Knowing the salmon semen
Atlantic salmon (Salmo salar) is a cultured fish species with a high economic value, which generates significant revenue from both wild catches and fish farming (Hindar et al.2006). However, the total annual catch of wild Atlantic salmon in the North Atlantic has shown a marked decline during recent decades from approximately 12 000 t in 1973 down to 1200 t in 2016 (NASCO 2016). The commercial response to this problem is in the conservation, restoration, enhancement and rational management of wild salmon in the North Atlantic. By focusing on biology, the aims of scientific projects are to increase fish survival and improve their behavioural adaptation to natural conditions. In a conservation program, it is essential to secure the revival of the species by ensuring genetic variability (Rurangwa et al.2004), using wild broodstock from the local habitat. The priority in reproduction for restocking is to ensure that all males contribute towards successful fertilisation. However, it is important to know the differences in sperm motility among males in order to maintain the genetic integrity of the broodstock used (McGinnity et al.1997, 2003; Rurangwa et al.2004). In addition, knowledge of sperm characteristics (velocity, optimal activation time and subpopulation structures) will help improve fertilization procedures (Bobe and Labbe´ 2010; Fauvel et al.2010).
In fish species with external fertilisation, the quest for reproductive success among competing males can lead to several adaptations, including behavioural, morphological and physiological, to enhance the competitiveness of their spermatozoa (Beatty et al.1969). The physiological quality of spermatozoa differs between competing males because of differences in investment in gametes and sperm production (Ball and Parker 1996). Sperm quality is generally correlated with fertilising ability, which is often used as a determining factor in studies on sperm competition (Levitan 2000). During spawning, the number of spermatozoa released and their swimming speed can affect the probability of fertilisation (Stoltz and Neff 2006). Faster swimming cells may be able to reach the egg first, increasing the probability of successful fertilization (Gage et al.2004; Stoltz and Neff 2006). Atlantic salmon is an anadromous species that migrates up rivers from the sea in order to breed. In the case of this species, spawning males are characterised by intense sexual competition, which confers strong selective pressure on their reproductive physiology (Vladic and Järvi 2001). The wild salmon population can exhibit a wide natural variation in sperm traits leading to sperm competition (Gage et al.2004). However, mature parr males also participate in the spawning, which means that they need to invest more in sperm production than anadromous males (Parker 1998).
Sperm quality can be assessed using simple methods, such as individual analysis of motility (Billard and Cosson 1992) and morphology (Holstein et al.1988), or sophisticated approaches involving molecular tools (Cabrita et al.2014). Motility is the parameter that is most used to assess sperm quality because it directly reflects the fertilising ability of the spermatozoa. Initially, the percentage of motile spermatozoa and/or the duration of their motility were evaluated using subjective visual estimates. However, computer-aided sperm analysis (CASA-Mot) systems were developed to provide more reliable, repeatable and objective measurements of sperm movement (Rurangwa et al.2004). Even though CASA technology was designed for mammalian species (Rurangwa et al. 2004), it is well adapted to fish spermatozoa (Gallego et al. 2013; Kime et al.2001), which are characterised by a short period of vigorous motility after activation. In general, fish spermatozoa remain active for less than 2 min in most aquatic species and, in the case of salmonids specifically, the duration of sperm motility is around 20–30 s (Kime et al.2001).
CASA-Mot systems provide a large amount of data based on the kinematic parameters of each spermatozoon. Applying subpopulation analysis to such data allows for the analysis of groups of spermatozoa with similar motility features and to estimate of sperm quality for each male (Soler et al. 2014). A subpopulation characterised by rapid linear movement has been proposed as an indicator of high-quality spermatozoa (Ferraz et al.2014). Variations in subpopulation distributions have been reported for several species, including boar (Flores et al.2009), bull (Valverde et al.2016), red deer (Martínez-Pastor et al.2005), stallion (Quintero-Moreno et al.2003), cat (Gutiérrez-Reinoso and García-Herreros 2016), dog (Núñez-Martínez et al.2006), fowl (García-Herreros 2016), rooster (García-Herreros 2016), human(Vásquez et al.2016), gilthead seabream (Beirao et al.2011) and steelhead (Kanuga et al.2012). This statistical methodology has improved knowledge regarding spermquality, although it is still used primarily as a research tool (Gil Anaya et al.2015).
