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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 Utero-tubal insufflation technique for diagnosis of tubal impatency in cows(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1981) Athman, K V; KAU; Neelakanta Iyer, C PThe objective of the present study was to design an apparatus for utero-tubal insufflation test and to standardize its use in both excised genitalia and in live animals for the diagnosis and treatment of tubal impotency. Perfect obturation of the cervix was obstained using a stainless steel catheter with 22 FR. Neo Foley’s latex catheter sleeved over. Unilateral and bilateral impotency were recorded in 8.70 and 19.25 per cent of the excised genitalia. The incidence of unilateral and bilateral impotency in 37 infertile animals were found to be 2.70 and 13.52 per cent respectively. The mean pressure at which air escaped from both the tubes in excised genitalia was 128.12 mm. Hg. In cows and 148.93 mm. Hg. In heifers. On the contrary in experimental animals the corresponding pressure in cows and heifers were 116.38 mm. Hg. And 127.50 mm. Hg. Respectively. It was observed that a higher pressure was required to open both the tubes in heifers than in cows. Blown up condition of the uterus took place in excised genitalia of cows and heifers between 100-280 mm. Hg. And 100-260 mm. Hg. Respectively. The pressure range for producing “blown up” condition in experimental animal was found to be 120-220 mm. Hg. In cows and 195-200 mm. Hg. In heifers. Since the safety margin was estimated to be 60.00 mm. Hg. In experimental cows, the test is considered to be a safe diagnostic test for tubal impotency. Although uterus might be “blown up” in a very few cases at a pressure lower than the higher pressure for escape of air from normally patent tubes, the test will not endanger the life of the animal. Since 69.56 percentage of cows with bilateral patency of salpinx on testing conceived after insemination, utero-tubal insufflation test could also be exploited for the treatment of temporary tubal blockage.ThesisItem Open Access Reproductive pattern and performance of nanny goats in Kerala(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1992) Krishna Kumar, G; KAU; Sudarsanen, VA study to elucidate the reproductive pattern and performance of nanny goats under the agroclimatic conditions of Kerala conducted on a flock of 154 nanny goats of mixed genotype, Malabari, Saanen x Malabari and Alpine x Malabari, maintained under the All India Co – ordinated Research Project on goats for milk, Kerala Agricultural University, has revealed the following results and conclusions. The method adopted was detection of oestrus twice a day using a vasectomised buck and by clinical examination. On detection, all animals of good reproductive health were bred artificially with freshly collected semen extended in Tris buffered yolk extender and preserved at low temperature. A group of animals were given a second insemination after seven hours of the first insemination for a comparison of the success rate to single and double inseminations during a heat. Following breeding, the duration of gestation, durations of stages of parturition, presentation of kids, incidence of multiple birth and secondary sex ratio were also recorded. Daily record of maximum and minimum temperature for a period of one year of the study was maintained. The period was divided into four quarters almost in agreement with the four natural seasons and the conceptions during each of the quarters were related with their secondary sex ratio and birth weight records to know the influence of the environmental temperature on them. Placental area weight records were also maintained and by applying linear regression equation they have been correlated with the birth weight to establish that the birth weight difference was due to the influence of environmental temperature on placental growth and development. Goats were found to be polyoestrous and 61. 54 per cent came into oestrus during the period from April to September when the day is long. Of the two peak breeding activities noticed the greater one occurred in July and the other during November. Breeding activity was found to be low during the months of January and February. The duration of oestrous cycle was found to be 18 – 23 days but the cycle length varied from 6 to 140 days. 45.8 per cent of the animals had an oestrus duration of less than 18 days. Duration of oestrus varied from 12- 48 h. Cessation of oestrus was noticed in 84.8 per cent of animals by 36 h and in 93.26 per cent of animals by 48 h. Conception percentage to first time single and double inseminations during a heat were 33.93 and 42.85 respectively. Overall conception percentages for the above were 71.42 and 82.14 respectively. Average gestation length was 145.62 ± 0.23 days while with singleton, twin and triplet, the durations were 146.05 ± 144.86 ± 0.32 and 145.25 ± 1.03 days. In twin pregnancies and in pregnancies with male foetus/foetuses in both singleton and homogamous twin, was found to have lesser duration of gestation. Mean durations of the first, second and third stages of labour were 57.47 ± 10.29 , 14.52 ± 1.34 and 128.87 ± 4.84 min respectively. Presentation of kids at the time of birth was 88.9 per cent in anterior and the rest in posterior presentation. Secondary sex ratios of singleton and multiple pregnancies were 52.05 and 54.08 per cent with an overall of 53.22 per cent. No significant variation could be observed in the secondary sex ratio of kids those were conceived between seasons having variable environmental temperature. Mean birth weight of kids born during the three trimesters with mean environmental maximum temperature 39.0, 33.4 and 36.30 C were respectively 1.64, 1.92 and 1.88 kg. Between the trimesters having 39. 00 C and 33.40 C there was a significant increase in birth weight. A definite correlation was found to exist between the placental weight and birth weight and the placental area and birth weight. The following conclusions were derived : 1. There is a reproductive pattern difference as could be seen from the above as 61.54 per cent of the oestruses occurred during the period of long days, contrary to the belief that breeding season of goats is the short days. Peak breeding activity was also noticed during the month of July. An instinctive attempt to reduce the number of births during the months of heavy rain which is not conducive for the survival of the young could be appreciated from the low breeding activity seen during January and February. 2. The duration of oestrous cycle, oestrus, gestation and the first, second and third stages of parturition were found to be in consonance with the already available informations. 3. Two inseminations during a heat was found to improve the conception percentage over single insemination. 4. Incidence of multiple pregnancy was found to be lower in the flock. 