Impact of supplementation of semen extender with antioxidants on quality of cooled or cryopreserved Marwari stallion spermatozoa

PRASHANT KUMAR; ASHOK KUMAR; J S MEHTA; G N PUROHIT; S K RAVI; YASH PAL; R A LEGHA; B N TRIPATHI; TR TALLURI

doi:10.56093/ijans.v90i10.111303

Authors

PRASHANT KUMAR College of Veterinary and Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner, Rajasthan.
ASHOK KUMAR ICAR-Central Island Agricultural Research Institute (CIARI), Port Blair
J S MEHTA Equine Production Campus, ICAR-NRC on Equines, Bikaner, Rajasthan
G N PUROHIT Deputy Director General (Animal Sciences), Krishi Bhawan, New Delhi
S K RAVI Equine Production Campus, ICAR-NRC on Equines, Bikaner, Rajasthan
YASH PAL Equine Production Campus, ICAR-NRC on Equines, Bikaner, Rajasthan
R A LEGHA Equine Production Campus, ICAR-NRC on Equines, Bikaner, Rajasthan
B N TRIPATHI Deputy Director General (Animal Sciences), Krishi Bhawan, New Delhi
TR TALLURI Equine Production Campus, ICAR-NRC on Equines, Bikaner, Rajasthan

https://doi.org/10.56093/ijans.v90i10.111303

Keywords:

Antioxidants, Ascorbic acid, Cryopreservation, Glutathione, ROS, Stallion semen

Abstract

The aim of the present study was to evaluate the effects of supplementation of semen extender with two antioxidants namely Ascorbic acid (AA @ 0.9 g/L), Glutathione (GSH @ 2.5 mM), and combination of both (AA @ 0.9 g/L + GSH @ 2.5 mM) either in alone or in combination on the quality of cooled or cryopreserved Marwari stallion spermatozoa. For this purpose, a total of 24 ejaculates were collected from four adult and fertile Marwari stallions (6 ejaculates from each stallion) using an artificial vagina. Each freshly ejaculated semen sample was investigated for the semen quality parameters, viz. colour, consistency, total volume, gel volume, gel free volume, pH, progressive sperm motility, sperm concentration, sperm viability, sperm plasma membrane integrity, acrosomal integrity and DNA integrity. In the freshly ejaculated semen, no significant variation was found among individual stallions for various semen quality parameters except in sperm concentration. Pre-freeze and post-thaw semen evaluation revealed that the values for the most of the semen quality parameters were significantly higher in the semen extender being treated with the combination (AA @ 0.9 g/L +GSH @ 2.5 mM) of antioxidants group rather than AA and GSH alone or control. Addition of AA (0.9 g/L) and GSH (2.5 mM) to the freezing extender improved equine pre-freeze and post-thaw semen quality with the superiority of control group which indicates the beneficial role of supplementation of antioxidants to the stallion semen during cryopreservation process.

Downloads

Download data is not yet available.

References

Aitken R J and Baker M A. 2004. Oxidative stress and male reproductive biology. Reproduction Fertility and Development 16(5): 581–88. DOI: https://doi.org/10.1071/RD03089

Ball B A, Medina V, Gravance C G and Baumber I. 2001. Effect of antioxidants on preservation of motility, viability and acrosomal integrity of equine spermatozoa during storage at 5°C. Theriogenology 56(4): 577–69. DOI: https://doi.org/10.1016/S0093-691X(01)00590-8

Bansal A K and Bilaspuri G S. 2010. Impacts of oxidative stress and antioxidants on semen functions. Veterinary Medicine International 686137. DOI: https://doi.org/10.4061/2011/686137

Baumber J, Ball B A and Linfor J J. 2005. Assessment of the cryopreservation of equine spermatozoa in the presence of enzyme scavangers and antioxidants. American Journal of Veterinary Research 66: 772–79. DOI: https://doi.org/10.2460/ajvr.2005.66.772

Bloom E. 1950. A one minute live-dead sperm stain by means of eosin-nigrosin. Fertility and Sterility 1: 176–77. DOI: https://doi.org/10.1016/S0015-0282(16)30125-X

Cueva F I, Caiza de la, Pojol M R, Rigau T, Bonet S, Miro J, Briz M and Rodriguez-Gill J E. 1997. Resistance to osmotic stress of horse spermatozoa. The role of tonic pumps and their relationship to any preservation success. Theriogenology 48: 947–68. DOI: https://doi.org/10.1016/S0093-691X(97)00322-1

Estrada E, Rodríguez-Gil J E, Rivera Del Alamo M M, Pena A and Yeste M. 2015. Effects of reduced glutathione on acrosin activity in frozen thawed boar spermatozoa. Reproduction Fertility and Development.

Gadea J, Gumbao D, Canovas S, Garcia-Vazquez F A, Grullon L A and Gardon J C. 2007. Supplementation of the dilution medium after thawing with reduced glutathione improves function and the in vitro fertilizing ability of frozen-thawed bull spermatozoa. International Journal of Andrology 31: 40–49. DOI: https://doi.org/10.1111/j.1365-2605.2007.00756.x

Gangwar C, Saxena A, Patel A, Singh S P, Yadav S, Kumar R and Singh V. 2018. Effect of reduced glutathione supplementation on cryopreservation induced sperm cryoinjuries in Murrah bull semen. Animal Reproduction Science 192: 171–78. DOI: https://doi.org/10.1016/j.anireprosci.2018.03.005

Hancock J L. 1951. A staining techniques for the study of temperature shock in semen. Nature (Lond.) 169: 323–26. DOI: https://doi.org/10.1038/167323b0

