Detection and Quantification of X and Y sperms in Enriched Sahiwal Semen diluted in PBS using Raman Spectroscopy
DOI:
https://doi.org/10.48165/ijar.2023.44.02.9Keywords:
Raman spectroscopy, Sahiwal, Semen enrichment, X Y SpermAbstract
The present study was undertaken to enrich X-sperms in Sahiwal bull semen by percoll density gradient method and to detect X and Y sperms in enriched semen by Raman spectroscopy. The experiment was performed at Semen Production Centre, CVASc, GBPUAT, Pantnagar, Uttarakhand. Ejaculates were collected by AV method from Sahiwal bull. X-sperm enrichment was done by the Percoll density gradient method. Three types of gradients were prepared i.e. 7 layers (70-10%), 7 layers (80-20%) and 3 layers (30%, 50% and 70%). Semen (1ml or 3ml) was placed over the gradient in a conical centrifuge tube. Centrifugation was done at 750 g (22-24°C) for 15 min. The pellets thus obtained were diluted in PBS medium. Raman spectroscopy was performed in different groups viz. fresh semen (Control), 3 layers (70%, 50% and 30% GI), 7 layers (70-10% GII), and 7 layers (80-20% GIII). Raman peaks (DNA specific) were not much clear for PBS diluted semen samples though intensities were highest for 7 layers (70-10%) among all groups. Thus semen enriched by the percoll-density gradient method (7 layers i.e. 70-10%) can be used to increase female calf birth after A.I.
References
Bhat, Y. and Sharma, M. (2020). X-sperm Enrichment of Bovine Semen by Percoll Density Gradient Method and Its Effect on Semen Quality, Sex Ratio and Conception Rate. Indi. J. Anim. Res., 54(10):1181-1187.
Boro, P., Naha, B.C., Madkar, A. and Chandra, P. (2016). Sexing of semen in bulls: A mini review. Int. J. Appl. Res. 2(4): 460- 462.
Chaudhary, D., Devlal, K. and Sharma, M. (2022). Study on Seminal Attributes of X- sperm Enriched Sahiwal Bull Semen. Theriogenology, 12(01): 11-16.
De Luca, A.C., Managó, S., Ferrara, M.A., Rendina, I., Sirleto, L., Puglisi, R., Balduzzi, D., Galli, A., Ferraro, P., Coppola, G. (2014). Non-invasive sex assessment in bovine semen by Raman spectroscopy. Laser Phys. Lett., 11:1-8.
H. Rodriguez-Martinez, JL Wallet and AJ Ziyech, ed. Nottingham University. Press, Nottingham, UK. 51-66.
Indian Ministry of Agriculture & Farmer’s Welfare (2018). National Action Plan for Dairy Development, New Delhi, Government of India. pp 13.
Lizuka, R., Kaneko, S., Aoki, R. and Kobayashi, T. (1987). Sexing of human sperm by discontinuous Percoll density gradient and its clinical application. Human Reprod., 2(7): 573-575.
Rath, D., Bathgate, R., Rodriguez-Martinez, H., Roca, J., Strzezek, J. and Waberski, D. (2009). Recent advances in boar semen cryopreservation. Control of pig Reproduction VIII, 66: 51-66.
Rawat., M. and Sharma, M. (2020). Effect of percoll density gra dient separation of X and Y sperm on buffalo bull semen quality. J. Exp. Zool., 23(1):623-630.
Sharma, M. and Sharma, N. (2016). Sperm sexing in animals. Adv. Anim. Vet. Sci.,4(10): 543-549
Sharma, M., Bhat, Y., Sharma, N. and Singh, A. (2018). Comparative study of seasonal variation in semen charac teristics of buffalo bull. J. Entomol. Zool. Stud., 6(1): 947- 951.
Sharma, N., Chand, D.K., Rawat, S., Sharma, M. and Verma, H. (2018). Effect of sexed semen on conception rate and sex ratio under field conditions. J. Entomo. Zool. Stud.,6(1): 702-705.
Sharma, V., Verma, A.K., Sharma, P., Pandey, D. and Sharma, M. (2022). Differential proteomic profile of X- and Y- sorted Sahiwal bull semen. Res. Vet. Sci.,144:181-189.
Singh, A., Sharma, M. and Prasad, S. (2017). Change in Viability, Motility and Membrane Integrity of Spermatozoa After Equilibration and Thawing in BHT Added Bovine Semen. Adv. Anim. Vet. Sci.,5(8): 329-333
Singh, A., Sharma, M., Prasad, S., Bhat, Y., Kumar, A., Pandey, D. And Shukla, S. (2017). Effect of Supplementation of Butylated Hydroxytoluene on Post-Thaw Viability, Motility and Membrane Integrity of Crossbred Bovine Spermatozoa. Int. J. Livestock Res., 7(4):93-101.
Downloads
Published
Issue
Section
How to Cite