5. Secondary sex ratio of the kids born was 53.22 per cent and it confirms well with the reports already available. Presentation of foetus was anterior in 88.9 per cent and the rest of posterior presentation. 6. Conception between periods of variable environmental temperature did not seem to influence the sex ratio, to significantly alter the secondary sex ratio. 7. Environmental temperature at the time of conception was found to significantly influence the birth weight of kids. From the correlation that could be established between placental weight and birth weight and between placental area and birth weight it could be inferred that the environmental temperature effect on birth weight is through its influence on placental development and growth.ThesisItem Open Access Observation on gestation and parturition in goat-Capra hircus(Department of Animal Reproduction, College of Veterinary and Animal Sciences,Mannuthy, 1981) Kuriakose, K K; KAU; Neelakanta Iyer, C PA systematic study involving the use of 85 does comprising 27 Malabari, 35 Alpine x Malabari and 23 Saanen x Malabari crosses reared and maintained at Mannuthy, Kerala, under All India Co – ordinated Research Project on Goats for Milk was carried out in order to ascertain the gestation length, process of parturition and placental details. In addition 583 breeding particulars gathered from the registers maintained at the Project also utilized for assessing the gestation length and frequency of occurrence of multiple births. The salient observations made and the valid inferences drawn are indicated below. The average gestation length of different genetic groups of goats was found to be 146.66 + 0.53 days. There was no significant variation in the gestation period of does of different genetic groups both in single and multiple births. Parity had no influence on the gestation length in does. The variation in gestation length due to difference in the sex of kid was also not significant. Gestation period was not found to vary between seasons. The frequency of occurrence of single, twin and multiple births was found to be 50.75 per cent, 45.05 per cent and 4.19 per cent respectively. Signs of approaching parturition was assessed based on the degree of manifestation of such symptoms as tumefaction of the vulva, udder engorgement and relaxation of the pelvic ligaments. The symptoms exhibited by the does were only moderate in the beginning but on nearing parturition there was conspicuous increase in the intensity of these changes. Colostrum was present in all the animals two days prior to parturition. Visible flow of cervical mucus was evident in 75 does two days prior to kidding and in all, 24 hours before parturition. There was no significant fluctuation of body temperature of does during prepartum period. The total duration of parturition was 360.91 + 30.85, 358.20 + 16.67 and 395.42 + 40.09 minutes in single births and 359.12 + 26.15, 357.20 + 32.23 and 368.73 + 24.03 minutes in multiple births in Malabari, Alpine x Malabari and Saanen x Malabari goats respectively. The duration of parturition was not variance between genetic groups both in single and multiple births. The weight of kid was positively correlated to the weight of placenta, total number of cotyledone and to the number of large and medium sized cotyledons. There was negative correlation between weight of kid and the number of small sized cotyledons. The number of cotyledons in the gravid horn was significantly higher than that in non gravid horn in all the genetic groups. Weight of the placenta as well as the total number of cotyledone was found to be more in multiple pregnancy than in single pregnancy.ThesisItem Open Access Investigation on the pathological conditions in the genitalia of female goats(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1980) Ramachandran, K; KAU; Neelakanta Iyer, C PA study was undertaken to assess the incidence and nature of pathological conditions affecting the genitalia of female goats. The material for the present study comprised of 950 genitalia, inclusive of 42 gravid ones, collected at random, from the slaughter house, Corporation of Cochin, Ernakulam. The organs which revealed gross lesions and those which were suspected to be affected were subjected to detailed histopathological studies. Pathological lesions of genital organs were observed in 48 (5.053%) genitalia. Thirty two (3.368%) genitalia revealed various pathological changes of the ovaries and pathological changes of the uterus were recorded in20 (2.105%) genitalia. The cervix, vagina and vulva did not reveal any pathological changes in any of the genitalia examined. The following pathological conditions were observed during the course of the present study: Ovarian hypoplasia (0.316%); cystic ovarian degeneration (0.947%); cystic corpus luteum (0.421%); chronic oophoritis (0.211%); multifollicular atresia (0.105%); par ovarian cyst (1.368%); Bursitis (0.842%); salpingitis (0.105%); macerated foetus (1.684%); endometritis (0.211%); cystic glandular hyperplasia (0.105%) and cyst in the endometrium with sub – epithelial haemorrhage (0.105%).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 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 Studies on post- partun oestrum in cross bred cows(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1979) Suresan Nair, S P; KAU; Neelakanta Iyer, C PThe object of the study was to find out the various reproductive parameters that influence the onset of post – partum oestrus in cross bred cows. With this object in view, a detailed investigation was undertaken in a herd of 115 cross bred cows of Jersey x Sindhi and Jersey x Local breeds belonging to the University Livestock Farm of the Kerala Agricultural University, Mannuthy during the period from February 1978 to March, 1979. After parturition, they were grouped into normal and abnormal calvers based on the normalcy of calving. All the cows were stall fed and were under identical conditions of feeding and management. The observations made and inferences drawn are summarised below: There were significant variations in the cessation of lochial discharge, regression of pregnancy corpus luteum, period of uterine and vulval involution, onset of first and second ovulations and the post – partum oestrus interval between normal and abnormal parturitions. Duration of lochial discharge, regression of pregnancy corpus luteum and onset of first ovulation were found to be significantly influenced by the season of calving. Parity of the cow was found to influence the period of lochia, involutions of uterus and also the post – partum oestrus interval. Milk yield of the cow significantly influenced regression of pregnancy corpus luteum and vulval involution. However, the breed of the cow and sex and weight of calf were not found to have any significant effect on the reproductive parameters studied. An incidence of 13.4 per cent of aberrations of oestrum was also observed.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 conditions
