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20165
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Subject: Animal Reproduction and Gynecology × End date: 2016 × Start date: 2016 × Type: Thesis ×
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    ThesisItemOpen Access
    EFFECT OF EXTENDER AND DIFFERENT SPERM NUMBERS PER STRAW ON QUALITY OF FROZEN SEMEN IN BEETAL AND SIROHI BUCKS
    (Assam Agricultural University, Khanapara, Guwahati, 2016-01) KALITA, MANOJ KUMAR; Sinha, S.
    A total of 80 pooled ejaculates collected from two Beetal and one Sirohi bucks maintained at Goat Research Station, Burnihat were used to study the effect of tris extender containing 20% egg-yolk, 1% soy-lecithin and 1.5% soy-lecithin, and that of 37.5, 50 and 75 x106 sperm / straw on the quality of frozen semen, and also the effect of number of spermatozoa in frozen semen straws on fertility. The freezing of semen was done in French mini straw by rapid horizontal vapour freezing technique using liquid nitrogen. The overall mean post-thaw per cent sperm motility, live sperm, intact acrosome and HOST-reacted sperm in tris extender containing 20% egg yolk, 1% soy-lecithin and 1.5 % soy-lecithin was 61.20 ± 0.45, 57.77 ± 0.54 and 60.20 ± 0.45; 72.32 ± 0.47, 65.40 ± 0.56 and 67.07 ± 0.56; 68.42 ± 0.43, 61.30 ± 0.74 and 63.80 ± 0.58; and 64.35 ± 0.63, 57.35 ± 0.5 and 60.17 ± 0.46 respectively. The post thaw values of tris extender with 20% egg yolk were significantly (P<0.01) higher than that of tris extender containing 1 % and 1.5% soy-lecithin for live sperm, intact acrosome and HOST-reacted sperm, However, the difference was not significant between 20% egg yolk and 1.5% soy-lecithin for sperm motility. The post thaw values were significantly (P<0.01) higher for 1.5% than that for 1% soy-lecithin in all the parameters studied. The overall mean post-thaw per cent sperm motility, live sperm, intact acrosome and HOST-reacted sperm for straws containing 37.5, 50 and 75 x106 sperm/ straw was 56.02 ± 0.47, 57.50 ± 0.41 and 65.57 ± 0.58; 67.42 ± 0.62, 70.55 ± 0.55 and 73.45 ± 0.57; 61.12 ± 0.69, 64.37± 0.66 and 68.25 ± 0.66; and 59.00 ± 0.62, 62.77± 0.52 and 65.57 ± 058 respectively. The post-thaw values of semen with 75 x106 sperm/ straw were significantly (P<0.01) higher than that of 50 and 37.5 x106 sperm / straw for all the sperm parameters studied. The post thaw values with 50 x106 sperm/ straw was significantly (P<0.01) higher than that with 37.5 x106 sperm /straw for sperm motility, intact acrosome and HOST-reacted sperm but not for live sperm. The fertility rate based on non-return rate was the highest at 75 x106 sperm /straw in both Beetal and Sirohi goats. It was revealed from the present study that the quality of frozen semen was superior in tris extender containing 20% egg yolk to that containing 1% or 1.5 % soy-lecithin. The post thaw quality and fertility of goat semen was significantly superior for 75 x106 sperm / straw to 50 x106 and 37.5 x106 sperm / straw.
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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.
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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.
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    ThesisItemOpen Access
    A STUDY ON REPRODUCTIVE DISORDERS IN CROSSBRED CATTLE WITH SPECIAL REFERENCE TO REPEAT BREEDING
    (Assam Agricultural University, Khanapara, Guwahati, 2016-07) ACHARYA, CHIRANJEEVI; Deka, B. C.
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    ThesisItemOpen Access
    EFFECT OF MEDIA ON IN-VITRO MATURATION OF BOVINE FOLLICULAR OOCYTES
    (Assam Agricultural University, Khanapara, Guwahati, 2016-07) DAS, ARUNIMA; BARUA, P. M.
    The present research was planned to study the effect of different concentrations of Vitamin E and Oestrous cow serum and their combination on in-vitro maturation of bovine follicular oocytes incubated in TCM-199 medium containing Follicular fluid, Oestradiol-17β, p-FSH, Gentamicin, Sodium Pyruvate, Cysteamine and Fetal Bovine Serum at 38.5 0C with 5% CO2 for 24 hours in a CO2 incubator. A total of 825 oocytes were recovered from slaughterhouse ovaries of cattle by aspiration cum slicing technique and graded as A, B, C and D with the recovery rates of 77.64 ± 0.02, 11.60 ± 0.01, 6.84 ± 0.01 and 3.91 ± 0.01 per cent, respectively. A total of 704 oocytes of grade A and B were used in the study. Maturation of oocytes was determined on the basis of rate of cumulus cell expansion and polar body formation. Vitamin E concentrations at 0, 100 and 150 µM levels in the maturation medium resulted in cumulus cell expansion rate of 74.46 ± 2.68, 74.28 ± 1.66 and 73.59 ± 0.78 per cent, respectively, while polar body formation rates were found to be 44.02 ± 4.37, 50.06 ± 2.40 and 45.43 ± 4.20 per cent, respectively. Oocytes maturation rates both in terms of cumulus cell expansion and polar body formation were not affected by levels of Vitamin E supplementation in the in-vitro maturation medium. Oestrous Cow Serum supplementation at 0, 20 and 30 per cent levels to the in-vitro maturation medium resulted in the cumulus cell expansion rate of 74.46 ± 2.68, 68.41 ± 2.40 and 63.90 ± 3.49 per cent, respectively; corresponding figures for polar body formation rate were 44.02 ± 4.37, 32.10 ± 2.84 and 29.50 ± 0.67 per cent. Addition of either 20 or 30 per cent Oestrous Cow Serum in the maturation medium did not improve the oocytes maturation rate both in terms of cumulus cell expansion as well as polar body formation. Cumulus cell expansion and polar body formation rates were recorded as 67.20 ± 3.45 and 38.26 ± 2.28 per cent, respectively when 100 µM Vitamin E and 20 per cent Oestrous cow serum combination was used in the in-vitro maturation medium. The corresponding rates for the control medium (without supplementation) were 74.46 ± 2.68 and 44.02 ± 4.37 per cent.