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ThesisItem Open Access EFFECT OF GONADOTROPIN RELEASING HORMONE AND PROSTAGLANDIN FOR IMPROVING REPRODUCTIVE EFFICIENCY IN GOATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2008) JULLIET; 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 lA and Group IB. Group II animals were those that exhibited weak oestrus signs and were divided into three subgroups namely Group llA, IIB and lie. Group IA animals were administered 0.0042 mg Buserelin (1 ml Reeeptal) 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 lA and IB was 19.33 ± 0.45 and 33 ± 0.58 h respectively. The conception rate in Group lA and IB was 50 per cent and 66.66 per cent respectively. The serum P4 level on day 0 in does in Group lA 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 Reeeptal) on day 0, 125 pg 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 pg 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 lA, IB, IIA, IIB and lie 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 signsThesisItem Open Access EFFECT OF BLOOD UREA NITROGEN, MINERAL STATUS AND UTERINE pH ON FERTILITY IN DAIRY COWS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2008) SEENA. N. S; K V ATHMANAn investigation was carried out with the objective of studying the effect of BUN and uterine pH on fertility in dairy cows under farm and field conditions and also for correlating the mineral status with fertility using 40 crossbred dairy cows selected at random during oestrus, 20 each from those belonging to University Livestock Farm, Mannuthy (Group 1) and those brought for insemination at Artificial Insemination Centres at Mannuthy and Kokkalai (Group 11). All the cows in Group 1 were found to be maintained in a relatively high nutritious diet computed as per the scientific feeding standards compared to Group II animals. Detailed elinico-gynaecological examination was carried out and blood samples and uterine mucus were collected from all the selected cows during oestrus for estimation of biochemical parameters and uterine pH respectively. They were inseminated during the most appropriate period of oestrus and were subjected to pregnancy diagnosis at 60 days post insemination. Conception rates in both the groups were compared in relation to each parameter. The mean duration of oestrus was slightly higher in Group 11 (30.00 ± 2.11 hours) compared to Group 1 (26.10 ± 1.74 hours). Intensity of oestrus was high, medium and low in 40, 45 and 15 per cent of animals respectively in Group 1 and 50. 40 and 10 per cent in Group 11. Physical changes of reproductive tract viz. degree of vulval oedema and hyperemia of vestibular mucous membrane were more pronounced in animals of Group II compared to Group I, where as degree of tonicity was high in Group 1 compared to Group II. Characteristics of cervical mucus were also found to be affecting fertility. Better conception rate was obtained in animals with clear and stringy cervical mucus exhibiting typical type of fern pattern. Spinnbarkeit value did not vary much between conceived and non conceived animals. Uterine pH did not show a marked variation between groups, even though a slightly higher value was recorded in Group 1. But, an inverse relationship could be obtained between BUN level in serum and uterine pH during oestrus. Correlation between uterine pH and BUN level was highly significant (P<0.01) with a correlation eo-efficient of r = -0.896 and r = - 0.753 in groups 1 and 11 respectively. The mean blood urea nitrogen level in animals of Group 1 was significantly higher (P<0.05) than that in Group 11. The BUN level also varied significantly (P<0.05) between conceived and non-conceived animals of both groups. A marginal increase in plasma glucose level could be noticed in conceived animals compared to non-conceived animals of Group 1. The mean serum total protein was significantly higher (P<0.01) in Group 1 compared to Group 11. The serum level of minerals viz. calcium, phosphorus, manganese, zinc and copper were also correlated with fertility. The mean level of serum calcium and manganese varied significantly between groups 1 and II, but there was no significant difference in serum phosphorus, zinc and copper between two groups. Also, a slightly higher mean value was observed for serum calcium, phosphorus, manganese and zinc in conceived animals compared to non-conceived. But serum copper level did not vary between conceived and non-conceived animals. In light of these findings, it can be concluded that the elevation in systemic concentration of urea is likely to impair fertility in dairy cows as a consequence of alterafions in uterine environment. The benefits of feeding excess dietary protein and urea to maintain peak milk production should be compared with potential negative effects on fertility. Hence, a good nutritional management is essential for improved fertility in dairy cows.ThesisItem Open Access