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Open Access
In vitro maturation of caprine follicular oocytes
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2010) Ambili, John; KAU; Joseph, Mathew
This study was designed to analyse the effect of three oocyte retrieval methods, aspiration, slicing and puncture on the yield of different quality grades of oocytes and to evaluate the in vitro maturation rate of different grades of caprine oocytes. One hundred and thirty eight ovaries of Malabari goats and its crossbreds collected from the slaughter house were subjected to the oocyte retrieval methods. The oocytes harvested were graded based on the number of cumulus cell layers and ooplasm character into A, B, C and poor quality grades. Oocytes of A, B and C grades were subjected to maturation for 24 h in TCM-199 medium under standard culture conditions. Average yield of COC per ovary by aspiration, slicing and puncture was 3.93 ± 0.11, 4.44 ± 0.06 and 3.59 ± 0.07 respectively. Yield was significantly higher in slicing method than aspiration and puncture. The percentage yield of A, B, C and poor quality grades of oocytes by aspiration method was 26.74 per cent, 26.62 per cent, 24.77 per cent and 21.87 per cent respectively. Mean yield of oocytes of each quality grade by the same method were 1.05 ± 0.05, 1.05 ± 0.08, 0.98 ± 0.07 and 0.86 ± 0.04 respectively. Slicing yielded 23.08 per cent A class, 28.15 per cent B class, 24.44 per cent C class and 23.96 per cent poor quality oocytes. Mean yield of oocytes per ovary in these classes by slicing method were 1.04 ± 0.04, 1.25 ± 0.05, 1.08 ± 0.07 and 1.06 ± 0.06 respectively. Percentage yield of A, B, C and poor quality oocytes by puncture was 29.01 per cent, 30.26 per cent, 22.07 per cent and 18.66 per cent respectively. Mean yield per ovary by puncture method was 1.04 ± 0.04, 1.08 ± 0.03, 0.79 ± 0.04 and 0.67 ± 0.03 for A, B, C and poor quality oocytes respectively. No significant difference was observed in the yield of A, B and C class oocytes between aspiration, slicing and puncture. Yield of poor quality oocytes were significantly more in slicing method. The cumulus expansion rate of A class oocytes obtained by aspiration, slicing and puncture was 77.75 per cent, 69.70 per cent and 71.49 per cent respectively. Class C oocytes exhibited a cumulus expansion rate of 63.48 per cent, 51.37 per cent and 63.39 per cent respectively when collected by aspiration, slicing and puncture method. Class C oocytes obtained by aspiration, slicing and puncture when subjected to in vitro maturation exhibited a cumulus expansion rate of 39.17, 32.57 and 37.29 per cent respectively. Retrieval method was found to have no significant effect on cumulus expansion potential of caprine oocytes, whereas the COC morphology had significant effect on cumulus expansion potential. Nuclear maturation rate of A class oocytes collected by aspiration, slicing and puncture method were respectively 40, 30 and 50 per cent and polar body extrusion rate was 30, 20 and 30 per cent respectively. Class B oocytes exhibited nuclear maturation rate of 20, 10 and 30 per cent and polar body extrusion rate of 10, 10 and 20 per cent respectively by aspiration, slicing and puncture. Ten, 10 and 20 per cent of C class oocytes retrieved by aspiration, slicing and puncture exhibited nuclear maturation and 10 per cent polar body extrusion was observed in C class oocytes retrieved by puncture. None of the C class oocytes collected by aspiration or slicing exhibited polar body extrusion. This study proved that slicing is a better method than aspiration or puncture for retrieval of oocytes from caprine ovaries as it yielded more number of oocytes per ovary. Retrieval methods had no significant effect, whereas COC morphology was found to have significant effect on cumulus expansion, nuclear maturation and polar body extrusion rates of different grades of oocytes
