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20053
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Subject: Animal Reproduction and Gynecology × End date: 2005 × Start date: 2005 × Type: Thesis × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    Comparison of different methods of pregnancy diagnosis in bitches
    (Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 2005) Asha Merina, Kuriakose; KAU; Joseph Mathew
    With the object of comparing different methods of pregnancy diagnosis and to determine the early reliable method, a study was undertaken to investigate the efficacy of transabdominal palpation, ultrasound scanning and haematological profile during pregnancy at various stages of gestation in bitches. Sixty six apparently normal healthy bitches were included in the study. The bitches subjected to pregnancy diagnosis were grouped based on gestational period as 20 to 30, 31 to 40 and 41 to 65 days post breeding. The data obtained were compiled and tabulated.
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    ThesisItemOpen Access
    Effect of ovum retrieval methods and cumulus-oocyte complex morphology on in vitro maturation of bovine oocytes
    (Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 2005) Magnus Paul, K; KAU; Sreekumaran, T
    This study was designed to evaluate the effect of different retrieval methods like aspiration, slicing and puncture on yield of different grades of oocytes and their in vitro maturation potential. The effect of cumulus oocyte complex morphology on in vitro maturation of bovine oocyte was also studied. Slaughterhouse derived bovine ovaries from South Indian breeds like Kangayam, Khillari, Hallikar and crossbred cattle of Kerala were subjected to three retrieval methods to yield different quality grades of oocytes. In this study, a total of one hundred and fifty ovaries were processed. Fifty ovaries were processed under each method of retrieval. Each quality grade of oocyte obtained through three retrieval method were subjected to maturation for 24 h in TCM-199 medium supplemented with LH, FSH, Estrogen, Pyruvate and Foetal calf serum. Culture environment was set as 38.5°C temperature, 5% carbondioxide tension and maximum humidity in standard CO2 incubator. Maturation changes were assessed by cumulus expansion, formation of M II plates and polar body extrusion. Total yield of COCs in slicing was 5.42 ± 0.56, which was significantly higher than aspiration and puncture which yielded 3.7 ± 0.43 and 3.66 ± 0.43 COCs respectively. This could be due to recovery of oocytes from a heterogeneous population of follicles in slicing method. Aspiration, slicing and puncture yielded A class oocytes at the rate of 44.86%, 37.2% and 43.17% respectively, wherein mean yield per ovary in same methods were 1.66 ± 0.25, 2.02 ± 0.25 and 1.58 ± 0.25 respectively. Yield of B class oocytes by aspiration, slicing and puncture were 32.97%, 29.89% and 31.69% respectively. Mean yield of B class oocytes per ovary was 1.22 ± 0.12, 1.62 ± 0.20 and 1.16 ± 0.16 respectively. Proportionate yield of D class oocytes by aspiration, slicing and puncture was 3.7%, 12.10% and 6.01% respectively. Aspiration, slicing and puncture did not differ significantly in yield of A, B and C class oocytes. But D class oocytes were proportionately more in slicing method. This finding could be attributed to the fact that the slicing recovered oocytes from a heterogeneous population of follicles, which included all kinds of growing follicles. When percentage yield of different classes of oocytes in same retrieval method was compared aspiration yielded A and B class oocytes in significantly higher proportion than C and D class oocytes. The same trend was observed in slicing and puncturing. In all the methods, the yield of D class oocytes was least. Cumulus expansion rate of A class oocytes were 83.08 per cent, 69.70 per cent and 70.37 per cent in aspiration, slicing and puncture method respectively. B class oocytes exhibited cumulus expansion rate of 68.29 per cent, 53.0 per cent and 62.07 per cent in aspiration, slicing and puncture method respectively. Retrieval method was found to have no significant effect on cumulus expansion potential of bovine COCs. Oocytes with more than three layers of cumulus cells exhibited better cumulus expansion rate than oocytes with less cumulus cell layers. Nuclear maturation rate of A class oocytes in aspiration, slicing and puncture was 81.08, 78.84 and 80.0% respectively, whereas in B class oocytes under same methods the percentage of nuclear maturation was 57.10, 41.67 and 52.63% respectively. D class oocytes failed to mature in all methods of retrieval except one oocytes in aspiration method. Aspiration, slicing and puncture did not differ significantly in the maturation percentage of oocytes. Polar body extrusion rate of A class oocytes in aspiration, slicing and puncture method was 45.5 per cent, 42.1 per cent and 44.0 per cent respectively. B class oocytes exhibited polar body extrusion rates of 28.6 per cent, 25.0 per cent and 26.3 per cent respectively in aspiration, slicing and puncture method respectively. Oocytes with more than three layers of cumulus cells exhibited better polar body extrusion rate irrespective of their retrieval method. Staining of 18 oocytes with fluorescent dye Hoechst 33342 as a method of cross checking, revealed 66.67 per cent Metaphase II rate. Ten oocytes tested for viability with FDA stain revealed 80 per cent viability after maturation. TCM-199 medium with 5 µg FSH/ml, 0.5 µg LH/ml, 1 µl Estradiol/ml, 0.2 mM pyruvate and 10 per cent serum was found to be ideal medium for maturation of bovine oocytes in vitro.. Oocytes with more than three layers of cumulus cells matured better than oocytes with less number of cumulus cell layers or denuded oocytes. These experiments proved that cumulus oocytes complex morphology have a very significant role in maturation of oocytes rather than retrieval methods.
