IMPROVING SEMEN QUALITY OF PERANAKAN ONGOLE CATTLE WITH MARIGOLD®-BASED ANTIOXIDANT IN TRIS-EGG YOLK DILUENT
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Abstract
This study aimed to determine the optimal concentration of Marigold® antioxidant suplemented in a Tris-egg yolk (TEY) extender to maintain the quality of liquid semen from Peranakan Ongole (PO) bulls. Semen samples were extended in TEY without antioxidant (P0, control) or supplemented with Marigold® extract at concentrations of 0.002% (P1), 0.004% (P2), and 0.006% (P3). Semen quality was evaluated based on macroscopic characteristics (volume, colour, pH, and consistency) and microscopic parameters (mass motility, progressive motility, viability, plasma membrane integrity (PMI), and morphological abnormalities). Fresh semen exhibited an average volume of 5.25±0.8 mL, creamy colour, pH 6.8±0.13, mass motility (++), abnormalities 4.1±0.8%, viability 86.12±6.2%, PMI 77.21±5.3%, and progressive motility 85.44±2.41%. Supplementation with Marigold® antioxidant significantly influenced microscopic parameters during storage at 4 °C. Sperm abnormalities increased with storage duration, particularly in P3 on day 2. Treatment with 0.004% Marigold® (P2) effectively maintained motility and PMI up to day 6, while 0.006% (P3) preserved viability and PMI up to day 10. The addition of Marigold® extract to the TEY extender improved the preservation of PO bull semen quality during 10 days of storage at 4 °C. A concentration of 0.004% was optimal for maintaining sperm quality in the short term (up to day 6), whereas 0.006% provided stronger protective effects during prolonged storage (up to day 10).
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References
[BPS] Badan Pusat Statistik. Penduduk, Laju Pertumbuhan Penduduk, Distri-busi Persentase Penduduk, Kepadatan Penduduk, Rasio Jenis Kelamin Penduduk Menurut Provinsi. Badan Pusat Statistik Republik Indo-nesia. (Diakses pada: 12 Agustus 2025).
[BSN] Badan Standarisasi Nasional. Standar Nasional Indonesia (2024) Semen Beku- Bagian 1: Sapi (RSNI3 4869-1:2024), Jakarta.
Arif AA, Maulana T, Kaiin EM, Purwantara B, Arifiantini RI (2022) The quality of frozen semen of Limousin bull in vari-ous semen diluents. Trop Anim Sci J 45:284–290. https://doi.org/10.5398/tasj.2022.45.3.284
Authaida S, Boonkum W, Chankitisakul V (2025) Enhancement of semen cryo-preservation from native Thai bulls through Moringa oleifera leaf extract supplementation. Animals 15(3):439. https://doi.org/10.3390/ani15030439
Baharun A, Arifiantini RI, Yusuf TL (2017) Freezing capability of Pasundan bull sperm using Tris-egg yolk, Tris-soy, and Andromed® diluents. Indonesian Journal of Veterinary Sciences 11(1):45-49. https://doi.org/10.21157/j.ked.hewan.v11i1.5810
Baharun A, Pertiwi PPP, Rahmi A, Rach-madanti NC, Handarini R, Iskandar H, Samsudewa D, Maulana T, Said S, Darussalam I, Nurcholis N, Arifiantini RI. 2025. Frozen semen quality of Pasundan bulls with different individ-ual variations. Open Vet J 15(1):244-251. https://doi.org/10.5455/OVJ.2025.v15.i1.22
Baharun A, Rahmi A, Handarini R, Maulana T, Said S, Iskandar H, Darussalam I, Nalley W, Arifiantini R (2024) Semen quality and frozen semen production in Pasundan bulls: A molecular weight perspective on seminal plasma and spermatozoa protein. J Adv Vet Anim Res 10(4):730-737. https://doi.org/10.5455/javar.2023.j728