Open Access
Effect of gonadotropin releasing hormone and prostaglandin for improving reproductive efficiency in goats
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Julliet; KAU; Joseph, Mathew
With the objective of studying the effect of GnRH and prostaglandin for improving reproductive efficiency in goats the study was carried out at University Sheep and Goat Farm, Mannuthy using 42 cycling goats. Based on the behavioural and physiological changes associated with oestrum the goats were divided into two groups viz., Group I and Group II. Group I animals were those that exhibited pronounced oestrus signs and were divided into two subgroups namely Group IA and Group IB. Group II animals were those that exhibited weak oestrus signs and were divided into three subgroups namely Group IIA, IIB and IIC. Group IA animals were administered 0.0042 mg Buserelin (1 ml Receptal) a potent GnRH analogue on day 0, and Group IB served as the Control. Blood was collected prior to GnRH administration and breeding from all does. The mean duration of oestrum in Group IA and IB was 19.33 ± 0.45 and 33 ± 0.58 h respectively. The conception rate in Group IA and IB was 50 per cent and 66.66 per cent respectively. The serum P4 level on day 0 in does in Group IA and IB was 0.43 ± 0.05 ng/ml and 0.40 ± 0.05 ng/ml respectively. Group IIA and Group IIB does were treated as per the CO-Synch protocol (i/m inj. of 0.0042 mg of Buserelin (1 ml Receptal) on day 0, 125 µg cloprostenol (0.5 ml clostenol) on day 7; 0.0042 mg of Buserelin and mating on day 9) and prostaglandin protocol respectively (two intramuscular injections of 125 µg cloprostenol (0.5 ml clostenol) 11 days apart followed by mating at 72 and 96 h), while Group IIC served as the control. The oestrus response, oestrus onset interval, duration of oestrum and conception rate in Group IIA was 90.9 per cent, 47.6 ± 0.45 h, 24.5 ± 0.63 h and 40 per cent respectively. The oestrus intensity score of induced oestrus ranged from 0 to 13. The serum P4 level in pregnant and non pregnant does was not significantly different on days 0, 7 and 9 (P>0.05). The oestrus response, oestrus onset interval, duration of oestrum and conception rate in Group IIB was 81.8 per cent, 54 ± 1.006 h, 39.77 ± 1.54 h and 66.66 per cent respectively. The oestrus intensity scores in induced oestrus ranged from 0 to 13. The serum progesterone level in does that became pregnant and those that were non pregnant were not significantly different on day 0, 11, and at 72 and 96 h. In Group II C the duration of oestrum and pregnancy rates was 40 ± 0.91 h and 33.33 per cent respectively. Pregnancy diagnosis was done at three months of gestation by abdominal palpation and the accuracy of the method was 90.9 per cent. Mean gestation length was 146.03 ± 0.76 days. Litter size at birth in Group IA, IB, IIA, IIB and IIC was 2, 2, 2, 1.83 and 2 respectively. Average birth weight of kids was 2.35 ± 0.164 kg and the mean birthweight of male and female kid was 2.42 ± 0.98 kg and 2.28 ± 0.36 kg respectively. Thus from the present study, it can be concluded that :- 1. Administration of GnRH on the day of oestrum in animals exhibiting pronounced oestrus signs failed to improve conception rate when compared to the control. 2. In animals exhibiting weak oestrus signs both CO-Synch and double prostaglandin protocols resulted in higher conception rate when compared to control group. 3. The double prostaglandin protocol was found to be more efficient in improving conception rate in animals exhibiting weak oestrus signs.
