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ThesisItem Open Access Effect of processing and freezing procedures on the acrosome morphology of buck spermatozoa(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1998) Ranjini, A; KAU; Prabhakaran Nair, KSix pooled semen samples (two ejaculates) of good quality from five Malabari crossbred bucks were processed and frozen in two different protocols to evaluate the effect of processing and freezing procedures on the acrosome morphology of buck spermatozoa. In protocol I, the samples were diluted 10 fold in Tris buffer before centrifuging twice and the final pellet was re-suspended in the non glycerolated fraction of Tris yolk diluent. The sample was glycerolated (six per cent), equilibrated (four hours), frozen (eight minutes), and thawed (250 C for 30 seconds). In protocol 11, centrifugation was done only once, after 15 fold dilution in Tris buffer. The re suspended pellet was glycerolated (seven per cent), equilibrated (three hours), frozen (10 minutes) and thawed (60° C for 10 seconds). The semen characters such as motility, live sperm, sperm abnormalities and acrosome abnormalities were evaluated at the end of washing and initial extension (stage I), cooling to 5° C (stage II), glycerolisation and equilibration (stage Ill) and freezing and thawing (stage IV). The results were compiled to evaluate the effect of different processing and freezing procedures on the semen characters in general and acrosome morphology in particular. The semen sample used for split sample dilution had a mean volume of 1.3282± 0.067 ml, creamy in colour, DDDD density, ++++ mass activity, pH of 7.275 2± 0.040 and a concentration of 2972 2± 293 millions per ml. No significant difference in the above semen characters were found between bucks. The initial sperm motility of 82.000 2± 0.606 was found to drop significantly during processing and freezing and the final post thaw motility obtained was 44.000 2± 0.790 in protocol I. Similarly in protocol II the initial motility dropped from 81.375 2± 1.089 to 44.750 2± 1.075 at the end of stage IV. Even though there was significant drop in motility between stages in both the protocols, there was no significant difference in the corresponding stages of the two protocols. It could be inferred that good post thaw motility was obtained in both the protocols. The fact that a single washing and centrifugation was only adopted in protocol II makes it a more acceptable procedure for buck semen freezing. The mean live sperm percentage of fresh semen was evaluated using both NE and NEG staining technique. The percentage of live sperms of 90.050 2± 0.801 was found to decrease to 54.250 2± 0.593 after freezing and thawing in protocol by NE staining. Similarly in protocol 11, the mean percentage of live sperms was found to reduce to 53.125 2± 0.793 with the same staining. Even though there was significant difference in the live sperm percentage between stages within protocol I and II no significant difference in the live sperm percentage between the corresponding stages of protocol I and I I . With NEG staining the initial live sperm percentage of 80.850 ± 1.494 was found to drop to 54.875 ± 0.677 in protocol I as against 53.400 ± 0.730 in protocol II. While there was significant difference in the live sperm percentage between stages within protocol I and II there was no variation between corresponding stages of the two protocols. A significantly lower percentage of live sperms was recorded with NEG staining when compared with NE staining probably on account of the fact that the differentiation of live and dead sperm was difficult in the former staining method as live sperms were stained light blue instead of colourless. The mean percentage of abnormal sperms of 3.050 ± 0.245 in fresh semen did not register any significant increase during processing. However, there was significant increase in the percentage of sperm abnormalities during freezing and thawing with the final abnormality percentage of 7.125± 0.706 in protocol I and 6.300± 0.36 in protocol II. The initial acrosomal abnormality of 8.825 in the fresh semen steadily rose to 23.375 in protocol I as against 19.825 in protocol II at the end of stage IV. There was no significant difference in the percentage of various acrosomal abnormalities between corresponding stages of the two protocols. However, there was significant increase in the acrosomal abnormalities during glycerolisation, equilibration, freezing and thawing under both the protocols. It was concluded that the processing and freezing under two different protocols did not significantly alter the post thaw motility, percentage abnormal and dead sperms and acrosomal abnormalities. A good post thaw motility and low acrosomal abnormality was obtained with a single washing of buck semen with 15 fold Tris buffer which was comparable with double washing with 10 