In a study in which one of our members participated, the objective of the work was to study sperm subpopulation structure and motility patterns in wild anadromous males and farmed male Atlantic salmon parr. Salmon sperm samples were collected from wild anadromous salmon (WS) and two generations of farmed parr males. Sperm samples were collected from sexually mature males and sperm motility was analysed at different times after activation (5 and 35 s). Differences among the three groups were analysed using statistical techniques based on Cluster analysis the Bayesian method. Atlantic salmon were found to have three sperm subpopulations, and the spermatozoa in ejaculates of mature farmed parr males had a higher velocity and larger size than those of WS males. This could be an adaptation to high sperm competition because salmonid species are naturally adapted to this process. Motility analysis enables us to identify sperm subpopulations, and it may be useful to correlate these sperm subpopulations with fertilisation ability to test whether faster-swimming spermatozoa have a higher probability of success.
Our CASA system works with many different fish species, such as sea bass, salmon, sole, sturgeon, eel, zebrafish, sea urchin, and many more. And if it's not adapted to the species you need, we can customize it accordingly.
RESEARCH ARTICLE:
https://doi.org/10.1071/RD17466
REFERENCES:
1. Ball, M. A., and Parker, G. A. (1996). Sperm competition games: external fertilization and ‘adaptive’ infertility. J. Theor. Biol. 180, 141–150. doi:10.1006/JTBI.1996.0090 |
2. Beatty, R. A., Bennett, G. H., Hall, J. G., Hancock, J. L., and Stewart, D. L. (1969). An experiment with heterospermic insemination in cattle. J. Reprod. Fertil. 19, 491–502. doi:10.1530/JRF.0.0190491 |
3. Beirao, J., Cabrita, E., Pérez-Cerezales, S., Martínez-Paramo, S., and Herráez, M. P. (2011). Effect of cryopreservation on fish sperm subpopulations. Cryobiology62, 22–31. doi:10.1016/J.CRYOBIOL.2010.11.005 |
4. Billard, R., and Cosson, M. P. (1992). Some problems related to the assessment of sperm motility in freshwater fish. J. Exp. Zool.261, 122–131. doi:10.1002/JEZ.1402610203 |
5. Bobe, J., and Labbe´, C. (2010). Egg and sperm quality in fish. Gen. Comp. Endocrinol. 165, 535–548. doi:10.1016/J.YGCEN.2009.02.011 |
6. Cabrita, E., Martı´nez-Pa´ramo, S., Gavaia, P. J., Riesco, M. F., Valcarce, D. G., Sarasquete, C., Herra´ez, M. P., and Robles, V. (2014). Factors enhancing fish sperm quality and emerging tools for sperm analysis. Aquaculture432, 389–401. doi:10.1016/J.AQUACULTURE.2014.04.034 |
7. Fauvel, C., Suquet, M., and Cosson, J. (2010). Evaluation of fish sperm quality. J. Appl. Ichthyol. 26, 636–643. doi:10.1111/J.1439-0426.2010. 01529.X |
8. Ferraz, M. A., Morato, R., Yeste, M., Arcarons, N., Pena, A. I., Tamargo, C., Hidalgo, C. O., Muino, R., and Mogas, T. (2014). Evaluation of sperm subpopulation structure in relation to in vitrosperm–oocyte interaction of frozen–thawed semen from Holstein bulls. Theriogenology81,1067–1072. doi:10.1016/J.THERIOGENOLOGY.2014.01.033 |
9. Flores, E., Fernandez-Novell, J. M., Pena, A., and Rodrı´guez-Gil, J. E. (2009). The degree of resistance to freezing–thawing is related to specific changes in the structures of motile sperm subpopulations and mitochondrial activity in boar spermatozoa. Theriogenology72, 784–797. doi:10.1016/J.THERIOGENOLOGY.2009.05.013 |
10. Gage, M. J. G., Macfarlane, C. P., Yeates, S., Ward, R. G., Searle, J. B., and Parker, G. A. (2004). Spermatozoa traits and sperm competition in Atlantic salmon: relative sperm velocity is the primary determinant of fertilization success. Curr. Biol. 14, 44–47. |
11. Gallego, V., Carneiro, P. C. F., Mazzeo, I., Vı´lchez, M. C., Pen˜aranda, D. S., Soler, C., Pe´rez, L., and Asturiano, J. F. (2013). Standardization of European eel (Anguilla anguilla) sperm motility evaluation by CASA software. Theriogenology79, 1034–1040. doi:10.1016/J.THERIOGENOLOGY.2013.01.019 |