Open Access
Comparative efficacy of bromocriptine, cabergoline and thyroxine in inducine oestrus in bitches
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Ajitkumar, G; KAU; Sreekumaran, T
Detailed survey on breeding, feeding and management of 1721 dogs belonging to 817 owners was conducted in different parts of Kerala State. Among the dogs surveyed, 518 (30.10%) were male and 1203 (69.90%) were female. Breed-wise details of dogs maintained by individual owners revealed German shepherd dog (20.74%) as the most popular breed of dog followed by Labrador retriever (18.13%), Dachshund (11.39%), Rottweiler (9.12%) and Spitz (5.52%). The percentage of local non-descript dogs was found to be 14.64 only. The prevalence, nature and magnitude of anoestrus among the female dogs were evaluated based on breeding details. Haematological, hormonal, mineral and cholesterol profile of anoestrous bitches were estimated. Data on breeding revealed that 92.93 per cent of bitches were allowed to breed and the remaining 7.07 per cent were kept as virgin throughout their lifetime. The age at first breeding in majority of the bitches (57.85%) was between 1 to 2 years with an average of 16.30 months. The average number of oestrus periods and the number of breeding per year were 2.14 ± 0.24 and 1.47 ± 0.32 respectively. The average duration of proestrual bleeding was found to be 10.87 ± 1.24 days. Most of the bitches (51.97 %) were bred twice in one oestrus and the period of acceptance varied from 4 to 22 days with an average of 8.94 ± 2.18 days. The incidence of failure of conception, abortion, stillbirth and pseudopregnancy were 18.87, 5.72, 15.56 and 13.69 per cent respectively. The neonatal problems encountered in 13.88 per cent of bitches included agalactia / hypogalactia / mammitis (5.29%), cannibalism (2.65%), puppy fading syndrome (2.09%), puerperal tetany (1.98%) and rejection of young one by dam / poor mothering ability (1.87%). Among the bitches surveyed, 134 (11.14%) were identified as in the stage of anoestrus, 65.67 per cent in primary and 34.33 per cent in secondary anoestrus and the average age was 22.68 and 34.34 months respectively. The average duration of secondary anoestrus was 11.71 months. Analysis of breed-wise prevalence revealed that it was high in breeds such as Dachshund (18.18%), Labrador retriever (16.74%), Great Dane (16.67%), Dalmatian (15.38%) and Doberman pinscher (14.00%) and was lowest (2.44%) among local non-descript bitches. Data on body condition revealed that 42 per cent of the anoestrous bitches were with the ideal body score of five, 46 per cent with body score more than five (obese) and 12 per cent with body score less than five (lean). Analysis of haematological profile of anoestrous bitches revealed normochromic normocytic anaemia. The mean serum progesterone, prolactin and thyroxine levels in anoestrous bitches were of 0.57 ± 0.03 ng/ml, 0.75 ± 0.05 ng/ml and 1.80 ± 0.06 µg/dl respectively. The mean serum calcium and phosphorus levels in anoestrous bitches were 8.94 ± 0.24 and 3.48 ± 0.12 mg per cent respectively. The mean serum iron, copper, cobalt, zinc, manganese and cholesterol levels were found to be within the normal range. Oestrus induction trials were carried out in forty anoestrous bitches and the data obtained were compared with that of control bitches. Out of 10 animals treated in each group, five (50%) in Group I (bromocriptine @ 50 μg/kg. body weight), nine (90%) in Group II (cabergoline @ 5 μg/kg. body weight), eight (80%) in Group III (thyroxine @10 μg/kg. body weight) and seven (70%) in Group IV (thyroxine @ 5 μg/kg. body weight) evinced proestrual bleeding. The mean treatment onset to proestrus in Groups I, II, III and IV was 28 ± 3.39, 13.44 ± 3.12, 24.50 ± 3.18 and 33 ± 2.21 days respectively. The average duration of proestrus in the treatment groups was 9.80 ± 0.86, 10.11 ± 0.68, 11.25 ± 0.88 and 10.71 ± 0.68 days respectively as against 9.70 ± 0.42 days in the control group. The average duration of oestrus in the four treatment groups and the control group was 7.60 ± 0.24, 8 ± 0.29, 8.5 ± 0.63, 7.85 ± 0.46 and 8 ± 0.39 days respectively. Fifty per cent of the animals treated with bromocriptine and 10 per cent of the animals treated with cabergoline exhibited nausea and vomiting as side effects, whereas none of the animals treated with thyroxine exhibited side effects. The mean serum progesterone and prolactin levels during anoestrus, on the first day of proestrus and on the day of first breeding did not differ significantly within and between the treatment groups and the control group. The mean serum thyroxine level during anoestrus and on the first day of proestrus did not differ significantly within and between the treatment and the control groups, whereas on the day of first breeding the mean thyroxine level recorded statistically significant difference between the treatment and control groups, the lowest being 1.56 ± 0.18 μg/dl in Group I and the highest being 2.10 ± 0.13 μg/dl in Group IV. Animals treated with levothyroxine (Group III & IV) exhibited a slight increase in the mean level of serum thyroxine on the first day of bleeding as well as on the day of first breeding, but the values were found to be within the normal limits. The haemoglobin level and total erythrocyte count were found to be increased on the first day of proestrus and on the day of first breeding in animals of Group III and IV. The mean serum phosphorus level on the first day of proestrus and on the day of first breeding in the treatment groups were found to continue at low levels as in the anoestrous stage. The mean serum copper level on the first day of proestrus exhibited statistically significant difference between the treatment and control groups. The mean serum level of calcium, iron, cobalt, zinc and manganese during anoestrus, on the first day of proestrus and on the day of first breeding did not reveal statistically significant difference between groups. The overall conception rate in the treatment groups were 40, 70, 50 and 40 per cent respectively as against 70 per cent in the control group. The average gestation length and litter size did not differ significantly between the treatment and control groups.
