Improvement of Sexed Sperm Quality of Bali Bulls by Adding Palmyra (Borassus flabellifer Linn.) Fruit Water to Citrate-Egg Yolk Extender

Thomas Mata Hine; Wilmientje Marlene Nalley; Aloysius Marawali; Johny Nada Kihe; Petrus Kune; Kirenius Uly

doi:10.14334/jitv.v29i3.3388

Improvement of Sexed Sperm Quality of Bali Bulls by Adding Palmyra (Borassus flabellifer Linn.) Fruit Water to Citrate-Egg Yolk Extender

Thomas Mata Hine, Wilmientje Marlene Nalley, Aloysius Marawali, Johny Nada Kihe, Petrus Kune, Kirenius Uly

DOI

https://doi.org/10.14334/jitv.v29i3.3388

Abstract

Sperm sexing is a technique designed to separate sperm carrying the X and Y chromosomes, which are then used for artificial insemination to generate offspring with the sex the breeder desires. However, the sexing process causes a decrease in sperm quality caused by oxidative stress due to an excessive increase in free radicals. Thus, to counteract the detrimental effects of free radicals on sperm life, antioxidants must be added to the diluent. This study was designed by adding palmyra fruit water (PFw) to citrate-egg yolk (CEy) diluent. This study aimed to evaluate the potency of PFw as a natural antioxidant supplement in CEy diluent to improve the quality of sexing sperm in Bali bulls. Sperm were collected using the artificial vagina method from three Bali bulls aged 3-4 years. Good quality sperm (motility ?70%, abnormality ?20%) were sexed with a three-layer albumin gradient method (5, 10, and 15 percent) for 20 minutes, and sperm that were on the bottom albumin layer were preserved in CEy, PFw, PFw-egg yolk (PFw-Ey), or CEy-PFw. The results showed that sperm preservation in the CEy-PFw diluent resulted in higher sperm quality (P<0.05) compared to the other three diluents, except for the sperm abnormality parameter (P>0.05). It was concluded that adding PFw into the CEy diluent could potentially improve the sexed sperm quality of Bali bulls.

Keywords

Bali Bulls; Citrate; Egg Yolk; Palmyra Fruit Water; Sperm Sexing

References

Abe-Inge V, Arthur C, Agbenorhevi JK, Kpodo FM. 2018. Mineral composition, antioxidant properties, phytochemical and anti-nutrient composition of African palmyra palm (Borassus aethiopum) fruit flour. AJFN. 6:143-152. DOI:10.12691/ajfn-6-5-2.

Alvarez-Rodríguez M, Alvarez M, Anel-López L, Martínez-Rodríguez C, Martínez-Pastor F, Borragan S, Anel L, de Paz P. 2013. The antioxidant effects of soybean lecithin-or low-density lipoprotein-based extenders for the cryopreservation of brown-bear (Ursus arctos) spermatozoa. Reprod Fertil Dev. 25:1185–1193. DOI:10.1071/rd12181.

Amirat-Briand L, Bencharif D, Vera-Munoz O, Pineau S, Thorin C, Destrumelle S, Desherces S, Anton M, Jouan M, Shmitt E. 2010. In vivo fertility of bull semen following cryopreservation with an LDL (low density lipoprotein) extender: Preliminary results of artificial inseminations. Anim Reprod Sci. 122:282–287. DOI:10.1016/j.anireprosci.2010.09.006.

Anzar M, Rajapaksha K, Boswall L. 2019. Egg yolk-free cryopreservation of bull semen. PLoS One. 14:e0223977. DOI:10.1371/journal.pone.0223977.

Arifiantini RI, Purwantara B. 2010. Motility and viability of friesian holstein spermatozoa in three different extender stored at 5oC. J Indones Trop Anim Agric. 35:222–226. DOI:10.14710/jitaa.35.4.222-226.

Arifiantini RI, Purwantara B, Riyadhi M. 2010. Occurrence of sperm abnormality of beef cattle at several artificial insemination centers in Indonesia. Anim Prod. 12.

Behera S, Nayak B. 2022. Phytochemical constituents and nutritional potential of palmyra palm: a review. Rev Contemp Sci Acad Stud. 2. DOI:10.55454/rcsas.2.12. 2022.003

Bhat Y, Sharma M. 2020. X-sperm enrichment of bovine semen by Percoll density gradient method and its effect on semen quality, sex ratio and conception rate. Indian J Anim Res. 54:1181–1187. DOI:10.18805/ijar.B-3823.

