Korean Journal of Agricultural Science (Korean J. Agric. Sci.; KJOAS)
Indexed in KCI (Korea Citation Index), Open Access, Peer Reviewed.
pISSN 2466-2402
eISSN 2466-2410

Shilajit improves sperm penetration on in vitro fertilization of pig oocytes



Yi YJ, Dissanayake WMN, Aganja RP, Lee SM. Shilajit improves sperm penetration on in vitro fertilization of pig oocytes. Korean Journal of Agricultural Science 50:131-139.

Korean Journal of Agricultural Science (Korean J. Agric. Sci.) 2023 March, Volume 50, Issue 1, pages 131-139. https://doi.org/10.7744/kjoas.20230008

Received on 18 January 2023, Revised on 20 January 2023, Accepted on 21 February 2023, Published on 30 March 2023.

Shilajit improves sperm penetration on in vitro fertilization of pig oocytes

Young-Joo Yi1,*, Wijesooriya Mudhiyanselage Nadeema Dissanayake1, Ram Prasad Aganja2, Sang-Myeong Lee3

1Department of Agricultural Education, College of Education, Sunchon National University, Suncheon 57922, Korea

2Division of Biotechnology, College of Environmental and Bioresource Sciences, Jeonbuk National University, Iksan 54596, Korea

3Laboratory of Veterinary Virology, College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea

*Corresponding author: yiyj@scnu.ac.kr


Shilajit is a dark-brown exudate found in the rock formations of the Himalayas and contains various minerals, vitamins, and enzymes. The present study was performed to examine sperm motility, sperm adenosine triphosphate (ATP) content, generation of reactive oxygen species (ROS), and calcium ion (Ca2+) levels after exposure of sperm to shilajit. In the presence of various concentrations of shilajit, in vitro fertilization (IVF) of pig oocytes occurred with a reduced incubation time (3 hrs) and low sperm number (1 × 105 sperm·mL-1). The percentage of hyperactive sperm significantly increased with 10 and 20 µg·mL-1 shilajit (p < 0.05). A significant number of intact acrosomal sperm were observed in sperm with 10 – 50 µg·mL-1 shilajit (p < 0.05). In addition, higher adenosine triphosphate (ATP) content was observed in sperm exposed to these concentrations of shilajit compared to sperm incubated at 100 µg·mL-1 shilajit or without shilajit (p < 0.05). When sperm were incubated with shilajit, the generation of ROS decreased compared to sperm without shilajit, and an increment of cytoplasmic Ca2+ was observed in sperm exposed to 10 and 20 µg·mL-1 shilajit (p < 0.05). In IVF trials, the sperm rapidly reached the oocytes in the presence of shilajit, and significantly higher total fertilization was observed in the IVF medium with 10 and 20 µg·mL-1 shilajit. In conclusion, shilajit was seen to have a beneficial effect on improving sperm penetration and fertilization, suggesting that it could be a potential agent to improve fertilization rates in cases of subfertility and infertility during assisted reproduction.


fertilization, IVF (in vitro fertilization), pig, shilajit, spermatozoa


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Authors Information

Young-Joo Yi, https://orcid.org/0000-0002-7167-5123

Wijesooriya Mudhiyanselage Nadeema Dissanayake, https://orcid.org/0000-0001-7130-7737

Ram Prasad Aganja, https://orcid.org/0000-0002-1945-7153

Sang-Myeong Lee, https://orcid.org/0000-0002-3624-3392


We would like to thank Professor Sang-Hoon Ha of the Division of Biotechnology, Jeonbuk National University for his contribution. This work was supported by a Research Promotion Program of Sunchon National University.

Conflicts of interest

No potential conflict of interest relevant to this article was reported.