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ThesisItem Open Access STUDIES ON ESTRUS SYNCHRONIZATION AND FOLLICULAR DEVELOPMENT PATTERN IN ONGOLE COWS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2012-07) VENKATA RAMANA, K; Sadasiva Rao, K(MAJOR); Chandrasekhar Reddy, K; Raghavender, K.B.P; Gopal Reddy, AABSTRACT: The present investigation was aimed to study the follicular dynamics, serum progesterone concentration during postpartum and effect of GnRH and PGF2α analogues to induce estrus in postpartum anoestrus lactating Ongole cows to reduce the service period. In the ultrasound monitoring of present study in thirteen cows, follicles measuring above 6 mm diameter and corpus luteum measuring 8-10 mm were detected by day 20-25 postpartum period In the present study, the first and second wave of the dominant follicle emerged on day 1.80 + 0.8 and 12.46 + 0.20 days. Where as in three wave cycles first, second and third waves emerged on 1.52 + 0.03, 9.60 + 0.40 and 15.14 + 1.13 days of the estrous cycle, respectively. The mean follicle diameter of the dominant follicles determined by ultrasonography of the ovaries. The first and second wave pattern revealed that 10.23 + 0.40 and 12.30 + 0.36 mm in the two wave cycle. It was observed in the present study that the first wave dominant follicle of three wave cycles attained a maximum size of 10.70+0.20 mm with a mean growth rate of 0.87+0.03 mm per day. In the second wave of the dominant follicle maximum size was 9.80+0.37 mm with growth rate of 0.77+0.06 per day and the third wave maximum diameter was 12.80+0.37 mm with a mean growth rate of 1.46+0.90 mm per day, which is significantly (P< 0.05) bigger and faster in growth rate than the first and second waves in the same estrous cycle. However, the growth rate of second wave and third wave were not significant (P<0.05). In the present study, in natural cycle, the corpus luteum grew to a mean value of maximum diameter of 15.93 + 0.37 and 17.8 + 0.37 mm on the day of 13.1+1.50, 14.6+0.56 and remained up to 14.6+ 1.50 and 15.9+0.45 days of the estrous cycle in two and three wave cycles, respectively. The wave emergence initiated by the GnRH administration was recorded in the recent study was 1.33+0.21 days and the dominant follicle grew to the maximum diameter of the 12.48 + 0.57 mm with a growth rate of 1.66 + 0.26 mm per day. In the present investigation about 22.85 per cent (16/70) of the postpartum lactating Ongole cows have shown estrus by day 60 out of seventy postpartum cows. The postpartum onset of behavioral estrus was highly variable and varied from 25 – 195 days with a mean value of 109.50 + 4.66. Estrus was exhibited by 32.50 and 62.50 per cent cows during the day and night time respectively. In the present study, estrus behaviour was classified as intense, normal and weak. Only 18.75 per cent of cows exhibited intense and 18.75 per cent cows exhibited weak estrus and the remaining (62.50 per cent) cows showed normal estrus. In the present study, the mean estrus duration recorded in Ongole cows was 18.23 + 4.20 hours and the mean estrous cycle length was 21.50+0.21 days. In the present study, 56.25, 57.14 and 100.00 per cent cows required one, two or more artificial inseminations per conception. The mean time required for conception after calving was recorded as 163.60+10.72 (75 -200) days. The serum progesterone concentration increased steadily from day 0 to 10 in cyclic cows and further increased in fertile cows from day 17 to 25 after breeding and there was a sharp decline in progesterone levels between day 17 to 25 (estrus). Among the postpartum anoestrus cows synchronized with GnRH + PGF2α + GnRH (Group A) and PGF2α at 12 days interval (Group B) the reproductive performance of group B was found to be better. The mean conception rate was recorded as 46.15+0.21, 54.54 + 0.36 and 67.00+0.26 in natural estrus , group A and group B, respectively. The conception rate in natural estrus was significantly (P<0.05) lesser than induced groups. The overall mean time interval from parturition to conception in natural postpartum lactating Ongole cows in control (group C) was 163.60+ 10.72 days with a range of 75-200. Where as the same in treatment groups was 84.95+3.79 with a range of 70-117 in group A and 89.40 + 3.89 with a range of 70-115 days in group B. The statistical analysis of the data revealed that the service period in treatment groups was significantly (P<0.01) shorter than control group (Group C). From this study, it may be concluded that, treatment of postpartum cows at day 60 with GnRH and PGF2α enhances the fertility significantly (P<0.01) and reduces the service period.ThesisItem Open Access CLINICAL STUDIES ON EVALUATION OF BREEDING TECHNIQUES IN BITCHES(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) CHANDRA