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20033
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Subject: Animal Reproduction and Gynecology × End date: 2003 × Start date: 2003 ×
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    ThesisItemOpen Access
    NUCLEAR TRANSFER IN RABBITS AND GOATS
    (Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2003-10) SAMBASIVA RAO, B; SESHAGIRI RAO, A (Major); SUBRAMANYAM NAIDU, K; RAO, V.H; CHANDRASEKHARA RAO, T.S
    A B S T R A C T A study was undertaken to standardize methods for the production of cloned rabbit and goat embryos using both blastomeres and somatic cells as nuclear donors. Superovulated (rabbit) and in vitro matured (goat) oocytes were enucleated and used as recipient cytoplasts. A total of 941 cloned rabbit embryos were produced using fresh (324) and frozen-thawed (647) blastomeres as nuclear donors. Ninety five per cent of frozen thawed and 94% of fresh blastomeres successfully fused to the enucleated oocytes. In both these groups of cloned embryos the rate of cleavage was 80%. Forty six and 44% of cloned and cleaved embryos developed to morula stage in the two groups respectively. Similarly 26 and 24% of the cloned and cleaved embryos developed into blastocysts in vitro in the two groups respectively. Two sources of blastomeres and four sources of oocytes were tested in different oocytes-blastomere combinations for their ability to support the in vitro development of cloned rabbit embryos. It was observed that when both the oocytes and the blastomeres originated from the same genetic strain, the rate of development of cloned embryos to blastocyst stage was significantly better. Super ovulated oocytes were collected from the donor does at 14, 17 or 19h phCG and electro fused with blastomeres at >18, >20 or >22h phCG. Though the oocytes collected at 14h phCG and fused to blastomeres at >20h phCG with four DC pulses of 2.5kV/cm exhibited relatively poor fusion rate, this group exhibited the best development to blastocyst stage along with the oocytes collected at 19h phCG and fused to blastomeres at >22h by a single DC pulse of 3.5kV/cm. It was concluded that it was necessary to adjust the electrical fusion / activation protocols to suit the age of oocytes at collection and fusion. Cloned rabbit embryos produced from fresh blastomeres only had developed to live offspring on transfer to recipient does. Further use of oocytes collected at 17h phCG along with fresh blastomeres only supported full term development of cloned rabbit embryos in recipient does. While, overall less than 1% of the cloned rabbit embryos developed to term, 1.7% of cloned rabbit embryos produced from oocytes collected at 17h phCG and fresh blastomeres developed to term. None of the cloned rabbit embryos produced from frozen-thawed blastomeres developed to live offspring in this study. DNA fingerprinting of the cloned rabbits was undertaken using three different primers. All the three primers indicated that the rabbit bunnies were genetically identical which confirmed their clonal nature. A total of 694 somatic cell cloned rabbit embryos were produced of which 279 were from cultured fetal fibroblasts and 415 from contemporary cumulus cells. Except that cumulus cell generated cloned embryos showed a significantly higher fusion rate, there were no differences in other development parameters studied. Though somatic cell cloned embryos developed to blastocyst stage more frequently than blastomeres cloned embryos, none of the somatic cell cloned embryos whether produced from fetal fibroblasts or cumulus cells developed to term in the recipient does. A total of 628 cloned goat embryos were produced of which, 402 were from blastomeres, 174 from fetal fibroblasts and 52 from contemporary cumulus cells. Blastomeres fused better than somatic cells to the in vitro matured and enucleated goat oocytes (76 vs 58%). While the cleavage rates were similar in all the three groups of cloned goat embryos, blastocyst development was significantly better in blastomere cloned (65%) than somatic cell cloned (32 and 41%) goat embryos. On transfer of 145 blastomere, 40 fetal fibroblast and 38 contemporary cumulus generated cloned embryos to recipient does, none of the recipients became pregnant and no cloned kids were produced.
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    ThesisItemOpen Access
    COMPARATIVE STUDIES ON MANAGEMENT OF RETENTION OF FETAL MEMBRANES WITH AND WITHOUT MANUAL REMOVAL IN CROSSBRED COWS
    (Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2003-09) CHANDRA PRASAD, B; MOULI KRISHNA, K
    ABSTRACT : The present investigation was carried out in Holstein Fresian crossbred cows at a commercial dairy farm located near Tirupati. A total of 30, consisting of ten calved normally (Group I) and 20 RFM affected crossbred cows maintained under uniform conditions were selected randomly. The RFM affected cows were divided into two groups of ten each (Group II and Group III). The Group II cows were subjected to manual removal of RFM and provided a variety of drug therapy. Whereas, Group III cows were not subjected to manual removal of RFM, however, 2-3 infusions of 20 ml diluted povidone - iodine intrauterine at weekly intervals starting at day 15-17 postpartum were placed. Cows in Group III were continuously watched for spontaneous expulsion of fetal membranes. Blood samples from each cow were collected at day 1, 8, 15, 25 and 35 postpartum in EDTA coated tubes. The TLC and lymphocytes, neutrophils in whole blood and haptoglobin, ceruloplasmin, arachidonic acid, nitrate and nitrite in blood plasma were determined. The reproductive tract of each cow was palpated per rectum at weekly intervals starting at day 15-17 postpartum until completion of uterine involution. Uterine biopsies were obtained after uterine involution and all cows were observed for first postpartum heat. During the period of experiment all cows were found to be healthy. The cows in Group III expelled retained fetal membranes spontaneously on an average 6-7 days (2-12 days). The haematological parameters studied did not differ significantly either between or within groups. The mean plasma haptoglobin concentrations did not differ between and within groups. Plasma ceruloplasmin concentrations differed (P<0.05) significantly between and within groups. The maximum and the minimum concentrations were recorded on day 8 and 35 postpartum respectively. Plasma concentrations of arachidonic acid differed (P<0.05) significantly within groups but not between groups. The concentrations gradually declined in Group I and Group II whereas, in Group III decreased after day 25 postpartum only. Nitrate and nitrite concentrations differed (P<0.05) significantly within and between groups. The concentrations of nitrate and nitrite declined in parallel with progress of uterine involution. The mean interval for uterine involution was found to be 33.10 ± 1.32, 35.20 ± 1.12 and 33.80 ± 1.67 days in Group I, Group II and Group III respectively. The mean days required for first postpartum observed heat were 75.30 ± 6.00, 83.20 ± 2.07 and 78.90 ± 5.89 days in Group I, Group II and Group III, respectively. The histopathological findings in uterine endometrium were almost similar in all the three treatment groups of crossbred cows. Based on the observations on spontaneous shedding of RFM,health, ceruloplasmin, nitrate, nitrite, uterine involution, interval to first postpartum observed heat and microscopic features on uterine endometrium revealed that the non-removal of RFM did not affect the subsequent performance of the animals. Infact it had slightest advantage over the manual separation. Hence, the concept of non-removal of RFM could be considered confidently for implementation.
