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Research (Published online: 27-05-2016)

15. Effect of washing on the post-thaw quality of cryopreserved ram epididymal spermatozoa - Touqeer Ahmed, Rafiqul Islam, Farooz Ahmad Lone and Asloob Ahmad Malik

Veterinary World, 9(5): 519-523

 

 

   doi: 10.14202/vetworld.2016.519-523

 

 

Touqeer Ahmed: Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shuhama, Alusteng, Srinagar, Jammu & Kashmir, India; touqeer_taz@yahoo.co.in

Rafiqul Islam: Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shuhama, Alusteng, Srinagar, Jammu & Kashmir, India; rafiqvet@gmail.com

Farooz Ahmad Lone: Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shuhama, Alusteng, Srinagar, Jammu & Kashmir, India; dr.farooz462@gmail.com

Asloob Ahmad Malik: Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shuhama, Alusteng, Srinagar, Jammu & Kashmir, India; malikasloob@gmail.com

 

Received: 01-02-2016, Accepted: 20-04-2016, Published online: 27-05-2016

 

Corresponding author: Touqeer Ahmed, e-mail: touqeer_taz@yahoo.co.in


Citation: Ahmed T, Islam R, Lone FA, Malik AA (2016) Effect of washing on the post-thaw quality of cryopreserved ram epididymal spermatozoa, Veterinary World, 9(5): 519-523.



Aim: The aim of the study was to evaluate the effect of washing on the post-thaw quality of ram cauda epididymal spermatozoa (P1: Unwashed, P2: Washed).

Materials and Methods: Fresh testicles of adult healthy slaughtered rams were collected and transported to the laboratory in an ice chest, where they were weighed, and cauda epididymides were separated. These cauda epididymides were used for recovery of spermatozoa in tris-citric acid fructose buffer by incision method. Spermatozoa samples showing ≥70% progressive motility were pooled and processed further. The mean values (standard error) of various parameters such as the percentage of sperm motility, live sperm, intact acrosome, and hypo-osmotic swelling test (HOST) rea cted spermatozoa were recorded.

Results: In this experiment, the percent sperm motility, live spermatozoa, and intact acrosome both at pre-freeze and postthaw were higher (p>0.05) in P1 than P2. However, the post-thaw percent HOST reacted spermatozoa was slightly higher (p>0.05) for P2 than P1.

Conclusion: Washing of cauda epididymal spermatozoa has no significant adverse effect on the quality during cryopreservation. Therefore, this processing method can be applied wherever necessary before the extension of the recovered spermatozoa sample in different ram extenders.

Keywords: cauda epididymis, centrifugation, processing, ram, spermatozoa, washing.



1. Behera, S., Harshan, H.M., Bhai, K.L. and Ghosh, K.N.A. (2015) Effect of cholesterol supplementation on cryosurvival of goat spermatozoa. Vet. World, 8: 1386-1391.
http://dx.doi.org/10.14202/vetworld.2015.1386-1391
PMid:27047048 PMCid:PMC4774814
 
2. Ali, A.T., Bomboi, G. and Floris, B. (2013) Replacing chicken yolk with yolks from other sources in ram semen diluents and their effects on fertility in vitro. Small Rum. Res., 113: 405-410.
http://dx.doi.org/10.1016/j.smallrumres.2013.01.017
 
3. Naing, S.W., Haron, A.W., Goriman, M.A.K., Yusoff, R., Abu Bakar, M.Z., Sarsaifi, K., Bukar, M.M., Thein, M., Kyaw, T. and San M.M. (2011) Effect of seminal plasma removal, washing solutions, and centrifugation regimes on boer goat semen cryopreservation. Pertanika J. Trop. Agric. Sci., 34(2): 271-279.
 
4. Forouzanfar, M., Ershad, S.F., Hosseini, S.M., Hajian, M., Hosseini, S.O., Abid, A., Tavalaee, M., Shahverdi, A., Dizaji, A.V. and Esfahani, M.H.N. (2013) Can permeable super oxide dismutase mimetic agents improve the quality of frozen–thawed ram semen. Cryobiology, 66: 126-130.
http://dx.doi.org/10.1016/j.cryobiol.2012.12.006
PMid:23276547
 
5. Azeredo, G.A., Esper, C.R. and Resende, K.T. (2001) Evaluation of plasma membrane integrity of frozen-thawed goat spermatozoa with or without seminal plasma. Small Rum. Res., 41(3): 257-263.
http://dx.doi.org/10.1016/S0921-4488(01)00189-4
 
6. Peterson, K., Kappen, M.A.P., Ursem, P.J.F., Nothling, J.O., Colenbrander, B. and Gadella, B.M. (2007) Microscopic and flow cytometric semen assessment of Dutch AI-bucks: Effect of semen processing procedures and their correlation to fertility. Theriogenology, 67(4): 863-871.
http://dx.doi.org/10.1016/j.theriogenology.2006.11.003
PMid:17126897
 
