The Quality of Frozen Buffalo Sperm Following Sexing using Bovine Serum Albumin (BSA) Column and Swim-Up (SU) Methods

https://doi.org/10.21059/buletinpeternak.v48i1.89394

Masrizal Masrizal(1), Tinda Afriani(2), Dwiki Wahyudi(3), Savira Saharani(4), Ananda Ananda(5*)

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

Abstract


This study assesses the impact of two sperm sexing techniques, the Bovine Serum Albumin (BSA) column method, and the swim-up (SU) method, on frozen buffalo spermatozoa quality.  A total of 50 straws of frozen buffalo semen were used in this study. Spermatozoa quality was evaluated before (post thawing) and after the spermatozoa sexing process. Spermatozoa trapped in BSA upper fraction, BSA lower fraction, SU upper fraction, and SU lower fraction were separately evaluated. The parameters measured consisted of  spermatozoa motility, viability, intact plasma membrane, intact acrosome cap, and spermatozoa DNA integrity. The results indicated that the quality of post-thawing buffalo spermatozoa remained relatively high, with motility at 41%, viability at 64.48%, intact plasma membrane at 55.42%, intact acrosome cap at 47.12%, and sperm DNA integrity at 74.94%. However, the use of the BSA column method significantly (p<0.05) decreased spermatozoa quality in both the upper and lower fractions, resulting in motility levels of 34% and 32%, viability rates of 49.36% and 44.71%, intact plasma membrane percentages of 44.78% and 37.13%, intact acrosome cap figures of 37.58% and 33.27%, and sperm DNA integrity levels of 74.76% and 72.45%, respectively. In contrast, the application of the SU method proved effective in preserving post-thawing spermatozoa quality, yielding motility rates of 42% and 41%, viability levels of 63.62% and 62.78%, intact plasma membrane percentages of 54.42% and 54.74%, intact acrosome cap figures of 46.94% and 45.74%, and sperm DNA integrity values of 70.57% and 70.01%, respectively. In summary, after freezing, the SU method excel the BSA column method in maintaining the quality of buffalo spermatozoa post-thawing.


Keywords


Buffalo; Sperm quality; Bovine Serum Albumin; Swim-Up

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References

Abou-haila, A. and D. R. P. Tulsiani. 2009. Signal transduction pathways that regulate sperm capacitation and the acrosome reaction. Arch. Biochem. Biophys. 485: 72–81. https://doi.org/10.1016/j.abb.2009.02.003

Afiati, F. 2004. Proporsi dan karakteristik spermatozoa X dan Y hasil separasi kolom albumen. Media Peternakan 27: 16–20.

Akhter, S., B. A. Rakha, R. Iqbal, and M. S. Ansari. 2014. Effect of bovine serum albumin on motility, plasmalemma, viability and chromatin integrity of buffalo bull spermatozoa. Pak. J. Zool. 46: 115–120.

Andrabi, S. M. H. 2009. Factors affecting the quality of cryopreserved buffalo (Bubalus bubalis) bull spermatozoa. Reprod. Domest. Anim. 44: 552–569. https://doi.org/10.1111/j. 1439-0531.2008.01240.x

Ansari, M. S., B. A. Rakha, N. Ullah, S. M. H. Andrabi, M. Khalid, and S. Akhter. 2011. Effect of L-cysteine in tris-citric egg yolk extender on post-thaw quality of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa. Pak. J. Zool. 43: 41–47.

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

Azizeddin, A., F. A. Ashkar, W. A. King, and T. Revay. 2014. Enrichment of Y-chromosome-bearing bull spermatozoa by swim-up through a column. Reprod. Domest. Anim. 49: 1–5. https://doi.org/10. 1111/rda.12252

Best, B. P. 2015. Cryoprotectant Toxicity: Facts, Issues, and Questions. Rejuvenation Res. 18: 422–436. https://doi.org/10.1089/rej. 2014.1656

Bhalakiya, N., N. Haque, D. Patel, A. Chaudhari, G. Patel, M. Madhavatar, P. Patel, S. Hossain, and R. Kumar. 2018. Sperm Sexing and Its Application in Livestock Sector. Int. J. Curr. Microbiol. App. Sci: 259–272.