Baharun A, Said S, Arifiantini RI, Karja NWK. Correlation between age, tes-tosterone and adiponectin concentra-tions, and sperm abnormalities in Simmental bulls. Vet World 14(8):2124-2130. https://doi.org/10.14202/vetworld.2021.2124-2130
Baharun A, Setiawan AB, Rahmi A, Iskan-dar H, Gunawan M, Anwar S, Maula-na T, Kaiin EM, Said S (2023) Frozen semen characteristics of Limousin bulls at different ages. Trop Anim Sci J 46(3):306-312. https://doi.org/10.5398/tasj.2023.46.3.306
Benko F, Palkovičová V, Ďuračka M, Árvay J, Lukáč N, Tvrdá E (2019) Antioxi-dant effects of marigold (Calendula of-ficinalis) flower extract on the oxida-tive balance of bovine spermatozoa. J Contemporary Agri 68(3-4):92-102. https://doi.org/10.2478/contagri-2019-0015
Diansyah AM, Yusuf M, Latief Toleng A, Ihsan Andi Dagong M (2022). Charac-teristic and kinematics of bali-polled bull sperms. J Adv Vet Anim Res 10(8):1787-1796. https://doi.org/10.17582/journal.aavs/2022/10.8.1787.1796
Iskandar H, Sonjaya H, Arifiantini RI, Hasbi H. The quality of fresh and frozen se-men and its correlation with molecular weight of seminal plasma protein in Bali cattle. Trop Anim Sci J 45(4):405-12. https://doi.org/10.5398/tasj.2022.45.4.405
Khoirunnisa A, Utomo S (2024) Pengaruh masa simpan dalam nitrogen cair ter-hadap kualitas sperma beku sapi Peranakan Ongole. J Anim Prod Tech 1:73-84.
Kudratullah, Arifiantini RI, Yuliani E, Pard-ede BP, Said S, Purwantara B (2025) Semen characterictics, freezability, and application of motility-based pro-tein markers (proAKAP4) in assessing the suitability of superior Bali bulls (Bos sondaicus) at the Regional AI Center. Reprod Breed 4(4):279-286. https://doi.org/10.1016/j.repbre.2024.09.005
Mangun M, Duma Y, Baharun A, Handarini R, Said S, Putra WPB, Kaiin EM, Pardede BP, Maulana T (2025) Sperm parameter variability in Dong-gala bulls: An objective-based analy-sis of frozen semen quality. J Adv Vet Anim Res 12(3):832-839. https://doi.org/10.5455/javar.2025.l945
Marawali Al, Abdullah MS, Jalaludin J (2019) Efektivitas suplementasi filtrat jambu biji dalam pengencer airkelapa-kuning telur terhadap kualitas semen cair sapi Bali. J Vet 20:20. https://doi.org/10.19087/jveteriner.2019.20.1.20
Maulana T, Agung PP, Gunawan M, Said S. Computer aided semen analysis (CASA) to determine the quality and fertility of frozen thawed Sumba On-gole sperm supplemented with amino acids. Livest Anim Res 20(2):194-201. https://doi.org/10.20961/lar.v20i2.58754
Merdana IM, Nandita IL, Elia P, Ida Ayu Pasti Apsari, Gorda IW, Sudimartini LM (2024) Phytochemical test and an-tioxidant activity of aqueous extract of marigold Flower. ATBES 8:101-107. https://doi.org/10.24843/ATBES.2024.v08.i02.p08
Mulu M, Moges N, Adane (2018) Review on process, advantages and disad-vantage of artificial insemination in cattle. Inter J Vet Sci Anim Husbandry 3(6):08-13.
Murphy EM, Kelly AK, O’maera C, Eivers B, Lonergan P, Fair S. influence of bull age, ejaculate number, and season of collection on semen production and sperm motility parameters in Holstein Friesian bulls in a commercial artificial insemination centre. J Anim Sci 96(6):2408-2418. https://doi.org/10.1093/jas/sky130
Nugraha CD, Pangestu DD (2023) Pengaruh konsentrasi antioksidan genisteine terhadap kualitas semen sapi peranakan ongole selama proses penyimpanan suhu dingin. Prosiding Seminar Nasional Teknologi dan Ag-ribisnis Peternakan X Fakultas Peter-nakan Universitas Jenderal Soedir-man 393-399.