Crespilho AM, Chiaradia L, Cortez A, Dinelli M, Papa FO, Gomes GM, Peixoto KC. 2017. Sensitivity evaluation of the computer-assisted sperm analysis (CASA) in the determination of frozen-thawed bull semen concentration. Braz J Vet Res Anim Sci. 54:247–252. DOI:10.11606/issn.1678-4456.bjvras.2017.127773.

Filho ICB, Pederzolli CD, Sgaravatti AM, Gregory RM, Filho CSD, Jobim MIM, Mattos RC. 2009. Skim milk-egg yolk based semen extender compensates for non enzymatic antioxidant activity loss during equine semen cryopreservation. Anim Reprod. 6:392–399.

Ford WCL. 2006. Glycolysis and sperm motility: does a spoonful of sugar help the flagellum go round? Hum Reprod Update. 12:269–274. DOI:10.1093/humupd /dmi053.

Haisya NBS, Utama BD, Edy RC, Aprilia HM. 2011. The potential of developing Siwalan palm sugar (Borassus flabellifer Linn.) as one of the bioethanol sources to overcome energy crisis problem in Indonesia. In: 2nd Int Conf Environ Eng Appl IPCBEE. 17.

Haris FZ, Ondho YS, Samsudewa D. 2020. Effect of vitamin E addition to frozen Simmental bull semen extender on post-thawing quality. E3S Web Conf. 142:02002. DOI:10.1051/e3sconf/202014202002.

Hine TM, Burhanuddin, Marawali A. 2014. Efektivitas air buah lontar dalam mempertahankan motilitas, viabilitas dan daya tahan hidup spermatozoa sapi Bali. J Vet. 15:263–273.

Huynh Thi Le D, Lu W-C, Li P-H. 2020. Sustainable processes and chemical characterization of natural food additives: Palmyra palm (Borassus flabellifer Linn.) granulated sugar. Sustainabil. 12:2650. DOI:10.3390/su12072 650.

Johnston SD, Zee YP, López?Fernández C, Gosálvez J. 2012. The effect of chilled storage and cryopreservation on the sperm DNA fragmentation dynamics of a captive population of koalas. J Androl. 33:1007–1015. DOI:10. 2164/jandrol.111.015248.

Kaka A, Yekti APA, Maylinda S, Rahayu S, Susilawati T. 2024. Effectiveness of nanoparticle-based young palmyra fruit water–egg yolk in liquid semen diluent of Sumba Ongole bulls. Int J Vet Sci. 14:16-24. DOI:10.47278/journal .ijvs/2024.205.

Kumar A, Vineeth MR, Sinha R, Singh RK, Thakur A, Gupta SK. 2016. Current status, scope, and constraints of sexed semen. An Indian perspective. Agric Rev. 37:240–244. DOI:10.18805/ar.v0i.11286.

Kurian A, Thiripuranathar G, Paranagama PA. 2017. Determination of total phenolic content and antioxidant activity of Borassus flabeliffer Linn. Fruit pulp was collected from several parts of Sri Lanka. Int J Pharm Sci Res. 8:2701–2705. DOI:10.13040/IJPSR.0975-8232.8(6).27 01-05.

Lagace TA, Ridgway ND. 2013. The role of phospholipids in the biological activity and structure of the endoplasmic reticulum. BBA-Molecular Cell Res. 1833:2499–2510. DOI:10.1016/j.bbamcr.2013.05.018.

Layek SS, Mohanty TK, Kumaresan A, Parks JE. 2016. Cryopreservation of bull semen: Evolution from egg yolk-based to soybean-based extenders. Anim Reprod Sci. 172:1–9. DOI:10.1016/j.anireprosci.2016.04.013.

Liu C-H, Dong H-B, Ma D-L, Li Y-W, Han D, Luo M-J, Chang Z-L, Tan J-H. 2016. Effects of pH during liquid storage of goat semen on sperm viability and fertilizing potential. Anim Reprod Sci. 164:47–56. DOI:10.1016/j. anireprosci.2015.11.011.

Mahfud A, Isnaini N, Yekti APA, Kuswati K, Susilawati T. 2019. Kualitas spermatozoa post thawing semen beku sperma Y hasil sexing pada Sapi Limousin. J Trop Anim Prod. 20:1–7. DOI:10.21776/ub.jtapro.2019.020.01.1.

Malik A, Jaelani A, Widaningsih N, Ni’mah GK, Raviani, Sakiman, Sasongko N. 2018. Effect of different concentration of fish oil in skim milk-egg yolk extenders on post- thawed semen qualities of Kalang swamp buffalo bull. Asian Pasific J Reprod. 7:139-142. DOI:10.4103/2305-0500.233576.

