Article history: Received: 21-12-2016, Accepted: 05-05-2017, Published online: 20-06-2017
Corresponding author: H. Wahid
E-mail: firstname.lastname@example.orgCitation: Tarig AA, Wahid H, Rosnina Y, Yimer N, Goh YM, Baiee FH, Khumran AM, Salman H, Assi MA, Ebrahimi M (2017) Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen, Veterinary World, 10(6): 672-678.
Aim: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL) and virgin coconut oil (VCO) in Tris-based extender on chilled and frozen-thawed bull semen quality parameters.
Materials and Methods: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test.
Results: The results showed that all the quality parameters assessed were significantly (p<0.05) improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+) in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups.
Conclusion: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.
Keywords: bull semen, cryopreservation, quality parameters, soybean lecithin, virgin coconut oil.
1. Singh, A., Singh, V., Narwade, B., Mohanty, T. and Atreja, S. (2012) Comparative quality assessment of buffalo (Bubalus bubalis) semen chilled (5° C) in egg yolk and soya milk-based extenders. Reprod. Domest. Anim., 47(4): 596-600. [Crossref] [PubMed]
3. Vishwanath, R. and Shannon, P. (2000) Storage of bovine semen in liquid and frozen state. Anim. Reprod. Sci., 62(1): 23-53. [Crossref]
4. Bucher, A., Kasimanickam, R., Hall, J.B., Dejarnette, J.M., Whittier, W.D., Kahn, W. and Xu, Z. (2009) Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-synch protocol in beef cows. Theriogenology, 71(7): 1180-1185. [Crossref] [PubMed]
5. Crespilho, A.M., Sa Filho, M.F., Dell'Aqua, J.A., Nichi, M., Monteiro, G.A., Avanzi, B.R., Martins, A. and Papa, F.O. (2012) Comparison of in vitro and in vivo fertilizing potential of bovine semen frozen in egg yolk or new lecithin based extenders. Livest. Sci., 149(1): 1-6. [Crossref]
6. Ansari, M.S., Rakha, B.A., Ullah, N., Andrabi, S.M.H. and Akhtar, S. (2012) Glutathione addition in Tris-citric egg yolk extender improves the quality of cooled buffalo (Bubalus bubalus) bull semen. Pak. J. Zool., 43: 49-55.
7. Thun, R., Hurtado, M. and Janett, F. (2002) Comparison of biociphos-plus® and Tris-egg yolk extender for cryopreservation of bull semen. Theriogenology, 57(3): 1087-1094. [Crossref]
8. Aires, V.A., Hinsch, K.D., Mueller-Schloesser, F., Bogner, K., Mueller-Schloesser, S. and Hinsch, E. (2003) In vitro and in vivo comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of bovine semen. Theriogenology, 60(2): 269-279. [Crossref]
9. Fukui, Y., Kohno, H., Togari, T., Hiwasa, M. and Okabe, K. (2008) Fertility after artificial insemination using a soybean-based semen extender in sheep. J. Reprod. Dev., 54(4): 286-289. [Crossref]
10. Zhang, S., Hu, J., Li, Q., Jiang, Z. and Zhang, X. (2009) The cryoprotective effects of soybean lecithin on boar spermatozoa quality. Afr. J. Biotechnol., 8(22): 6476-6480. [Crossref]
11. Trotta, M., Pattarino, F. and Ignoni, T. (2002) Stability of drug-carrier emulsions containing phosphatidylcholine mixtures. Eur. J. Pharm. Biopharm., 53(2): 203-208. [Crossref]
12. Memon, A.A., Wahid, H., Rosnina, Y., Goh, Y.M., Ebrahimi, M., Nadia, F.M. and Audrey, G. (2011) Effect of butylated hydroxytoluene on cryopreservation of Boer goat semen in Tris egg yolk extender. Anim. Reprod. Sci.,129: 44-49. [Crossref] [PubMed]
13. Memon, A.A., Wahid, H., Rosnina, Y., Goh, Y.M., Ebrahimi, M. and Nadia, F. (2012) Effect of antioxidants on post-thaw microscopic, oxidative stress parameter and fertility of Boer goat spermatozoa in Tris egg yolk glycerol extender. Anim. Reprod. Sci., 136(1): 55-60. [Crossref] [PubMed]
14. Nasiri, A.H., Towhidi, A. and Zeinoaldini, S. (2012) Combined effect of DHA and a tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition. Andrologia, 44(1): 550-555. [Crossref] [PubMed]
15. Bucak, M.N., Atessahin, A., Varisli, O., Yuce, A., Tekin, N. and Akcay, A. (2007) The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen: Microscopic and oxidative stress parameters after freeze-thawing process. Theriogenology, 67(5): 1060-1067. [Crossref] [PubMed]
17. Dosumu, O., Duru, F., Osinubi, A., Oremosu, A. and Noronha, C. (2010) Influence of virgin coconut oil (VCNO) on oxidative stress, serum testosterone and gonadotropic hormones (FSH, LH) in chronic ethanol ingestion. Agric. Biol. J. N. Am., 6: 1126-1132. [Crossref]
18. Nevin, K.G. and Rajamohan, T. (2006) Virgin coconut oil supplemented diet increases the antioxidant status in rats. Food Chem., 99(2): 260-266. [Crossref]
19. Dickinson, E. (1993) Towards more natural emulsifiers. Trends Food Sci. Technol., 4(10): 330-334. [Crossref]
