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  • ThesisItemOpen Access
    SEASONAL INFLUENCE ON SEXUAL BEHAVIOUR AND PROTEIN PROFILING OF MAJOR HSP (HSP 70, 90) IN RELATION TO SEMEN QUALITY AND FERTILITY OF GIR BULL 2893
    (JAU, JUNAGADH, 2019-07) SOLANKI GAJENDRASINGH BHAWARSINGH; F. S. KAVANI
    For successful AI in cattle and buffalo, bull fertility is essential to assure optimal reproductive efficiency. Routinely used traditional semen quality tests and even CASA provide limited information about the potential fertility of bulls. Several studies have been conducted in the past decades in the search of exact and potential fertility markers in bulls. So determining the seminal proteomic pattern of bulls with high semen freezability and low semen freezability, we can use this technique as a prediction tool to choose the semen donor bulls. Studies on sexual behaviour, semen characteristics, fertility and potential biomarkers of fertility, viz., heat shock proteins (HSP 70, 90) in semen as such, and the documented reports on the influence of seasons on these aspects are meager in Gir bulls, the world famous milch purpose cattle breed of India. These indices could be used in selection and culling of Gir bulls in the breeding program. Hence these investigations were undertaken at Cattle Breeding Farm, JAU, Junagadh during two seasons (summer and winter) from Nov 2017 to Oct 2018. The study was carried out to know the seasonal variation and correlations amongst and between the sexual behaviour, semen quality traits, conception rates and major heat shock proteins expression in six mature Gir bulls. The effect of season on individual bulls for these parameters was also studied. A total of 96 observations were availed at fortnightly interval for sexual behaviour study, while 384 semen samples at twice-weekly interval during each season from six bulls were utilized. The first AI conception rate was found by inseminating a total of 1200 Gir cows, equally dividing for each bull and season. The protein profiling of major HSP proteins (70, 90) in sperm from neat as well as post-thawed semen was carried out. The mean temperament (0-5 scale), libido (0-9 scale), penile erection (0-4 scale), protrusion (0-4 scale), intensity of thrust (0-4 scale), reaction time (sec.) and Flehmen reaction (0, 1) during summer were 1.77±0.08, 5.23±0.22, 2.73±0.07, 2.68±0.07, 3.19±0.11, 153.54±4.82 and 0.46±0.07, respectively, while the respective values in winter season were 2.08±0.06, 5.96±0.22, 3.02±0.06, 2.96±0.43, 3.40±0.39, 128.09±4.24 and 0.56±0.07. There were significant differences among bulls for these traits in both the seasons. However, in general, summer significantly exerted an adverse effect on the sexual behaviour of Gir bulls, while winter was the favoured season.Abstract ii The neat seminal characteristics of Gir bulls, viz., volume (ml), colour (score), concentration (million/ml), mass activity (score), initial motility (%), live sperm (%), abnormal sperm (%), HOST reactive sperm (%) and acrosomal integrity (%) observed in summer season were 5.02±0.27, 3.06±0.11, 1.070.46±49.56, 3.83±0.05, 79.58±0.05, 87.23±0.75, 6.73±0.47, 76.02±0.57, 90.35±0.35, respectively, whereas the respective values in winter were 5.81±0.26, 3.35±0.11, 1282.98±58.08, 3.94±0.25, 82.40±0.56, 90.17±0.71, 5.13±0.51, 78.23±0.49, 92.42±0.31. The percentages of post-thaw motility, post-thaw live sperm, post-thaw abnormal sperm, post-thaw HOST reactive sperm, post-thaw acrosomal integrity and first AI conception rate for semen cryopreserved during summer were 53.19±0.32, 65.90±0.92, 9.94±0.57, 55.27±0.82, 74.38±0.38 and 49.33±0.32, respectively, whereas the corresponding values for semen cryopreserved during winter were 55.29±0.39, 69.06±1.09, 8.19±0.54, 58.58±0.86, 75.17±0.35 and 55.04±0.35. There were significant differences among bulls for all of these traits in both the seasons. Despite the fact that the sperm motility, concentration, viability, abnormality, membrane integrity and acrosomal integrity were within permissible limit for semen processing throughout the year, the quality was significantly better during the winter than the summer season under Junagadh Agro-climatic conditions. Temperament, libido, penile erection, penile protrusion, and intensity of thrust were significantly and positively correlated among each other, and negatively with reaction time and Flehmen reaction. Sperm motility traits like mass activity, initial motility and post-thaw motility showed significant positive correlations among themselves (r=0.562 to 0.689) and with first AI conception rates (r=0.133 to 0.357). Sperm viability was positively correlated with post-thaw motility (r=0.164) and conception rate (r=0.266). Membrane integrity (r=0.188) and acrosomal integrity (0.373) were positively