DETECTION OF SERUM RELAXIN AS A DIAGNOSTIC TOOL FOR EARLY PREGNANCY DIAGNOSIS IN BITCHES(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2007) DEEPTHI. L; T. SreekumaranWith 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 EFFECT OF PROSTAGLANDIN - PREGNANT MARE SERUM GONADOTROPIN (PMSG) COMBINATION FOR ENHANCING PROLIFICACY IN MALABARI GOATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES Mannuthy - Thrissur, 2002) P. SENTHILKUMAR; P. P. BalakrishnanThe object of present investigation was to evaluate the efficacy of prostaglandin-PMSG combination treatment at different dose levels in order to enhance the fertility and prolificacy of Malabari does. The material used for the study consisted of 48 cycling nulliparous Malabari does of eight to ten months age and body weight 18 to 20 kg, belonging to Kerala Agricultural University Goat farm, Mannuthy. All the experimental does were administered with two doses of cloprostenol (SYNCHROMATE) at the rate of 0.5ml intramuscularly 11 days apart. One day prior to the second prostaglandin administration the does were randomly divided into four groups viz. Group I, II, III and IV with 12 in each group. On the same day group I, II and III were administered PMSG (FOLLIGON) intramuscularly at the rate of 200, 400 and 600 lU respectively. Group IV was maintained as control with the prostaglandin treatment alone. After the second dose of prostaglandin all does in group I, II and III (100%) showed oestrus and in group IV only II does (91.67%) exhibited oestrus. The mean time taken for onset of oestrus in group I, II, III and IV was 28.00±2.70, 30.00±4.3I, 24.00, 43.64±4.36 h respectively. Group IV was significantly different from prostaglandin-PMSG group I, II and III (P < 0.01). The mean duration of oestrus in group I, II, III and IV was 84.00±6.94, 64.00±7.44, 86.00±752 and 34.91 ±4.97 h respectively. Group IV was significantly different from group I, II and 111 (P < 0.01).Mean intensity oestrus score was 11.50±0.49, 12.25±0.33, 14.25±0.72 and 8.82±1.59 respectively in group I, II, III and IV. Group IV was statistically significant from group I, II and III (P < 0.01). All prostaglandin-PMSG treated does exhibited common oestrus signs like wagging of tail, standing to be mounted, vulval redness, vulval oedema and vulval discharge whereas in control group only wagging of tail, vulval redness and vulval oedema noticed.The percentage of conception rate in group I, II, III and IV was 41.67, 50.00, 33.33 and 45.45 respectively. In prostaglandin-PMSG groups I, II and III mean litter size was 1.60±0.25, 1.5010.43 and 1.50+0.65 respectively but in group IV the same was 1.2010.20. There was no significant difference between the groups in litter size. However, more litter size with twins and triplets was noticed in prostaglandin- PMSG groups than the control group. Ingroup I, II and III mean birth weight was 1.4510.14, 1.2410.13 and 1.2710.18 kg respectively whereas in group IV it was 1.6210.24 kg. There was no significant difference among the groups with respect to the birth weight of kids. The percentage of preweaning mortality of kids in group I, II, III and IV was 50.00, 44.44, 50.00 and 33.33 respectively. The causes of preweaning mortality were pneumonia, enteritis and other etiological factors such as sudden death of weak born kids. Analysis of the results of present investigation indicatedthat prostaglandin double dose combined with PMSG at low dose regimen of 200 lU treatment can be used for enhancing the litter size without affecting thereproductive efficiency of nulliparous young does. For enhancing the litter size of goat, though requires further detailed investigation, it appears to offer a clear indication on the possibility of hormonally modulated for enhancement of litter size among goats. This might find in potential commercial application in intensive goat production systemThesisItem Open Access C(College of Veterinary and Animal Sciences, Mannuthy., 2005) ASHA MERINA KURIAKOSE; Dr. JOSEPH MATHEWWith the object of comparing different methods of pregnancy diagnosis and to determine the early and 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. At 20 to 30 days post breeding the accuracy of abdominal palpation were 61.9, 60 and 75 per cent, respectively, in large, medium and small breeds and by ultrasound scanning it was 90.4, 80 and 75 per cent in that order. Accuracy of abdominal palpation in large, medium and small breeds at 31 to 40 days of gestation were 66.6, 100 and 80 respectively and it was 100 per cent for ultrasound scanning. At 41 to 65 days post breeding the accuracy of diagnosis by abdominal palpation in large, medium and small breeds were 81.25, 75 and 60 per cent respectively, while, by ultrasound scanning it was 100 per cent in all animals. Foetal heart beat could be observed in all pregnant bitches from 33 days of gestation. There was a high correlation between predicted value and actual