Enhancing Fertility Through Induction Of Ovulation In Mares
Material type: Book ; Format:
; Literary form:
Publisher: 2014 Dissertation note: Abstract
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Comparison of The Effect of Ovsynch and G6G Synchronization Protocols on Ovulation and Pregnancy Rate in Nili-Ravi Buffaloes
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Publisher: 2015 Dissertation note: Thesis submitted with blank CD
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Effect Of Alpha Lipoic Acid On Post Thaw Quality Of Jack Semen
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Publisher: 2016 Dissertation note: Improvement of post-thaw quality of Donkey (Equus asinus) semen is essential to augment the in-vivo and in-vitro fertilization rate and to be used for mule production. By the help of cryopreservation sperm cells can be stored for the long time but it causes lethal sub-lethal damage to the sperm. In most species including Donkey and horses sperm cryosurvival rates are not optimal because of its plasma membrane composition. One of the major cryopreservation damage is produced by Reactive oxygen species (ROS) generating oxidative stress caused by ROS are important for normal sperm function but in normal concentration. When they are produced in more quantity they cause damage to Acrosome, DNA and plasma membrane. . Alpha lipoic acid is a non-vitamin coenzyme that helps in significant metabolic and antioxidant functions in the body. Alpha lipoic acid has been reported to have extra functions by which they are able to synthesize vitamin C from its reduced form in the presence of glutathione. It is matchless among biological antioxidants, because it is equally lipid and water soluble. This allows it to nullify free radicals almost everywhere in the body, inside as well as outside the cells. Therefore, the objective of present study was to determine the effect alpha lipoic acid on post thaw quality of jack semen. Alpha lipoic acid scavenge on reactive oxygen species formed in semen during the process of cryopreservation, so it maintained good semen quality during post thaw. Two adult donkeys (Equus asinus) (4-6 years old) kept at animal shed Ravi campus pattoki were used in the study. All the animals were managed under optimal condition of feeding and management. Donkeys were offered green fodder with ad libitum supply of water. Semen collection was done twice a week (one ejaculate/collection) using an equine artificial vagina having temperature of 45-50 ºC. Five collection from each donkey were done (n=10). Ejaculates were filtered with muslin cloth to remove gel. Semen volume was measured by collecting semen in a graduated collection tube after
filtration and the sperm concentration was measured by using a phase contrast microscope (40 x, Nikon) and was scored with a coverslip and then immediately was kept in water bath having 37 ºC temperature after collection until evaluation and processing. Semen quality parameters like volume, concentration and motility were recorded. After initial evaluation, semen samples were extended with centrifugation extender in 1:1 and seminal plasma was removed after centrifugation. Supernatant was removed so that seminal plasma up to 20% will remain with sperm pellet and was maintained at 37 ºC temperature in water bath and was extended with extender having different concentrations of Alpha lipoic acid (0mM, 0.5mM, 1mM, 1.5mM, and 2mM) and cooled for 2 hours and then equilibrated for 2 hrs at 4oC. Then, French semen straws of 0.5ml capacity were filled with semen (100x106/straw). All semen straws were arranged on a rack and then placed at 4cm height above liquid nitrogen vapors for 10 minutes. Finally semen straws was put in liquid nitrogen, Total five replicates per donkey were performed. Now post thaw quality was checked in which various tests were performed, like %age motility, Acridine orange assay for DNA integrity, HOST for plasma membrane integrity, FITC-PNA/PI for viability and acrosomal integrity. It was expected that Alpha lipoic acid shown positive effect on post thaw quality of donkey semen, in the meaning of increased percentage motility, Less DNA damage during cryopreservation and incubation, Increased acrosomal and plasma membrane integrity. So alpha lipoic acid shown positive effect by counter acting on ROS during cryopreservation by reducing melondialdehyde production as indicated by MDA test carried out in this study. Results acquired from this study shown that an increase in sperm motility, plasma membrane integrity, DNA integrity, Acrosomal integrity, viability and survival was caused by ALA competences in energy production and anti-oxidant properties, when used at the concentration of 1.5mM. In summary, based on the results
of our study, it can be concluded that an optimal concentration (1.5mM) of ALA improved PMI, sperm motility and viability, minimize DNA damage and improved sperm survival.