Open Access
Early pregnancy diagnosis using ultrasonography in goats
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2009) Sreejith, J R; KAU; Athman, K V
The objective of the study was to evaluate the efficacy of B-mode ultrasonography in early pregnancy diagnosis in goats and to identify the optimum stage of gestation for early pregnancy diagnosis using of transrectal and transabdominal ultrasonography. Thirty apparently healthy does with the history of breeding were selected for the study and these goats were randomly divided into three groups consisting of ten animals each. Group I consisted of ten goats which were scanned between third and fourth week (15- 28 days) post-breeding. Ten goats scanned between fifth and sixth week (29- 42 days) post-breeding, were included in group II and group III consisted of ten goats which were scanned between seventh and eighth week (43- 56 days) post-breeding. These animals were subjected to B-mode real-time ultrasound scanning transrectally (7.5 MHz probe)) and transabdominally (3.5 MHz probe). The accuracy of transrectal scanning in group I, II and III was 90, 100 and 100 per cent respectively and the accuracy for corresponding weeks was 50, 100 and 100 per cent respectively for transabdominal scanning. The embryonic vesicle was detected earliest on day 19 of gestation by transrectal scanning and on day 26 by transabdominal scanning. The embryo was first observed on day 22 and day 28 by transrectal and transabdominal scanning respectively. The foetal heartbeat which was an indication of foetal viability was detected earliest by transrectal scanning on day 24 of gestation. But by transabdominal ultrasonography, it could be detected only on day 34 of gestation. Placentomes and foetal skeleton were observed on day 42 and 54 of gestation respectively using both methods of scanning. The mean diameter of gestational sac recorded was 5.1 mm on day 19 and 27 mm on day 36 of gestation by transrectal scanning. The mean diameter of gestational sac recorded was 15.7 mm on day 26 and 34.4 mm on day 36 of gestation by transabdominal scanning. The mean crown-rump length ranged from 7.2 mm on day 22 to 34.4 mm on day 43 of gestation using transrectal scanning. By transabdominal scanning, the mean crown-rump length recorded was 16.7 mm on day 28 and 32.7 mm on day 43 of gestation. The diameter of the placentomes recorded by transrectal and transabdominal scanning on day 42 of gestation was 8.4 mm and 8.5 mm respectively. All the foetal measures by transrectal and transabdominal scanning were highly correlated (r > 0.9) with gestational age. The overall accuracy for the prediction of foetal numbers by transrectal and transabdominal scanning was 80 and 50 per cent respectively. The accuracy for prediction of foetal number using transrectal scanning in pregnant animals of group I and III was 71.43 per cent. The accuracy for the prediction was 100 per cent in pregnant animals of group II. By transrectal scanning, the accuracy for the prediction of singletons, twins and triplets by transrectal scanning was 100, 83.33 and 50 per cent respectively. By transabdominal ultrasonography, it was not possible to predict foetal number accurately in pregnant animals group I. The accuracy for the prediction of foetal number in group II by transabdominal scanning was 66.66 per cent while it was 85.71 per cent in group III. . The accuracy for the prediction of singletons, twins and triplets by ultrasonography was 50, 58.33 and 25 per cent respectively. In conclusion, transrectal scanning was accurate for pregnancy diagnosis from fourth week of gestation and transabdominal from fifth week of gestation and that real-time ultrasound scanning by both transrectal and transabdominal approaches was found to be reliable, safe and accurate for the diagnosis of pregnancy in goats from fifth week of gestation.