value of gestational age by measuring the gestational sac diameter and the coefficient of correlation was 0.792, 0.855 and 0.953 in large, medium and small breeds, respectively. However, the accuracy of estimating the foetal number by ultrasound scanning was only 23.52 per cent. There was significant variation (P<0.05) in haemogram of pregnant bitches compared to non pregnant. Total erythrocyte count at 20 to 30, 31 to 40 and 41 to 65 in pregnant bitches were 7.62 ± 0.17, 6.99 ± 0.2 and 6.70 ±0.19 million/cmm and that of non pregnant was 8.32 ± 0.28 million/cmm. Haemoglobin concentration at 20 to 30, 31 to 40 and 41 to 65 days post breeding were 11.73 ± 0.18, 10.9 ± 0.2 and 10.69 ± 0.2 g/dl and that of non pregnant was 12.37 ± 0.28 g/dl. There was significant variation observed in haemoglobin concentration between pregnant and non pregnant dogs. Erythrocyte sedimentation rate at 20 to 30, 31 to 40 and 41 to 65 days post breeding were 12.03 ± 0.97, 17.62 ± 1.11 and 17.07 ± 1.09 mm/hr respectively and that of non pregnant was 2.5 ± 1.57 mm/hr. There was significant variation observed in ESR between pregnant and non pregnant bitches. The packed cell volume at 20 to 30, 31 to 40 and 41 to 65 days post breeding in pregnant animals were 39.83 ± 0.72, 37.70 ± 0.82 and 36.88 ± 0.8 per cent respectively. In non pregnant one it was 45 ± 1.15 per cent There was significant variation observed in PCV between pregnant and non pregnant. There was no significant variation in leucogram and serum biochemistry between pregnant and non pregnant. There was significant variation of plasma fibrinogen between pregnant and non pregnant animals. The plasma fibrinogen concentration at 20 to 30, 31 to 40 and 41 to 65 days post breeding were 321.10 ± 20.92, 251.33 ±21.49 and 210.52 ± 22.11 mg/dl respectively and that of non pregnant it was 156.33 ± 37.22 mg/dl. The mean plasma fibrinogen concentration was found to be highest between 20 to 30 days of gestation. Hence estimation of plasma fibrinogen could be taken as an additional index for early pregnancy diagnosis in bitches. Results of this study indicates that ultrasound scanning could be used as a more accurate and scientific method of diagnosing pregnancy even during early stages especially to confirm the foetal viability. However, for routine clinical practice an approach of transabdominal palpation combined with ultrasound scanning could be used for confirmed pregnancy diagnosis in bitches. Moreover, increase in plasma fibrinogen, by haematological studies could be taken as a supportive index for pregnancy diagnosis in bitches.ThesisItem Open Access EVALUATION OF BOAR SEMEN EXTENDED IN BELT'S VILLE THAW SOLUTION(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2001) Kantharaj., S.; Athman, K.V.An experiment was conducted to study the effect of natural service and artificial insemination on reproductive performance of pigs and to assess the fertility and preservability of boar semen extended in Belt's ville thaw solution. Twenty-four Large White Yorkshire (LWY) gilts, twenty-four sows and six LWY boars were selected for the study. Out of six boars, three boars were used for natural service and the other three maintained for artificial insemination purpose. A total of 52 ejaculates were collected from boars maintained for artificial insemination purpose at weekly intervals by the gloved hand technique. Among these boars, no significant differences were found for most of the semen characteristics except proximal and distal protoplasmic droplets, which were significantly different between boars (P<0.05). Negative correlation between total volume of ejaculate and sperm concentration was observed. No significant correlation between total sperm abnormalities and live sperm count could be detected. Twenty ejaculates from these boars were used for preservation studies. No significant variation in the preservability of spermatozoa could be detected between boars, whereas, there was a highly significant difference (p<0.01) in the preservability of spermatozoa between hours of preservation. Twelve LWY gilts and twelve sows (Group-I) and three boars were used for natural service. Each gilt and sow was mated twice during the oestrus. Twelve gilts and twelve sows (Group-II) were artificially inseminated twice during the oestrus with the semen diluted in BTS. There was a significant difference (P<0.05) in the duration of oestrus between gilts and sows. There was a marginal difference in the conception rates between naturally served and artificially inseminated gilts and sows. The farrowing rate was 100 per cent for both naturally served and artificially inseminated gilts and sows. There was a significant difference in the gestation length between artificially inseminated gilts and sows. There was a significant variation in the litter size between naturally served and artificially inseminated gilts and sows. It can be concluded from the study that