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Title of Thesis
DEVELOPMENT OF A HOMOLOGOUS IN VITRO FERTILIZATION TEST TO PREDICT FERTILITY OF BUFFALO SEMEN |
Author(s)
Abid Mehmood |
Institute/University/Department Details
Department of Biochemistry/ University of Arid Agriculture Rawalpindi |
Session
2007 |
Subject
Biochemistry |
Number of Pages
159 |
Keywords (Extracted from title, table of contents and abstract of thesis)
fertilization, fertility, buffalo, semen, artificial insemination, reproductive disorder, farm animals, fertility rates, in vitro fertilization, vesicle oocytes, fresh oocytes, sperm functional test, in vitro capacitation, vitrified-thawed occtyes |
Abstract
Artificial insemination technique is an effective tool for rapid improvement in the genetic potential and control of reproductive disorder in farm animals. Use of AI is still limited in buffalo, the main dairy animal of South Asia. Identification of high fertility semen donor buffalo bulls for AI would increase its popularity among farmers. The potential fertility of bulls can be measured with field fertility rates, but this procedure is expensive and time consuming. Therefore in the present study in vitro fertilization was tested to predict the in vivo fertility of buffalo bulls. Studies were carried out to standardize protocols of in vitro fertilization in buffalo for in vitro testing of bull fertility. Overall recovery rate of good quality oocytes from buffalo ovaries was 1.3 COCs/ovary. Oocytes recovered by aspiration method gave a better in vitro maturation rate (expansion = 86.4% and GVBD = 85%) than those recovered by slicing method. There was a trend for better cleavage rate with static culture system supplemented with oestrus buffalo serum, although the difference was non significant compared to static systems with FCS, and flux culture systems with either OBS or FCS. The capacitation of buffalo spermatozoa with higher heparin concentrations (50-100 µg/ml) and 30 min incubation time improved in vitro fertilization rates in buffalo (cleavage rate 75-80% and cleavage index 62-74%). Better cleavage rate and cleavage index was observed in the oocytes inseminated with spermatozoa separated with swimup method as compared to Percoll gradient (P<0.001). There were differences among bulls in the cleavage rate and cleavage index of oocytes following IVF with spermatozoa harvested by swim-up method (P<0.05). In vitro fertilization rates did not differ between vitrified "germinal vesicle stage oocytes€and fresh oocytes. Vitrified oocytes inseminated with frozen-thawed semen of 4 buffalo bulls showed different cleavage indices (P<0.05). Among the spermatological and IVF parameters, cleavage rate was identified as being most predictive of conception rate and accounted for 75% of variation (P = 0.0001). The predictive equation formed by spermatological parameters accounted for 52% and that for IVF parameters 81 % of the variation. The results suggest that the in vitro fertilization of homologous, zona intact oocytes is potentially an informative method for assessing in vivo fertilizing ability of buffalo bulls.
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Chapter |
Title of the Chapters |
Page |
Size (KB) |
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| 1 |
0 |
Contents |
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149.6 KB |
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| 2 |
1 |
Introduction |
1 |
39.94 KB |
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| 3 |
2 |
Review Of Literature |
5 |
391.25 KB |
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2.1 |
Criteria For A Bull Fertility Test |
5 |
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2.2 |
Prediction Of Bull Fertility |
7 |
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2.3 |
Conventional Parameters To Predict Bull Fertility |
7 |
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2.4 |
Sperm Functional Test |
16 |
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2.5 |
In Vitro Fertilization To Predict Bull Fertility |
25 |
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2.6 |
Status Of In Vitro Production Of Buffalo Embryos |
30 |
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2.7 |
Freezing Of Oocytes |
34 |
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| 4 |
3 |
Materials And Methods |
42 |
188.16 KB |
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3.1 |
Comparison Of Oocyte Recovery Methods From Ovaries |
42 |
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3.2 |
In Vitro Maturation Of Oocytes In Different Systems |
46 |
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3.3 |
In Vitro Capacitation Of Frozen Thawed Buffalo Semen |
49 |
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3.4 |
Effectiveness Of Sperm Separation Methods |
53 |
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3.5 |
Use Of Vitrified-Thawed Occytes For In Vitro Bull Fertility Test |
56 |
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| 5 |
4 |
Results And Discussion |
61 |
476.95 KB |
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4.1 |
Comparison Of Occtye Recovery Methods From Ovaries |
61 |
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4.2 |
In Vitro Maturation Of Oocytes In Different Systems |
67 |
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4.3 |
In Vitro Capacitation Of Frozen Thawed Buffalo Semen |
70 |
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4.4 |
Effectiveness Of Sperm Separation Methods |
78 |
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4.5 |
Use Of Vitrified-Thawed Occtyes For In Vitro Bull Fertility Test |
90 |
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4.6 |
Relationship Of Spermatological And IVF Parameters With In Vivo Bull Fertility |
95 |
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| 6 |
5 |
Summary |
110 |
409.54 KB |
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5.1 |
Literature Cited |
114 |
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