Open Access
Preservability of bovine preantral follicles in situ
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Harinarayanan, P M; KAU; Vijayakumaran, V
The study was designed and conducted with the objectives of assessing: (1) efficiency of two different temperatures, media and duration of storage for short term preservation of bovine preantral follicles in situ and (2) efficiency of conventional vitrification (CV) and solid surface vitrification (SSV), for long term cryopreservation of bovine preantral follicles in situ. Ovaries of six freshly slaughtered adult crossbred cows were collected and transported to the laboratory within one hour in normal saline at room temperature. The ovarian cortex from both ovaries of each animal was separated, divided further into fifteen pieces. One hour after the slaughter a cortical piece was selected at random from each animal and fixed in Bouin’s fluid for histological processing later on (control). Twelve cortical pieces from each animal were randomly allotted for short term preservation, of which, six pieces each were placed in vials containing normal saline and remaining in Dulbecco’s phosphate buffered saline (DPBS). Three vials from normal saline group and DPBS group were stored at room temperature (25ºC) and the remaining three vials from each group were stored at refrigeration temperature (3-5ºC). One cortical piece was taken out, from each media, from both the storage temperatures, at four, eight and twelve hours of storage and fixed for histological analysis. The remaining two cortical pieces were further divided into smaller fragments of approximately 1mm3 size. The fragments from one piece were vitrified using conventional vitrification (CV) and the rest by solid surface vitrification (SSV) and stored in liquid nitrogen for ten days. The fragments were warmed and fixed after the storage period. The quality of preantral follicles in the fixed ovarian tissues was evaluated based on morphology in histological sections. The preantral follicles were counted, classified as primordial, primary and secondary. Storage at refrigeration temperature preserved the preantral follicles very much better than the storage at room temperature. The amount of normal preantral follicles reduced progressively with passage of time. Primordial follicles were better adapted to preservation than primary and secondary follicles. The preantral follicles were best preserved at refrigeration temperature up to four hours of storage. The morphologically normal preantral follicles in normal saline (55.17 %) and DPBS (56.17 %) at this temperature and storage period did not show significant difference from that of control (59 %). The primordial follicles were preserved in both normal saline (61.42 %) and DPBS (61.15 %) at refrigeration temperature up to four hours, at levels similar to control (62.63 %). However, only DPBS at refrigeration temperature could ideally preserve primary (51.13 % vs. 53.06 % in control) and secondary follicles (42.10 % vs. 48.83 % in control). The number of morphologically normal preantral follicles in situ was significantly reduced after both CV (25.5 %) and SSV (37.17 %). But, SSV was significantly better than CV for preserving the quality of preantral follicles. According to this study DPBS at refrigeration temperature is ideal for preserving all the three classes of bovine preantral follicles in situ up to four hours of storage. Though vitrification of bovine ovarian tissue could not preserve preantral follicles at ideal levels, SSV was found to be far superior to CV in preserving bovine preantral follicles in situ.
Open Access
Synchronization of ovulation and timed artificial insemination to improve fertility in postpartum dairy cows
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Rajeswari, T; KAU; Aravinda Ghosh, K N
