000 04966nam a22002177a 4500
005 20151020124824.0
008 151020b xxu||||| |||| 00| 0 eng d
041 _aeng
082 _a2318-T
100 _aFarhan Younas (2007-VA-495)
110 _cProf. Dr. Mian Abdul Sattar
245 _aEffect Of L-Cysteine And Glutathione On Post Thaw Quality Of Sahiwal Bull Spermatozoa
260 _c2015
300 _a52p.;
502 _aFreezing and thawing of semen leads to production of reactive oxygen species (ROS) due to plasma membrane lipid peroxidation. Because of this semen quality can be compromised. To overcome this problem, antioxidants have been used in cryopreservation medium. Glutathione and cysteine have thiol groups which penetrate into the cell and protect it from oxidative stress. In this study, effect of different concentrations of cysteine and glutathione on post thaw quality of Sahiwal bull spermatozoa was determined. Semen was collected with artificial vagina from five mature regular donor Sahiwal bulls kept at the Semen Production Unit Qadirabad, Sahiwal. Semen samples possessing >60% motility and >500x10 6 sperm/ml were included in study. After collection, semen samples from five bulls were pooled, divided into seven equal aliquots and kept at 37 ºC in water bath. After that dilution was done with Tris citric egg yolk extender having different concentrations of cysteine and glutathione as Con (0.0 mM), C1 (1.0 mM cystein), G1 (1.0 mM glutathione), CG0.5/1(0.5 mM Cysteine+1.0 mM glutathione), CG1/0.5 (1.0 mM cysteine+0.5 mM Glutathione), CG0.5/0.5 (0.5 mM cysteine+0.5 mM glutathione) and CG1/1 (1.0 mM cysteine+1.0 mM glutathione). Diluted samples were cooled to 4ºC in two hours and equilibrated for 4 hours at 4 o C. After that they were packaged into 0.5 ml French semen straws (20x10 6 sperm/straw). All semen straws were placed 4cm above liquid nitrogen surface in vapors for 10 minutes. Then, semen straws were plunged into liquid nitrogen for freezing and stored until post thaw analysis. The experiment was repeated for five times (replicates = 5). Four semen straws/treatment were thawed for 30 seconds in water bath at 37ºC and evaluated for visual motility, plasma membrane integrity (PMI), acrosome integrity, mitochondrial trans membrane potential and CASA motility parameters and kinematics. 42 Summary PMI in group CG0.5/0.5 was significantly higher (40.00±1.42 %) as compared to Con 26.67±0.80 (P<0.5). Plasma membrane integrity in groups CG1/1, CG0.5/1, G1 and C1 was significantly higher (36.00±1.88 %, 36.20±1.07 %, 33.60±1.21 % and 32.80±0.80 % respectively) as compared to Con (26.67±0.80 %) (P<0.05). There was no significant difference in C1 (32.80±0.80 %) and G1 (33.60±1.21 %) (P>0.05). In case of acrosome integrity, NAR value of group CG0.5/0.5 was significantly higher (71.40±1.08 %) as compared to Con (59.67±0.37 %) (P<0.05). All other groups also showed significant differences as compared to Con (P<0.05). CG0.5/0.5 also showed significantly higher NAR value (71.40±1.08 %) as compared to C1 (64.40±1.40 %) and G1 (67.60±2.07 %) (P<0.05). CG0.5/0.5 had significantly higher value (71.40±1.08 %) as compared to CG1/0.5 and CG1/1 (65.60±0.81 % and 68.80±0.97 % respectively) (P<0.05). CG0.5/0.5 had significantly higher subjective motility (54.00±1.88) as compared to Con (36.66±0.92) Mitochondrial transmembrane potential of CG0.5/0.5 was significantly higher (37.00±0.71 %) as compared to Con (25.33±1.28 %) (P<0.05). All the other treatment groups also had higher mitochondrial transmembrane potential as compared to Con (P<0.05). In groups of combination of cysteine and glutathione, CG0.5/0.5 showed significant difference (37.00±0.71 %) as compared to CG1/1 and CG1/0.5 (29.00±1.00 % and 33.80±0.86 %) respectively (P<0.05). CASA results showed that CG1/1 had significantly higher motility as compared to the control. But the percentage of progressive spermatozoa was significantly higher in CG0.5/0.5. VSL of group CG0.5/0.5 was significantly higher (53.33±2.90 %) as compared to Con (45.10±0.50 %). However, VSL, VCL, ALH and BCF did not vary significantly among groups. STR and LIN of group CG0.5/0.5 were significantly higher as compared to the control group. 43 Summary In conclusion, addition of cysteine and glutathione in tris citric egg yolk extender improved the post thaw quality of Sahiwal bull spermatozoa. In case of additive effect of cysteine and glutathione, CG0.5/0.5 showed higher plasma membrane integrity, acrosome integrity, mitochondrial transmembrane potential, progressive and rapid spermatozoa as compared to CG0.5/1, CG1/0.5 and CG1/1. 44
650 _aDepartment of Theriogenology
700 _aDr. Syed Murtaza Hasan Andrabi
700 _aProf. Dr. Nasim Ahmad:
700 _aProf. Dr. Aneela Zameer Durrani
942 _cTH
999 _c6373