correlated with the conception rate. Reaction time was significantly negatively correlated with mass activity (r=-0.203) and initial motility (r=-0.181). Conception rate was significantly positively correlated with temperament (r=0.261), penile erection (r=0.253), penile protrusion (r= 0.277) and thrust (r=0.324), while negatively with reaction time (r=-0.293). The reaction time was also negatively correlated with initial live sperm, acrosomal integrity and post-thaw motility. The mean protein concentrations (ng/mg of protein) of major heat shock protein 70 (HSP70) were significantly higher during summer as compared to winter season both at a pre-freeze ((56.68 ±43.19 vs. 40.25±3.07) and post-thaw stage (118.24±6.44 vs. 80.81±5.11). While the concentration (ng/mg of protein) of HSP90P at the pre freeze stage was non-significantly higher during summer than winter (1.27±0.05 vs. 1.19±0.07) season, while the respective values at the post-freeze stage were 2.36±0.08 and 2.37±0.13. This was also confirmed qualitatively by western blotting. The HSP70P in neat semen had a significant positive correlation with mass activity (r=0.326), initial motility (r=0.463), sperm viability (0.565), acrosomal integrity (r=0.330), first AI conception rate (r=0.431) and HSP70P in post-thaw semen (r=0.474). The HSP90P had a significant positive correlation with sperm motility (r=0.322), viability (r=0.301) and conception rate (r=0.409). It could be concluded from the present study that the Gir bulls had good sexual behaviour, better semen quality and quantity throughout the year. However, semen quality was better during winter as compared to the summer season. The protein expression of major HSPs (70, 90) may be utilized as markers for heat tolerance, sexual behaviour, semen quality and conception rate for the Gir bull.
  • ThesisItemOpen Access
    CLINICAL ASSESSMENT OF NUTRITIONAL SUPPLEMENT AND DOUBLE OVSYNCH PROTOCOL FOR ESTRUS INDUCTION, SYNCHRONIZATION AND PREGNANCY RATE IN ANESTRUS JAFFRABADI BUFFALOES 2855
    (jau,junagdh, 2019-06) Raval Rupeshkumar J.; Dr. F.S. Kavani
    The effect of nutritional supplementation on estrus induction and fertility/pregnancy rate; and efficacy of double Ovsynch protocol for induction and synchronization of estrus/ovulation and fertility/pregnancy rate were assessed in Jaffrabadi buffaloes. Fourty eight buffaloes equally divided into 4 groups were utilized for this study. Group I included pubertal heifers (mean age 30±2 months) supplemented with fixed quantity of chelated mineral mixture, bypass fat and protein over the routine farm feeding practices; and body weight gain, ovarian dynamics, blood biochemical and endocrine profiles during six months were recorded at month interval. Group II consisted of post-pubertal acyclic heifers (age 42±2 months), Group III comprised of post-pubertal acyclic heifers (age 48±2 months) and Group IV covered post-partum acyclic buffaloes. Animals of all the four groups were randomly subdivided into two equal subgroups (treatment and control) of 6 each. Animals of treatment subgroup of group II, III and IV were treated with established double Ovsynch protocol followed by fixed time artificial insemination (FTAI) on day 26 and 27 using Jaffrabadi bull semen. Animals were monitored for ovarian changes by trans-rectal ultrasonography, biochemical and hormonal profiles on the day of injections and day 27 (FTAI), and on day 12, 21 and 35 post-FTAI; and at weekly interval up to five weeks in control subgroups. Ultrasonography and per-rectal examination in inseminated animals were used to confirm the pregnancy on day 35 and 70, respectively. Data collected were compared between and within treatment and control subgroups of II, III and IV groups. In group I animals, the overall difference in plasma total protein level and plasma total cholesterol concentration was significant (p<0.05) between subgroups. Ovarian dynamics revealed significant (p<0.05) increase in ovarian follicles especially in right ovary in treatment subgroup as compared to control. FSH and LH levels were significantly (p<0.01) higher in animals of treatment subgroup as compared to control from 3rd to 6th month and at 4th month, respectively. Insulin level in treatment subgroup was significantly (p<0.01) higher as compared to control. None of these animals exhibited behavioural signs of estrus. However, monthly per-rectal palpation and ultrasonography revealed establishment of cyclicity with the presence ii of developed follicles/corpus luteum during the last three months of study period. Therefore, none of these animals was bred or used for further study. In animals of treatment subgroup II and III a significant (p<0.05) increase in numbers of small follicles was observed on day 26. Number of large sized follicles was significantly (p<0.05) higher