SHEKAR REDDY, K; SUBRAMANYAM NAIDU, K(MAJOR); SADASIVA RAO, K; RAGHAVENDER, K.B.P.; VAIKUNTA RAO, VABSTRACT : The exfoliative vaginal cytology, vaginoscopic examination of vaginal mucosa and endocrine profiles during different phases of estrus cycle in induced as well as in normal estrus cycle were recorded in the present study. Higher percentage of superficial cells (89.94 ± 0.64) and lower percentage of intermediate (7.30 ± 0.77) and parabasal cells (2.76 ± 0.30) were characteristic vaginal cytological changes in bitches during estrus. Vaginoscopic examination revealed that the vaginal mucus was creamy and paper white with angular shrinkage during estrus. The estrus was induced with Cabergoline (CABG) and Pregnant Mare Serum Gonadotropin (PMSG) in two groups of bitches. The progesterone, estrogen and LH levels were recorded before and after the treatment. Progesterone and estradiol levels were significantly lower before the treatment and progressively increased after the treatment. During late proestrus the serum progesterone levels increased progressively towards LH surge and during estrus, progesterone levels were significantly higher on each day of estrus. Significantly higher estradiol levels were noticed during proestrus and these levels were reduced at the time of LH surge. The LH concentration was recorded as 380.58 + 38.73 ng/ml at the time of LH surge. The LH surge was maintained for two days and thereafter significantly reduced during late estrus and early diestrus. The days taken for induction of estrus were 9.20 + 1.32 in Cabergoline followed by 13.83 + 2.65 in PMSG and 70.83 + 12.42 in Control group of bitches. The inter estrus interval in control group of bitches were longer than the Cabergoline and PMSG treated groups. The duration of proestrus was shorter in Cabergoline treated bitches than Control and PMSG group of bitches. There was no significant difference in the duration of estrus among Control, Cabergoline and PMSG groups of bitches. The therapeutic response to the treatment was 83.33 and 100 per cent in Cabergoline and PMSG treated bitches respectively. The conception rates were 66.66, 83.33 and 83.33 per cent in Control (mated based on vaginal cytology), Cabergoline (mated based on vaginoscopy) and PMSG groups (mated based on progesterone level), respectively. The litter size varied from 3.50 + 1.12 to 4.83 + 0.83 in three groups. Vomiting was observed as a side effect in Cabergoline treated group.ThesisItem Open Access STUDIES ON ESTRUS SYNCHRONIZATION IN SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA, 2012-04) MURALI MOHAN, K; SADASIVA RAO, K (Major); RAM CHANDRA REDDY, K; RAGHAVENDER, K. B. P; GOPALA REDDY, A; RAGHUNANDAN, TABSTRACT : A study was undertaken on synchronization of estrus in sheep. The ewes were synchronized with vaginal sponges containing 30 mg of Flurogestone acetate (FGA) and Controlled internal drug releasing (CIDR) device containing 300 mg of progesterone. A total of 240 post partum, parous, healthy ewes aged about 2 to 5 years were selected. Out of 240 ewes, 120 ewes were treated with vaginal sponge another 120 ewes treated with CIDR during breeding and non breeding season. Ewes treated with vaginal sponge and CIDR were divided into 5 groups each group consists of 24 animals. Each group was further subdivided into 2 groups consists of 12 animals and were studied during breeding and non breeding seasons. Group I ewes were considered as untreated control. Ewes in group II were treated with intravaginal sponge/CIDR and left in place for 12 days followed by intramuscular injection of 400 IU of PMSG at the time of sponge removal. Ewes in group III were treated with vaginal sponge/CIDR and 600 IU of PMSG was given intramuscularly at the time of removal of implant. Ewes in group IV were treated as in group II except 200 IU of hCG injection at the time of mating. Ewes in group V were treated as in group III except 200 IU of hCG injection at the time of mating. The haematological and biochemical parameters were studied on day 0, day 6 and day 12 treatment with either vaginal sponge or CIDR during breeding and non breeding seasons. The progesterone profiles were also recorded on day 0, 3. 