Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.50

  • SJR 0.263

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Indian Journal of Animal Research, volume 52 issue 8 (august 2018) : 1134-1140

Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen

P.J. Chaudhary, A.J. Dhami, D.V. Chaudhari, K.K. Hadiya, J.A. Patel
1Department of Animal Reproduction Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand-388 001, Gujarat, India
Cite article:- Chaudhary P.J., Dhami A.J., Chaudhari D.V., Hadiya K.K., Patel J.A. (2018). Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen. Indian Journal of Animal Research. 52(8): 1134-1140. doi: 10.18805/ijar.B-3453.
This study was undertaken on three mature bulls each of Gir cattle and Surti buffalo breeds to evaluate the comparative efficacy of egg yolk based standard TFYG (Tris-citrate-fructose-yolk-glycerol) extender and egg yolk free soybean based commercial extenders Optixcell® (IMV, France) and Andromed® (Minitube, Germany) under split-sample technique. The ejaculates (9/bull) were extended @ 100×106 sperm ml-1 with three extenders and frozen using biofreezer following 4 hr of equilibration. The pooled means of progressively motile sperm observed (irrespective of extenders) at initial, pre-freeze and post-thaw stage in Gir bulls semen were 76.53±0.53, 71.11±0.53 and 39.86±0.90% and in Surti buffalo 80.76±0.39, 74.65±0.45 and 40.35±1.07%, respectively. The corresponding values for live sperm were 75.64±0.76, 69.01±0.97 and 47.99±1.11 % for Gir and 80.90±0.45, 75.76±0.48 and 52.33±0.86 % for Surti buffalo; and those of intact acrosome 94.29±0.25, 90.29±0.27 and 79.29±0.33 % for Gir bulls, and 93.94±0.21, 89.94±0.23 and 78.95±0.26 % for Surti buffalo semen, respectively. The HOS reactive sperm at initial, pre-freeze and post-thaw stage were 76.18±0.74, 71.04±0.76 and 27.90±0.70 % for Gir, and 81.83±0.35, 76.47±0.39 and 27.83±0.68 % for Surti bulls, respectively. The overall mean post-thaw incubation (37°C) survival of spermatozoa observed at 60, 120 and 180 min were 28.40±0.91, 17.78±0.86 and 9.44±0.72% for Gir bulls semen, and 28.01±0.99, 18.40±1.01 and 10.51±0.93% for Surti buffalo semen, respectively. Optixcell was proved superior, and at par with TFYG, than the Andromed in maintaining greater motility, viability, morphology, acrosomal/plasma membrane integrity including post-thaw sperm longevity of cattle and buffalo spermatozoa with significant differences only in sperm motility and post-thaw longevity. The motile, live and HOST reactive sperm were significantly higher in buffalo semen than cattle at initial and pre-freeze stage, but not at post-thaw stage. The results showed that egg yolk free commercial Optixcell extender and egg yolk based TFYG extender were at par in terms of most of the sperm quality traits, hence any one of them can be preferred over Andromed for successful routine cryopreservation of cattle and buffalo semen. 
  1. Aires, V.A., Hinsch, K.D., Schloesser, F.M., Bogner, K., Schloesser, S.M. 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: 269-279.
  2. Akhter, S., Ansari, M.S., Andrabi, S.M.H., Rakha, B.A., Ullah, N. and Khalid, M. (2012). Soya-lecithin in extender improves the freezability and fertility of buffalo (bubalus bubalis) bull spermatozoa. Reprod. Dom. Anim., 47(5), 815-819
  3. Akhter, S., Ansari, M.S., Rakha, B.A., Andrabi, S.M.H., Iqbal, S. and Ullah, N. (2010). Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell® extender. Theriogenology, 74: 951-955.
