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Subject: Animal Reproduction and Gynecology × Author: BHAJONI, MADHURIMA × End date: 2017 × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    INFLUENCE OF ADDITIVES ON IN-VITRO MATURATION OF BOVINE OOCYTE
    (Assam Agricultural University, Khanapara, Guwahati, 2016-07) BHAJONI, MADHURIMA; Bhuyan, D.
    A study was conducted to find an effective in-vitro culture system based on in-vitro maturation of bovine oocytes. Ovaries from slaughter house were utilized to study ovarian biometry, follicular biometry and performance of in-vitro maturation of oocyte. Significantly higher weight, length, width and thickness were recorded in ovary with CL than that without CL. The number of large, medium and small follicles was more in ovary without CL than with CL group. The mean number of medium size follicles was significantly (P<0.01) higher in ovary without CL (6.32±0.75) than with CL (3.33±0.18). The recovery rates of grade A (47.58%) and B (37.42%) oocytes were higher than that of grade C (8.82%) and D (6.12%) by aspiration method. In the present study in-vitro maturation of oocytes was done at 38.5 0C in humidified atmosphere of 5% CO2 for 24 hours and matured in-vitro in medium-I or control (TCM-199+10% FBS+L-glutamine+sodium pyruvate+Gentamicin+pFSH+ Estradiol 17-β), medium-II (control+5% ECS), medium-III (control+100µM/ml cysteamine), medium-IV (control+10ng/ml EGF) and medium-V (control+ 5% ECS+ 100µM/ml cysteamine+10ng/ml EGF). The mean diameter of oocytes with cumulus cells for grade A oocytes varied significantly (P<0.01) after in-vitro maturation (IVM) in different media. The medium having either epidermal growth factor or cysteamine as additives showed higher diameter of oocytes after IVM as compared to medium with estrous cow serum or foetal bovine serum or combination of all three additives. The mean diameter of oocyte without cumulus cells before and after IVM did not differ significantly between different media. The increase in diameter of oocytes with cumulus cell for grade A was significantly (P˂0.05) higher in medium-III than that of I, II and V and in medium-IV than that of I, II and V. There was no significant difference in increase in diameter of oocyte without cumulus cells for grade A and oocyte with and without cumulus cells for grade B between different media. The rates of maturation based on cumulus cell expansion and nuclear maturation were the highest in the medium-IV (86.92% and 62.36%) containing epidermal growth factor (EGF) followed by medium-III (82.24% and 56.82%) containing cysteamine among all the media, the difference between media III and IV being non-significant.