on day 26 in comparison to day 0 and 7 in group IV animals. Total number of follicles in both ovaries of group II animals was significantly (p<0.01) higher on day 26. Large and subordinate follicular diameter on ovaries of group II, III and IV animals increased gradually, but the differences between periods were significant (p<0.001) only in group IV with highest recorded diameter of large follicle on day 17 ; and of subordinate follicle on day 26. The right ovary was found relatively more active as compared to left ovary. Overall total number of follicles, diameters of large and subordinate follicles were significantly (p<0.05) higher in control as compared to treatment subgroup of group II animals. The conception rates at FTAI in group II, III and IV was 66.66, 83.33 and 16.66 %, respectively; whereas the corresponding values in control sub-groups were 0.00, 16.66 and 16.66 %, respectively. The differences in conception rate in treated and control heifers were significant (p<0.05), however, benefit of double Ovsynch treatment over control group in multiparous buffaloes was not observed. Plasma FSH concentration values differed significantly (p<0.001) among different groups at all-time intervals. Plasma FSH concentration obtained in control subgroup of group IV was significantly (p<0.001) higher than group II and III. Plasma LH concentration values differed significantly (p<0.001) among groups at various time periods. Plasma LH concentrations in group II and IV differed significantly (p<0.001). Plasma estrogen concentration obtained in animals of group III was significantly (p<0.05) higher than group IV. Plasma estrogen concentration obtained in control subgroups II and III were significantly (p<0.01) higher than control subgroup IV. Plasma insulin value obtained on all days for group IV animals were significantly (p<0.01) lower than group II and III animals. In group II, there was significantly (p<0.001) increased level of plasma FSH in treatment subgroup as compared to control subgroup; whereas, LH level was significantly (p<0.05) higher in control subgroup as compared to treatment subgroup. In group III, plasma FSH and LH levels were significantly (p<0.001) higher in treatment subgroup as compared to control subgroup. In group IV, there was significantly (p<0.01) increased level of FSH in treatment subgroup as compared to control; whereas, plasma LH level was significantly higher (p<0.05) in control subgroup as compared to treatment subgroup. The period-wise mean estrogen and insulin levels were significantly (p<0.05) higher in treatment subgroup as compared to control subgroup. Blood glucose level was significantly (p<0.001) higher in group II on day 17 as compared to group III and IV. Plasma total cholesterol values were significantly (p<0.05) higher in treatment subgroup IV as compared to subgroup II and III. Plasma total protein values were significantly (p<0.001) lower in group II heifers than in post-partum buffaloes of group IV, and group III heifers being intermediate. In group III, blood glucose level was significantly (p<0.05) lower in treatment subgroup as compared to control subgroup. On the basis of ovarian dynamics and results, it is concluded that double Ovsynch protocol could be better choice for improving conception rate in post-pubertal acyclic Jaffrabadi buffalo heifers as compared to multiparous acyclic
  • ThesisItemOpen Access
    SEASONAL INFLUENCE ON SEXUAL BEHAVIOUR AND PROTEIN PROFILING OF MAJOR HSPS (HSP 70, 90) IN RELATION TO SEMEN QUALITY AND FERTILITY OF JAFFARABADI BULLS
    (jau.junagadh, 2019-08) PARMAR KIRANKUMAR H.; Dr. F. S. KAVANI
    For successful Artificial Insemination (AI) in buffaloes, quality of semen, superior germplasm, congenial agro-climatic conditions and bull fertility are of vital importance to the bovine industry. The thermotolerance at the cellular levels is mediated by family of heat shock proteins (HSPs) that include several isoforms, viz., HSP70 and HSP90. These proteins are the important molecular chaperone that can protect cells from thermal damage, apoptosis and help in maintaining the intracellular homeostasis. In view of meager literature on these aspects in Jaffarabadi the world famous buffalo breed, the present investigation was undertaken at Cattle Breeding Farm, JAU, Junagadh during two seasons (summer and winter) from Nov 2017 to Oct 2018. The study was carried out to know the seasonal variation and correlations amongst and between the sexual behaviour, semen quality traits, conception rates and major heat shock proteins expression in four mature Jaffarabadi bulls. A total of 64 observations were availed at fortnightly interval for sexual behaviour study, while 192 semen samples at thrice-fortnightly interval during each season were utilized for the study. The first AI