Manjunath P. 2018. New insights into the understanding of the mechanism of sperm protection by extender components. Anim Reprod. 9:809–815.

Mohamed MY, AM AE-H, Shaarawy AM. 2019. Influence of adding different energy sources to the bull and ram spermatozoa exposed to different refrigerating times. Egypt J Sheep Goats Sci. 14:1–18. DOI:10.21608 /ejsgs.2019.56661.

Morrell JM, Valeanu AS, Lundeheim N, Johannisson A. 2018. Sperm quality in frozen beef and dairy bull semen. Acta Vet Scand. 60:1–10.

Mousavi SM, Towhidi A, Zhandi M, Amoabediny G, Mohammadi-Sangcheshmeh A, Sharafi M, Hussaini SMH. 2019. Comparison of two different antioxidants in a nano lecithin-based extender for bull sperm cryopreservation. Anim Reprod Sci. 209:106171. DOI:10.1016/j.anireprosci.2019.106171.

Mughal DH, Ijaz A, Yousaf MS, Rehman H, Aleem M, Zaneb H, Wadood F. 2013. Assessment of optimal osmotic pressure of citrate egg yolk extender for cryopreservation of buffalo bull (Bubalus bubalis) semen. J Anim Plant Sci. 23:964–968.

Mukai C, Okuno M. 2004. Glycolysis plays a major role for adenosine triphosphate supplementation in mouse sperm flagellar movement. Biol Reprod. 71:540–547. DOI:10. 1095/biolreprod.103.026054.

Naz S, Umair M, Iqbal S. 2018. Comparison of Tris egg yolk?based, Triladyl® and Optixell® extender on post?thaw quality, kinematics and in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Andrologia. 50:e13063. DOI:10.1111/and.13063.

Panche AN, Diwan AD, Chandra SR. 2016. Flavonoids: an overview. J Nutr Sci. 5:e47. DOI:10.1017/jns.2016.41.

du Plessis SS, Agarwal A, Mohanty G, Van der Linde M. 2015. Oxidative phosphorylation versus glycolysis: what fuel do spermatozoa use?. Asian J Androl. 17:230. DOI:10.4103/1008-682X.135123.

Purwoistri RF, Susilawati T, Rahayu S. 2013. Kualitas spermatozoa hasil sexing menggunakan pengencer andromed dan cauda epididymal plasma-2 (CEP-2) ditambah kuning telur 10%. J Kedokt Hewan-Indonesian J Vet Sci. 7:116-119. DOI:10.21157/j.ked.hewan.v7i2. 922.

Raheja N, Choudhary S, Grewal S, Sharma N, Kumar N. 2018. A review on semen extenders and additives used in cattle and buffalo bull semen preservation. J Entomol Zool Stud. 6:239–245.

Rahman MS, Pang M-G. 2020. New biological insights on X and Y chromosome-bearing spermatozoa. Front cell Dev Biol. 7:388. DOI:10.3389/fcell.2019.00388.

Rai J. 2018. Sperm sexing of dairy cattle: economics, animal welfare, and technological challenges. Curr Sci. 114:1438–1442. DOI:10.18520/cs/v114/i07/1438-1442.

Ramu S, Jeyendran RS. 2013. The hypo-osmotic swelling test for evaluation of sperm membrane integrity. Spermatogenes Methods Protoc. 927:21–25. DOI:10. 1007/978-1-62703-038-0_3.

Rath D, Barcikowski S, De Graaf S, Garrels W, Grossfeld R, Klein S, Knabe W, Knorr C, Kues W, Meyer H. 2013. Sex selection of sperm in farm animals: status report and developmental prospects. Reprod. 145:R15–R30. DOI:10.1530/rep-12-0151.

Ratnawati D, Luthfi M, Pamungkas D, Affandhy L. 2020. Motility characterization of albumin sexed spermatozoa in two different diluents and additional antioxidants. J Indones Trop Anim Agric. 45:277–286. DOI:10.14710/jitaa.45.4.277-286.

Razmkabir M. 2018. A field study on the reproductive efficiency of sex-sorted semen in Holstein heifers. J Livest Sci Technol. 6:41–46. DOI:10.22103/jlst .2018.12446.1239.

Renuka K, Devi VR, Subramanian SP. 2018. Phytochemical screening and evaluation of in vitro antioxidant potential of immature palmyra palm (Borassus flabellifer Linn.) fruits. Int J Pharm Pharm Sci. 10:77–83. DOI:10.22159 /ijpps.2018v10i8.27162

Saranya P, Vijayakumar TP. 2016. Preliminary phytochemical screening of raw and thermally processed Palmyra palm (Borassus flabellifer Linn.) fruit pulp. J Innov Pharm Biol Sci. 3:186–193.