Carvalho, J. O., R. Sartori, G. M. Machado, G. B. Mourão, and M. A. N. Dode. 2010. Quality assessment of bovine cryopreserved sperm after sexing by flow cytometry and their use in in vitro embryo production. Theriogenology 74: 1521–1530. https://doi.org/10.1016/j.theriogenology.2010.06.030

Celeghini, E. C. C., R. P. de Arruda, A. F. C. de Andrade, J. Nascimento, C. F. Raphael, and P. H. M. Rodrigues. 2008. Effects that bovine sperm cryopreservation using two different extenders has on sperm membranes and chromatin. Anim. Reprod. Sci. 104: 119–131. https://doi.org/ 10.1016/j.anireprosci.2007.02.001

Check, J. H., B. S. Shanis, S. O. Cooper, and A. Bollendorf. 1989. Male sex preselection: Swim-up technique and insemination of women after ovulation induction. Syst. Biol. Reprod. Med. 23: 165–166. https://doi.org/ 10.3109/01485018908986839

Dixon, K. E., J. E. A. Songy, D. M. Thrasher, and J. L. Kreider. 1980. Received for publication: Theriogenology 13: 437–444. https://doi. org/10.1016/0093-691X(80)90070-9

Donnelly, E. T., E. Kristine Steele, N. McClure, and S. E. M. Lewis. 2001. Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation. Hum. Reprod. 16: 1191–1199. https://doi.org/ 10.1093/humrep/16.6.1191

Dow, M. P. D. and B.D. Bavister. 1989. Direct contact is required between serum albumin and hamster spermatozoa for capacitation in vitro. Gamete Res. 23: 171–180. https://doi.org/10.1002/mrd.1120230204

Ducha, N., T. Susilawati, Aulanni`am, S. Wahyuningsih, and M. Pangestu. 2012. Ultrastructure and Fertilizing Ability of Limousin Bull Sperm after Storage in Cep-2 Extender with and Without Egg Yolk. Pakistan J. Biol. Sci. 15: 979–985. https://doi.org/10.3923/pjbs.2012.979.985

Forman, D. P., J. D. Kirby, and J. A. Proudman. 2013. Reproductive Physiologi In Poultry, in: Hafez, E.S.E., Hafez, B. (Eds.), Reproduction in Farm Animals. Wiley-Blackwell, Philadelphia.

Garner, D. L. and E. S. E. Hafez, E. S. E. 2000. Spermatozoa and Seminal Plasma, in: B. Hafez, and E.S.E. Hafez (Eds.), Reproduction in Farm Animals. Lippincott Williams & Wilkins, Philadelphia, pp. 96–109. https://doi.org/10.1002/978111926 5306.ch7

Gillan, L., T. Kroetsch, W. M. Chis Maxwell, and G. Evans. 2008. Assessment of in vitro sperm characteristics in relation to fertility in dairy bulls. Anim. Reprod. Sci: 103, 201–214. https://doi.org/10.1016/j.anireprosci.2006.12.010

Gunawan, M., E. M. Kaiin, and R. Ridwan. 2017. Peningkatan produktivitas sapi Bali melalui inseminasi buatan dengan sperma sexing di Techno Park Banyumulek, Nusa Tenggara Barat. Sem. Nas. Masy. Biodiv. Indon. pp. 216–219. https://doi.org/10.13057/ psnmbi/m030209

Gunawan, M., E. M. Kaiin, and S. Said. 2015. Aplikasi inseminasi buatan dengan sperma sexing dalam meningkatkan produktivitas sapi di peternakan rakyat. Sem. Nas. Masy. Biodiv. Indon. pp. 93–96. https://doi.org/10.13057/psnmbi/m010115

