuertes, B., Murphy, C., Whelan, H., O’Gorman, A., Brennan, L., … S. Fair. (2017). Relationship between in vitro sperm functional assessments, seminal plasma composition, and field fertility after AI with either non-sorted or sex-sorted bull semen. Theriogenology, 87, 221-228.
Januskauskas, A., Johannison, A., & Rodriguez-Martinez. H. (2003). Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure, and field fertility. Theriogenology, 60(4), 743-758.
Khan, M. I. R., & Ijaz, A. (2008). Effects of osmotic pressure on motility, plasma membrane integrity and viability in fresh and frozen-thawed buffalo spermatozoa. Animal, 2(4), 548-553.
Komariah, I., Arifiantini, & Nugraha, F. W. (2013). Kaji banding kualitas spermatozoa sapi simmental, limousine, dan Fresien holstein terhadap proses pembekuan. Buletin Peternakan, 37(3), 143-147.
Kostaman, T., & Sutama, I. K. (2006). Studi motilitas dan daya hidup spermatozoa kambing boer pada pengencer tris-sitrat-fruktosa. J. Sain Vet., 24(1), 56-63.
Kumar, R., Mohanarao, G. J., Arvind & Atreja, S. K. (2011). Freeze–thaw induced genotoxicity in buffalo (Bubalus bubalis) spermatozoa in relation to total antioxidant status. Molecular Biology Reports, 38, 1499-1506.
Kuswahyuni, I. S. (2009). Pengaruh lingkar skrotum dan volume testis terhadap volume semen dan konsentrasi sperma jantan simmental, limousin, dan brahman. Seminar Nasional Teknologi Peternakan dan Veteriner. Bandung.
Leao A. P. A., A. V. de Souza, N. F. Mesquita, L. J. Pereira, & Zangeronimo, M. G. (2021). Antioxidant enrichment of roster semen extender – A systematic review. Research in Veterinary Science, 136, 111-118.
Lemma, A. (2011). Effect of cryopreservation on sperm quality and fertility Artificial Insemination in Farm Animals. IntechOpen, 191-215.
Madyawati, S. P., Srianto, P., Dewi, C. P., Ismudiono, & Sianita, N. (2011). Pengaruh penambahan tirosin kinase dalam bahan pengencer skim kuning telur dan Tris kuning telur terhadap persentase motilitas dan spermatozoa hidup sap Friesian Holstein post-thawing. J. Veterina Medika, 4(2), 87-90.
Mansour, N., Lahnsteiner, F., & Patzner, R. A. (2010). Motility and cryopreservation of spermatozoa of European common frog. Rana temporaria. Theriogenologi, 74, 724-732.
Margaret, E., Shailaja, A. M., & Rao, V. (2015). Evaluation of Antioxidant Activity in Different Parts of Syzygium cumini (Linn.). International Journal of Current Microbiology and Applied Sciences, 4(9), 372-379.
Mohamed, A. A., Ali, S. I., & El-Baz. F. K. (2013). Antioxidant and antibacterial activities of crude extracts and essential oils of Syzygium cumini leaves. PLoS ONE, 8(4), e60269. 10st August 2021
Nagy, S., Hallap, T., Johannisson, A., & Rodriguez-Martinez, H. (2004). Changes in plasma membrane and acrosome integrity of frozen-thawed bovine spermatozoa during a 4 h incubation as measured by multicolor flow cytometry. Animal Reproduction Science, 80(3-4), 225-235.
Nugraha, C. D., Herwijanti, E., Novianti, I., Furqon, A., Septian, W. A., Busono, W., & Suyadi, S. (2019). Correlations between age of Bali bull and semen production at National Artificial Insemination Center, Singosari – Indonesia. J. Indonesian Trop. Anim. Agric., 44(3), 258-265.
Ozkavukcu, S., Erdemli, E., Isik, A., Oztuna, D., & Karahuseyinoglu, S. (2008). Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. Journal of Assisted Reproduction and Genetics, 25, 403-411.
Pagl, R., Aurich, J. E., Muller-Schlosser, F., Kankofer, M., & Aurich, C. (2006). Comparison of an extender containing defined milk protein fractions with a skim milk-based extender for storage of equine semen at 5 °C. Theriogenology, 66(50), 1115-1122.
Pesch, S., & Bergmann, M. (2006). Structure of mammalian spermatozoa in respect to viability, fertility and cryopreservation. Micron, 37(7), 597-612.
Petyim, S., Neungton, C., Thanaboonyawat, I., Laokirkkiat, P., & Choavaratana, R. (2014). Sperm preparation before freezing improves sperm motility and reduces apoptosis in post-freezing-thawing sperm compared with post-thawing sperm preparation. Journal of Assisted Reproduction and Genetics, 31, 1673-1680.