Schulze M, Nitsche-Melkus E, Hensel B, Jung M, Jakop U. 2020. Antibiotics and their alternatives in Artificial Breeding in livestock. Anim Reprod Sci. 220:106284. DOI:10.1016/j.anireprosci.2020.106284.

Seidel GE. 2014. Update on sexed semen technology in cattle. Anim. 8:160–164. DOI:10.1017/S1751731114000202.

Setiadi DR, Mulia BH, Widianti A, Manansang J, Hastuti YT, Yoelinda VT, Agil M, Arifiantini RI, Nugraha TP. 2022. Successful semen collection and cryopreservation of Banteng semen (Bos javanicus) using an artificial vagina. IOP Conf. Ser.: Earth Environ. Sci. 1109:12095. DOI:10.1088/1755-1315/1109/1/012095.

Spinaci M, Perteghella S, Chlapanidas T, Galeati G, Vigo D, Tamanini C, Bucci D. 2016. Storage of sexed boar spermatozoa: limits and perspectives. Theriogenology. 85:65–73. DOI:10.1016/j.theriogenology.2015.05.018.

Stevenson J, Luu W, Kristiana I, Brown AJ. 2014. Squalene mono-oxygenase, a key enzyme in cholesterol synthesis, is stabilized by unsaturated fatty acids. Biochem J. 461:435–442. DOI:10.1042/bj20131404.

Sun L, Fan W, Wu C, Zhang S, Dai J, Zhang D. 2020. Effect of substituting different concentrations of soybean lecithin and egg yolk in tris-based extender on goat semen cryopreservation. Cryobiol. 92:146–150. DOI:10.101 6/j.cryobiol.2019.12.004.

Sylla L, Palombi C, Stradaioli G, Vagniluca A, Monaci M. 2015. Effect of semen collection by transrectal massage of accessory sexual glands or artificial vagina on the outcome of breeding soundness examinations of Italian yearling beef bulls. Theriogenol. 83:779–785. DOI:10.1016/j.theriogenology.2014.11.011.

Tarig AA, Wahid H, Rosnina Y, Yimer N, Goh YM, Baiee FH, Khumran AM, Salman H, Ebrahimi M. 2017. Effect of different concentrations of egg yolk and virgin coconut oil in Tris-based extenders on chilled and frozen-thawed bull semen. Anim Reprod Sci. 182:21–27. DOI:10.1016/j.anireprosci.2017.03.024.

Tariq A, Ahmad M, Iqbal S, Riaz MI, Tahir MZ, Ghafoor A, Riaz A. 2020. Effect of carboxylated poly l-Lysine as a cryoprotectant on post-thaw quality and in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull semen. Theriogenol. 144:8–15. DOI:10.1016/j.theriogenology. 2019.12.012.

Tourmente M, Villar-Moya P, Rial E, Roldan ERS. 2015. Differences in ATP generation via glycolysis and oxidative phosphorylation and relationships with sperm motility in mouse species. J Biol Chem. 290:20613–20626. DOI:10.1074/jbc.m115.664813.

Vishwanath R, Moreno JF. 2018. Semen sexing–current state of the art with emphasis on bovine species. Anim. 12:s85–s96. DOI:10.1017/s1751731118000496.

Wijewardana R, Nawarathne SB, Wickramasinghe I, Gunawardane CR, Wasala W, Thilakarathne B. 2016. Retention of physicochemical and antioxidant properties of dehydrated bael (Aegle marmelos) and palmyra (Borassus flabellifer) fruit powders. Procedia food Sci. 6:170–175. DOI:10.1016/j.profoo.2016.02.041.

Yadav HP, Sahu SK, Lone SA, Shah N, Singh A, Verma UK, Baithalu RK, Mohanty TK. 2018. Advances in sperm sexing in bovines. J Exp Zool India. 21:1–9.

Yi YJ, Li ZH, Kim ES, Song ES, Kim HB, Cong PQ, Lee JM, Park C-S. 2008. Comparison of Motility, Acrosome, Viability, and ATP of Boar Sperm with or without Cold Shock Resistance in Liquid Semen at 17oC and 4oC, and Frozen-thawed Semen. Asian-Australasian J Anim Sci. 21:190–197. DOI:10.5713/ajas.2008.70351.

Zubair M, Lodhi LA, Ahmad E, Muhammad G. 2013. Hypo osmotic swelling test as screening for evaluation of semen of bull. J Entomol Zool Stud. 1:124–128.