Harshan, H. M., S. Sankar, L. P. Singh, M. K. Singh, S. Sudharani, M. R. Ansari, S. K. Singh, A. C. Majumdar, and P. Joshi. 2009. Identification of PDC-109-like protein(s) in buffalo seminal plasma. Anim. Reprod. Sci. 115: 306–311. https://doi.org/10.1016/j. anireprosci.2008.11.007

Henkel, R. R. and W. B. Schill. 2003. Sperm preparation for ART. Reprod. Biol. Endocrinol. 1: 1–22. https://doi.org/10. 1186/1477-7827-1-108

Inaudi, P., S. Petrilli, A. Joghtapour, P. Trusso, and F. Petraglia. 2002. Reduction of steps in the preparation of motile sperm for intrauterine insemination does not reduce efficacy of the procedure: Simplified one-step swim-up method versus classic swim-up. Hum. Reprod. 17: 1288–1291. https://doi.org/10. 1093/humrep/17.5.1288

Kaiin, E. M. and M. Gunawan. 2017. Kualitas sperma sapi hasil sexing setelah kapasitasi secara in vitro. Pros. [D1] Sem. Nas. Masy. Biodiv. Indon. 3: 466–470. https://doi.org/10. 13057/psnmbi/m030328

Kaiin, E. M., S. Said, and B. Tappa. 2008. Kelahiran Anak Sapi Hasil Fertilisasi secara in Vitro dengan Sperma Hasil Pemisahan. Media Peternak. 31: 22–28.

Khatamee, M. A., S. R. Horn, A. Weseley, T. Farooq, S. B. Jaffe, and R. Jewelewicz. 1991. A controlled study for gender selection. Fertil. Steril. 56: 254–258. https://doi.org/10.1016/S0015-0282(16)54481-1

Lopulalan, F., T. Saili, and L. O. Ba’a. 2018. Kualitas Dan Fertilitas Spermatozoa Sapi Bali Hasil Sexing Dengan Menggunakan Metode Swim-Down. J. Ilmu dan Teknol. Peternak. Trop. 5: 24. https://doi.org/ 10.33772/jitro.v5i2.4665

Lusignan, M. F., A. Bergeron, M. Lafleur, and P. Manjunath. 2011. The major proteins of bovine seminal plasma interact with caseins and whey proteins of milk extender. Biol. Reprod. 85: 457–464. https://doi.org/10. 1095/biolreprod.110.089961

Manjunath, P. 2012. New insights into the understanding of the mechanism of sperm protection by extender components. Anim. Reprod. 9: 809–815.

Matsuoka, T., H. Imai, H. Kohno, and Y. Fukui. 2006. Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J. Reprod. Dev. 52: 675–683. https://doi.org/10.1262/jrd.18033

Maxwell, W. M. C., G. Mendoza, and I. G. White. 1984. Post-thawing survival of motile ram sperm after isolation by layering on protein columns. Theriogenology 21: 601–606. https://doi.org/10.1016/0093-691X(84)90445-X

Meryman, H. T. 2007. Cryopreservation of living cells: Principles and practice. Transfusion 47: 935–945. https://doi.org/10.1111/ j.1537-2995.2007.01212.x

Navarro-Serna, S., E. París-Oller, O. Simonik, R. Romar, and J. Gadea. 2021. Replacement of albumin by preovulatory oviductal fluid in swim-up sperm preparation method modifies boar sperm parameters and improves in vitro penetration of oocytes. Animals 11. https://doi.org/10.3390/ ani11051202

Okabe, M. 2013. The cell biology of mammalian fertilization. Development 140: 4471–4479. https://doi.org/10.1242/dev.090613

Oldenhof, H., M. Gojowsky, S. Wang, S. Henke, C. Yu, K. Rohn, W.F. Wolkers, and H. Sieme. 2013. Osmotic stress and membrane phase changes during freezing of stallion sperm: Mode of action of cryoprotective agents. Biol. Reprod. 88: 1–11. https://doi.org/ 10.1095/biolreprod.112.104661

