Effect of preservation of spermatozoa at sub-zero temperature on DNA integrity by Comet assay
Nandre, Rahul (2007) Effect of preservation of spermatozoa at sub-zero temperature on DNA integrity by Comet assay. Masters thesis, College of Veterinary Science and Animal Hsbandry, Anand Agricultural University, Anand, Gujarat, India.
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The present investigation of “Effect of Preservation of Spermatozoa at Sub-zero Temperature on DNA Integrity by Comet Assay” was carried out in summer and winter seasons at Department of Genetic Engineering and Biotechnology in collaboration with Department of Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Anand, Gujarat during the year 2006-07. The study included evaluation of seminal characteristics and Comet percentage in fresh semen diluted with TFYG and preserved at refrigeration temperature and total protein content in seminal plasma at above stages. Based on 36 ejaculate (6 ejaculates × 3 bulls × 2 period). The mean values for various seminal characteristics were: ejaculate volume, 4.11 + 0.21 ml, colour and consistency were score were milky white and 1.83 + 0.08, Density was 2.20 + 0.06, mass activity 2.96 + 0.18, individual motility 77.36 ± 1.04%, sperm Concentration 593.66 + 30.98 million/ml, viable percentage 95.05 ± 0.70%, abnormal sperm 3.11 ± 0.43% and comet percentage 5.30 ± 0.94%. Significant effect of interaction of season and period was observed on individual motility, abnormal sperm count being higher in winter than summer. In Comet percentage also significant effect of season was observed. Comet percentage was higher in summer than winter. Effect of season was significant on motility of spermatozoa being higher in winter than summer. Average seminal plasma total proteins contain in neat semen was 0.55 ± 0.04 g%. In winter it was 0.73 ± 0.07 g% and in summer 0.38 ± 0.03 g%. Season had significant effect, being higher in winter as compared to summer. After studying observations of three Surti buffalo bulls seminal plasma, it was concluded that during winter season, the more protein spots were expressed in the range of pH 5.0 to 6.7 and molecular weight 66.20 to 97.40 kDa as compared to summer season. These protein spots were also having variation in intensity of expression. Based on differentially expressed proteins characterization; seminal plasma was having better energy source and modulates sperm motility in winter season. Based on semen evaluation tests of freshly diluted semen, frozen thawed semen, DNA integrity of spermatozoa, protein percent and protein expression of seminal plasma; it could be concluded that quality of semen was found better during winter as compared to summer season in Surti buffalo bulls.
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