Lowering the Level of Egg Yolk in Diluent or Removal of Seminal Plasma with Standard Egg Yolk Level do not Help in the Efficient Freezing of Buck Semen

Authors

  • Ghulam Rasool Bhat Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India
  • Farooz Ahmad Lone Goat Research and training center, Uttar Pradesh Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go-Anusandhan Sansthan (DUVASU), Mathura, UP, India
  • Arjuma Khatun Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India
  • Mukul Anand Goat Research and training center, Uttar Pradesh Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go-Anusandhan Sansthan (DUVASU), Mathura, UP, India
  • Rahul Dhariya Goat Research and training center, Uttar Pradesh Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go-Anusandhan Sansthan (DUVASU), Mathura, UP, India
  • Sanchi Rana Division of Animal Reproduction, Gynaecology and Obstetrics,Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India
  • Riaz Ahmad Shah Division of Animal Biotechnology,Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India
  • Haidar Ali Ahmad Division of Animal Nutrition, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India
  • Ishraq Hussain Division of Veterinary Biochemistry Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir University of Agricultural Science and Technology of Kashmir, Sri nagar 190006, UT Jammu and Kashmir, India

DOI:

https://doi.org/10.48165/ijar.2025.46.01.7

Keywords:

Buck, CASA, Egg yolk, Flow Cytometry, Seminal plasma, Washing

Abstract

The aim of this investigation was to compare the sperm quality in removal or intact seminal plasma buck semen frozen in tris diluent containing 20% and 2.5% egg yolk and to test whether lowering egg yolk level of diluent or removal of seminal plasma with standard egg yolk level helps in efficient freezing of buck semen. A total of 18 fresh ejaculates were divided into three aliquots of 500µl each corresponding to three treatment groups designated as BT1 (tris with 20% egg yolk, washed), BT2 (tris with 20% egg yolk, non-washed) and BT3 (tris with 2.5% egg yolk, non-washed). Semen aliquots of BT1 (washed), BT2 (unwashed) and BT3 (unwashed) were diluted and frozen with standard procedures. Post thaw semen quality was carried out with phase contrast microscopy, fluorescent microscopy, Computer Assisted Sperm Analysis (CASA) and flow cytometry. Majority of fertility associated with post-thaw sperm quality parameters evaluated by CASA, Flow Cytometry and subjective methods were significantly (p<0.05) lower in BT2 compared to BT1 and BT3 counterparts. In conclusion, the removal of seminal plasma or reduction in egg yolk concentration did not result in any real advantage for efficient post-thaw sperm survival in goats. An alternate diluent needs to be explored for buck semen which would contain both seminal plasma and egg yolk factors for better cryo-survival of sperm.

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Sarıozkan, S., Bucak, M.N., Tuncer, P.B., Tas¸demir ,U., Kinet, H. and Ulutas, P.A. (2010). Effects of different extenders and centrifugation/washing on post thaw microscopic-oxida tive stress parameters and fertilizing ability of Angora buck sperm. Theriogenology, 73: 316–323.

Schjenken, J.E. and Robertson, S.A. (2020). The Female Response to Seminal Fluid. Physiological Reviews, 100: 1077–1117. Sen, C. C., Tekin, K. and Akcay, E. (2015). Effect of Egg Yolk and Removal of Seminal Fluid on Semen Cryopreservation in Norduz Goat. Harran Üniv. Vet. Fak. Derg., 4(2): 64-67. Shah, S.A.H., Haider, M. S., Ahmed, H., Fayyaz, M. H. and Andrabi, S.M.H. (2023). Cryopreservation protocol resolv ing the "temperature" challenges of long-distance transpor tation of Beetal buck (Capra hircus) sperm. Small Rum. Res., 226: 107030.

Singh, R. K., Kumaresan, A., Chhillar, S., Rajak, S. K., Tripathi, U. K., Nayak, S. and Malhotra, R. (2016). Identification of suitable combinations of in vitro sperm function test for the

prediction of fertility in buffalo bull. Theriogenology, 86: 2263–2271.

Tabarez, A., Garcia, W. and Palomo, M.J. (2017). Effect of the type of egg yolk, removal of seminal plasma and donor age on buck sperm cryopreservation. Small Rum Res., 149: 91-98.

Van de Hoek, M., Rickard, J. P. and de Graa, S. P. (2022). Motility Assessment of Ram Spermatozoa. Biology, 11: 1715. Vasquez, J., Florentini, E.A., Camarago, L.A., Gonalez, J. and Valdivia, M. (2013). Hypo osmotic swelling test in ram (Ovis aries) spermatozoa. Livestock Science, 157: 618-622. Yimer, N., Noraisyah, A.H., Rosnina, Y. and Hafizal, A.M. (2014). Comparison of Cryopreservative Effect of Different Levels of Omega-3 Egg-Yolk in Citrate Extender on the Quality of Goat Spermatozoa. Pak. Vet. J., 34(3): 347-350.

Zou, J., Wei, L., Li, D., Zhang, Y., Wang, G., Zhang, L., Cao, P. and Li, G. (2022). Study on cryopreservation of Guanzhong dairy goat semen with bovine semen seminal plasma. Theriogenology, 189: 113-117.

Zou T, Liu X, Ding S and Xing J. 2010. Evaluation of sperm mito chondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia. J. Biomed. Res., 24(5): 404-10.

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Published

2025-04-14

How to Cite

Bhat, G.R., Lone, F.A., Khatun, A., Anand, M., Dhariya, R., Rana, S., … Hussain, I. (2025). Lowering the Level of Egg Yolk in Diluent or Removal of Seminal Plasma with Standard Egg Yolk Level do not Help in the Efficient Freezing of Buck Semen . The Indian Journal of Animal Reproduction, 46(1), 39–48. https://doi.org/10.48165/ijar.2025.46.01.7