Study of effect of adding horsmint ethanol extract and cysteine into extender on fertility features of frozen Baluchi ram semen after thawing

Authors

1 University of Gonbad Kavoos

2 Department of animal science, Faculty of agriculture and natural resources

3 University of Mohaghegh Ardabili

Abstract

Background and goal: According to an increasing interest for using artificial insemination by sheep producers, finding methods to maintain frozen semen fertility would be important and applicable. One of the freezing semen disadvantages is increasing of free radicals led to enhanced sperm membrane peroxidation then consequently causes lowered semen fertility after thawing. Some plants bearing high levels of plant antioxidants can help to decrease semen peroxidation. Cysteine, in addition, is a precursor of glutathione peroxidase being the most important antioxidant property in semen. Therefore, the aim of the study was to investigate the effect of adding different levels of ethanol extract of Mentha Longifolia (0, 100 and 200 microL/ml) and cysteine (0, 5 and 10 mmol/l) into ram semen on fertility features after freezing and thawing processes.
Materials and methods: In the study, semen samples were collected from four mature, fertile Baluchi rams (> 2y-old) by artificial vagina and after initial evaluation were pooled together. Then, samples were extended in an extender based on Tris-egg yolk. Treatments were arranged as a 3×3 factorial experiment including nine treatments and six straws were assigned to each as replicate. After treating, the straws were filled by the equal amount of semen and after chilling in refrigerator (4 °C) for two hours were frozen in liquid nitrogen container. After thawing the straws at day 10, sperm motility parameters were assayed by CASA. Furthermore, the percentages of viability, plasma membrane integrity and normal morphology of sperms were evaluated by microscope. Data were analyzed by SAS software using GLM procedure and the means comparison was performed by Tukey-Kramer test at 5% error.
Results: Results of the study showed that the effects of adding horsemint extract and cysteine into the extender and the interaction between them on the all motility and viability parameters of frozen ram sperm after thawing were significant (p<0.05), exempt for sperm morphology where only the effect of adding cysteine was significant. Means comparison showed that the greatest sperm total motility, plasma membrane integrity, linearity and curvilinear velocity were observed in cysteine10 treatment (without horsemint extract), while the greatest sperm viability, straight line velocity and average path velocity were seen in cysteine5 treatment (without horsemint extract). The greatest sperm progressive motility percentage was observed in horsemint200 group (without cysteine).
Conclusion: In general, considering to the beneficial effect of adding cysteine into the extender based on tris- egg yolk on the most of fertility parameters of ram frozen semen after thawing, using levels of 5 and 10 mmol cysteine per ml extender in order to maintain fertility of frozen ram semen is recommended.

Keywords

References

  1. 1.Alvarez, J.G., Touchstone, J.C., Blasco, L. and Storey, B.T. 1987. Spontaneous lipid peroxidation and production of hydrogen peroxide and superoxide in human spermatozoa: Superoxide dismutase as major enzyme protectant against oxygen toxicity. Journal of Andrology. 8: 338-348.

    2.Armstrong, J.S., Rajasekaran, M., Chamulitrat, W., Gatti, P., Hellstrom, W.J. and Sikka, S.C. 1999. Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radical Biology and Medicine, 26(7): 869-880.

    3.Bailey, J.L., Bilodeau, J.F. and Cormier, N. 2000. Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. Journal of Andrology. 21: 1-7.

    4.Bansal, A.K. and Bilaspuri, G.S. 2011. Impacts of oxidative stress and Antioxidants one semen functions. Veterinary Medicine International. 686137: 1-7.

    5.Bilodeau, J.F., Chatterjee, S., Sirard, M.A. and Gagnon, C. 2000. Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Molecular Reproduction and Development. 55: 200-202.

    6.Baumber, J., Ball, B.A., Gravance, C.G., Medina, V. and Davies-Morel M.C. 2000. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential and membrane lipid peroxidation. Journal of Andrology. 21: 895-902.

    7.Craig, J.W. 1999. Health – promoting properties of common herbs. American Journal of Clinical Nutrition. 70 (S3): 491-499.

    8.Dolti, P., Moghaddam,Gh. and Ahmadian, H. 2016. Evaluation of the effects of cysteine and trehalose on longterm cryopreservation of ram semen. Journal of Agri-Food and Applied Sciences. 4 (1): 1-6.

    9.Hancock, J. 1956. The morphology of boar spermatozoa. Journal of the Royal Microscopical Society. 76: 84-97.

    10.DaghighKia, H. Sadeghi SadeghAbad, F. Mohamadzadeh, H. Vaseghi Dodran, H. and Ashrafi I. 2017. The effect of Origanum Vulgare extract as natural antioxidant on quality cryopreserved ram sperm. Journal of Animal Science Research.26(4): 111-120.( In Persian)

    11.Farhadi, R. DaghighKia, H. and Ashrafi I. 2016. The Effect of Salvia Sahendica Ethanolic Extract as Natural Antioxidant on Quality Parameters of Cryopreserved Holstein Bull Sperm. Research on Animal Production. 12: 79-86.