The present research work was undertaken to evaluate different oestrus synchronization protocols and to recommend a better and more consistent timed artificial insemination (TAI) protocol for improving fertility in postpartum dairy cows. The study was performed in 30 crossbred cows at day 40 postpartum belonging to University Livestock Farm, Mannuthy during the period from July 2007 to May 2008. Efficacy of various synchronization protocols for inducing oestrus and ovulation and conception rate in the experimental and control groups were determined. In Group I, 20µg of GnRH analogue (Buserelin) was administered intramuscularly on day 40 postpartum followed by 500µg of PGF2α analogue (Cloprostenol) on day 7 intramuscularly and a second dose of 10µg GnRH was administered on day 9 followed by TAI at 24th and 32nd hours. In Group II, 20µg of GnRH analogue was administered intramuscularly on day 40 postpartum followed by 500µg of PGF2α analogue intramuscularly on day 11 and a second dose 10µg GnRH analogue was administered on day 13 followed by TAI at 24th and 32nd hours. In Group III, induction of oestrus was done by administering PGF2α analogue 500µg intramuscularly on day 40 postpartum. A second dose of PGF2α analogue was administered on day 11, followed by TAI at 72nd and 80th hour. In Group IV, cows with a palpable functional CL on day 40 postpartum were administered 500µg PGF2α analogue and were inseminated at observed oestrus. Cows inseminated during first natural post partum oestrus formed the control group (Group V). In the experimental and control groups serum progesterone was estimated on day 40 postpartum, on days of hormone administration and during oestrum. Pregnancy diagnosis was done by rectal palpation of genitalia at 60 days after AI in all groups and the data were subjected to statistical analysis. Response to oestrus synchronization was 83.33, 33.33, 66.67 and 100 per cent in Group I to IV respectively. The time taken for induction of oestrum was 52.50 ± 0.99, 52.33 ± 0.71, 52.83 ± 1.40 and 53 ± 0.97 h respectively in Group I to IV but there was no significant difference between the groups. The duration of oestrus in Groups I to V were 37.33 ± 0.71, 35.67 ± 0.88, 40.50 ± 0.76, 38.83 ± 0.83 and 39.83 ± 0.48 h respectively. The percentage of animals showing high, medium and low intensities of oestrum respectively were 0, 33.33 and 50 in Group I, 0, 33.33 and 0 in Group II, 33.33, 33.33 and 0 in Group III, 83.33, 16.66 and 0 in Group IV, 83.33,16.66 and 0 in Group V. The mean serum progesterone level on day 0, 7, 9 and observed oestrum for those conceived in Group I were1.56 ± 0.69, 5.00 ± 0.94, 0.55 ± 0.09 and 0.36 ± 0.06 ng/ml. For those animals in Group I that did not conceive, the corresponding values for the same days were 0.36 ± 0.01, 0.76 ± 0.61, 0.40 ± 0.11 and 0.32 ± 0.08 ng/ml respectively. In Group II, the mean serum progesterone level on day 0, 11, 13 and observed oestrum for the conceived were 0.49 ± 0.23, 1.59 ± 0.59, 0.35 ± 0.13 and 0.88 ± 0.13 ng/ml respectively and those that did not conceive had mean serum progesterone level 0.99 ± 0.44, 0.35 ± 0.20, 0.31 ± 0.07 and 0.88 ± 0.21 ng/ml respectively. In Group III, the mean serum progesterone level on day 0, 11 and observed oestrum for the conceived were 0.83 ± 0.19, 3.05 ± 0.38 and 0.33 ± 0.10 ng/ml respectively and for the non conceived the corresponding values were 0.20 ± 0.05, 0.34 ± 0.11 and 1.39 ± 0.01 ng/ml respectively. The conceived animals in Group IV had mean serum progesterone level 2.77 ± 0.38 and 0.46 ± 0.12 ng/ml respectively on day 0 and observed oestrum while for those that did not conceive the corresponding values were 1 and 0.5ng/ml respectively. In the control group those that conceived had serum progesterone levels 1.30 ± 0.29 and 0.55 ± 0.08 ng/ml on day 40 and observed oestrum and the corresponding values for those that did not conceive were 1.6 ± 0.85, 0.685 ± 0.235 ng/ml respectively. The conception rates after synchronization in Groups I to V were 66.66, 33.33, 66.66, 83.33 and 66.66 per cent respectively. The overall conception rate in Groups I to V were 83.33, 83.33, 66.66, 83.33 and 66.66 per cent respectively. The mean calving to conception interval for the experimental groups was 53.84 ± 2.31 days whereas the corresponding values for the control and herd were 95 ± 6.19 and 200.78 ± 15.97 days respectively. The present study revealed that treatment with GnRH and PGF2α during early post partum period was useful for reducing the intercalving interval in a herd. Similarly single PGF2α administration on confirmation of a functional CL by clinical examination was useful and more economical for individual animals and small herds. Hence it is recommended that Ovsynch and PGF2α protocol could be suitably employed for reducing the intercalving period in post partum dairy cows.