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    ThesisItemOpen Access
    HORMONAL APPROACH FOR AUGMENTING THE FERTILITY IN REPEAT BREEDING CROSSBRED COWS
    (SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517 502 (A.P) INDIA, 2003-08) ABHILASH, R S; SESHAGIRI RAO, A (Major); MOULI KRISHNA, K; SURESH KUMAR, R.V
    ABSTRACT : The present investigation "Hormonal approach for augmenting the fertility in repeat breeding crossbred cows" was carried out in 80 repeat breeding and 40 normal breeding crossbred cows to find out incidence of repeat breeding, to study the various aspects of estrous cycle viz., interestrous interval, duration of estrus and ovulation time in normal and repeat breeders and for comparing the trace mineral levels in normal and repeat breeders to detect the possible causes of repeat breeding. The treatments with GnRH, hCG and progesterone were exercised in repeat breeder cows. The incidence of repeat breeding in crossbred cows was observed to be 10.17 per cent. The mean values of copper, manganese and zinc were found to be 0.86 ± 0.07 Vs 1.22 ± 0.10, 0.22 ± 0.01 Vs 0.37 ± 0.09 and 1.17 ± 0.10 Vs 1.87 ± 0.15 ppm respectively in repeat and normal breeders and noticed to be significantly lower level in repeat breeders. Whereas, the iron level did not vary much (2.57 ± 0.13 Vs 2.44 ± 0.14). The length of estrous cycle (inter estrous interval) was observed to be short, normal and long in 10.00, 55.00 and 35.00 per cent respectively for normal breeders and 17.50, 48.75 and 33.75 per cent respectively for repeat breeders. In the present study the duration of estrus was observed to be longer in 70.00 per cent of repeat breeders and 50.00 per cent of normal breeders. The time of ovulation was found to be normal in 50.00 per cent of repeat breeders and 80.00 per cent of normal breeders, delayed in 38.75 per cent of repeat breeders and 15.00 per cent of normal breeders and ovulation failure was noticed in 11.25 per cent of repeat breeders and 5.00 per cent of normal breeders. In the present study more per cent of animals exhibited estrus with in 24 hrs after treatment when compared to before treatment in GnRH treated (75.00 Vs 40.00 per cent) and in hCG treated (85.00 Vs 45.00 per cent) repeat breeding crossbred cows. Similarly the number of animals exhibited longer duration of estrus (24-36 hrs and > 36 hrs) were decreased after treatment in GnRH and hCG (20.00 Vs 35.00, 5.00 Vs 25.00 and 15.00 Vs 35.00, 0.00 Vs 20.00 per cent) treated cows. However, much difference could not be observed in the progesterone treated cows. More per cent of animals were observed to ovulate within the normal ovulation time of <16 hrs after treatment with GnRH (80.00 Vs 35.00 per cent) and with hCG (85.00 Vs 45.00 per cent) when compared to before treatment. Similarly, the number of animals ovulated in an abnormal period 16-48 hrs and more than 48 hrs were decreased after treatment in these groups (20.00 Vs 45.00, 0.00 Vs 20.00, 15.00 Vs 40.00 and 0.00 Vs 15.00 per cent). However, not much difference was observed in the cows treated with progesterone. The conception rates of repeat breeding crossbred cows to first insemination, second insemination and overall were observed to be 45.00, 9.10 and 50.00 per cent in GnRH treated group; 60.00, 12.50 and 65.00 per cent for hCG treated group; 35, 15.38 and 45.00 per cent for progesterone treated group and 10.00, 11.11 and 20.00 per cent for control groups. To conclude in the present study, delayed ovulations and early embryonic mortality arose as the root cause for repeat breeding in crossbred cows. Both GnRH and hCG exhibited the effect on synchronization of duration of estrus and ovulation time. Improved conception rates were observed in all the three hormone treated crossbred cows when compared to control animals