7. Kozdrowski, R., Dubiel, A., Bielas, W. and Dzieciol, M. (2007) Two protocols of cryopreservation of goat semen with the use of computer-analysis system. Acta Vet. Brno., 76: 601-604.
http://dx.doi.org/10.2754/avb200776040601
 
8. Sarıozkan, S., Bucak, M.N., Tuncer, P.B., Tasdemir, U., Kinet, H. and Ulutas, P.A. (2010) Effects of different extenders and centrifugation/washing on post thaw microscopic-oxidative stress parameters and fertilizing ability of Angora buck sperm. Theriogenology, 73: 316-323.
http://dx.doi.org/10.1016/j.theriogenology.2009.09.015
PMid:19880169
 
9. Carvajal, G., Cuello, C., Ruiz, M., Vazquez, J.M., Martinez, E.A. and Roca, J. (2004) Effect of centrifugation on Boar sperm. J. Androl., 25: 389-396.
http://dx.doi.org/10.1002/j.1939-4640.2004.tb02805.x
PMid:15064317
 
10. Webb, G.W. and Dean, M.M. (2009) Effect of centrifugation technique on post-storage characteristics of stallion spermatozoa. J. Equine Vet. Sci., 29(9): 675-680.
http://dx.doi.org/10.1016/j.jevs.2009.07.016
 
11. Purdy, P.H. (2006) A review on goat sperm cryopreservation. Small Rum. Res., 63(3): 215-225.
http://dx.doi.org/10.1016/j.smallrumres.2005.02.015
 
12. Lone, F.A., Islam, R., Khan, M.Z. and Sofi, K.A. (2011b) Effect of collection methods on the quality and quantity of spermatozoa recovered from the cauda epididymidis of slaughtered ram. Indian Vet. J., 88(9): 46-48.
 
13. Lone, F.A., Islam, R., Khan, M.Z. and Sofi, K.A. (2012) Effect of different egg yolk-based extenders on the quality of ovine cauda epididymal spermatozoa during storage at 4C. Reprod. Domest. Anim., 47: 257-262.
http://dx.doi.org/10.1111/j.1439-0531.2011.01847.x
PMid:21883509
 
14. Zemjanis, R. (1970) Diagnostic and Therapeutic Techniques in Animal Reproduction. 2nd ed. Williams and Willkins Co., Baltimore.
PMCid:PMC2047459
 
15. Foote, R.H. (1970) Influence of extender, extension rate and glycerolating technique on fertility of frozen bull semen. J. Dairy Sci., 53: 1478-1482.
http://dx.doi.org/10.3168/jds.S0022-0302(70)86418-9
 
16. Blom, E. (1977) Sperm morphology with reference to bull infertility. In: Some Papers Contributed to the 1st All India Symposium on Animal Reproduction. Punjab Agricultural University, Ludhiana, India, p66-81.
 
17. Watson, P.F. (1975) Use of Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. Microsc. Res. Tech., 61: 1-6.
http://dx.doi.org/10.1136/vr.97.1.12
 
18. Vasquez, J., Florentini, E.A., Camargo, L.A., Gonzales, J. and Valdivia, M. (2013) Hypo osmotic swelling test in epididymal ram (Ovis aries) spermatozoa. Livest. Sci., 157: 618-622.
http://dx.doi.org/10.1016/j.livsci.2013.08.023
 
19. Hoogewijs, M., Rijsselaere, T., De Vliegher, S., Vanhaesebrouck, E., De Schauwer, C., Govaere, J., Thys, M., Hoflack, G., Soom, A.V. and de Kruif, A. (2010) Influence of different centrifugation protocols on equine semen preservation. Theriogenology, 74(1): 118-126.
http://dx.doi.org/10.1016/j.theriogenology.2010.01.022
PMid:20207406
 
20. Islam, R., Ahmed, K. and Deka, B.C. (2006) Effect of holding and washing on the quality of goat semen. Small Rum. Res., 66: 51-57.
http://dx.doi.org/10.1016/j.smallrumres.2005.06.034
 
21. Jones, R.C. and Holt, W.V. (1974) The effect of washing on the ultra structure and cytochemistry of ram spermatozoa. J. Reprod. Fertil., 41(1): 159-167.
http://dx.doi.org/10.1530/jrf.0.0410159
 
22. Islam, R. and Ahmed, K. (2003) Effect of seminal plasma and of its removal on the quality of goat semen during preservation at 5C. Indian Vet. J., 80: 1302-1303.
 
23. Lone, F.A., Islam, R., Khan, M.Z. and Sofi, K.A. (2011a) Effect of transportation temperature on the quality of cauda epididymal spermatozoa of ram. Anim. Reprod. Sci., 123: 54-59.
http://dx.doi.org/10.1016/j.anireprosci.2010.10.012
PMid:21131148