  4. Amirat, L., Anton, M., Tainturier, D., Chatagnon, G., Battut, I. and Courtens, J. (2005). Modifications of bull spermatozoa induced by three extenders: Biociphos, low density lipoprotein and Triladyl, before, during and after freezing and thawing. Reproduction, 129: 535-543.
  5. Ansari, M.S., Rakha, B.A., Akhter, S. and Ashiq, M. (2016). Optixcell improves the post-thaw quality and fertility of buffalo bull sperm. Theriogenology, 85(3): 528-532.
  6. Asr, S.J., Beheshti, R. and Kohram, H. (2011). The evaluations of Tris-citrate-egg yolk or Bioxcell extenders on the post-thawed buffalo sperm parameters. Annals Biol. Res., 2(4): 360-365. 
  7. Beran, J., Stadnik, L., Bezdicek, J., Louda, F., Citek, J. and Duchacek, J. (2012). Effect of sire and extender on sperm motility and share of live or dead sperm in bulls’ fresh ejaculate and in AI doses after thawing. Archiv Tierzucht, 55(3): 207-218. 
  8. Beura, D., Mishra, P.C., Das, P., Mohanty, D.N., Barik, A.K. and Biswal, S.S. (2014). Effect of Bioxcell and Tris citric egg yolk extender on post thaw semen quality and in vivo fertility in cross bred Jersey bull. Indian J. Anim. Reprod., 35(1): 29-32.
  9. Bousseau, S., Billard, J.P., Marquan-Le, Guienne, B., Gurien, B., Camus, A. and Lechat, M. (1998). Comparison of bacteriological qualities of various egg yolk sources and the in vitro and in vivo fertilizing potential of bovine semen frozen in egg yolk free or lecithin based diluents. Theriogenology, 50: 699-706. 
  10. Celeghini, E.C.C., Arruda, R.P., Andrade, A.F.C., Nascimento, J., Raphael, C.F. and Rodrigues, P.H.M. ( 2008). Effects that bovine sperm cryopreservation using two different extenders has on sperm membranes and chromatin. Anim. Reprod. Sci., 104: 119-131.
  11. Chaudhari, D.V., Dhami, A.J., Hadiya, K.K. and Patel, J.A. (2015). Relative efficacy of egg yolk and soya milk-based extenders for cryopreservation of buffalo semen. Vet. World, 8(2): 239-244.
  12. Chaudhari, D.V., Dhami, A.J., Patel, J.A. and Hadiya, K.K. (2017). Evaluation of comparative efficacy of egg yolk and soya based extenders for refrigeration preservation (5ºC) of buffalo semen. Indian J. Anim. Res., 51(1): 21-24. DOI:10.18805/ijar.v0i0f.3808.
  13. Crespilho, A.M., Filho, M.F., Aqua, J.A., Nichi, M., Monteiro, G.C., 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. Livestock Sci., 149: 1-6.
  14. Dhami, A.J., Jani, V.R., Mohan, G. and Sahni, K.L. (1994). Effect of extenders and additives on freezability, post-thaw thermo resistance and fertility of frozen Murrah buffalo semen under tropical climate. Buffalo J., 10(1): 34-45. 
  15. Gil, J., Januskauskas, A., Haard, M., Johanisson, A., Soderquist, L. and Rodriguez-Martinez, H. (2000). Functional sperm parameters and fertility of bull semen extended in Biociphos-Plus and Triladyl. Reprod. Dom. Anim., 35: 69-77.
  16. Hinsch, E., Hinsch, K., Boehm, J., Schill, W. and Mueller-Schloesser, F. (1997). Functional parameters and fertilization success of bovine semen cryopreserved in egg yolk containing extenders. Reprod. Dom. Anim., 32: 143-149.
  17. Jayendran, R.S., Van der Ven, H.H., Perez-Pelaez, M., Crabo, B.G. and Zaneveld, L.J.D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil., 70: 219-228.
  18. Jimenez, F., Puchades, S., Moce, E., Cartro, M.P., Vicente, J.S. and Rodriguez, M. (2004). Use of powdered egg yolk vs. fresh egg yolk for the cryopreservation of ovine semen. Reprod. Dom. Anim., 39: 438-441.
