Tayyab Munir

Use Of Butylated Hydroxytoluene (Bht) As Antioxidant In Buffalo Semen Extender - 2011

The Buffalo is the major milk producing animal in Pakistan. Nili-Ravi breed of buffalo is the one of the best breeds of the world. Buffalo contributes seventy percent of total milk production in Pakistan. The excellent buffalo bulls are few in number and their semen volume and sperm concentration is also low as compared to cattle. The animal has lower fertility particularly when inseminated with frozen semen. There are many reasons for poor fertility of buffalos and one of the most important reasons may reasons for poor fertility of buffalos and one of the most important reasons may be the production of Reactive Oxygen Species (POS) from the plasma membrane of spermatozoa causing damage to the membrane integrity due to lipid peroxidation. The addition of optimum level of antioxidants like BHT to buffalo semen extender before freezing may be answer to this problem. The main objective of the present study was to determine the level of total of antioxidants in the semen and to optimize the quantity of BHT in the semen extender of the Nili Rave Buffalo bulls.
In this study semen from Nili-Ravi buffalo bulls ( n=10) was collected twice per week and diluted with Tris egg yolk citrate extender. Antioxidant BHT was added to the extended semen at the rate of 1.0mM, 1.0 mM and 2 mM while Control was containing no BHT. The semen was visually analyzed under phase contrast microscope for motility, vitality (Eosin/Nigrosin staining), plasma membrane (HOS assay) and acrosomal integrity (NAR), at three stages i.e. after dilution, before freezing and after freezing. For the determination of total antioxidant capacity (TAC), semen samples were transported under ice to Spectophotometric Laboratary, GC. University Lahore. Data collected was presented as mean= SEM. Treatment groups were compared using one way ANOVA and Results were compared by using Duncan Multiple Range Test using SPSS (version 16.0) ; SPSS Chicago IL.
Result of this study show significant (P< 0.05) improvement in semen motility, vitality, plasma membrane and acrosomal integrity at 1.5 mM BHT concentration as compared to Control, however, insignificant difference was noticed with in treated groups and with in bulls. Maximum TEAC value (2.42) was observed in the extended semen containing two mM BHT and minimum TEAC values (0.58mM) in the post thaw semen sample with BHT.
It is concluded that addition of antioxidants in the buffalo semen improve the semen quality before and after freezing. Fortification of buffalo semen extender with BHT (@ of 1-1.5 mM) improved post thaw semen quality and hoped to increase the fertility of buffalo. However, field fertility trials need to be carried out for confirmation.



Department of Theriogenology

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