Nugraha CD, Widodo N, Kuswati K, Suyadi S (2022) The real potential of semen production of Bali bull: over year ob-servation at Singosari National Artifi-cial Insemination Center (SNAIC), Singosari-Indonesia. E3S Web Conf 335:00045. https://doi.org/10.1051/e3sconf/202233500045
Qamar AY, Naveed MI, Raza S, Fang X, Roy PK, Bang S, et al. (2023) Role of antioxidants in fertility preservation of sperm- A narrative review. Animal Bi-oscience 36(3):385-403. https://doi.org/10.5713/ab.22.0325
Said S, Diansyah AM, Rahayu JD, Maulana T, Gunawan M, Kaiin EM. A compara-tive of semen quality traits and sperm kinematic parameters in relation to fer-tility prediction in Murrah buffaloes. J Adv Vet Anim Res 12(3):751-759. https://doi.org/10.5455/javar.2025.l938
Santoso, Sitaresmi PI, Mahari DA, Anwar RI, Lupitasaria FB, Herdis, Arifiantini RI (2024) Sperm kinematic character-istics of pasundan bulls during frozen semen processing. IOP Conf Ser: Earth Environ Sci 1341:012005. https://doi.org/10.1088/1755-1315/1341/1/012005
Sarastina S, Susilawati T, Ciptadi G (2020) Analisa beberapa parameter motilitas spermatozoa pada berbagai bangsa sapi menggunakan computer assisted semen analysis (CASA). J Ternak Trop 6(2):1-12.
Sukirman I, Sukmawati E, Rasad SD, Solihati N (2019) The influence of breed and type of extender on the quality of bull semen. AnimProd 21(2):64-70.
Sumadiasa IWL, Yuliani E, Lukman L (2022) Effect of guava filtrate supple-mentation in tris and citrate-based ex-tenders on spermatozoa quality of Brangus bull after sex-rest. Adv Anim Vet Sci 11(1):150-158. https://doi.org/10.17582/journal.aavs/2023/11.1.150.158
Sun WS, Jiang J, Sun J, Zhang X, Bao R, Ding P, et al (2020) The effect of cry-opreservation on the acrosome struc-ture, enzyme activity, motility, and fer-tility of bovine, ovine, and goat sperm. Anim Reprod 17(4):e20200219. https://doi.org/10.1590/1984-3143-ar2020-0219
Wajdi SA, Utomo B, Rimayanti R, Safitri E, Suprayogi TW, Wurlina W (2021) Suppementation of kelor leaf (Moringa oleifera) aqueous extract increase on post-thawed Limousin bull sperm quality. J Med Vet 4:249. https://doi.org/10.20473/jmv.vol4.iss2.2021.249-255
Wang Y, Fu X, Li H (2025) Mechanisms of oxidative stress-induced sperm dys-function. Front Endocrin 16:1520835. https://doi.org/10.3389/fendo.2025.1520835
Wijayanti A, Suprayogi TW, Prastiya RA, Hernawati T, Sardjito T, Saputro AL, Amaliya A, Sulistyowati D (2023) Ef-fect of addition of green tea extract (Camellia sinensis) in egg yolk tris di-luter on spermatozoa quality in Bali cattle (Bos sondaicus) after freezing. J Med Vet 6(1):66-74. https://doi.org/10.20473/jmv.vol6.iss1.2023.66-74
Yuniar RM, Kusumawati A, Setyawan EMN (2024) Efek penambahan antioksidan selenium, kurkumin dan kombinasinya terhadap motilitas, recovery rate dan viabilitas spermatozoa pada kriopres-ervasi semen sapi Peranakan Ongole. J Sain Vet 42(3):389. https://doi.org/10.22146/jsv.85255