Open Access
Diagnostic and therapeutic approaches for enhancing reproductive efficiency in female dogs
(College of Veterinary and Animal Sciences, Mannuthy, 2010) Deepthi, L; KAU; Aravinda Ghosh, K N
The present study was undertaken for finding optimal breeding time in bitches, early pregnancy diagnosis and tackling pseudopregnancy using antiprolactin drugs. Experiment consisted of 55 apparently healthy bitches which were brought to the Small Animal Obstetrics and Gynaecology unit of Veterinary College Hospital, Mannuthy and University Veterinary Hospital, Kokkalai for getting breeding advice. Fertility rate was assessed in two different groups viz., Group I (n=30) and Group II (n=25). The conception rate was 73.3 per cent in group I and 88.0 per cent in group II. Conception rate was more in those animals, which were bred, based on vaginoscopy than those bred based upon clinico-gynaecological examination and vaginal exfoliative cytology. This supported the view that performing vaginoscopy will augment conception rate in bitches. Clinico-gynaecological examination together with vaginal exfoliative cytology was found to be useful for finding optimal breeding time in bitches under field conditions. Vaginoscopy, in addition to being a non-invasive method, had the added advantage of predicting the fertilization period, thereby breeding advice could be more accurate as it helps in the visual inspection of vaginal mucosal folds. Pregnancy diagnosis was carried out in 20 animals at 20th, 35th and 50th day of gestation. Abdominal palpation at 20 days after breeding was non-confirmatory of pregnancy but at 35 days of gestation foetus could be appreciated as tense distinct uterine swellings. By ultrasound scanning at 20 days of gestation the image of embryonic vesicles appeared as spherical structures with anechoic consistency. No foetus or foetal membranes could be visualized at this stage. By day 25 to 27, echogenic foetal mass and heart beat became detectable. By day 28 to 33, head and body of the foetus were similar in size and revealed flickering heart beats. Anatomical features of the foetus became more obvious by about 34 to 39 days of gestation. Shape and size of the foetal head became distinguishable from the body at this stage. The percentage accuracy for transabdominal palpation at 20 days of gestation was found to be 20 per cent which improved to 75 per cent by about 35 days. The percentage accuracy at 50 days was 60 per cent. This study suggested that transabdominal palpation was not reliable in early and late gestation. For ultrasound scanning, the percentage accuracy at 20 days of gestation was found to be 15 per cent which improved to 90 per cent by 35 days. The percentage accuracy improved to 100 per cent by 50 days of gestation. Thus ultrasound scanning could be used as reliable tool for assessing the foetal viability. The level of serum alkaline phosphatase at 20th, 35th and 50th day of gestation was found to be 67.90 + 2.98, 91.85 + 2.10 and 139.65 + 6.84 U/L respectively. The level of haptoglobin at 20th day of gestation was 53.10 + 3.22 mg/dl where as the level elevated to 81.12 + 3.40 and 119.44 + 3.16 mg/dl by 35 days and 50 days of gestation respectively. The level of serum globulin at 20th, 35th and 50th day of gestation was 2.43 + 0.12, 3.05 + 0.11 and 3.74 + 0.15 g/dl respectively. Statistical analysis revealed significant difference between day 20 and day 50 (P<0.05). Thus the level of serum alkaline phosphatase, serum haptoglobin and serum globulin was found to be increasing as the pregnancy advanced and this could be used as indicators of healthy pregnancy. Total erythrocyte count (TEC) at 20, 35 and 50 days of gestation was 6.07 + 0.16, 5.58 + 0.15 and 5.12 + 0.16 million/cmm respectively. Haemoglobin concentration at day 20, 35th day and 50 days of gestation were found to be 11.86 + 0.28, 10.45 + 0.27 and 9.17 + 0.30 g/dl respectively. Packed cell volume values were 40.45 + 0.83, 37.30 + 0.87and 33.40+ 1.10 percent respectively at day 20, day 35 and 50 days of gestation. Erythrocyte sedimentation rate at day 20, day 35 and 50th day of gestation were found to be 10.31 + 0.73, 15.41 + 0.54 and 21.85 + 1.04 mm/hr respectively. Haemogram studies showed significant decrease in the total erythrocyte count, haemoglobin and packed cell volume at different gestational age (P<0.05). These changes were attributed by haemodilution and increased plasma volume. But erythrocyte sedimentation rate has shown an increase which could be attributed to the endometrial implantation by the embryo. The total leucocyte count at day 20, day 35 and day 50 of gestation was found to be 13844.90 + 539.90, 15449.00 + 569.86 and 17502.50 + 780.21 cells/cmm respectively. The neutrophil count at 20th, 35th and 50th day of gestation was 67.30 + 1.11, 70.30 + 4.95 and 75.35 + 1.27 per cent respectively. The lymphocyte count was found to be 27.80 + 0.87, 31.55 + 0.88 and 36.95 + 1.03 per