Ionata L M, Anderson T M, Pickett B W, Heird J C and Squires E L. 1991. Effect of supplementary sexual preparation on semen characteristics of stallion. Theriogenology 36(6): 923–57. DOI: https://doi.org/10.1016/0093-691X(91)90318-8

Jayendran R S, Vander-Ven H, Perez-Pelaez M, Crabo B G and Zanevld L J D. 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characters. Journal of Reproduction and Fertility 70: 219–28. DOI: https://doi.org/10.1530/jrf.0.0700219

Khlifaoui M, Battut I B, Chatagnon J F, Trimeche G A and Tainturier D. 2005. Effects of glutamine on post-thaw motility of stallion spermatozoa: an approach of the mechanism of action at spermatozoa level. Theriogenology 63: 138–49. DOI: https://doi.org/10.1016/j.theriogenology.2004.04.012

Maia M S and Bicudo S D. 2009. Radicais livres, antioxidantes e funçãoespermáticaemmamíferos: umarevisão. Rev Bras Reprod Anim. 33: 183–93.

Oliveira R A, Wolf C A, Oliveira M A V and Gambarini M L. 2013. Addition of glutathione to an extender for frozen equine semen. Journal of Equine Veterinary Sciences 33(12): 1148–52. DOI: https://doi.org/10.1016/j.jevs.2013.05.001

Oliveira R A de, Viu M A O and Gambarini M L. 2015. Cooling of equine semen at 16°C for 36 hours with addition of different glutathione concentrations. Ciências Agrárias, Londrina 36(6): 3699–3704. DOI: https://doi.org/10.5433/1679-0359.2015v36n6p3699

Oliveira J V, Oliveira P V L, Melo C M, Guasti P N, Silva Y F R S, Monteiro G A, Silva Y F R S, Papa P, Alvarenga M A, Dell’Aqua J A and Papa F O. 2016. Strategies to improve the fertility of fresh and frozen donkey semen. Theriogenology 85(7): 1267e73. DOI: https://doi.org/10.1016/j.theriogenology.2015.12.010

Pal Y, Legha R A and Tendon S A. 2009. Comparative assessment of seminal characteristics of horse and donkey stallions. Indian Journal of Animal Sciences 79(10): 1028–29.

Papa F O. 2016. Effect of BHT (Butylated Hydroxytoluene) addition in cooling and freezing extenders on equine sperm freezability. Faculdade de MedicinaVeterinária e Zootecnia (FMVZ). UniversidadeEstadualPaulista (UNESP). Campus de Botucatu. Botucatu, SP, Brazil.

Phetudomsinsuk K, Sirinarumitr K, Choothesa A, Suthanmapinunt P, Kornkaewrat K, Laikul A, et al. Freezability of Thai native crossbred horse semen in different extenders. Thai Journal of Veterinary Medicine 39: 105–14.

Pickett B W and Shiner K A. 1994. Recent developments in AI in horses. Livestock Production Science 40: 31–36. DOI: https://doi.org/10.1016/0301-6226(94)90263-1

Pickett B W, Faulkner L C, Seidel G E, Berndtson W E and Voss J L. 1976. Reproductive physiology of stallions VI. Seminal and behavioural characteristics. Journal of Animal Sciences 43: 617–25. DOI: https://doi.org/10.2527/jas1976.433617x

Prashant K, Rabindra K, Jitendra S M, Ashok K C, Sanjay K R, Sharat C M, Mohd. M A, Ram A L, B N Tripathi, T R Talluri. 2019. Ameliorative effect of ascorbic acid and glutathione in combating the cryoinjuries during cryopreservation of exotic jack semen. Journal of Equine Veterinary Science 81: 102796. DOI: https://doi.org/10.1016/j.jevs.2019.102796

Ravi S K, Legha R A, Pal Y and Sharma R C. 2013. Characteristics and freezability of Kathiawari horse semen. Indian Journal of Animal Sciences 83(11): 1146–48.

Sakkas D, Urner F and Bizzaro D. 1998. Sperm nuclear DNA damage and altered chromatin structure: effect on fertilization and embryo development. Human Reproduction 13(4): 11– 19. DOI: https://doi.org/10.1093/humrep/13.suppl_4.11

Silva K M G, Moraes T A P, Silva E C B, Gamboa S C and Guerra M M P. 2009. Efeito da adição de trolox e pentoxifilinanamotilidade, integridade do acrossoma e do DNA de espermatozoidesequinosapósdescongelamento. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 61: 42–49. DOI: https://doi.org/10.1590/S0102-09352009000100007

Soni Y. 2016. Study on cryopreservation of stallion semen using glycerol and dimethylformamide cryoprotectants. PG Thesis, College of Veterinary and Animal Science, Bikaner, Rajasthan. Watson P F. 1975. Use of Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. Veterinary Record 97: 12–15. DOI: https://doi.org/10.1136/vr.97.1.12

Yogesh S, Talluri T R, Ashok K, Ravi S K, Mehta J S, Tripathi B N. 2018. Effects of different concentration and combinations of cryoprotectants on sperm quality, functional integrity in three Indian horse breeds. Cryobiology 86: 52–57. DOI: https://doi.org/10.1016/j.cryobiol.2018.12.005

Gibb Z, Grupen C G, Maxwell W M and Morris L H. 2017. Field fertility of liquid stored and cryopreserved flow cytometrically sex-sorted stallion sperm. Equine Veterinary Journal 49: 160– 66. DOI: https://doi.org/10.1111/evj.12583

Zhandi M and Ghadimi V. 2014. Effect of glutathionesupplemented INRA82 extender on miniature caspian stallion sperm quality during storage at 5°C. Journal of Equine Veterinary Science 34(5): 606–10. DOI: https://doi.org/10.1016/j.jevs.2013.11.009