conception rate was calculated by inseminating a total of 800 Jaffarabadi buffaloes, equally dividing for each bull and season. The protein profiling of major HSP proteins (70, 90) in sperm from neat as well as post-thawed semen was also carried out. The mean values/scores for temperament (0-5 scale), libido (0-9 scale), penile erection (0-4 scale), protrusion (0-4 scale), intensity of thrust (0-4 scale), reaction time (sec.) and Flehmen reaction (0, 1) during winter were 1.28±0.08, 6.75±0.19, 3.22±0.74, 2.88±0.59, 3.15±0.65, 125.00±6.92 and 0.91±0.43, respectively, while the respective values in summer season were 2.22±0.07, 5.93±0.43, 2.84±0.65, 2.75±0.59, 2.93±0.13, 143.5±7.07 and 0.88±0.06. There were significant differences among season for temperament, libido, and penile erection. The neat seminal characteristics of Jaffarabadi bulls, viz., ejaculate volume (ml), colour/density (score), sperm concentration (million/ml), mass activity (score), initial motility (%), live sperm (%), abnormal sperm (%), HOST reactive sperm (%) and acrosomal integrity (%) observed during winter season were 5.19±0.18, 2.38±0.10, 1253.36±24.75, 3.73±0.05, 80.31±0.05, 86.20±0.64, 5.00±0.40, 85.75 Abstract ii ±0.43, 93.56±0.56, respectively, whereas the respective values in summer were 5.07±0.13, 2.00±0.00, 1186.60±24.78, 3.72±0.11, 78.75±0.46, 82.31±0.74, 7.13±0.52, 84.93±0.45, 90.56±0.23. The mean percentages of post-thaw motility, live sperm, abnormal sperm, HOST reactive sperm, acrosomal integrity and first AI conception rate for semen cryopreserved during winter were 58.85±0.56, 67.06±0.77, 8.62±0.43, 58.52±0.67, 75.22±0.27 and 45.75±0.21, respectively, whereas, corresponding values for semen cryopreserved during summer were 56.35±0.43, 65.62±0.76, 9.08±0.51, 55.43±0.60, 74.29±0.22 and 43.50±0.11. The temperament was significantly (P<0.05) and positively correlated with penile erection (r=383), penile protrusion score (r=0.352), intensity of thrust (r=0.378), and reaction time (r=0.358). Libido score had significant (P<0.05) positive correlations with penile erection, penile protrusion score and intensity of thrust. Penile erection score had a highly significant (P<0.01) correlations with penile protrusion (r=0.512), thrust(r=0.378), and Flehmen reaction (r= -0.362). Protrusion score showed a highly significant (P<0.01) positive correlation with thrust (r=0.419). The seminal traits like semen volume and concentration showed highly significant positive correlations with post-thaw sperm motility (r=0.249; r=0.409, respectively). Sperm motility traits like mass activity (r=0.463), initial motility (r=0.508) and post thaw motility showed the significant positive correlations among themselves. Sperm viability was highly significant positively correlated with post-thaw motility (r=0.367) and conception rate (0.466). Acrosome integrity (r=0.456) and sperm plasma membrane integrity (r=0.403) were significantly and positively correlated with conception rate. While, post-thaw motility (PTM) had highly significant (P<0.01) positive correlation with first AI conception rate (r=0.475). Temperament, libido, and erection score didn’t show significant correlation with other semen characteristics. Reaction time was significantly and negatively correlated with mass activity (r=-0.360), initial motility (r=-0.391) and conception rate (r=-0.393). The mean concentration (ng/mg of protein) of major heat shock protein 70 (HSP70) was non-significantly higher during summer as compared to winter season at a pre-freeze (146.50±8.48 vs. 138.10±4.41) and significantly higher at post-thaw stage (345.90±15.52 vs. 280.90±14.74), while the concentration (ng/mg of protein) of HSP90P was non-significantly higher during summer than winter both at the pre-freeze (2.49±0.43 vs. 1.63±0.31) and at the post-freeze stage (7.13±0.73 vs. 5.50±0.74). This was also confirmed qualitatively by western blotting. The HSP70P in neat semen had a significant positive correlations with mass activity (r=0.397), sperm viability (r=0.478), acrosomal integrity (r=0.373), while in post-thaw semen it had highly significant correlation with first AI conception rate (r=0.477). The HSP90P had a significant positive correlation with sperm motility (r=0.372) in neat semen, while in post-thaw it had highly significant correlation with conception rate (r=593) and it was significantly correlated with HSP90 in neat semen(r=0.398). From the present study, it could be concluded that the Jaffarabadi bulls had good sexual behaviour, better semen quality and quantity throughout the year. However, semen quality was better during winter as compared to the summer season. The protein expression of major HSPs (70, 90) may be utilized as markers for heat tolerance, sexual behaviour, semen quality and conception rate for the Jaffarabadi bulls.