6, 9 and 12 of the treatment, respectively. The haemoglobin, serum glucose, calcium and phosphorus levels were significantly higher during breeding season compared to non breeding season in ewes treated either with vaginal sponge or CIDR. In vaginal sponge group, the progesterone levels before insertion were 1.23±0.12 ng/ml during breeding season. The progesterone levels were 2.46±0.11, 3.40±0.13 and 4.71±0.14 ng/ml on day 3, 6 and 9 of treatment, respectively in breeding season. In non breeding season, the progesterone levels were 0.96±0.12 ng/ml before insertion of vaginal sponges. During treatment, the progesterone levels were 1.62±0.13 ng/ml on day 3, 2.32±0.11 ng/ml on day 6, 3.35 ±0.14 ng/ml on day 9 and 3.19±0.12 ng/ml on day 12 of treatment (at the time of removal). The progesterone levels were significantly (P<0.01) increased from day 0 to day 9 of treatment and thereafter it was significantly (P<0.01) decreased on day 12 of treatment. Significantly, higher progesterone levels were recorded in all groups of ewes inserted with vaginal sponges (2.76±0.06 to 2.84±0.07 ng/ml) compared to control group of ewes (1.67±0.08 ng/ml). In CIDR treatment ewes, the progesterone levels were 1.44±0.13, 2.64±0.11, 3.63±0.10, 5.11±0.16 and 3.34±0.12 ng/ml on day 0, 3, 6, 9 and 12 of treatment, respectively during breeding season. While in non breeding season, the corresponding values were 0.84±0.11, 1.84±0.14, 2.33±0.15, 3.01±0.18 and 2.55±0.10 ng/ml on day 0, 3, 6, 9 and 12, respectively. The progesterone levels were significantly (P<0.01) increased from day 0 to day 9 of treatment and thereafter decreased on day 12 of treatment during breeding and non breeding seasons. When compared the vaginal devices with CIDR, the progesterone levels were significantly (P<0.01) higher in CIDR treatment than the vaginal sponges treatments in the present study. The percentage of ewes responded for synchronization of estrus was 50.00, 83.33, 100.00, 91.67 and 100.00 in control, VS 4, VS 6, VS 4h and VS 6h, respectively in breeding season. In non breeding season, the estrus response was 25.00, 83.33, 100.00 83.33 and 100.00 per cent in control, VS 4, VS 6, VS 4h and VS 6h, respectively. Likewise the ewes treated with CIDR, the estrus response rates were 50.00 Vs. 16.67; 83.33 Vs. 83.33; 100.00 Vs. 100.00; 91.67 Vs. 83.33 and 100.00 Vs. 100.00 per cent in control, CIDR 4, CIDR 6, CIDR 4h and CIDR 6h in breeding Vs. non breeding seasons, respectively. The estrus response was significantly (P<0.01) higher in treated ewes than that of control groups in both treatments and both seasons. Estrus response were significantly (P<0.01) higher in VS6/CIDR6 and VS6h/CIDR6h groups of ewes followed by VS4/CIDR4, VS4h/CIDR4h and control groups of ewes in ewes synchronized with vaginal sponge and CIDR. The estrus response was significantly (P<0.01) differed during non breeding season in both vaginal sponge and CIDR synchronization protocol. The time taken for induction of estrus in vaginal sponges group was 46.58±2.28, 41.24±1.56, 46.95±2.66 and 40.62±2.73 h during breeding season and 48.01±2.81, 46.66±3.77, 48.55±3.03, and 46.69±3.89 h during non breeding season in control, VS 4, VS 6, VS 4h and VS 6h, respectively. Among the CIDR group of ewes, the time taken for induction of estrus was 35.56±2.36 Vs. 38.53±3.21 h in CIDR4, 34.78±2.32 Vs. 36.82±2.14 h in CIDR6, 35.63±2.84 Vs. 38.77±2.86 h in CIDR 4h and 34.80±2.21 Vs. 36.27±1.85 h in CIDR 6h group of ewes during breeding Vs. non breeding season, respectively. The time taken for induction of estrus was significantly (P<0.01) long in the vaginal sponge treatment than the CIDR treatment in the present study.ThesisItem Open Access STUDIES ON EVALUATION OF BREEDING SOUNDNESS AND AUGMENTING THE FERTILITY OF LOW GRADE EJACULATES IN ONGOLE BULLS (Bos indicus)(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2010-05) SRINIVAS, MANDA; SUBRAHMANYAM NAIDU, K (Major); BABU RAO, K; SREENU, MAKKENA; SRILATHA, ChABSTRACT: The present investigation “Studies on Evaluation of Breeding Soundness and Augmenting the Fertility of Low Grade Ejaculates in Ongole Bulls (Bos indicus)” was undertaken to study the biometry of testes, accessory sex glands and evaluate fresh, post dilution and post thaw semen of various grades and compare their fertility rates. In the present study, the overall mean body condition score and scrotal circumference were 6.63 ± 0.18 and 34.63 ± 0.21 cm. All the testicular measurements by vernier showed significant positive correlations with ultrasonographic measurements of the testes, accessory sex glands, ejaculate volume and live sperm percent (P<0.05). Seminal vesicle dimensions revealed significant positive correlation with ejaculate volume (P<0.01) and live sperm percent (P<0.05). Ampulla showed a significant positive correlation with dimensions of prostrate body. A significant difference (P<0.01) was noticed in the mean mass activity between normal ejaculates (2.74 ± 0.05) and low grade ejaculates (1.40 ± 0.55). The sperm concentration had a highly significant positive correlation with total sperm per ejaculate (P<0.01). The live sperm percent in fresh normal and low grade ejaculates was 80.20 ± 0.59 and 53.90 ± 0.38, respectively The overall mean abnormal sperm count was 14.73 ± 2.19 and 29.58 ± 0.43 percent in fresh semen of normal and low grade quality. The