20. Khumran, A.M., Yimer, N., Rosnina, Y., Ariff, M.O., Wahid, H., Kaka, A., Ebrahimi, M. and Sarsaifi, K. (2015) Butylated hydroxytoluene can reduce oxidative stress and improve quality of frozen-thawed bull semen processed in lecithin and egg yolk based extenders. Anim. Reprod. Sci., 163: 128-134. [Crossref]
21. Amirat-Briand, L., Bencharif, D., Vera-Munoz, O., Pineau, S., Thorin, C., Destrumelle, S., Desherces, S., Anton, M., Jouan, M., Shmitt, E. and Tainturier, D. (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(3): 282-287. [Crossref] [PubMed]
22. Kaka, A., Wahid, H., Rosnina, Y., Yimer, N., Khumran, A.M., Behan, A.A. and Ebrahimi, M. (2015b) Alpha-linolenic acid supplementation in tris extender can improve frozen-thawed bull semen quality. Reprod. Domest. Anim., 50(1): 29-33. [Crossref] [PubMed]
23. Yildiz, C., Kaya, A., Aksoy, M. and Tekeli, T. (2000) Influence of sugar supplementation of the extender on motility, viability and acrosomal integrity of dog spermatozoa during freezing. Theriogenology, 54(4); 579-585. [Crossref]
24. Kaka, A., Wahid, H., Yimer, N., Khumran, A.M., Kazhal, S., Behan, A.A., Ebrahimi, M. and Ubedullah, K. (2015a) a-linolenic acid supplementation in BioXcell® extender can improve the quality of post-cooling and frozen-thawed bovine sperm. Anim. Reprod. Sci., 153: 1-7. [Crossref] [PubMed]
25. De Paz, P., Esteso, M.C., Alvarez, M., Mata, M., Chamorro, C.A. and Anel, L. (2010) Development of extender based on soybean lecithin for its application in liquid ram semen. Theriogenology, 74(4): 663-671. [Crossref] [PubMed]
26. Akhter, S., Ansari, M., Rakha, B., Ullah, N., Andrabi, S. and Khalid, M. (2011) In vitro evaluation of liquid-stored buffalo semen at 5°C diluted in soya lecithin based extender (Bioxcell®), Tris-citric egg yolk, skim milk and egg yolk-citrate extenders. Reprod. Domest. Anim., 46(1): 45-49. [Crossref] [PubMed]
27. Rehman, F., Qureshi, M. and Khan, R. (2014) Effect of soybean-based extenders on sperm parameters of Holstein-Friesian bull during liquid storage at 4°C. Pak. J. Zool., 46(1): 185-189.
28. Graham, J.K. and Foote, R.H. (1987) Effect of several lipids, fatty acyl chain length, and degree of unsaturation on the motility of bull spermatozoa after cold shock and freezing. Cryobiology, 24(1): 42-52. [Crossref]
29. Trimeche, A., Anton, M., Renard, P., Gandemer, G. and Tainturier, D. (1997) Quail egg yolk: Anovel cryoprotectant for the freeze preservation of Poitou Jackass sperm. Cryobiology, 34: 385-393. [Crossref] [PubMed]
30. De Leeuw, F., De Leeuw, A., den Daas, J., Colenbrander, B. and Verkleij, A. (1993) Effects of various cryoprotective agents and membrane-stabilizing compounds on bull sperm membrane integrity after cooling and freezing. Cryobiology, 30(1): 32-44. [Crossref] [PubMed]
31. Hafez, E.S.E. (1993) Reproduction in Farm Animals. Vol. 6. Lea and Febiger, Philadelphia. p330-343, 405-424.
32. Badens, E., Magnan, C. and Charbit, G. (2001) Microparticles of soy lecithin formed by supercritical processes. Biotechnol. Bioeng., 72(2): 194-204. [Crossref]
33. Crespilho, A.M., Papa, F.O., Santos, M.D.P. and Sa Filho, M.F.D. (2012) Use of cooled bull semen as a strategy to increase the pregnancy rate in fixed-time artificial insemination programs-case report. Am. J. Anim. Vet. Sci., 7(4): 175-179. [Crossref]
34. Papa, F.O., Felicio, G.B., Melo, C.M., Vita, B., Avanzi, B.R. and Aqua, J.A. (2010) Effect of substituting soybean lecithin for egg yolk in an extender used for the cryopreservation of stallion semen. Anim. Reprod. Sci., 121: 71-72.
35. van Wagtendonk-de Leeuw, A.M., Haring, R.M., Kaal-Lansbergen, L.M.T. and den Daas, J.H.G. (2000) Fertility results using bovine semen cryopreserved with extenders based on egg yolk and soy bean extract. Theriogenology, 54(1): 57-67. [Crossref]
36. Ustuner, B., Alcay, S., Nur, Z., Sagirkaya, H. and Soylu, M.K. (2014), Effect of egg yolk and soybean lecithin on tris-based extender in post-thaw ram semen quality and in vitro fertility. Kafkas Univ. Vet. Fak. Derg., 20(3): 393-398. [Crossref]
37. Salmani, H., Towhidi, A., Zhandi, M., Bahreini, M. and Sharafi, M. (2014) In vitro assessment of soybean lecithin and egg yolk based diluents for cryopreservation of goat semen. Cryobiology, 68(2): 276-280. [Crossref] [PubMed]
38. Bucak, M.N., Tuncer, P.B., Sariozkan, S. and Ulutas, P.A. (2009) Comparison of the effects of glutamine and an amino acid solution on post-thawed ram sperm parameters, lipid peroxidation and antioxidant activities. Small Rumin. Res., 81(1): 13-17. [Crossref]
39. Coyan, K., Baspinar, N., Bucak, M.N. and Akalin, P.P. (2011) Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters. Cryobiology, 63(1): 1-6. [Crossref] [PubMed]40. Atessahin, A., Bucak, M.N., Tuncer, P.B. and Kizil, M. (2008) Effects of antioxidant additives on microscopic and oxidative parameters of Angora goat semen following the freeze-thawing process. Small Rumin. Res., 77(1): 38-44. [Crossref]