  • ThesisItemOpen Access
    SEASONAL INFLUENCE ON SEXUAL BEHAVIOUR AND PROTEIN PROFILING OF MAJOR HSP (HSP 70, 90) IN RELATION TO SEMEN QUALITY AND FERTILITY OF GIR BULL 2893
    (JAU, JUNAGADH, 2019-07) SOLANKI GAJENDRASINGH BHAWARSINGH; F. S. KAVANI
    For successful AI in cattle and buffalo, bull fertility is essential to assure optimal reproductive efficiency. Routinely used traditional semen quality tests and even CASA provide limited information about the potential fertility of bulls. Several studies have been conducted in the past decades in the search of exact and potential fertility markers in bulls. So determining the seminal proteomic pattern of bulls with high semen freezability and low semen freezability, we can use this technique as a prediction tool to choose the semen donor bulls. Studies on sexual behaviour, semen characteristics, fertility and potential biomarkers of fertility, viz., heat shock proteins (HSP 70, 90) in semen as such, and the documented reports on the influence of seasons on these aspects are meager in Gir bulls, the world famous milch purpose cattle breed of India. These indices could be used in selection and culling of Gir bulls in the breeding program. Hence these investigations were undertaken at Cattle Breeding Farm, JAU, Junagadh during two seasons (summer and winter) from Nov 2017 to Oct 2018. The study was carried out to know the seasonal variation and correlations amongst and between the sexual behaviour, semen quality traits, conception rates and major heat shock proteins expression in six mature Gir bulls. The effect of season on individual bulls for these parameters was also studied. A total of 96 observations were availed at fortnightly interval for sexual behaviour study, while 384 semen samples at twice-weekly interval during each season from six bulls were utilized. The first AI conception rate was found by inseminating a total of 1200 Gir cows, equally dividing for each bull and season. The protein profiling of major HSP proteins (70, 90) in sperm from neat as well as post-thawed semen was carried out. The mean temperament (0-5 scale), libido (0-9 scale), penile erection (0-4 scale), protrusion (0-4 scale), intensity of thrust (0-4 scale), reaction time (sec.) and Flehmen reaction (0, 1) during summer were 1.77±0.08, 5.23±0.22, 2.73±0.07, 2.68±0.07, 3.19±0.11, 153.54±4.82 and 0.46±0.07, respectively, while the respective values in winter season were 2.08±0.06, 5.96±0.22, 3.02±0.06, 2.96±0.43, 3.40±0.39, 128.09±4.24 and 0.56±0.07. There were significant differences among bulls for these traits in both the seasons. However, in general, summer significantly exerted an adverse effect on the sexual behaviour of Gir bulls, while winter was the favoured season.Abstract ii The neat seminal characteristics of Gir bulls, viz., volume (ml), colour (score), concentration (million/ml), mass activity (score), initial motility (%), live sperm (%), abnormal sperm (%), HOST reactive sperm (%) and acrosomal integrity (%) observed in summer season were 5.02±0.27, 3.06±0.11, 1.070.46±49.56, 3.83±0.05, 79.58±0.05, 87.23±0.75, 6.73±0.47, 76.02±0.57, 90.35±0.35, respectively, whereas the respective values in winter were 5.81±0.26, 3.35±0.11, 1282.98±58.08, 3.94±0.25, 82.40±0.56, 90.17±0.71, 5.13±0.51, 78.23±0.49, 92.42±0.31. The percentages of post-thaw motility, post-thaw live sperm, post-thaw abnormal sperm, post-thaw HOST reactive sperm, post-thaw acrosomal integrity and first AI conception rate for semen cryopreserved during summer were 53.19±0.32, 65.90±0.92, 9.94±0.57, 55.27±0.82, 74.38±0.38 and 49.33±0.32, respectively, whereas the corresponding values for semen cryopreserved during winter were 55.29±0.39, 69.06±1.09, 8.19±0.54, 58.58±0.86, 75.17±0.35 and 55.04±0.35. There were significant differences among bulls for all of these traits in both the seasons. Despite the fact that the sperm motility, concentration, viability, abnormality, membrane integrity and acrosomal integrity were within permissible limit for semen processing throughout the year, the quality was significantly better during the winter than the summer season under Junagadh Agro-climatic conditions. Temperament, libido, penile erection, penile protrusion, and intensity of thrust were significantly and positively correlated among each other, and