abnormal sperm count (%) showed significant negative correlation and significant positive correlation with HPAP sperm percent and HNAN sperm percent respectively (P<0.05). The overall means of HOS-G sperm was 56.50 ± 0.50 and 30.35 ± 0.66 percent for HPAP, 17.30 ± 0.54 and 19.48 ± 0.66 percent for HPAN in fresh semen of normal and low grade ejaculates, respectively. The overall mean sperm penetration distance was 19.75 ± 0.19 and 14.40 ± 0.19 mm/20 min in normal and low grade ejaculates. Sperm penetration distance of normal fresh semen had a significant positive correlation with fertility (P<0.05), while it had a non significant negative correlation in low grade ejaculates. The overall mean post dilution sperm motility was 68.11 ± 0.89, 48.26 ± 0.34 and 67.66 ± 0.50 percent in the normal, low grade unfiltered and filtered semen respectively . Post dilution sperm concentration of low grade unfiltered and filtered semen had a significant negative correlation with post diluted HPAN sperm percent (P<0.05). Live sperm percent of diluted low grade unfiltered and filtered semen had a significant negative correlation with post diluted HPAN sperm percent (P<0.05) while, the post diluted abnormal sperm percent had a significant negative correlation with post diluted sperm penetration distance (P<0.05). Post dilution abnormal sperm in normal semen had a significant negative correlation with fertility (P<0.05). Post dilution HPAP sperm percent of low grade filtered semen had a significant positive correlation with post dilution sperm penetration distance (P<0.05). Post thaw sperm motility of low grade filtered semen had a significant positive correlation with live sperm percent by fluorogenic assessment, high mitochondrial membrane potential and sperm nuclear morphology (P<0.05). Post thaw live sperm percent of low grade filtered semen had a significant negative correlation with post thaw abnormal sperm percent (P<0.05) and a highly significant positive correlation with high mitochondrial membrane potential (P<0.01). The overall mean post thaw abnormal sperm count in the normal, low grade unfiltered and filtered semen were 22.10 ± 0.36, 35.56 ± 0.50 and 18.10 ± 0.34 percent respectively. Post thaw HPAP sperm percent of normal semen had a significant negative correlation with post thaw HNAN sperm percent and percent linearity assessed by Sperm Class Analyzer (P<0.05). Post thaw HPAP sperm percent of low grade filtered semen had a highly significant negative correlation with post thaw HNAN sperm percent (P<0.01) and a significant negative correlation with percent linearity (P<0.05). Post thaw sperm penetration distance of low grade filtered semen had a significant positive correlation with high mitochondrial membrane potential (P<0.05). The overall mean post thaw progressive motility percent estimated by Sperm Class Analyzer (SCA) was 33.35 ± 0.70, 17.99 ± 0.56 and 28.79 ± 0.72 in the normal, low grade unfiltered and filtered semen respectively. The overall mean post thaw curvilinear velocity (Fm/sec) was 98.54 ± 1.60, 88.43 ± 1.70 and 90.79 ± 1.67 in the normal, low grade unfiltered and filtered semen respectively. The post thaw linearity percent of normal and low grade filtered semen had a non significant negative correlation with fertility; while low grade unfiltered semen had a non significant positive correlation with fertility. The overall mean post thaw straightness percent was 74.52 ± 1.31, 72.50 ± 1.47 and 74.28 ± 1.27 in the normal, low grade unfiltered and filtered semen respectively. In low grade filtered semen the functional membrane as well as acrosomal integrities revealed a significant positive correlation with high mitochondrial membrane potential and sperm nuclear morphology (P<0.05). In the low grade filtered semen the parameter showed a significant positive correlation with sperm nuclear morphology (P<0.05). The overall mean post thaw sperm nuclear morphology was 77.56 ± 0.81, 55.69 ± 0.68 and 73.95 ± 0.83 percent in the normal, low grade unfiltered and filtered semen respectively. The overall mean malondialdehyde (MDA) concentration of post thaw semen was 0.61 ± 0.01, 1.12 ± 0.01 and 0.69 ± 0.01 Fmol/ml in the normal, low grade unfiltered and filtered samples respectively. The overall mean fertility rate post insemination was 66.88 ± 4.22, 20.00 ± 1.89 and 60.63 ± 4.27 with frozen thawed semen of normal, low grade unfiltered and filtered ejaculates, respectively. It is concluded from the present study that the fertility rates of normal and low grade filtered semen differed non significantly suggesting that filtration has considerably improved the quality of low grade ejaculates; hence the filtration procedure could be used practically to improve low grade ejaculates from superior bulls to obtain optimum fertility.