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    ThesisItemOpen Access
    Fertility trials on induced oestrum in repeat breeding cattle with prolonged oestrum
    (Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 2005) Jeba Sujana, Dhas; KAU; Aravinda Ghosh, K N
    The objective of the present study is to evaluate the fertility in repeat breeding cattle with prolonged oestrum after oestrus induction and subsequent treatment with GnRH and hCG. Detailed data of cross bred cows and heifers belonging to University Livestock Farm, Mannuthy and those presented at Artificial Insemination Centre, attached to the Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy were collected. Repeat breeding animals with a history of prolonged oestrum were randomly selected and allotted into four groups of ten animals in each. Group I repeat breeding animals were subjected to induction of oestrus using 25 mg dinoprost, a PGF2 alpha analogue (Lutalyse) and insemination was done twice at an interval of 24 h on detection of proper signs of oestrum. In group II, oestrus induction was done using 25 mg Dinoprost on tenth day of cycle and 10 μg Buserelin a GnRH analogue (Receptal) was administered 48 h after PGF2 alpha administration. In group III, oestrus was induced using 25 mg Dinoprost on tenth day of cycle and 1500 IU hCG (Chorulon) was administered 48 h after PGF2 alpha administration. Both group II and III repeat breeding animals were inseminated twice at an interval of 24 h on detection of proper signs of oestrum. Group IV repeat breeding animals were subjected to insemination during natural oestrum. A total of 817 animals were screened of which 21.91 per cent animals were repeat breeders, out of which 26.26 per cent animals showed prolongation of oestrual signs. Among the 193 farm animals investigated, 19.17 per cent were repeaters out of which 21.62 per cent of animals showed prolonged oestrum. The duration of prolonged oestrum ranged from 32 to 96 h with a mean of 65.65  2.57 h. Out of 40 repeat breeding animals showing prolonged oestrum selected for the study, 35 per cent animals exhibited oestrus length between 30-48 h, 45 per cent between 48-72 h, and 20 per cent between 72 and 96 h. The intensity of oestrum was found to be higher in 77.5 per cent, medium in 20 per cent and low in 2.5 per cent animals. All the 30 repeat breeding animals in group I, II, III subjected to PGF2 alpha administration responded to the treatment by exhibiting oestrus signs, and hence the efficacy was 100 per cent. The time taken for induction of oestrus in group I, II and III were 52.7  2.99 h, 51.7  2.68 h and 52.0  2.68 h respectively. The duration of oestrus in groups I, II and III were 68.6  3.75 h, 38.6  3.75 h and 37.4  3.75 h respectively. Physical changes in the reproductive tract of repeat breeding animals showing prolonged oestrum were more pronounced during natural oestrum than during induced oestrum. The conception rate during induced oestrum in group I, II, III and control group were 50, 40, 40 and 30 per cent respectively. Overall conception rate for three consecutive oestrus in group I, II, III and control group were 60, 60, 50 and 40 percentage respectively. It can be recommended that induction of oestrus using prostaglandin could be employed for enhancing the conception rate in repeat breeding animals with the history of prolonged oestrum.