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Effect Of Age On Lipid Peroxidation Of Fresh And Frozen-Thawed Semen Of Nili-Ravi Buffalo Bulls
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Publisher: 2016 Dissertation note: Buffalo spermatozoa are rich in polyunsaturated fatty acids and prone to lipid peroxidation. Malondialdehyde (MDA) is a byproduct of lipid peroxidation and causes irreversible damage to sperm structure and function. In buffalo, blood plasma MDA level increases with age. Therefore, we hypothesized that MDA level in buffalo bull semen will increase with age and will affect the semen quality. The objective of the study was to compare MDA level and quality of fresh and frozen-thawed semen in aged vs. young Nili-Ravi buffalo bulls. Single ejaculate was collected on weekly basis for four weeks from aged (13.6±1.0 years; n=3) and young (3.4±0.3 years; n=3) Nili-Ravi buffalo bulls. MDA level was estimated through thiobarbituric acid assay (TBA) in fresh and frozen-thawed semen. The quality of fresh and frozen-thawed semen was estimated through sperm motility, viability, DNA and acrosome integrity. MDA level (nmol/ml) did not differ (P>0.05) between aged vs. young bulls in fresh (2.3±0.2 vs. 2.9±0.7) and frozen-thawed (53.1±2.8 vs. 48.4±2.6) semen, respectively. In fresh semen, sperm motility and concentration did not differ (P>0.05) in aged vs. young bulls; however, the volume of fresh semen increased (P<0.05), while sperm viability and DNA integrity decreased (P<0.05) in aged vs. young bulls. In frozen-thawed semen, sperm motility, viability, and DNA integrity decreased (P<0.05) in aged vs. young bulls. In frozen-thawed vs. fresh semen, MDA level (nmol/ml) increased within young (48.4±2.6 vs. 2.3±0.2) and aged bulls (53.1±2.8 vs. 2.9±0.7), while motility and viability decreased (P<0.05) within the age groups. In conclusion, 1) lipid peroxidation (MDA) does not increase due to age in buffalo bull semen, and 2) freezing causes increase in lipid peroxidation irrespective of age and deteriorates semen quality of Nili-Ravi bulls.
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Effect Of Insemination Timing Following 5 Vs. 7 Day Cidr + Co-Synch In Nili Ravi Buffalo Heifers
Material type: Book Publisher: 2017 Dissertation note: Pakistan’s economy is agricultural based in which livestock has a major contribution. Livestock contributes 11.8% to total national GDP. Nili Ravi is established breed of Pakistan. Nili Ravi has high lactation yields (1800-2500 liters with a 6.5% fat) and their males are more suited to ploughing and drafting on dry plane land. Increased calving interval and estrus detection are the biggest bottleneck in achieving the goal “a calf per 13 months” and high conception rate in dairy animals ultimately this leads to in economic losses. To overcome the problem of poor breeding in buffalo various exogenous hormonal intervention have been made to control estrous cycle of buffalo. Several ovulation synchronization protocols offer to bypass the estrus detection and artificially inseminate the animal at pre-established time. Co-synch is modified form of ovulation synchronization. Incorporation of CIDR in Co-synch improves the efficacy of protocol. It is easier in management and cost effective.
Present study was designed to compare the efficacy of 5 or 7 day CIDR + Co-synch in Nili Ravi buffalo and find appropriate time of A.I in Nili Ravi using 5 or 7 day CIDR + Co-synch protocol. Study was conducted on Military Dairy Farm Okara. Selected animals (N = 80) were divided into two treatments. Treatment 1) 7 day CIDR + Co-synch (n =40): The animals received an intravaginal CIDR insert containing 1.38 g of progesterone (P4) for 7 days. On the day of CIDR removal, 150 µg of PGF2α was injected intramuscularly (IM); Half of these animals (n=20) received 100 µg of GnRH IM and TAI after 72 hrs of CIDR removal/ PGF2α administered, the remaining half (n=20) received GnRH and TAI after 84 hrs of CIDR removal/ PGF2α administration. Treatment 2) 5 day CIDR + Co-synch (n=40): The animals received an intravaginal CIDR insert containing 1.38 g of P4 for 5 days. The animals were administered 150 µg of PGF2α IM at the time of CIDR removal. Half of these animals (n=20) received 100 µg of GnRH IM and TAI after 72 hrs of CIDR removal/ PGF2α administration, the other half (n=20) received 100 µg GnRH and FTAI after 84 hrs of CIDR removal/ PGF2α administration. Data is analyzed using IBM SPSS Statistics 20 software program. Follicular growth rate, pre-ovulatory follicular size is analyzed through independent t-test. Interval from CIDR removal to ovulation and interval from GnRH to ovulation is analyzed by Mann Whitney U test. Pregnancy rate is compared by binary linear regression. Ovulation rate and estrus response is analyzed through Chi square. A probability level of (P < 0.05) is considered as significant.
The results of this study showed the pregnancy rates were high (P < 0.05) in buffalo heifers of subgroups of both treatments inseminated after 84 hrs of CIDR removal (50% in 5 day CIDR + Co-synch and 65% in 7 day CIDR + Co-synch) than the subgroup inseminated after 72 hrs interval of CIDR removal (30% in 5 day CIDR + Co-synch and 25% in 7 day CIDR + Co-synch).