fold Tris buffer.ThesisItem Open Access Management of Oestrous Cycle in Crossbred Cattle Using Prostaglandin(Department of Animal Reproduction, College of Veterinary and Animal Sciences,Mannuthy, 1994) Ajit Kumar, G; KAU; Madhavan, EWith the object of studying the efficacy of administration of prostaglandin F2 alpha in the management of oestrous cycle and fertility of fixed time insemination in induced oestrus, 96 heifers and cows which were cycling, selected from Kerala Agricultural University Livestock Farms, were allotted to three different groups. Sixteen heifers and 16 cows in group I were given 25 mg PGF2 alpha (Lutalyse) intramuscularly when they had functional corpora lutea, while in group II, 16 heifers and 16 cows were administered two injections of PGF2 alpha 25 mg each 13 days apart. Sixteen heifers and 16 cows were treated as control (Group III). Twelve heifers (75%) and 16 cows (100%) in group I and all animals in group II responded to treatment. Mean time taken for induction of oestrus was 56.64 and 65.44 h respectively in heifers and cows. This difference was statistically significant. Parity of cows significantly influenced the time taken for induction of oestrus. Duration of oestrus was significantly different between heifers (21.86 h) and cows (31.13h) of the experimental group. Significant difference in the duration of oestrus was observed among cows between experimental and control groups. All experimental animals which responded to PGF2 alpha showed marginal increase in physical characters of the reproductive tract like vulval oedema, hyperaemia of vaginal mucosa, vulval discharge and tonicity of uterine horns. Majority of the experimental animals showed medium to high intensity of oestrus compared to natural oestrus. The first insemination and overall conception rates of heifers in group I, inseminated 72 h post-treatment, were 33.33 and 66.67 per cent respectively as against 33.33 and 50 per cent when inseminated 96 h post-treatment. Cows in groupI recorded 25 and 62.50 per cent and 37.50 and 62. 50per cent first insemination and overall conception rates respectively when inseminated 72 h and 96 h post-treatment. When heifers and cows in group II were inseminated 72h after the administration of the second dose of PGF2 alpha, 12.50 and 25 per cent conceived at first insemination with overall conception rate of 62.50 and 37.50 per cent respectively. The corresponding values in group II, when inseminated 96 h, were 25 and 12.5 per cent and 50 and 12.5 per cent respectively. Among control 6.25 per cent of heifers and 18.75 per cent of cows conceived at first insemination. The first insemination conception rate of heifers of experimental (25%) and control groups (6.25%) were significantly different. However, overall conception rate did not differ between the three groups. There was no influence of parity or season on the conception rate. Similarly the time taken for induction of oestrus, duratioh of oestrus, physical changes of the reproductive tract during oestrus and intensity of oestrus did not affect the conception rate. Heifers and cows required 1.88 and 1.93 inseminations respectively per conception when treated with PGF2 alpha as against 3.14 and 3.33 in the control group. Though, the present investigation revealed that PGF2 alpha can be used in the management of oestrous cycle in crossbred heifers and cows with fixed time insemination, the high cost of the drug and the conception rate obtained warrant further studies on fertility in relation to detected oestrus and fixed time insemination after the administration of PGF2 alpha.ThesisItem Open Access Superovulatory response, embryo collection and transfer in crossbred cows(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1996) Unnikrishnan, M P; KAU; Suresan Nair, S POut of eleven crossbred cows superovulated with 2000 IU of PMSG on day eleven and 25 mg PGF2 alpha 48 h later, nine animals (81.8%) exhibited oestrum after an average interval of 39.44 ± 2.44 h. Average duration of oestrum was 40.00 ± 2.00 h with 66.7 per cent of them exhibiting intense heat signs and 33.3 per cent exhibiting only moderate heat signs. Average number of corpora lutea and unovulated follicles in both the ovaries put together was 6.67 ± 0.50 and 6.22 ± 0.57 respectively. Ovarian response was more in right ovary than in left ovary, though not statistically significant. Average fluid recovery on flushing was 79.5 per cent which was comparatively low. The average embryo recovery and percentage of embryo recovery were 3.38 ± 0.70 and 53.11 ± 10.18 respectively, which was comparatively lower. The reason for poor recovery of embryo was attributed to poor fluid recovery on flushing. The average number and percentage of transferrable embryo recovered were 1.50 ± 0.53 and 33.34 ± 10.14. Reason for these lower rates were attributed to loss of embryo quality, due to prolonged action of