    12.Khalili, B., Jafaroghli, M. Farshad, A. and Paresh- Khiavi, M. 2010. The Effects of Different concentrations of Glycine and cysteine on the Freezability of Moghani Ram spermatozoa. Asian-Australian Journal of Animal Sciences. 23(3): 318-325.

    13.Krzyzosiak, J., Evenson, D., Pitt, C., Jost, L., Molan, P. and Vishwanath, R. 2001. Changes in susceptibility of bovine sperm to in situ DNA denaturation during prolonged incubation at ambient temperature under conditions of exposure to reactive oxygen species and nuclease inhibitor. Reproduction, Fertility and Development. 12(6): 251-261.

    14.Lenzi, A., Picardo, M., Gandini, L., Lombardo, F., Terminali, O., Passi, S. and Dondero, F. 1994.Glutathione treatment of dyspermia: effect on the lipoperoxidation process. Journal of Human Reproduction, 9: 2044-2050.

    1. Linford, E., Glover, F.A., Bishop, C., and Stewart, D.L. 1976. The relationship between semen evaluation methods and fertility in the bulls. Reproduction and Fertility. 47: 283-291.
    2. Malo, C., Gil, L., Cano, R., Martínez, F. and Galé, I. 2011. Antioxidant effect of rosemary (Rosmarinus officinalis) on boar epididymal spermatozoa during cryopreservation. Theriogenology. 75: 1735-1741.

    17.Moreno, J.S., Coloma, M.A., Diaz, A.T., Brunet, A.G., Pastor, A.P., Soria, A.Z., Carrizosa, J.A., Urritia, B., and Sebastian, A.L. 2008. A comparison of the protective action of chicken and quail egg yolk in the cryopreservation. Cryobiology. 57(1): 25-29.

    1. Meister, A.M.E.A. and Anderson, M.E. 1983. Glutathione. Annual Review of Biochemistry, 52(1): 711-760.
    2. Osawa, T. 1994. Novel natural antioxidants for utilization in food and biological systems. In Postharvest Biochemistry of plant Food- Materials in the Tropics; Uritani, I, Garcia, V, Mendoza V. Japan Scientific Societies Press: Tokyo, Japan; 241−251.
    3. Perumal, P., Selvaraju, S., Selvakumar, S., Barik, A.K., Mohanty, D.N., Das, S., Das, R.K. and Mishra, P.C. 2011. Effect of pre-freeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates. Reproduction in Domestic Animals, 46(4): 636-641.

    21.Rice-Evans, C., Miller, N.J. and Paganga, G. 1997. Antioxidant properties of phenolic compounds. Trends in Plants Science. 2: 152-159.

    22.Revell, S., and Mrode, R. 1994. An osmotic resistance test for bovine semen. Animal Reproduction Science. 36: 1-2.77-86.

    23.Rahbari, F., Moslemipour, F., Mostafalou, Y. and Rahchamani, R. 2017. Effect of cysteine addition and semen extender type on fertility of bovine semen stored at refrigerator temperature. The Third national conference on innovative achievements. 1-12.

    24.Roca, J., Gil, M.A., Parrilla, I., Vazquez, J.M. and Martinez, E.A. 2004. Survival and fertility of boar spermatozoa after freeze-thawing in extender supplemented with butylated hydroxytoluene. Journal of Andrology, 25: 397-405.

    25.Sikka, S.C. 2001. Relative impact of oxidative stress on male reproductive function. Current Medicinal Chemistry; 8: 851-862.

    26.Safdarian, M. 2007. Artificial insemination of sheep and goats. The first edition. Institute of Applied Science Agriculture, 232p. (In Persian)

    27.Topraggaleh, T.R., Shahverdi, A., Rastegarnia, A., Ebrahimi, B., Shafiepour, V., Sharbatoghli, M., Esmaeili, V. and Janzamin, E. 2014. Effect of cysteine and glutamine added to extender on post-thaw sperm functional parameters of buffalo bull. Andrologia. 46(7): 777-783.

    1. Uysal, O., Bucak, M.N., Yavas, I. and Varisli, O. 2007. Effect of various antioxidants on the quality of frozen-thawed bull semen. Journal of Animal and Veterinary Advances. 6(2): 1362-1366.
    2. Yanishlieva, N.V. and Marinova, E.M. 1996. Antioxidative effectiveness of some natural antioxidants in su nflower oil.Z Lebensm Unters Forsch; pp: 220–223.
    3. Zavatti, M., Zanoli, P., Benelli, A., Rivasi, M., Baraldi, C. and Baraldi, M. 2011. Experimental study on Satureja Montana as a treatment for premature ejaculation. Journal of Ethnopharmacology. 133: 629-633.
    4. Zhandi, M. and Sharafi, M. 2015. Negative effect of combined cysteine and glutathione in soy lecithin-based extender on post-thawed ram spermatozoa. Cell and Tissue Banking. 16(3): 443-448.
Journal of Ruminant Research
Volume 8, Issue 1
June 2020
Pages 49-62

Files

Share

How to cite

Statistics