الفهرس 
IntroductionOnly 14 pages are availabe for public view
 |  | from | 110 |  |  |
| | | from | 110 | | |
Abstract
Buffalo is the main farm animal in Egypt, it is consider the backbone of agriculture economy producing high quality milk and meat even raring in poor conditions. Chitosan nanoparticles characterization was done by transmission electron microscopy: A DROP of chitosan dissolved in distal water was placed on the carbon coated copper grids, air dried at room temperature. The mean diameter of CNPs was 16.7 nm. Ovaries were collected from abattoir; medium sized follicles were aspirated and pooled in a 15 ml conical tube. The tubes were left 15 minutes until the settlement of follicular cells.Equilibration and Vitrification solutions were prepared by using TCM 199 medium supplemented with 10% FCS as a basic media. Five immature oocytes were placed in 50μl drops of basic medium for one minute then transferred to equilibrated solution (100μl drops of basic media with add 10% serum+ 10% EG+10% DMSO) for 15 minutes in room temperature.Then equilibrated oocytes were transferred to vitrification solution VS (100μl drops of basic media with add 10% serum + 20%EG + 20% DMSO). During vitrification CNPs was added at different concentrations (control group, 25 ,50 ,100μg/mL CNPs) for 30 sec.We study the effect of CNPs addition during vitrification on morphology, viability, cleavage, blastocyste rate of vitrified/warmed buffalo oocytes as follows:In First Experiment: The immature oocytes were divided into 4 groups according to the concentration of CNPs (control group;25 ,50 ,100μg/mL CNPs) added to vitrification solution to assess its effect on oocytes morphology.The highest morphology score was reported when using 50 μg/mL CNPs when compared to control (84.44 ±3.49 vs:73.53±4.39 respectively).Addition of 25μg/mL CNPs not significantly differ from control group (75.25±4.31 vs 73.53±4.39). Moreover, addition of 100 μg/mL CNPs significantly decreased normal morphology % when compared to control group (66.67±4.69 vs 73.53±4.39).IN Second Experiment: The immature oocytes were divided into 4 groups according to the concentration of CNPs (control group, 25 ,50 ,100μg/mL CNPs) added to vitrification solution to assess its effect on oocytes viability. The highest viability rate was reported when using 50 μg/mL CNPs (77.67±4.12 ) which is significantly different than control and 25μg/mL CNPs (64.71±4.75 and 64.36±4.78). Addition of 100 μg/mL significantly decreased viability when compared to control group ( 59.80±4.87 vs 64.71±4.7).IN Third Experiment: The immature oocytes were divided into 4 groups according to the concentration of CNPs (control group, 25 ,50 ,100μg/mL CNPs) added to vitrification solution to assess its effect on oocytes cleavage rate. The highest cleavage rate was reported when using 50 μg/mL CNPs (21.36±4.05) which was significantly increased when compared to control group (10.78±3.08). Addition of 25 μg/mL significantly increased cleavage rate when compared to control group (11.88±3.23 vs 10.78±3.08). Addition of 100 μg/mL CNPs significantly decreased cleavage rate when compared to control group (7.84±2.67 vs 10.78±3.08).IN Forth Experiment: The immature oocytes were divided into 4 groups according to the concentration of CNPs (control group, 25 ,50 ,100μg/mL CNPs) added to vitrification solution to assess its effect on blastocyste rate. The highest blastocyte rate was reported when using 50 μg/mL CNPs (6.80±2.49 )which significantly increased blastocyste formation rate when compared to control (0.98±0.98). Addition of 25 μg/mL CNPs significantly increased this parameter when compared to control group (2.97±1.69 vs 0.98±0.98), but addition of 100 μg/mL result did not differ from control (0.98±0.98 vs 0.98±0.98).