The effect of L-carnitine on ram epididymal sperm motility parameters during preservation in HTF medium with CASA
Subject Areas : Theriogenology and Reproductive Medicine
1 - Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia Branch, Islamic Azad University, Urmia, Iran
2 - Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia Branch, Islamic Azad University, Urmia, Iran
Keywords: Ram, Sperm, L-carnitine, HTF, CASA,
Abstract :
L-carnitine, an endogenous amino acid, is highly effective in preventing programmed cell death. As an antioxidant, it also protects DNA structure against reactive oxygen species and increases the activity and amount of antioxidant enzymes. In this study, 20 pairs of mature ram testicles were collected immediately after Islamic slaughter from the Urmia industrial slaughterhouse. The samples were transferred to the faculty laboratory in a cool box. After cleaning and drying the testes from blood, a cut was made in areas without capillaries on the tail of the epididymis. The sperm were transferred to microtubes containing HTF (Human Tubal Fluid) culture medium with 10% bovine serum albumin. After preparing a dilution of 30 to 50 million sperm per ml of HTF, five levels of L-carnitine (0, 2, 4, 6, 8, 10 mM) were added to the medium. The samples were then kept in a refrigerator at 5 degrees Celsius for up to 36 hours. At 1, 12, 24, and 36 hours, the sperm motility pattern was evaluated with CASA. Statistical results showed that L-carnitine at levels of 6, 8, and 10 mM and at 24 and 36 hours caused a clear and significant increase in the motility indices of ram epididymal sperm compared to the control group (p<0.05).
[1] Zhao J, Meng P, Jin M, Ma X, Ma H, Yang H. et al. Combined addition of L-carnitine and L-proline improves cryopreservation of dairy goat semen. Animal Reproduction Science. 2023; 257:107325. doi:10.1016/j.anireprosci.2023.107325
[2] Mohammadi V, Sharifi SD, Sharafi M, Mohammadi-Sangcheshmeh A, Shahverdi A, Alizadeh A. et al. Manipulation of fatty acid profiles in roosters’ testes, alteration in sexual hormones, improvements in testicular histology characteristics and elevation sperm quality factor by L-carnitine. Theriogenology, 2021. Theriogenology; 161: 8-15. doi.org/10.1016/j.theriogenology.2020.10.005
[3] Yang K, Wang N, Gou HT, Wang JR, Sun HH, Sun LZ. et al. Effect of L-carnitine on sperm quality during liquid storage of boar semen. Asian-Australasian Journal of Animal Sciences. 2020; 33(11): 1763-1769.
doi:10.5713/ajas.19.0455
[4] Pedreira AC, Malacarne AM, Dalmaso ACS, Carvalho KIFZ, Chagas TV, Gambetta MRI. et al. L-carnitine solution used on Rhamdia quelen thawed sperm activation boosts sperm movement, maintains larval quality, and permits to optimize the sperm use. Animal Reproduction Science, 2022. 245: 107054. doi:10.1016/j.anireprosci.2022.107054
[5] Arshadi N, Abdy K. The effects of Xylose monosaccharide on Water Buffalo (Bubalus bubalis) epididymal sperm kinetic parameters at 37 ˚ C. Journal of Basic and Clinical Veterinary Medicine. 2023; 4(1): 1-11. doi:10.30495/jbcvm.2023.1979108.1037
[6] Salinas MBS, Lertwichaikul T, Khunkaew C, Boonyayatra S, Sringarm K, Chuammitri P. et al. Freezability biomarkers in the epididymal spermatozoa of swamp buffalo. Cryobiology, 2022. 106: 39-47. doi:10.1016/j.cryobiol.2022.04.005
[7] Dziekońska A, Niedźwiecka E, Niklewska ME, Koziorowska-Gilun M, Kordan W. Viability longevity and quality of epididymal sperm stored in the liquid state of European red deer (Cervus elaphus elaphus). Animal Reproduction Science. 2020; 213: 106269. doi:10.1016/j.anireprosci.2019.106269
[8] Miró J, Morató R, Vilagran I, Taberner E, Bonet S, Yeste M. Preservation of epididymal stallion sperm in liquid and frozen states: effects of seminal plasma on sperm function and fertility. Journal of Equine Veterinary Science. 2020; 88: 102940. doi:10.1016/j.jevs.2020.102940
[9] Souza, CV, Brandão FZ, Santos JDR, Alfradique VAP, Dos Santos VMB, Morais MC. Effect of different concentrations of l-carnitine in extender for semen cryopreservation in sheep. Cryobiology. 2019; 89: 104-108. doi:10.1016/j.cryobiol.2019.05.009
[10] Ibănescu I, Leiding C, Ciornei SG, Roșca P, Sfartz I, Drugociu D. Differences in CASA output according to the chamber type when analyzing frozen-thawed bull sperm. Animal Reproduction Science. 2016; 166: 72-79.
doi:10.1016/j.anireprosci.2016.01.005
[11] Palacín I, Vicente-Fiel S, Santolaria P, Yániz JL. Standardization of CASA sperm motility assessment in the ram. Small Ruminant Research. 2013; 112(1-3): 128-135. doi:10.1016/j.smallrumres.2012.12.014
[12] Sarıözkan S, Ozdamar S, Türk G, Cantürk F, Yay A. In vitro effects of l-carnitine and glutamine on motility, acrosomal abnormality, and plasma membrane integrity of rabbit sperm during liquid-storage. Cryobiology. 2014; 68(3): 349-353. doi:10.1016/j.cryobiol.2014.04.006
[13] Agarwal A, Said TM. Carnitines and male infertility. Reproductive Biomedicine Online. 2004; 8(4): 376-84.
[14] Fattah A, Sharafi M, Masoudi R, Shahverdi A, Esmaeili V, Najafi A. l-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm. Cryobiology. 2017; 74:148-153. doi:10.1016/j.cryobiol.2016.10.009
[15] Abd El-baset SA, Abd El-Wahab SM, Mansour AMA, Mohamed EAH. Light and electron microscopic study of the effect of L-carnitine on the sperm morphology among sub fertile men. Middle East Fertility Society Journal. 2010; 15(2): 95-105. doi:10.1016/j.mefs.2010.04.005
[16] Tatemoto H, Osokoshi N, Hirai M, Masuda Y, Konno T, Yamanaka K. Addition of l-carnitine to the freezing extender improves post-thaw sperm quality of Okinawan native Agu pig. Theriogenology. 2022; 188: 170-176. doi:10.1016/j.theriogenology.2021.12.030
[17] Zahmel J, Simonsen KS, Stagegaard J, Palma-Vera SE, Jewgenow K. Current state of in vitro embryo production in African Lion (Panthera leo). Animals. 2022; 12(11): 1424. doi:10.3390/ani12111424