PMSG. Donors of parity over three performed better on superovulation and flushing, than those below three. Animals of age group six to nine years produced more transferrable embryos than cows of age group ten and above. A conception rate of 30 per cent was achieved after transfer of embryos to naturally synchronised recipients. Heifers appeared to be better recipients than cows. Although a high incidence of abortion was encountered, birth of an embryo transfer male calf was also recorded in the study. All the donor cows came into regular oestrous cycle within four months of superovulation treatment.ThesisItem 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, MathewThis 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 oocytesThesisItem 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, MathewWith 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.ThesisItem 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, TDetailed 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.ThesisItem Open Access Detection of serum relaxin as a diagnostic tool for early pregnancy diagnosis in bitches(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2007) Deepthi, L; KAU; Sreekumaran, TWith the object of fioding a suitable and reliable method of early pregnancy diagnosis in bitches, the study was undertaken to investigate the efficacy of trans abdominal palpation, ultrasound scanning and relaxin detection was conducted. The study consisted of 45 apparently healthy bitches whieh were brought to the clinics for finding the optimal breeding time. Out of this, ten animals were selected at random for pregnancy diagnosis and were subjected to different methods of pregnancy diagnosis at different gestational age-16 to 20 days, 21 to 24 days and 25 to 30 days post breeding. Blood samples were collected for the estimation of haemoglobin, packed eell volume and erythrocyte sedimentation rate at the day of breeding and also at the above gestation periods. Body weights were reeorded at the day of breeding and also at different gestation periods. In the present study, it was found that abdominal palpation was difficult m diagnosing pregnancy between 16 to 20 days of gestation. When palpation was done in between 21 to 24 and 25 to 30 days post breeding, the accuracy obtained was 50% and 70% respeetively. This study suggests that trans abdominal palpation was not useful in diagnosing early pregnancy. By ultrasound scanning, the percentage accuracy at 16 to 20 days was 50%, which improved to 80 percent and 100 percent at 21-24 and 25-30 days post breeding respeetively. Foetal heartbeat could be observed in all the positive cases from 24 days of gestation. Pseudo-pregnancy, pyometra and abortion could be easily identified by this method. The earliest positive result obtained for serum relaxin detection was obtained at 20" day post breeding and the percentage accuracy was 50% at this period, as against 100% at 21-30 days of gestation. In the present study, it was found that serum relaxin test was not influenced by pseudo-pregnancy and uterine pathological conditions like pyometra. There was significant variation in haemogram (P <0.01) at the day of breeding and at different gestational age. Haemoglobin concentration at 16-20, 21-24 and 25-30 days of gestation were 10.88+0.31, 10.24+0.22, 8.77+0.28g/dl, which was lower than the value 11.56+0.27 obtained prior to breeding. The packed cell volume values were 34.66+0.9, 30.77+0.94, 28.22+1.02 and 26±0.94 percent at day 0, 16-20, 21-24, 25-30 days post breeding. There was significant variation in the values before and after conception. There was significant variation in erythrocyte sedimentation rate between day zero and at different gestational age. The values obtained varied significantly and recorded as 4.6±0.33, 14.3±1.09, 17.8±1.28 and 21.76±1.47mm/hr at day 0, 16-20, 21-24 and 25-30 days of gestation respectively. The body weight of all the ten animals varied significantly (P<0.01). It was observed that the body weight had shown a steady and progressive increase as the pregnancy advanced. The study revealed that abdominal palpation was not very useful in diagnosing early pregnancy. By ultrasound scanning, uterus as well as foetus could be visualized after 23 days of gestation. Serum relaxin detection could be used as an early tool for pregnancy diagnosis in bitches from 20 days post breeding. Results of the present study suggest that the relaxin test was accurate in diagnosing early pregnancy and its advantage being that it could be conducted and interpreted easily by a dog breeder or a dog owner. It could be concluded that detection of serum relaxin is a quick, simple and accurate tool for diagnosing early pregnancy under field conditionsThesisItem Open 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 MathewWith 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.ThesisItem Open Access