COMPARISON OF THREE DIFFERENT METHODS FOR THE ANALYSIS OF RAM SPERM CONCENTRATION

Authors

  • Andrej Baláži NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic
  • Jaromír Vašíček NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic; University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Nitra, Slovak Republic
  • Andrea Svoradová NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic
  • Marián Macháč University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Nitra, Slovak Republic
  • Rastislav Jurčík NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic
  • Ján Huba NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic
  • Ivan Pavlík NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic
  • Peter Chrenek NPPC – Research Institute for Animal Production Nitra, Lužianky, Slovak Republic; University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Nitra, Slovak Republic

Keywords:

ram sperm, concentration, flow cytometry, CASA, cell counter

Abstract

Determination of sperm concentration is a critical component of semen analysis. Traditionally, the haemocytometer has been the standard tool for calibrating other techniques used to estimate sperm concentration, including photometry, Coulter counters, flow cytometry and computer-automated semen analysis (CASA). In the present study, fresh ram sperm samples (n = 7) from the Native Wallachian (NW) Slovak sheep breed were collected from one male by electro-ejaculation (EE) and analysed for sperm concentration using flow cytometer FACS Calibur, CASA Sperm VisionTM and using EVETM Automatic Cell Counter. Our results showed no significant (P ≥ 0.05) differences in the sperm concentration when analysed by these three methods. Thus, it is possible to use a cell counter to determine the approximate sperm concentration directly at the place of semen collection. This is a very practical finding since instruments such as flow cytometer or CASA are not suitable for transport to the place of semen collection.

References

Anzar, M., Roetsch, T. & Buhr, M. M. (2009). Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen. Journal of Andrology, 30(6), 661–668. https://doi.org/10.2164/jandrol.108.007500

Amann, R. P. & Katz, D. F. (2004). Reflections on CASA after 25 years. Journal of Andrology, 25, 317–325. https://doi.org/10.1002/j.1939-4640.2004.tb02793.x



Amann, R. P. & Waberski, D. (2014). Computer-assisted sperm analysis (CASA): capabilities and potential developments. Theriogenology, 81, 5–17. https://doi.org/10.1016/j.theriogenology.2013.09.004



Ax, R. L., Dally, M., Didion, B. A., Lenz, R. W., Love, C. C., Varner, D. D., Hafez, B. & Bellin, M. E. (2016). Semen Evaluation. In Hafez, B. & Hafez, E. S. E. (Ed.), Reproduction in Farm Animals [Online PDF] (pp. 363–375). https://doi.org/10.1002/9781119265306.ch25



Bailey, E., Fenning, N., Chamberlain, S., Devlin, L., Hopkisson, J. & Tomlinson, M. (2007). Validation of sperm counting methods using limits of agreement. Journal of Andrology, 28(3), 364–373. https://doi.org/10.2164/jandrol.106.002188



Boe-Hansen, G. B & Satake, N. (2019). An update on boar semen assessments by flow cytometry and CASA. Theriogenology, 137, 93–103. https://doi.org/10.1016/j.theriogenology.2019.05.043



Brito, L., Beckman, B., Cardwell, B., DeJarnette, J. M., Hutchens, L., Kaya, A., Krieger, K. E., Lenz, R., Mitchell, J. R. & Siddiqui, A. (2012). NAAB-CSS semen quality control program minimum guidelines. NAAB Technical Conference AI and Reproduction, Vancouver, 37–41.



Brito, L. F. C., Althouse, G. C., Aurich, C., Chenoweth, P. J., Eilts, B. E., Love, C. C., Luvoni, G. C., Mitchell, J. R., Peter, A. T., Pugh, D. G. & Waberski D. (2016). Andrology laboratory review: evaluation of sperm concentration. Theriogenology. https://doi: 10.1016/j.theriogenology.2016.01.002.



Hansen, C., Christensen, P., Stryhn, H., Hedeboe, A. M., Rode, M. & Boe-Hansen, G. (2002). Validation of the FACSCount AF System for Determination of Sperm Concentration in Boar Semen. Reproduction in Domestic Animals, 37, 330–334. https://doi.org/10.1046/j.1439-0531.2002.00367.x



Iguer-ouada, M. & Verstegen, J. P. (2001) Evaluation of the "Hamilton Thorn computer-based automated system" for dog semen analysis. Theriogenology, 55, 733–749. https://doi.org/10.1016/S0093-691X(01)00440-X



Kubovičová, E., Makarevič, A. V., Špaleková, E. & Hegedušová, Z. (2011). Motility and fertilizing ability of frozen-thawed ram sperm from two sheep breeds. Slovak Journal of Animal Science, 44(4), 134–139. Retrieved from https://sjas.ojs.sk/sjas/article/view/316/304



Kulíková, B., Baláži, A., Tóthová, J., Jurčík, R., Huba, J. & Chrenek, P. (2018). Dilution factor affects the ability of ram sperm to survive cryopreservation. Slovak Journal of Animal Science, 51(1), 41–44. Retrieved from https://sjas.ojs.sk/sjas/article/view/64/53



Morrell, J. M., Johannsson, A., Juntilla, L., Rytty, K., Bäckgren, L., Dalin, A. M. & Rodriguez-Martinez H. (2010). Stallion sperm viability, as measured by Nucleocounter SP-100, is affected by extender and enhanced by single layer centrifugation. Veterinary Medicine International, 2010:659862. https://doi.org/10.4061/2010/659862



Prathalingam, N. S., Holt, W. W., Revell, S. G., Jones, S., & Watson, P. F. (2006). The precision and accuracy of six different methods to determine sperm concentration. Journal of Andrology, 27(2), 257–262. https://doi.org/10.2164/jandrol.05112



Rijsselaere, T., Van Soom, A., Maes, D. & de Kruif, A. (2003) Effect of technical settings on canine semen motility parameters measured by the Hamilton-Thorne analyzer. Theriogenology, 60, 1553–1568. https://doi.org/10.1016/S0093-691X(03)00171-7



Zinaman, M. J., Uhler, M. L., Vertuno, E., Fisher, S. G. & Clegg, E. D. (1996). Evaluation of computer-assisted semen analysis (CASA) with IDENT stain to determine sperm concentration. Journal of Andrology, 17(3), 288–292. https://doi.org/10.1002/j.1939-4640.1996.tb01784.x



World Health Organization. (2010). WHO laboratory manual for the examination and processing of human semen. WHO Press, 2010.

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2020-06-25

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