Effect of Modified Freezing Diluent on Post-Thaw Sperm  Characteristics of Poor Freezable Sahiwal Bull

Authors

  • Manisha Sethi Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Nadeem Shah Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Tushar K Mohanty Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Mukesh Bhakat Division of Livestock Production Management, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Dileep K Yadav Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Pratyush Kumar Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India
  • Nishant Kumar Animal Reproduction, Gynaecology and Obstetrics, Artificial Breeding Research Centre (ABRC), ICAR-National Dairy Research Institute, Karnal-132001, Haryana, India

DOI:

https://doi.org/10.48165/ijvsbt.20.3.17

Keywords:

Bull, Glycerol, Modified freezing, Sperm, Trehalose

Abstract

The study aimed to enhance the cryo-survivability and fertilization potential of poor freezable Sahiwal bull semen by devising a modified  extender and freezing protocol. This protocol incorporated a combination of trehalose and glycerol as cryoprotectants and freezing  with an aluminium block. The semen samples were split into three; the split 1 was extended in routine tris fructose egg yolk glycerol  (TFEG) extender and underwent the conventional vapour freezing (Control group). The other split samples considered as treatment  samples were extended with diluent having 75 % of TFEG and 25 % of trehalose (280 mOsm/l both) with glycerol percentage being 5, and  frozen using a conventional vapour freezing (T1) and modified freezing protocol with an aluminium block (T2). After freezing-thawing,  the progressive sperm motility, viability, HOS response and acrosome integrity were significantly (p ≤ 0.05) improved in treatment 2  compared to the control and treatment 1 groups. Along with this, the degree of oxidative stress (MDA production) was substantially  lower in treatment groups (T1 and T2) compared to control group. The results highlight the promising effectiveness of the modified  freezing method incorporating trehalose and glycerol as cryoprotectants and using an aluminium block. The significance of these  findings lies in the potential application of this technique for freezing-thawing semen, particularly from bulls with poor freezability. 

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References

Ahmed, H., Jahan, S., Salman, M.M., & Ullah, F. (2020). Effect of different concentrations of fructose and glycerol in tris citric acid extender on post-thaw quality and fertility of buffalo bull spermatozoa. CryoLetters, 41(2), 106-114.

Aisen, E., Quintana, M., Medina, V., Morello, H., & Venturino, A. (2005). Ultramicroscopic and biochemical changes in ram spermatozoa cryopreserved with trehalose-based hypertonic extenders. Cryobiology, 50(3), 239-249.

Badr, M.R., Mary, G., & Hassan, M.H. (2010). Effect of trehalose on cryopreservation, oxidative stress and DNA integrity of buffalo spermatozoa. Journal of Reproduction and Infertility, 1(2), 50-57.

Bakas, L.S., & Disalvo, E.A. (1991). Effect of Ca2+ on the cryoprotective action of trehalose. Cryobiology, 28, 347-353.

Baracaldo, M.I., Barth, A.D., & Bertrand, W. (2006). Steps for freezing bovine semen: from semen collection to the liquid nitrogen tank. In: IVIS Reviews in veterinary medicine, International Veterinary Information Service- IVIS (eds), Ithaca, NY.

Baruah, K.K., Dhali, A., Mech, A., Bora, B., Das, J., Bora, R., Mondal, M., Sarmah, B.C., Deka, B.C., & Rajkhowa, C. (2013). Effect of concentration and addition method of glycerol on the quality of cryopreserved Mithun (Bosfrontalis) spermatozoa. Journal of Animal Physiology and Animal Nutrition, 97(6), 1051-1058.

Bucak, M.N., Ateşşahin, A., Varışlı, Ö.,Yüce, A., Tekin, N., & Akçay, 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.

Chen, Q., & Haddad, G.G. (2004). Role of trehalose phosphate synthase and trehalose during hypoxia: From flies to mammals. Journal of Experimental Biology, 207(18), 3125-3129.

Chhillar, S., Singh, V.K., Kumar, R., & Atreja, S.K. (2012). Effects of taurine or trehalose supplementation on functional competence of cryopreserved Karan Fries semen. Animal Reproduction Science, 135(1-4), 1-7.

Consuegra, C., Crespo, F., Dorado, J., Diaz-Jimenez, M., Pereira, B., Ortiz, I., & Hidalgo, M. (2019). Vitrification of stallion sperm using 0.25 mL straws: Effect of volume, concentration and carbohydrates (sucrose/trehalose/raffinose). Animal Reproduction Science, 206, 69-77.

DAHD (2022-23), Department of Animal Husbandry and Dairying, Annual Report, Government of India, New Delhi.

El-Badry, D.A., El-Maaty, A.M.A., & Gamal, A. (2017). The effect of trehalose supplementation of INRA-82 extender on quality and fertility of cooled and frozen-thawed stallion spermatozoa. Journal of Equine Veterinary Science, 48, 86-92.

El-Sheshtawy, R.I., Sisy, G.A., & El-Nattat, W.S. (2015). Effects of different concentrations of sucrose or trehalose on the post thawing quality of cattle bull semen. Asian Pacific Journal of Reproduction, 4(1), 26-31.

