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20161
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Subject: Animal Reproduction and Gynecology × Author: BORAH, DHARITRI × Start date: 2000 × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    EFFECT OF ADDITIVES IN MEDIUM ON IN-VITRO MATURATION AND FERTILIZATION OF GOAT OOCYTES
    (Assam Agricultural University, Khanapara, Guwahati, 2016-07) BORAH, DHARITRI; Biswas, R. K.
    A total of 2539 oocytes were recovered from 712 goat ovaries obtained from slaughter house soon after sacrifice and the mean recovery rate of oocytes per ovary was 3.43 ± 0.06, 4.21 ± 0.08 and 3.24 ± 0.78 by aspiration, slicing and puncture techniques respectively, being significantly higher (P<0.01) in slicing as compared to other two techniques. The recovery of good quality oocytes with two or more cumulus cells layers around the oocytes was significantly higher (P<0.01) in puncture (73.92 ± 0.92%) than that in aspiration (66.27 ± 0.68%) and slicing (64.76 ± 0.92%) techniques. The effect of addition of 10ng/ml EGF + 50 ng/ml IGF-1, 10ng/ml EGF + 600µM/ml cysteine and 10ng/ml EGF + 0.2mM/ml sodium pyruvate in TCM-199 + 100µl/ml foetal bovine serum + 100µM/ml cysteamine + 1µg/ml 17β-Oestradiol + 5µg/ml pFSH + 5µg/ml oLH + 10 per cent follicular fluid and 10 per cent estrous goat serum (control medium) was studied for in-vitro maturation (IVM) of goat oocytes on incubation at 38.50C for 24 hours in a CO2 incubator maintaining 5 per cent CO2 under humidified condition. The IVM rate of oocytes on the basis of cumulus cell expansion and polar body extrusion was found to be significantly higher (P<0.01) with EGF + IGF-I (88.74± 1.85% and 61.71 ± 1.61%) than that with EGF + sodium pyruvate (82.86 ± 0.97% and 54.62 ± 1.88%), EGF + cysteine (78.58 ± 1.45% and 49.02 ± 1.52%) and control medium (75.27 ± 1.58% and 43.03 ± 1.48%). The oocytes matured in the IVM media used were fertilized in-vitro in Fert-TALP using swimmed-up sperm capacitated in sperm TALP. The incidences of in-vitro fertilization of oocytes on the basis of two polar bodies and 2-cell stage were also higher when oocytes were matured in-vitro using EGF + IGF-I (44.67 ± 8.86 and 15.39 ± 4.48%) than that with EGF + sodium pyruvate (25.51 ± 7.31 and 11.56 ± 4.72%), EGF + cysteine (22.46 ± 8.37 and 11.56 ± 4.72%) and control medium (20.48 ± 4.27 and 8.10 ± 3.84%) although the differences were not found to be significant.