negatively with reaction time and Flehmen reaction. Sperm motility traits like mass activity, initial motility and post-thaw motility showed significant positive correlations among themselves (r=0.562 to 0.689) and with first AI conception rates (r=0.133 to 0.357). Sperm viability was positively correlated with post-thaw motility (r=0.164) and conception rate (r=0.266). Membrane integrity (r=0.188) and acrosomal integrity (0.373) were positively correlated with the conception rate. Reaction time was significantly negatively correlated with mass activity (r=-0.203) and initial motility (r=-0.181). Conception rate was significantly positively correlated with temperament (r=0.261), penile erection (r=0.253), penile protrusion (r= 0.277) and thrust (r=0.324), while negatively with reaction time (r=-0.293). The reaction time was also negatively correlated with initial live sperm, acrosomal integrity and post-thaw motility. The mean protein concentrations (ng/mg of protein) of major heat shock protein 70 (HSP70) were significantly higher during summer as compared to winter season both at a pre-freeze ((56.68 ±43.19 vs. 40.25±3.07) and post-thaw stage (118.24±6.44 vs. 80.81±5.11). While the concentration (ng/mg of protein) of HSP90P at the pre freeze stage was non-significantly higher during summer than winter (1.27±0.05 vs. 1.19±0.07) season, while the respective values at the post-freeze stage were 2.36±0.08 and 2.37±0.13. This was also confirmed qualitatively by western blotting. The HSP70P in neat semen had a significant positive correlation with mass activity (r=0.326), initial motility (r=0.463), sperm viability (0.565), acrosomal integrity (r=0.330), first AI conception rate (r=0.431) and HSP70P in post-thaw semen (r=0.474). The HSP90P had a significant positive correlation with sperm motility (r=0.322), viability (r=0.301) and conception rate (r=0.409). It could be concluded from the present study that the Gir bulls had good sexual behaviour, better semen quality and quantity throughout the year. However, semen quality was better during winter as compared to the summer season. The protein expression of major HSPs (70, 90) may be utilized as markers for heat tolerance, sexual behaviour, semen quality and conception rate for the Gir bull.
  • ThesisItemOpen Access
    CLINICAL ASSESSMENT OF NUTRITIONAL SUPPLEMENT AND DOUBLE OVSYNCH PROTOCOL FOR ESTRUS INDUCTION, SYNCHRONIZATION AND PREGNANCY RATE IN ANESTRUS JAFFRABADI BUFFALOES By
    (JAU, JUNAGADH, 2019-07) RAVAL RUPESHKUMAR JAGDISHCHANDRA; Dr. F.S. Kavani
    The effect of nutritional supplementation on estrus induction and fertility/pregnancy rate; and efficacy of double Ovsynch protocol for induction and synchronization of estrus/ovulation and fertility/pregnancy rate were assessed in Jaffrabadi buffaloes. Fourty eight buffaloes equally divided into 4 groups were utilized for this study. Group I included pubertal heifers (mean age 30±2 months) supplemented with fixed quantity of chelated mineral mixture, bypass fat and protein over the routine farm feeding practices; and body weight gain, ovarian dynamics, blood biochemical and endocrine profiles during six months were recorded at month interval. Group II consisted of post-pubertal acyclic heifers (age 42±2 months), Group III comprised of post-pubertal acyclic heifers (age 48±2 months) and Group IV covered post-partum acyclic buffaloes. Animals of all the four groups were randomly subdivided into two equal subgroups (treatment and control) of 6 each. Animals of treatment subgroup of group II, III and IV were treated with established double Ovsynch protocol followed by fixed time artificial insemination (FTAI) on day 26 and 27 using Jaffrabadi bull semen. Animals were monitored for ovarian changes by trans-rectal ultrasonography, biochemical and hormonal profiles on the day of injections and day 27 (FTAI), and on day 12, 21 and 35 post-FTAI; and at weekly interval up to five weeks in control subgroups. Ultrasonography and per-rectal examination in inseminated animals were used to confirm the pregnancy on day 35 and 70, respectively. Data collected were compared between and within treatment and control subgroups of II, III and IV groups. In group I animals, the overall difference in plasma total protein level and plasma total cholesterol concentration was