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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 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 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 statusThesisItem Open Access Superovulation and embryo recovery in rabbits(Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 1997) Sathesh Kumar, S; KAU; Suresh Nair, S PSuperovulation was induced in Newzealand White and Sovira. Chinchilla breeds of rabbits by administration of a single dose of 150 IU PMSG followed by double mating at induced cycle and 150 IU HCG soon after second mating to induce ovulation. The onset and intensity of oestrum, number of ovulations, embryo recovery and quality of embryos were studied and compared with those of the controls of the respective breeds. The mean interval from PMSG administration to onset of oestrus in both the breeds was 56.0 + 5.1 h. It was further observed that most of the treated animals showed intense oestrum when compared to controls. The ovulation rate based on the number of corporalutea in control animals of Newzealand White breed was 4.7 + 1.2 as against 22.0 ± 1.35 in the treated group. There was significant difference (P<0.01) in the ovulation rate between the groups. The percentages of embryo recovery, fertile embryos and transferrable embryos in the control group were 47.22, 86.7 and 92.3 while those of the treatment group were 31.67, 100 and 87.8 respectively. There was no statistically significant difference between the groups. While the control animals in Soviet Chinchilla breed has an ovulation rate of 6.7 ± 0.65, the treated rabbits showed a higher ovulation rate of 20.0 ± 3.2. There was significantly higher ovulation rate (P<0.01) in treated group when compared to controls. The embryo recovery rate, fertilized embryos and transferrable embryos in the control group were 53.33 per cent, 77.8 per cent and 93.8 per cent respectively. The corresponding values in the treatment group were 36.27 per cent, 69.82 per cent and 93.1 per cent respectively. There was no significant difference between the groups. No breed influence on the above parameters could also be noticed in this study It may be concluded that superovulation could be successfully induced in Newzealand White and Soviet Chinchilla breeds of rabbits with single dose of 150 IU PMSG, followed by 150 IU HCG soon after second mating. Eventhough there was superovulation, the embryo recovery rate was comparatively lower in the treated group probably on account of an altered oestrogen-progesterone profile interfering with the transport of the zygote, however the fertilization rate and the quality of the embryos were unaffected with the superovulation treatment.
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