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Research (Published online: 20-03-2016)

13. Effect of different antioxidant additives in semen diluent on cryopreservability (−196C) of buffalo semen - Hardik A. Patel, G. M. Siddiquee, Dinesh V. Chaudhari and Vishal S. Suthar

Veterinary World, 9(3): 299-303



   doi: 10.14202/vetworld.2016.299-303



Hardik A. Patel: Sabarmati Ashram Gaushala, Ahmedabad, Gujarat, India;

G. M. Siddiquee: Department of Gynaecology and Obstetrics, College of Veterinary Sceicne and Animal Husbandry, Sardarkrushinagar Dantiwada Agricultural University, Deesa, Banaskanth, Gujarat, India;

Dinesh V. Chaudhari: Pashupalan Sansodhan Kendra, Veterinary College, Anand Agricultural University, Anand, Gujarat, India;

Vishal S. Suthar: Directorate of Research, Kamdhenu University, Gandhinagar, Gujarat, India;


Received: 04-10-2015, Revised: 30-01-2016, Accepted: 06-02-2016, Published online: 20-03-2016


Corresponding author: Vishal S. Suthar, e-mail:

Citation: Patel HA, Siddiquee GM, Chaudhari DV, Suthar VS (2016) Effect of different antioxidant additives in semen diluent on cryopreservability (−196C) of buffalo semen, Veterinary World, 9(3): 299-303.

Aim: The aim of this study was to evaluate the effect of different antioxidant additives in standard tris-fructose-egg yolkglycerol (TFYG) extender on the cryopreservability of buffalo semen.

Materials and Methods: Semen collection using artificial vagina, twice weekly for 5 weeks from three pedigreed health breeding bulls of Mehsani breed, aged between 6 and 8 years. Immediately after initial evaluation all 30 qualifying ejaculates (10/bull) were split into three aliquots and diluted at 34C keeping the concentration of 100 million spermatozoa/ml with standard TFYG extender as control and TFYG having two antioxidant additives - Cysteine HCl at 1 mg/ml and ascorbic acid at 0.2 mg/ml to study their comparative performance. Semen filled in French Mini straws using IS-4 system and gradually cooled to 4C and equilibrated for 4 h in cold handing cabinet. After completion of equilibration, straws were cryopreserved in LN2 by Programmable Bio-freezer. Semen was examined at post-dilution, post-equilibration, and post-thaw stages for sperm quality parameters, and at each stage plasma was separated for enzymatic analysis of aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (AKP).

Results: The mean percentage of sperms in TFYG, TFYG + cysteine HCl and TFYG + ascorbic acid diluents at postthaw stage in terms of progressive motility (52.830.52, 57.830.52, 57.830.52), livability (78.700.21, 82.330.23, 81.730.22), and abnormality (5.430.21, 5.030.17, 5.230.18) varied significantly (p<0.05) between control TFYG and TFYG having antioxidant additives. The mean U/L activities of AST (78.700.47, 72.800.48, 73.300.54), LDH (172.700.41, 155.780.42, 156.330.41), and AKP (103.610.34, 90.200.34, 91.030.34) in semen diluted with TFYG, TFYG + cysteine HCl and TFYG + ascorbic acid diluents at post-thaw stage, respectively, which showed significantly (p<0.05) higher leakage of enzymes in control TFYG than TFYG incorporated with additives.

Conclusion: Incorporation of antioxidant additives such as cysteine HCl and ascorbic acid in standard TFYG diluents improves sperm quality parameters, reduces enzyme leakage, and ultimately advances cryopreservability of buffalo semen.

Keywords: antioxidants, cryopreservation, diluent, egg yolk, enzyme, semen.

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