significant (p<0.05) between subgroups. Ovarian dynamics revealed significant (p<0.05) increase in ovarian follicles especially in right ovary in treatment subgroup as compared to control. FSH and LH levels were significantly (p<0.01) higher in animals of treatment subgroup as compared to control from 3rd to 6th month and at 4th month, respectively. Insulin level in treatment subgroup was significantly (p<0.01) higher as compared to control. None of these animals exhibited behavioural signs of estrus. However, monthly per-rectal palpation and ultrasonography revealed establishment of cyclicity with the presence ii of developed follicles/corpus luteum during the last three months of study period. Therefore, none of these animals was bred or used for further study. In animals of treatment subgroup II and III a significant (p<0.05) increase in numbers of small follicles was observed on day 26. Number of large sized follicles was significantly (p<0.05) higher on day 26 in comparison to day 0 and 7 in group IV animals. Total number of follicles in both ovaries of group II animals was significantly (p<0.01) higher on day 26. Large and subordinate follicular diameter on ovaries of group II, III and IV animals increased gradually, but the differences between periods were significant (p<0.001) only in group IV with highest recorded diameter of large follicle on day 17 ; and of subordinate follicle on day 26. The right ovary was found relatively more active as compared to left ovary. Overall total number of follicles, diameters of large and subordinate follicles were significantly (p<0.05) higher in control as compared to treatment subgroup of group II animals. The conception rates at FTAI in group II, III and IV was 66.66, 83.33 and 16.66 %, respectively; whereas the corresponding values in control sub-groups were 0.00, 16.66 and 16.66 %, respectively. The differences in conception rate in treated and control heifers were significant (p<0.05), however, benefit of double Ovsynch treatment over control group in multiparous buffaloes was not observed. Plasma FSH concentration values differed significantly (p<0.001) among different groups at all-time intervals. Plasma FSH concentration obtained in control subgroup of group IV was significantly (p<0.001) higher than group II and III. Plasma LH concentration values differed significantly (p<0.001) among groups at various time periods. Plasma LH concentrations in group II and IV differed significantly (p<0.001). Plasma estrogen concentration obtained in animals of group III was significantly (p<0.05) higher than group IV. Plasma estrogen concentration obtained in control subgroups II and III were significantly (p<0.01) higher than control subgroup IV. Plasma insulin value obtained on all days for group IV animals were significantly (p<0.01) lower than group II and III animals. In group II, there was significantly (p<0.001) increased level of plasma FSH in treatment subgroup as compared to control subgroup; whereas, LH level was significantly (p<0.05) higher in control subgroup as compared to treatment subgroup. In group III, plasma FSH and LH levels were significantly (p<0.001) higher in treatment subgroup as compared to control subgroup. In group IV, there was significantly (p<0.01) increased level of FSH in treatment subgroup as compared to control; whereas, plasma LH level was significantly higher (p<0.05) in control subgroup as compared to treatment subgroup. The period-wise mean estrogen and insulin levels were significantly (p<0.05) higher in treatment subgroup as compared to control subgroup. Blood glucose level was significantly (p<0.001) higher in group II on day 17 as compared to group III and IV. Plasma total cholesterol values were significantly (p<0.05) higher in treatment subgroup IV as compared to subgroup II and III. Plasma total protein values were significantly (p<0.001) lower in group II heifers than in post-partum buffaloes of group IV, and group III heifers being intermediate. In group III, blood glucose level was significantly (p<0.05) lower in treatment subgroup as compared to control subgroup. On the basis of ovarian dynamics and results, it is concluded that double Ovsynch protocol could be better choice for improving conception rate in post-pubertal acyclic Jaffrabadi buffalo heifers as compared to multiparous acyclic