Park, C. H., S. G. Lee, D. H. Choi, and C. K. Lee. 2009. A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes. Anim. Reprod. Sci. 115: 169–181. https://doi.org/10.1016/j. anireprosci.2008.12.004

Pegg, D. E. 2015. Principles of cryopreservation, in: J. G. Day and G. N. Stacey. (Eds.), Cryopreservation and Freeze-Drying Protocols. Humana Pr., New Jersey (US), pp. 39–57. https://doi.org/10.1007/978-1-59745-362-2_3

Said, S., E. M. Kaiin, and B. Tappa. 2005. Produksi anak sapi potong dan perah berjenis kelamin sesuai harapan., in: Prosiding Seminar Nasional Industri Peternakan Modern. Puslit Bioteknologi, Mataram, p. 435.

Saili, T. 1999. Efektifitas penggunaan albumin sebagai medium separasi dalam upaya mengubah Rasio Alamiah Spermatozoa Pembawa kromosom X dan Y pada Sapi. Tesis. Program Pasca Sarjana Institut Pertanian Bogor.

Shamsuddin, M., H. Rodriguez-Martinez, and B. Larsson. 1993. Fertilizing capacity of bovine spermatozoa: Selected after swim-up in hyaluronic acid-containing medium. Reprod. Fertil. Dev. 5: 307–315. https://doi.org/10.1071/RD9930307

Sieme, H., H. Oldenhof, and W. F. Wolkers. 2015. Sperm membrane behaviour during cooling and cryopreservation. Reprod. Domest. Anim. 50: 20–26. https://doi.org/ 10.1111/rda.12594

Srivastava, N., A. Jerome, S. K. Srivastava, S. K. Ghosh, and A. Kumar. 2013. Bovine seminal PDC-109 protein: An overview of biochemical and functional properties. Anim. Reprod. Sci. 138: 1–13. https://doi. org/10.1016/j.anireprosci.2013.02.008

Suarez, S. S. and H. Ho. 2003. Hyperactivated motility in sperm Introduction : Functions of hyperactivation initiation of hyperactivation in vivo. Reprod Dom Anim 124: 119–124. https://doi.org/10.1046/j.1439-0531.2003.00397.x

Susilawati, T. 2014. Sexing Spermatozoa. UB Press, Malang.

Tanphaichitr, N., A. Agulnick, M. Seibel, and M. Taymor. 1988. Comparison of the in vitro fertilization rate by human sperm capacitated by multiple-tube swim-up and Percoll gradient centrifugation. J. Vitr. Fertil. Embryo Transf. 5: 119–122. https://doi.org/10.1007/BF01131172

Visconti, P. E., H. Galantino-Homer, X. Ning, G. D. Moore, J. P. Valenzuela, C. J. Jorgez, J. G. Alvarez, and G. S. Kopf. 1999. Cholesterol Efflux-mediated signal transduction in mammalian sperm. J. Biol. Chem. 274: 3235–3242. https://doi.org/10.1074/jbc. 274.5.3235

Wang, W. H., B. N. Day, and G. M. Wu. 2003. How does polyspermy happen in mammalian oocytes? Microsc. Res. Tech. 61: 335–341. https://doi.org/10.1002/jemt.10346

Welch, G. R. and L. A. Johnson. 1999. Sex preselection: Laboratory validation of the sperm sex ratio of flow sorted X- and Y-sperm by sort reanalysis for DNA. Theriogenology 52: 1343–1352. https://doi.org/10.1016/S0093-691X(99)00221-6

Yadav, S. K., D. K. Gangwar, J. Singh, C. K. Tikadar, V. V. Khanna, S. Saini, S. Dholpuria, P. Palta, R. S. Manik, M. K. Singh, and S. K. Singla. 2017. An immunological approach of sperm sexing and different methods for identification of X- and Y-chromosome bearing sperm. Vet. World 10: 498–504. https://doi.org/10. 14202/vetworld.2017.498-504



DOI: https://doi.org/10.21059/buletinpeternak.v48i1.89394

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