Growth and reproductive performance of crossbred heifers in selected areas(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1998) Rajeev, R; KAU; Aravinda Ghosh, K NGrowth and reproductive status of crossbred heifers under field condition were assessed and the role of calcium, phosphorus, copper, zinc and manganese with reproductive performance was evaluated with the aim of evolving suitable corrective measures in cases of those with impaired reproductive performance due to subnormal serum mineral status. One hundred and twelve heifers were subjected to repeated gynaecoclinical examination. It was observed that there were 36.6 per cent true anoestrum, 19.6 per cent under developed genitalia, 29.5 per cent normally cycling, 9.8 per cent repeat breeders, 3.6 per cent suboestrum and 0.9 per cent bilateral ovarian hypoplasia. From the above heifers 89 were randomly selected and classified based on the breeding history and repeated gynaecological examination as 15 normally cycling (control), 41 true anoestrous heifers, 22 under developedgenitalia and 11 repeat breeders. The daily weight gain obtained was 55.05 ± 4.2 g, 32.26 ± 2.49 g, 27.33 ± 3.4 g and 24.1 ± 4.8 g. The above result gave significant difference in weight gain between control animals and other groups. The growth rate of heifers might have influenced the normal reproductive performance. Serum samples drawn from 89 heifers were analysed for calcium, inorganic phosphorus and trace elements namely copper, zinc and manganese. Serum calcium and phosphorus were estimated by employing spectronic-20, i&hile trace elements were estimated through atomic absorption spectrophotometer. The serum calcium level obtained was 11.1 ± 0.31 mg%, 10.74 ± 0.13 mg%, 10.8 ± 0.2 mg% and 10.8 ± 0.42 mg% in normally cycling, true anoestrous, under developed genitalia and repeat * breeding heifers respectively. The serum levels of all the four groups were well within the normal range and no significant variation among the groups. Hence the influence of calcium on reproduction could not be established. The serum inorganic phosphorus was 4.87 ± 0.13 mg% in normally cycling heifers (control) as against 3.83 ± 0.09 mg% for true anoestrous heifers, 3.52 ± 0.1 mg% for underdeveloped genitalia and 4.7 ± 0.15 mg% for repeat breeders. The level was significantly lower (<0.05) in true anoestrous and underdeveloped genitalia compared to control group. It can be summarised that hypophosphataemia might be one of the cause for true anoestrum and under developed genitalia. Among the trace elements estimated the serum level of copper only was found to be significantly varying among normally cycling, true anoestrous and heifers with under developed genitalia. The serum copper in control group heifers registered a value of 1,26 ± 0.07 ppm which was significantly higher (P<0.01) than those recorded for true anoestrous heifers (O’. 9 ± 0.04 ppm) and heifers with under developed genitalia (0.71 ± 0.05), ajhile no statistical significant variation obtained between serum value of repeat breeders (1.27 ± 0.08 ppm) and the control group. It is therefore reasonable to assume that hypocupraemia as evidenced by lower serum value might have contributed to true anoestrum and under developed genitalia condition and not with that of repeat breeding condition. The serum zinc and manganese levels of control group were 1.71 ± 0.05 ppm and 0.04 ± 0.002 ppm respectively. The corresponding values for the true anoestrum heifers were 1.61 ± 0.03 ppm and 0.04 ± 0.002 ppm and for heifers with under developed genitalia group were 1.6 ± 0.05 ppm and 0.04 ± 0.002 ppm respectively. These values did not vary significantly from those of control group. The corresponding values for repeat breeders were recorded to be 1.73 ± 0.06 ppm and 0.04 ± 0.002 ppm which did not differ: significantly from the values obtained for control group. The result of supplementation with dicalcium phosphate and copper sulphate to the respective mineral deficient heifers with true anoestrum and under developed genitalia showed that the mineral supplementation could induce oestrum. The serum mineral status comparison at different level of feeding showed significant difference (P<0.05) in the serum phosphorus level as well as copper level of moderate plane group with that of low and poor plane groups. Hence the effect of plane of nutrition on serum mineral status could be established in case of serum phosphorus and copper. The soil level of calcium, phosphorus, copper, zinc and manganese found to be well within the normal range. The level of exchangeable calcium and available phosphorus were ranged 0.11-0.12 per cent and 0.05-0.06 per cent respectively. The available copper, zinc and manganese levels obtained were ranged 4.43-4.5 ppm, 5.3-5.44 ppm and 96.34-99.7 ppm respectively. The result showed that the soil mineral content did not influence: the serum mineral status