artificial insemination in pigs could well be performed under commercial farming conditions for effective disease control and increased productivity.ThesisItem Open Access EFFECT OF NORGESTAMET AND PROSTAGLANDINF^LPHA IN CROSSBRED HEIFERS FOR AUGMENTING FERTILITY(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2001) GOPIKRISHNAN., f.; VAHlDA, A. M.With the objective of studying the effect of norgestamet-oestradiol and PGF7a treatment on oestrus synchronisation and to enhance the fertility rate in crossbred heifers at fixed time double insemination, forty-five cycling heifers, selected from Kerala Agricultural University Livestock Farms, were randomly allotted to three treatment groups, Group 1,11 and III. Fifteen heifers of Group I were given 3 mg norgestamet implant kept in situ for nine days and an injection containing a combination of 3 mg norgestamet and 5 mg oestradiol valerate on the day of insertion of implant. While the fifteen heifers of Group II were given double dose regimen of 15 mg PGF2CX (luprostiol) at 11 days apart. Group III consisted of fifteen heifers that were kept as control. In both Groups I and II, 14 heifers (93.33 per cent) each responded to the treatments. The time taken for the induction of oestrus in the Group I was 50.1 ± 4.58 hours as against 69.50 ± 1.34 hours in the Group II. There was significant difference on the time of induction of oestrus between Group I and Group II. The duration of oestrus in the Group I was 27.8 ± 1.09 hours, as against 28 ± 1.07 and 19 ± 0.33 hours respectively in Group II and Group III. Though analysis of variance showed no significant difference between Group I and Group 11, pairwise comparison showed no significant difference between Group I and Group II. A higher percentage of heifers in Group I and II showed medium to high intensity of oestrus than Group III. The cervicovaginal mucus examination revealed an increase in typical fern pattern in Group 1 and II than Group III. The first, second and third service pregnancy rate in Group I were 21.42. 28.57 and 14.28 per cent respectively at fixed time artificial insemination, 48 and 72 hours after the removal of the implant, as against 42.86, 21.43 and 14.28 per cent at fixed time artificial insemination, 72 and 96 hour after the second injection or PGF^a in Group 11. In Group III heifers 20 per cent each required one and three inseminations and 13.44 per cent required two inseminations to effect pregnancy. The overall pregnancy rate obtained for Group 1, II and III were 64.27, 78.57 and 53.33 per cent respectively. The AI indices for Group I and II were 1.7_ and 1.63 as against 2.13 of Group III. The present study reveals that though norgestamet oestradiol and PGF.a could be effectively used for synchronisation of oestrus, use of PGF2a is the better alternative for augmenting fertility m crossbred heifers.ThesisItem Open Access EFFECT OF DIFFERENT FREEZING RATES ON CANINE SPERMATOZOA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES Mannuthy - Thrissur, 2000-12) GEETHA, R.; Sreekumaran, TThe obiective of the study was to find out the effect of different freezing rates on post thaw motility, livability and acrosomal damage of dog spermatozoa. A total of 36 ejaculates of good quality collected from six Dachshund dogs by digital manipulation were processed for freezing in Tris citric acid fructose egg yolk diluent containing four per cent glycerol. The processed semen samples were subjected to three different freezing protocols such as 4cm height above the liquid nitrogen level for 10 minutes (Fast freezing), 8cm for 15 minutes (Moderate freezing) and 12cm for 20 minutes (Slow freezing). The mean volume of sperm rich fractions was 0.68±0.03ml. The colour and consistency of sperm rich fractions were thin milky. The mean density of sperm rich fraction was DD(D) and mean pH was 6.63±0.02. The mean concentration of sperm rich fraction was 221 ±7.36 millions per ml and the average initial motility was found to be 75±0.93 per cent. The mean percentage of live sperm count, sperm abnormalities and acrosomal damage of spermatozoa was 81.17±0.73, 5.23±0.29 and 2.32±0.25 respectively. Significant (P<0.05) variation in livability, sperm abnormalities and acrosomal damage of spermatozoa was found between dogs. The average percentage of motility, live sperm count, sperm abnormalities and acrosomal damage of spermatozoa was 70.41±1.22, 75.63±0.65, 7.28±0.43 and 5.34±0.31 after dilution, 58.75±1.34, 63.60±0.89, 10.04±0.32 and 10.13±0.41 after chilling and 47.78±1.59, 50.65±1.31, 11.79±0.36 and 16.20±0.57 after equilibration period respectively. There was significant (P<0.01) reduction in sperm motility and livability and increase in sperm abnormalities and acrosomal damage of spermatozoa after dilution, chilling and equilibration period. Significant (P<0.01) difference was found between dogs for the above parameters. The percentage of post thaw motility of spermatozoa