48-Title: Effect of buffalo specific semen extenders on hos test positive sperms at different stages of cryopreservation in Murrah buffalo bulls

48-Title: Effect of buffalo specific semen extenders on hos test positive sperms at different stages of cryopreservation in Murrah buffalo bulls

Authors: Satish, GN Purohit, RK Sharma, Pradeep Kumar, A Jerome, JS Mehta, Devender Kumar,  Renu Bala, Nisha Verma and Sajjan Singh

Source: Ruminant Science (2022)-11(1):235-237.

How to cite this manuscript: Satish, Purohit GN, Sharma RK, Kumar Pradeep, Jerome A, Mehta JS, Kumar Devender,  Bala Renu, Verma Nisha and Singh Sajjan (2022). Effect of buffalo specific semen extenders on hos test positive sperms at different stages of cryopreservation in Murrah buffalo bulls. Ruminant Science 11(1):235-237.


With an objective to evaluate the best buffalo specific extender, semen was collected from Murrah buffalo bulls (n=6) by artificial vagina and subjected to dilution using buffalo specific extenders Tris egg yolk, CIRB (LDL), GVE (TEYC with 1 mg/mL tocopherol) and GSE (TEYC with 0.25% sericin). Semen was cryopreserved and HOS test-positive sperms were evaluated immediately after dilution, at the equilibration stage and in cryopreserved semen thawed after 24 h. A comparison of HOST reactive sperms at three different stages revealed that the overall HOST reactive sperms were non-significantly different for the different extenders although slightly higher for the CIRB extender. At the equilibration stage and frozen-thawed semen the HOST reactive sperms were significantly higher (P<0.001) for the CIRB extender as compared to TEYC, GSE, and the GVE extender. It was concluded that the LDL-based extender (CIRB) is optimum for the dilution and cryopreservation of buffalo semen.


Amann RP and Graham JK (1993). Spermatozoal function. In: Equine Reproduction. Eds: AO Mcinnon and JL Voss, Lea and Febiger, London, UK. 715-745.

Batool A,  Mehboob K, Qadeer S, Ansari MS, Rakha BA, Ullah N, Andrabi SKM and Akhter S (2012). Effect of á-tocopherol acetate and ascorbic acid in extender on quality of zebu bull spermatozoa. Pakistan Journal of Zoology 44(6):1487-1491.

Cabrita E, Alvarez R, Anel E and Herraez MP (1999). The hypo-osmotic swelling test performed with coulter counter: A method to assay functional integrity of sperm membrane in rainbow trout. Animal Reproduction Science 55(3-4):279-287.

Dalal J, Chandolia RK, Pawaria S, Kumar A, Kumar D, Selokar NL, Jerome A, Yadav PS and Kumar P (2020). Low density lipoproteins protect sperm during cryopreservation in buffalo: Unraveling mechanism of action. Molecular Reproduction Development 1-14.

Jeyendran RS, Vander-Ven HH, Perez-Pelaez M, Crabo BG and Zanevld LJD (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characters. Journal of Reproduction Fertility 70:219-228.

Kharde SD, Ingole SD, Kekan PM, Bharucha SV and Nagvekar AS (2021). Seasonal evaluation of semen in Holstein Friesian cross breed bulls. Ruminant Science 10(2):329-332.

Kumar D, Kumar P and Yadav PS (2016). Quantitative evaluation of buffalo semen by CASA during cryopreservation process. Indian Journal of Animal Reproduction 37(1):15-18.

Kumar P, Saini M, Kumar D, Balhara AK, Yadav SP, Singh P and Yadav PS (2015). Liposome-based semen extender is suitable alternative to egg yolk-based extender for the cryopreservation of buffalo (Bubalus bubalis) semen. Animal Reproduction Sciences 159:38 45.

Kumar S, Anand M and Yadav S (2014). Effect of dietary vitamin e supplementation on cryopreserved semen quality in Haryana bulls. Ruminant Science 3(1):25-28.

Neild D, Chaves G, Flores M, Mora N, Beconi M and Aguero A (1999). The hypo-osmotic swelling test in equine spermatozoa. Theriogenology 51:721-727.

Patil S, Kumar P, Singh G, Bala R, Jerome A, Patil CS, Kumar D, Singh S and Sharma RK (2020). Semen dilution effect on sperm variables and conception rate in buffalo. Animal Reproduction Sciences 214:106304.

Perez-Llano B, Lorenzo JL, Yenes P, Trejo A and Garcia-Casado P (2001). A short hypo-osmotic swelling test for the prediction of boar sperm fertility. Theriogenology 56(3): 387-398.

Purohit GN, Singh B, Vyas SK and Yadav SBS (1998). Studies on sexual behaviour and seminal quality characteristics of Surti buffalo bulls and their interrelationships. Indian Journal Agricultural Research 19:62-63.

Rodriguez-Gil JE, Monserrat A and Rigau T (1994). Effects of hypo-osmotic incubation on acrosome and tail structure of canine spermatozoa. Theriogenology 42(5): 815-829.

Rota A, Penzo N, Vincenti L and Mantovani R (2000). Hypo-osmotic swelling as a screening assay for testing in vitro fertility of bovine spermatozoa.Theriogenology 53(7):1415-1420.

Shukla MK, Misra AK and Gupta HP (2012). Caffeine as semen additive to improve Murrah buffalo (Bubalus bubalis) semen cryopreservation. Ruminant Science 1(2):121-124.

Singh Vipin, Kumar Anuj, Yadava CL, Patel Akhil, Yadav Brijesh, Panday Vijay and Saxena Atul (2018). Effect of season on certain seminal plasma enzymes profile at pre-freeze and post thaw period in the Haryana bull. Ruminant Science 7(2):279-282.