Open Access
Factors affecting conception rate on artificial insemination in goats
(College of Veterinary and Animal Sciences, Mannuthy, 2010) Remya Rajan, V; KAU; Metilda, Joseph
With the objective of evaluating the factors affecting conception rate on artificial insemination in goats, a study was carried out at Artificial Insemination Centre, under the department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, Thrissur using 22 ejaculates collected thrice weekly from an adult Malabari crossbred buck. Semen samples for chilling were diluted in Tris-yolk buffer and preserved at 3-5oC for 72 hours. Cryopreservation was done in Tris yolk glycerol extender after the removal of seminal plasma. The semen was diluted and packed in 0.25 ml straws and each dose contained 200 million progressively motile sperms before processing.A total of 44 adult healthy does brought to the AI Centre for insemination were selected for the study after detecting heat using buck jar technique. Selected does were at random allotted to four groups with eleven animals in each group. Does belonging to Group I, II and III were inseminated using chilled semen having motility over 35 per cent and stored for 0-24 h, 24-48 h and 48-72 h respectively. Animals of Group IV were inseminated using frozen semen having a minimum of 35 per cent post thaw motility. Transcervical insemination was carried out in the does in oestrus by speculum method. The depth of penetration of AI gun was assessed using another sheath as yardstick. Pregnancy diagnosis was performed at two months of gestation by ultrasonography or at three months of gestation by abdominal palpation.Average volume, density and mass activity of buck semen were 1.25 ± 0.97 ml, DDDD and ++++ respectively. Colour of the semen was creamy with a yellowish tinge. Average semen pH and sperm concentration were 6.89 ± 0.21 and 2781.82 ± 51.69 millions per ml respectively. The mean percentage of initial motility, sperm viability, abnormality and intact acrosomes was 82.77 ± 0.33, 90.14 ± 0.53, 2.28 ± 0.12 and 92.27 ± 0.21 respectively.The percentage of sperm motility at 24, 48 and 72 h of preservation at refrigeration temperature was 70.25 ± 0.60, 61.13 ± 0.72 and 42.81 ± 0.95 respectively. The live sperm percentage dropped to 83.42 ± 1.27 at 24 h, 78.84 ± 1.47 at 48 h and 74.57 ± 1.53 at 72 h of preservation by chilling. The percentage of sperm abnormalities increased to 2.97 ± 0.13 at 24 h, 4.12 ± 0.15 at 48 h and 5.34 ± 0.11 at 72 h of preservation. The percentage of intact acrosomes was 90.27 ± 0.18 at 24 h, 87.71 ± 0.37 at 48 h and 85.09 ± 0.44 at 72 h of refrigeration storage. There was significant difference between sperm motility, viability, abnormality and acrosome integrity at various storage periods.The percentage of sperm motility of 78.50 ± 0.50 at the end of initial extension decreased to 37.67 ± 0.49 after cryopreservation. The mean live sperm percentage was 84.66 ± 0.91 at the end of initial extension which after freezing and thawing dropped to 60.36 ± 0.77. The mean percentage of sperm abnormalities increased from 3.82 ± 0.12 to 6.34 ± 0.22 at the end of cryo preservation. The mean percentage of intact acrosomes was 85.68 ± 0.72 at the end of initial extension which showed a decrease to 76.06 ± 1.41 after cryopreservation. There was significant difference (p<0.01) between semen quality of frozen semen and chilled semen at various storage periods. Predominant behavioural signs observed using “Buck jar” technique were bleating, wagging of tail, frequent urination and flehmen reaction with an average intensity of heat score 2.91 ± 0.10. The major clinical signs were vulval oedema, moistness and hyperaemia of vagina, mucus discharge and opening of cervical os. Average depth of penetration of AI gun was 20.07 ± 1.35 mm. Overall conception rate in does inseminated using chilled semen was 72.73 per cent. Conception rate in Group I, II and III were 81.82, 63.64 and 72.73 per cent respectively, which did not differ significantly. The conception rate in does inseminated using frozen semen was 27.27 per cent. The study indicated that progressive motility and fertility of buck semen decrease on freezing causing a significant (p<0.01) reduction in conception rates compared to chilled semen. Intensity of heat and depth of penetration of AI gun have significant correlation with pregnancy status of does inseminated using frozen semen. The study revealed that liquid storage of buck semen under refrigeration is a viable alternative for propagation of germplasm of superior bucks with low freezability as it ensures better conception rates.

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