  • ThesisItemOpen Access
    TAURINE AS AN ADDITIVE IN ANDROMED EXTENDER FOR CRYOPRESERVATION OF GIR BULL SEMEN
    (JAU,JUNAGADH, 2018-06) CHIKHALIYA PIYUSH S; DR. A. R. AHLAWAT
    ABSTRACT The present investigation on the “Taurine as an additive in andromed extender for cryopreservation of Gir bull semen” was undertaken at Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Junagadh Agricultural University, Junagadh,Gujarat. The study was carried out on four Gir bull (4-6 years) located at Cattle Breeding Farm J.A.U. Junagadh for a period of 6 weeks. Semen was collected using artificial vagina method once weekly for six weeks from each of the bulls. The semen was evaluated for physical characteristics and oxidative stress at, neat semen (pre-dilution), per-freeze and post-thaw stages of cryopreservation using different Taurine concentrations viz. 25mM Taurine, 50 mM Taurine, 75 mM Taurine and control was kept. Onevaluation of 24 semen ejaculates (6 ejaculates from each bull) of neat semen, ejaculate volume (ml), sperm concentration (million/ml), mass motility (0-5 scale), individual motility (%), sperm viability (%), sperm abnormality (%), HOST reactive spermatozoa (%), and acrosome integrity were found to be 5.80± 0.41, 1267.63± 106.32, 3.88 ± 0.07, 85.83 ± 1.94,87.58 ± 0.42,10.87 ± 0.41, 83.00 ± 0.84 and 91.87 ± 0.32, respectively. There was a significant difference in volume and concentration of semen between the bulls while the difference was non-significant for the mass motility, individual motility, viability, abnormality, HOST reactive spermatozoa and acrosomal integrity. The levels of oxidative stress parameters viz. lipid peroxidation and glutathione reductase were 51.14 ± 0.18 μmol/ml and 32.24± 0.64 U/L, respectively. The difference was non-significant among the bulls for the glutathione reductase, while significant (P<0.05) difference was found for the lipid peroxidation. Individual motility, mass motility, viability,HOST reactive spermatozoa, glutathione reductase and acrosomal integrity were positively correlated with each other. At pre-freeze stages of cryopreservation, 50mM Taurine group had value of 70.20 ± 1.22, 69.87 ± 3.09, 69.5 ± 1.14 and 86.54 ± 0.81 percent for individual motility, sperm viability, HOST reactive sperm and acrosomal integrity respectively, value were significantly (P<0.05) higher as compared to that of the 25 mM Taurine, 75 mM Taurine and control groups.Mean per cent sperm abnormality was 15.37 ± 0.89 in 50mM Taurine group at pre-freeze stages of cryopreservation, which were significantly (P<0.05) lower as compared to that of the othergroups. Mean Malondialdehyde (MDA) levelwere30.74 ± 0.55μmol/ml in 50mM Taurine group at post-thaw stages of cryopreservation, and were significantly (P<0.05) lower as compared to that of the other groups.Mean glutathione reductase levels in 50mM Taurine group were 64.76 ± 0.78 U/L at pre-freeze stages of cryopreservation, and were significantly (P<0.05) higher as compared to that of the other groups. At post-thaw stages of cryopreservation, 50mM Taurine group had value of 62.70 ± 0.42, 65.66 ± 0.58, 61.87 ± 0.76,and 84.50 ± 0.64 percent for individual motility, sperm viability, HOST reactive sperm and acrosomal integrity respectively, value were significantly (P<0.05) higher as compared to that of the 25 mM Taurine, 75 mM Taurine and control groups.Mean per cent sperm abnormality was 20.29 ± 0.73 in 50mM Taurine group at post-thaw stages of cryopreservation, which were significantly (P<0.05) lower as compared to that of the othergroups. Mean Malondialdehyde (MDA) levelwas 15.95 ± 0.74μmol/ml in 50mM Taurine group at post-thaw stages of cryopreservation, and were significantly (P<0.05) lower as compared to that of the other groups.Mean glutathione reductase levels in 50 mM Taurine group were 80.25 ± 4.85U/L at post-thaw stages of cryopreservation, and were significantly (P<0.05) higher as compared to that of the other groups. All of above physico-morphological and bio-chemical parameters were better in 50 mM Taurine group at pre-freeze and post-thaw stages of cryopreservation. In conclusion, it was found that 50 mM is the most appropriate concentration of taurine as an additive in andromed extender to improve the semen quality parameters in Gir bull semen.