  19. Kumar, P., Saini, M., Kumar, D., Balhara, A.K., Yadav, S.P., Singh, P. and Yadav, P.S. (2015). Liposome-based semen extender is suitable alternative to egg yolk-based extender for cryopreservation of buffalo (Bubalus bubalis) semen. Anim. Reprod. Sci., 159: 38-45.
  20. Layek, S.S., Mohanty, T.K., Kumaresan, A. and Parks, J.E. (2016). Cryopreservation of bull semen: evolution from egg yolk based to soybean based extenders. Anim. Reprod. Sci., http://dx.doi.org/10.1016/j.anireprosci. 2016.04.013
  21. Leeuw, A.M., Haring, R.M., Lansbergen, L.M. and Daas, J.H. (2000). Fertility results using bovine semen cryopreserved with extenders based on egg-yolk and soybean extract. Theriogenology, 54: 57-67.
  22. Leite, T.G., Filhoa, V.R., Arrudab, R.P., Andradeb, A.F., Emericka, L.L., Zaffalonb, F.G., Martinsa, J.A. and Andradec, V.J. (2010). Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry. Anim. Reprod. Sci., 120: 31-38.
  23. Meena, G.S., Raina, V.S., Gupta, A.K., Mohanty, T.K. and Bishist, R. (2010). Comparative performance of Biociphos and Tris-egg yolk based extenders for buffalo semen preservation. Indian J. Anim. Sci., 80: 414-417.
  24. Muino, R., Fernandez, M. and Pena, A.I. (2007). Post-thaw survival and longevity of bull spermatozoa frozen with an egg yolk-based or two egg yolk-free extenders after an equilibration period of 18 hours. Reprod. Dom. Anim., 42: 305-311. 
  25. Muralinath, E., Murthy, A.S.N., Rao, A.V.N., Harnath, G.B., Reddy, B.B. and Rao, V.H. (1990). Effect of supplementation of prostaglandins on post-thaw motility and in vitro motility in cervical mucus of frozen buffalo spermatozoa. Indian J. Anim. Reprod., 11(1): 10-13.
  26. Rana, C.M., Dhami, A.J., Zalu, G.K., Vibhakar K.V. and Landani, N.G. (2003). Preservation of Gir (Bos indicus) and Jafarabadi 
  27. (Bubalus bubalis) semen at 5°C and ultralow temperature using Sephadex filtration technique. Indian J. Anim. Sci., 73: 858-863.
  28. Rastegarnia, A., Shahverdi, A., Rezaei, T. and Shafiepour, V. (2013). In vitro comparison of soybean lecithin-based extenders for 
  29. cryopreservation of buffalo (Bubalus bubalis) semen. Comp. Clin. Path., 22: 1-10. 
  30. Sharma, S. and Atreja, S.K. (2014). A comparative evaluation of spermatozoa quality after cryopreservation of bovine semen in conventional Egg Yolk Tris Citrate (EYTC) versus “in house” Soya Milk Tris (SMT) extender. Compendium, National Seminar on Biotechnological Approaches to Challenges in Animal Health and Production, DUVASU, Mathura, India, 6-7 March, pp. 88-89. 
  31. Singh, V.K., Singh, A.K., Kumar, R., and Atreja. S.K. (2013). Development of soya milk extender for semen cryopreservation of Karan Fries (crossbreed cattle). CryoLetters, 34(1): 52-61.
  32. Snedecor, G.W. and Cochran, W.G. (1994). Statistical Methods. 8th edn. Iowa State University Press, Ames, Iowa, USA.
  33. Taraphder, S., Gupta, A.K., Raina, V.S. and Tomar, S.S. (2001). Effect of incubation (37°C) on post-thaw motility of Murrah buffalo bull spermatozoa. Indian J. Dairy Sci., 54(2): 80-83.
  34. Thun, R., Hurtado, M. and Janett, F. (2002). Comparison of Biociphos-Plus and Tris egg yolk extender for cryopreservation of bull semen. Theriogenology, 57: 1087-1094.
  35. Veerabramhaiah, K., Rao, A.S., Rao, V.H., Naidu, K.V. and Rao, S.T.V. (2011). Efficacy of the Tris and Biociphos-Plus extenders on the freezability of Punganur bull semen. Indian J. Anim. Reprod., 32(2): 1-4.
  36. Watson, P.F. (1975). Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa. Vet. Rec., 97: 12-15. 

Editorial Board

View all (0)