CRYOPRESERVATION OF ROOSTER SEMEN USING N-METHYLACETAMIDE AS CRYOPROTECTIVE AGENT

K. PRANAY KUMAR¹*, B. SWATHI², M. SHANMUGAM^3
1Department of Veterinary Physiology, College of Veterinary Science, P.V. Narsimha Rao Telangana Veterinary University, Rajendranagar, Hyderabad, 500030
²Department of Veterinary Physiology, College of Veterinary Science, P.V. Narsimha Rao Telangana Veterinary University, Rajendranagar, Hyderabad, 500030
³ICAR-Directorate of Poultry Science, Rajendranagar, Hyderabad, 500030
* Corresponding Author : pranaykonda28@gmail.com

Received : 06-02-2018     Accepted : 10-02-2018     Published : 15-02-2018
Volume : 10     Issue : 3       Pages : 5123 - 5126
Int J Agr Sci 10.3 (2018):5123-5126
DOI : http://dx.doi.org/10.9735/0975-3710.10.3.5123-5126

Keywords : cryopreservation, fertility, genetic resources, N-methylacetamide, rooster, semen
Academic Editor : Rajashri Madishetti, Dr B. Vasanthi
Conflict of Interest : None declared
Acknowledgements/Funding : Authors are thankful to ICAR-Directorate of Poultry Science, Rajendranagar, Hyderabad, India for providing necessary facilities to carry out the present study.
Author Contribution : All author equally contributed

An attempt has been made to cryopreserve PD6 rooster semen using N-methylacetamide (MA) as a cryoprotective agent (CPA). Semen was collected from roosters, and pooled semen sample was diluted such that the concentration was arrived at 4 million/µl, followed by equilibration at 5°C for 30 minutes. Then the samples were diluted in 1:1 proportion with diluent containing 12%, 18% and 24% MA so that the final concentration of MA was 6%, 9% and 12% respectively and the final sperm concentration was 2 million/µl in each treatment. Immediately semen was loaded into 0.5 ml straws, sealed and kept 4.5 cm above liquid nitrogen (LN2) and exposed to nitrogen vapours for 30 minutes and subsequently immersed into LN2and stored in LN2 container. Frozen semen was thawed in ice water. Semen was evaluated before and after cryopreservation for progressive sperm motility, live and abnormal sperm percent and seminal plasma lipid peroxidation. Fertility trial was conducted by inseminating PD-3 line hens per vagina (200 million sperm in 0.1 ml semen dose) six times at three days interval. The progressive sperm motility percent in 6% MA, 9% MA and 12% MA treatments were 30.91, 34.55 and 27.73 respectively. The live sperm percent in 6% MA (31.82), 9% MA (35.63) and 12% MA (29.91) was similar between the treatments. Abnormal sperm percent was comparable between treatments. The percent fertility achieved with 6%, 9% and 12% MA cryopreserved semen were 1.5%, 0% and 5.3%, respectively, while control group inseminated with fresh semen produced 94.3% fertility. The percent hatchability on fertile eggs set (FES) for 12% MA treatment was 62.5% that was significantly (P<0.05) lower than the control group 93.82%. There was zero hatchability on FES in 6% MA and 9% MA groups.

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