Follicular growth rate (mm/day) was tended to be high in 7 day CIDR treatment as compared to 5 day CIDR treatment (1.5±0.3 vs. 1.3±0.4, P=0.06). Pre-ovulatory follicle size in 7 day CIDR + Co-synch was significantly high in animals inseminated after 84 hrs as compared to subgroup inseminated after 72 hrs interval of CIDR removal (12.29mm vs. 10.74mm, P < 0.05). Similar trend was found in 5 day CIDR + Co-synch, the subgroup inseminated after 84 hrs of CIDR removal had larger pre-ovulatory follicular size than subgroup inseminated at 72 hrs interval (10.63mm vs. 11.73mm, P < 0.05). Interval from PG/CIDR removal to ovulation in 7 day CIDR treatment was 99 ± 0.9 hrs while in 5 day CIDR treatment it was 96 ± 1.6 hrs which did not differ significantly (P = 0.14). The interval from A.I/GnRH administration to ovulation in 7 day CIDR + Co-synch was significantly lower in animals inseminated after 84 hrs as compared to subgroup inseminated after 72 hrs interval of CIDR removal (15±1.2 vs. 27.±1.4, P < 0.05). Similar trend was found in 5 day CIDR + Co-synch, the subgroup inseminated after 84 hrs of CIDR removal/PG administration had lower A.I/GnRH administration to ovulation interval than subgroup inseminated at 72 hrs interval (16.2±1.3 vs. 21.6±1.6, P < 0.05). Ovulation rate was significantly high (P = 0.05) in 7 day CIDR treatment (95 %) than the 5 day CIDR treatment (80%). The estrus response in buffalo heifers was not significantly different between the treatments (90% in 7 day CIDR vs. 80% in 5 day CIDR treatments, P > 0.05).
In conclusion higher pregnancy rates were achieved in 7 day CIDR + Co-synch when animals were inseminated after 84 hrs interval of CIDR removal/PG administration. Acceptable pregnancy rates were resulted when timed AI was done after 84 hr of CIDR removal/PG administration using 5 day CIDR + Co-synch regimen. However further studies with larger sample size may be carried out to establish a strong statistical analysis between 5 and 7 day CIDR + Co-synch while inseminating animals after 84 hrs of CIDR removal/PG administration. Results of this study suggest that high pregnancy rate is primarily attributed to larger pre-ovulatory follicular size and appropriate time of AI in relation to time of ovulation. This study provides a way out to large dairy farmer to increase fertility with optimized management.
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Relationship Of Estradiol And Progesterone With Standing Estrus, Size Of Preovulatory Follicle And Interval To Ovulation In Beetal Goats
Material type: Book ; Literary form:
Publisher: 2017 Dissertation note: The objective of the current study was to determine follicular dynamics and plasma
concentrations of ovarian steroid in response to single PGF2α, given randomly during the luteal
phase in Beetal goat. A total of seven Beetal goats were given a single dose of PGF2α at the
unknown day of luteal phase upon confirmation of corpus luteum via ultrasonography during the
breeding season (November). Follicular dynamics were monitored using 7.5MHz transrectal
transducer at every 12 h following PGF2α (0 h) until ovulation. Plasma samples for estradiol 17β
and progesterone were collected at 0, 24, 36, 48, 60, 72, 84 and 96 h. An apronized buck was
used at every 6 h to detect standing estrus. Relative to PGF2α, all goats (n = 7) exhibited onset of
standing estrus at 50.6 ± 4.8 h and ovulation occurred at 82.3 ± 4.0h. However, the onset of
standing estrus and ovulation staggered (P < 0.05) among the goats. The onset of standing estrus
after PGF2α varied (P < 0.05) among early (n = 3), intermediate (n = 2) and late (n = 2)
responding goats i.e., 44 ± 2.0 vs. 51 ± 3.0 vs. 60 ± 0 h, respectively. The ovulation time among
early, intermediate and late responding goats was 72 vs. 84 vs. 96 h, respectively. The peak
plasma concentration of estradiol 17β was observed 12 h prior to ovulation in the goats. Mean
diameter of ovulatory follicle and duration of standing estrus were similar among the groups.
The corpus luteum regressed rapidly following PGF2α in early responding goats followed by
intermediate and late responding goats. Although plasma concentration of progesterone did not
differ (P = .065) among early, intermediate and late responding goats, but the change in
progesterone concentration over time differed (P < 0.05) among the groups. In conclusion, this
study indirectly shows that the onset of standing estrus and interval to ovulation following PGF2α
may vary in Beetal goats due to follicular and hormonal dynamics during the luteal phase.
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