Fernández-Santos, M.R., Martínez-Pastor, F., García-Macías, V., Esteso, M.C., Soler, A.J., De Paz, P., Anel, L., & Garde, J.J. (2007). Extender osmolality and sugar supplementation exert a complex effect on the cryopreservation of Iberian red deer (Cervuselaphus hispanicus) epididymal spermatozoa. Theriogenology, 67(4), 738-753.

Foote, R.H. (1970). Fertility of bull semen at high extension rates in Tris-buffered extenders. Journal of Dairy Science, 53(10), 1475-1477.

García, B.M., Ferrusola, C.O., Aparicio, I.M., Miró-Morán, A., Rodriguez, A.M., Bolaños, J.G., Fernandez, L.G., da Silva, C.B., Martinez, H.R., Tapia, J.A., & Peña, F.J. (2012). Toxicity of glycerol for the stallion spermatozoa: effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential. Theriogenology, 77(7), 1280-1289.

Harrison, R.A.P., & Vickers, S.E. (1990). Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. Reproduction, 88(1), 343-352.

Hu, J.H., Zan, L.S., Zhao, X.L., Li, Q.W., Jiang, Z.L., Li, Y.K., & Li, X. (2010). Effects of trehalose supplementation on semen quality and oxidative stress variables in frozen-thawed bovine semen. Journal of Animal Science, 88(5), 1657- 1662.

Iqbal, S., Naz, S., Ahmed, H., & Andrabi, S.M.H. (2018). Cryoprotectant effect of trehalose in extender on post‐thaw quality and in vivo fertility of water buffalo (Bubalusbubalis) bull spermatozoa. Andrologia, 50(1), e12794.

Isachenko, E., Isachenko, V., Katkov, I.I., Dessole, S., & Nawroth, F. (2003). Vitrification of mammalian spermatozoa in the absence of cryoprotectants: From past practical difficulties to present success. Reproductive Biomedicine Online, 6(2), 191-200.

Jeyendran, R.S., Vander-Ven, H.H., Perez-Pelaez, M., Crabo, B.G., & Zanevld, L.J.D. (1984). Development of an assay to assess the functional

integrity of the human sperm membrane and its relationship to other semen characters. Reproduction, 70(1), 219-228.

Mathur, A.K., Tyagi, S., Mukherjee, S., & Singh, S.P. (2002). Semen quality attributes in Frieswal bulls. Indian Journal of Animal Sciences, 72(1), 59-60.

Mukhopadhyay, C.S., Gupta, A.K., Yadav, B.R., Khate, K., Raina, V.S., Mohanty, T.K., & Dubey, P.P. (2010). Subfertility in males: An important cause of bull disposal in bovines. Asian-Australasian Journal of Animal Sciences, 23(4), 450-455.

Najafi, A., Zhandi, M., Towhidi, A., Sharafi, M., Sharif, A.A., Motlagh, M.K., & Martinez- Pastor, F. (2013). Trehalose and glycerol have a dose-dependent synergistic effect on the post thawing quality of ram semen cryopreserved in a soybean lecithin-based extender. Cryobiology, 66(3), 275-282.

Perumal, P., Selvaraju, S., Selvakumar, S., Barik, A.K., Mohanty, D.N., Das, S., Das, R.K., & Mishra, P.C. (2011). Effect of pre-freeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates. Reproduction in Domestic Animals, 46(4), 636-641.

Reddy, N.S.S., Mohanarao, G.J., &Atreja, S.K. (2010). Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalusbubalis) sperm quality following cryopreservation. Animal Reproduction Science, 119(3-4), 183-190.

Shaikh, K.Q., Nakhashi, H.C., Suthar, B.N., Sutaria, P.T., & Sharma, V.K. (2016). Trehalose as semen diluent additive for cryopreservation of Kankrej bulls semen. Life Sciences Leaflets, 71, 29-40.

Starciuc, T., Malfait, B., Danede, F., Paccou, L., Guinet, Y., Correia, N.T., & Hedoux, A. (2020). Trehalose or Sucrose: Which of the two should be used for stabilizing proteins in the solid state? A dilemma investigated by in situ micro-raman and dielectric relaxation spectroscopies during and after freeze drying. Journal of Pharmaceutical Sciences, 109(1), 496-504.

Suleiman, S.A., Ali, M.E, Zaki, M.S., Malik, E.M.E.A., & Nast, M.A. (1996). Lipid peroxidation and human sperm motility: protective role of vitamin E. Journal of Andrology, 17(5), 530-537.

Swain, D.K., Yadav, S., & Singh, S.K. (2017). Effect of four different in vitro incubation temperatures on functional dynamics, process of capacitation and apoptosis in goat spermatozoa. Small Ruminant Research, 147, 120-124.

Published

2024-05-10

How to Cite

Sethi, M., Shah, N., Mohanty, T.K., Bhakat, M., Yadav, D.K., Kumar, P., & Kumar, N. (2024). Effect of Modified Freezing Diluent on Post-Thaw Sperm  Characteristics of Poor Freezable Sahiwal Bull. Indian Journal of Veterinary Sciences and Biotechnology, 20(3), 87–91. https://doi.org/10.48165/ijvsbt.20.3.17