was significantly (P<0.01) higher in fast freezing rate (34.31±1.66) when compared to moderate (25.83±1.66) and slow (24.44±1.27) freezing rates. There was significantly (P<0.01) higher percentage of live sperms and lower percentage of sperm abnormalities in fast freezing rate than in moderate and slow freezing rates. Eventhough the percentage of acrosomal damage was not statistically significant among fast, moderate and slow freezing rates, lower percentage of acrosomal damage was recorded in fast freezing rate. From this study it could be inferred that fast freezing in which the straws were frozen at to 4cm height above the liquid nitrogen level for 10 minutes was superior to moderate (8cm for 15 minutes) and slow (12 cm for 20 minutes) freezing rates.ThesisItem Open Access MORPHOLOGY AND VIABILITY OF BOVINE EMBRYOS FROZEN IN MEDIA CONTAINING BSA AND PROPANEDIOL(COLLEGE OF VETERINARY AND ANIMAL SCIENCES Mannuthy - Thrissur, 2000-10) RAMACHANDRAN, K.; . Suresan Nair, S.PThe observation of the study was to compare the effect of Bovine Serum Albumin (BSA) and 1,2 Propanediol on the morphology and viability of bovine embryos frozen under two freezing and thawing protocols, A total of sixteen crossbred cows, kept under identical conditions, maintained in the Network Project on Embryo Transfer attached to the Department of Animal Reproduction, College of Vetennary and Animal Sciences, Thrissur. The animals were superovulated by Folltropm-V and Prosolvm, starting on the day 11 of the cycle. Of the 16 cows superovxilated 13 showed good response. While two cows did not show any response, there were multiple follicles in both the ovanes without any evidence of ovulation in the third animal. A total of 85 (76 transferable and 9 non transferable) embryos were recovered from a total of 24 flushings from 13 cows, non-surgically on day 7. A total of 56 embryos (mean 3.50 ± 0.822) were recovered m the first treatment, from 13 flushings. In one cow, though, there was 80 per cent flushing, no embryos could be recovered. While 22 embryos (mean 2.75 ± 0.861) were recovered m the second treatment from 8 flushings, only 7 (mean 1.4 ± 0.4) embryos were recovered in the third treatment, from 5 flushings. A total of 72 transferable (60 morulae and 12 blastocysts) were sleeted for the freezing tnals. The embryos were divided into three groups with 24 (20 morulae and 4 blastocysts) embryos and assigned to three media. The first medium was FBS with !0 per cent glycerol, second PBS containing 10 per cent glycerol and 1 per cent BSA; the third medium was with a composition of 10% glycerol and 20% 1,2 propanediol in PBS. Two freezing protocols were used for freezing of the embryos. In the first protocol, with 12 embryos (10 morulae and 2 blastocysts), the initial cooling was at a rate of 1 °C/min from room temperature to -6°C and then at a rate of 0.3°C/min to -35°C, while in the second protocol the initial rate of cooling was at 5°C/mm to -7°C and then at 0.3°C/mm to -30°C before transferring to liquid nitrogen. Thawing was earned out at 37°C for 20 sec after 30 days of preservation. Cryoprotectants were removed by two methods, a four step-wise using decreasing concentrations of cryoprotectants m the first method and one step using 1M sucrose phosphate buffered saline in the second. Thirty four embryos (26 morulae and 8 blastocysts) found normal after freezing and thawing were subjected to culture for 24 h in PBS enriched with 4 per cent BSA at 37°C and 5 per cent CO2 tension. Sixteen embryos (13 morulae and 3 blastocysts) were transferred to 15 recipient cows. While one cow was confirmed pregnant on examination 60 days after transfer, eleven cows returned to heat subsequently, two cows came to oestrus on days 34 and 35 respectively, after the transfer. The third showed oestrum on 45*'' day of transfer. The glucose, acid phosphatase and alkaline phosphatase values showed a normal range of 86.2 to 195.2 mg/100 ml; 14.17 KA to 22.3 KA and 119.02 to 129.00 KA (mean 128.075 ± 9.019, 18.675 ± 0.667 and 122.67 ± 0.788) respectively in the lummal fluid of the recipient animals. The average serum progesterone levels on day 0, 14 and 28 days after oestrus in 11 cows which showed subsequent heat after the transfer were 0.357 ±2.140, 3.053 ± 0.420 and 2.572 ± 0.627 ng/ml and that of the animals which failed to show oestrum were 0.157 ± 0.166, 3.793 ± 0.406 and 3.867 ± 0.362 ng/ml respectively. While significant difference was seen between the freezing media I and II and II and III respectively on the morphology of embryo after the freezing and thawing, no significant differences were seen between the media I and III, between the freezing protocols and cryoprotectant removal procedures on the morphology of embryos frozen. No significant differences were noticed on the effect of the freezing media, freezing protocols and the cryoprotectant removal procedure after the culture on the morphology of the embryos.