Effects of partial deoxygenation of extender on plasma membrane integrity and enzyme activity in frozen-thawed crossbred bull semen

Title: Effects of partial deoxygenation of extender on plasma membrane integrity and enzyme activity in frozen-thawed crossbred bull semen

Authors: AR Mustapha, YA Beigh, K Abhishek, K Rahul, D Omer, A Kumar, JK Prasad, S Bhure, N Srivastava and SK Ghosh

Source: Ruminant Science (2017)-6(2):327-331.

Cite this reference as: Mustapha AR, Beigh YA, Abhishek K, Rahul K, Omer D, Kumar A, Prasad JK, Bhure S, Srivastava N  and Ghosh SK (2017). Effects of partial deoxygenation of extender on plasma membrane integrity and enzyme activity in frozen-thawed crossbred bull semen. Ruminant Science 6(2):327-331.

Abstract

Present study was conduted to evaluate the effect of partial deoxygenation of extender using nitrogen- degasification method on freezability of crossbred bull semen. After the initial semen evaluation (progressive motility >70%), it was extended with the degasified egg yolk-citrate extender of different oxygen concentrations: 11.7 ppm (Control-Group I), 2 ppm (Group II), 4 ppm (Group III) and 8 ppm (Group IV). An aliquot from each group was centrifuged at 3000 g for 10 minute at 4 oC to separate the seminal plasma for estimation of enzymes {Lactate dehydrogenase (LDH) and alkaline phosphatase (ALP)}. Plasma membrane integrity and lipid peroxidation status were estimated in all the three stages. The progressive motility was significantly (p<0.05) higher at both prefreeze and post thaw stages in 4 ppm group as compared to all other groups. The average seminal plasma content of lactate LDH was the lowest in Group-III indicating low LDH leakages over the control as well as in 2 and 8 ppm groups. Also, the level of activities of ALP was highest in Group-III than the rest of treatment and control groups. Highest percentage (P<0.05) of intact plasma membrane was observed at both prefreeze and post thaw stages in 4 ppm than the control and other two groups. The lipid peroxidation (LPO) in 4 ppm level of dissolved oxygen extender was also found to be significantly (p<0.05) lower than the remaining groups. The present study indicate that dissolved oxygen level of extender at 4 ppm level is beneficial in maintaining post-thaw motility, optimum functions of sperm plasma membrane and seminal plasma enzymes by minimizing enzyme leakages in sperm cell during cryopreservation.

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