Pratiwi, R. I., Suharyati, S., & Hartono, M. (2014). Analisis kualitas semen beku sapi simmental menggunakan pengencer Andromed® dengan variasi waktu pre-freezing. Jurnal Ilmiah Peternakan Terpadu, 2(3), 8-15.
Pratiwi, W. C., Affandy, L., & Ratnawati, D. (2006). Pengaruh Lama Thawing terhadap Kualitas Semen Beku Sapi Limousin dan Brahman. Animal Production, 11(1), 48-52.
Rasul, Z., Ahmad, N., & Anzar, M. (2001). Changes in motion characteristic, plasma membrane integrity, and acrosome morphology during cryopreservation of buffalo spermatozoa. Journal of Andrology, 22(2), 278-283.
Ratnawati, D., Affandhy, L., Pratiwi, W. C., & Prihandini, P. W. (2008). Pengaruh pemberian suplemen tradisional terhadap kualitas semen pejantan sapi bali. Loka Penelitian Sapi Potong. Semarang.
Rodriguez-Martinez, H., Saravia, F., Wallgren, M., Roca, J., & Pena, F. J. (2008). Influence of seminal plasma on the kinematics of boar spermatozoa during freezing. Theriogenology, 70(8), 1242-1250.
Ruan, Z. P., Zhang, L. L., & Lin, Y. M. (2008). Evaluation of the antioxidant activity of Syzygium cumini leaves. Molecules, 13(10), 2545-2556.
Samsudewa, D., & Suryawijaya, A. (2008). Pengaruh Berbagai Methode Thawing terhadap Kualitas Semen Beku Sapi. Seminar Nasional Tekhnologi Peternakan dan Veteriner. Fakultas Peternakan Universitas Dipenogoro. Semarang.
Santoso, H., Arifiantini, R. I., Gunawan, A., & Sumantri, C. (2021). Characteristics and Potential Production of Frozen Semen of Pasundan Bull. Tropical Animal Science Journal, 44(1), 24-31.
Sarma, N., Begum, T., Pandey, S. K., Gogoi, R., Munda, S., & Lal., M. (2020). Chemical Composition of Syzygium cumini (L.) skeels leaf essential oil with respect to its uses from North East Region of India. Journal of Essential Oil Bearing Plants, 23(3).
Savitri, F. K., Suharyati, S., & Siswanto. (2014). Kualitas semen beku sapi bali dengan penambahan berbagai dosis vitamin C pada bahan pengencer skim kuming telur. Jurnal Ilmiah Peternakan Terpadu, 2(3), 30-36.
Schafer-Somi, S., Kluger, S., Knapp, E., Klein, D., & Aurich, C. (2006). Effects of semen extender and semen processing on motility and viability of frozen-thawed dog spermatozoa. Theriogenologi, 66(2), 173-182.
Sholikah, N., Isnaini, N., Puspita Anugra Yekti, A., & Susilawati, T. (2016). Pengaruh penggantian Bovine Serum Albumin (BSA) dengan putih telur pada pengencer CEP-2 terhadap kualitas semen sapi Peranakan Ongole pada suhu penyimpanan 3- 5 ºC. Jurnal Ilmu-Ilmu Peternakan, 26(1), 7–15.
Sukmawati, E., Arifiantini, R. I., & Purwantara, B. (2014). Daya tahan spermatozoa terhadap proses pembekuan pada berbagai jenis sapi pejantan unggul. Jurnal Ilmu Ternak dan Veteriner, 19(3), 168-175.
Sunami, S., Isnaini, N., & Wahjuningsih, S. (2017). Kualitas semen segar dan recovery rate (RR) sapi limousine pada musim yang berbeda. J. Ternak Tropika, 18(1), 36-50.
Thuwanut, P., Chatdarong, K., Techakumphu, M., & Axner, E. (2008). The effect of antioxidants on motility, viability, acrosome integrity and DNA integrity of frozen-thawed epididymal cat spermatozoa. Theriogenology, 70(2), 233-240.
Umar, S., & Maharani, M. (2005). Pengaruh berbagai waktu ekuilibrasi terhadap daya tahan sperma Sapi Limousin dan uji kebuntingan. Jurnal Agribisnis Peternakan, 1(1), 17-21.
Zahn, F. S., Papa, F. O., & Melo, C. M. (2006). Blood serum, seminal plasma and sperm membrane protein profiles in stallions; are they corelated to semen freezability? Anim. Reprod. Sci., 94, 64-66.
Zulyazaini, Dasrul, S., Wahyuni, Akmal, M., & Abdullah, M. A. N. (2016). Karakteristik semen dan komposisi kimia plasma seminalis sapi aceh yang dipelihara di BIBD Sare Aceh Besar. Jurnal Agripet, 16(2), 121-130.