12. García-Herreros, M. (2016). Sperm subpopulations in avian species: a comparative study between the rooster (Gallus domesticus) and Guinea fowl (Numida meleagris). Asian J. Androl. 18, 889–894. |
13. Gil Anaya, M. C. G., Calle, F., Pe´rez, C. J., Martın-Hidalgo, D., Fallola, C., Bragado, M. J., Garcıa-Marın, L. J., and Oropesa, A. L. (2015). A new Bayesian network-based approach to the analysis of sperm motility: application in the study of tench (Tinca tinca) semen. Andrology3, 956–966. doi:10.1111/ANDR.12071 |
14. Gutiérrez-Reinoso, M. A., and García-Herreros,M. (2016).Normozoospermic versus teratozoospermic domestic cats: differential testicular volume, sperm morphometry, and subpopulation structure during epididymal maturation. Asian J. Androl. 18, 871–878. |
15. Hindar, K., Fleming, I. A., McGinnity, P., and Diserud, O. (2006). Genetic and ecological effects of salmon farming on wild salmon: modelling fromexperimental results. ICES J.Mar. Sci.63, 1234–1247. doi:10.1016/ J.ICESJMS.2006.04.025 |
16. Holstein, A. F., Roosen-Rumge, E. C., and Schirren, C. (1988). ‘Illustrated Pathology of Human Spermatogenesis.’ (Groose: Berlin.) |
17. Kanuga, M. K., Drew, R. E., Wilson-Leedy, J. G., and Ingermann, R. L. (2012). Subpopulation distribution of motile sperm relative to activation medium in steelhead (Oncorhynchus mykiss). Theriogenology77, 916–925. doi:10.1016/J.THERIOGENOLOGY.2011.09.020 |
18. Levitan, D. R. (2000). Sperm velocity and longevity trade off each other and influence fertilization in the sea urchin Lytechinus variegatus. Proc. Biol. Sci. 267, 531–534. doi:10.1098/RSPB.2000.1032 |
19. Martínez-Pastor, F., Garcia-Macias, V., Alvarez, M., Herra´ez, P., Anel, L., and de Paz, P. (2005). Sperm subpopulations in Iberian red deer epididymal sperm and their changes through the cryopreservation process. Biol. Reprod.72, 316–327. doi:10.1095/BIOLREPROD.104.032730 |
20. McGinnity, P., Stone, C., Taggart, J. B., Cooke, D., Cotter, D., Hynes, R., McCamley, C., Cross, T., and Ferguson, A. (1997). Genetic impact of escaped farmed Atlantic salmon (Salmo salar L.) on native populations: use of DNA profiling to assess freshwater performance of wild, farmed, and hybrid progeny in a natural river environment. ICES J. Mar. Sci. 54, 998–1008. doi:10.1016/S1054-3139(97)80004-5 |
21. North Atlantic Salmon Conservation Organization (NASCO) (2016). Report of the twenty-second annual meeting of the Council of the North Atlantic Salmon Conservation Organization. Document no. CNL(05)50, NASCO, Edinburgh. |
22. Núñez-Martínez, I., Moran, J. M., and Peña, F. J. (2006). A three-step statistical procedure to identify sperm kinematic subpopulations in canine ejaculates: changes after cryopreservation. Reprod. Domest. Anim.41, 408–415. doi:10.1111/J.1439-0531.2006.00685.X |
23. Parker, G. A. (1998). Sperm competition and the evolution of ejaculates: towards a theory base. In ‘Sperm Competition and Sexual Selection’. (Eds T. R. Birkhead and A. P. Möller.) pp. 3–54. (Academic Press: London.) |
24. Quintero-Moreno, A., Miro, J., Rigau, A. T., and Rodriguez-Gil, J. E. (2003). Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology |
25. Rurangwa, E., Kime, D. E., Ollevier, F., and Nash, J. P. (2004). The measurement of sperm motility and factors affecting sperm quality in cultured fish. Aquaculture 234, 1–28. doi:10.1016/J.AQUACULTURE. 2003.12.006 |
26. Soler, C., Garcı´a-Molina, A., Contell, J., Segarvall, J., and Sancho, M. (2014). Kinematics and subpopulations structure definition of blue fox (Alopex lagopus) sperm motility using the ISAS_v1 CASA system. Reprod. Domest. Anim.49, 560–567. doi:10.1111/RDA.12310 |
27. Stoltz, J. A., and Neff, B. D. (2006). Sperm competition in a fish with external fertilization: the contribution of sperm number, speed and length. J. Evol. Biol.19, 1873–1881. doi:10.1111/J.1420-9101.2006.01165.X |
28. Valverde, A., Arena´n, H., Sancho, M., Contell, J., Ya´niz, J., Ferna´ndez, A., and Soler, C. (2016). Morphometry and subpopulation structure of Holstein bull spermatozoa: variations in ejaculates and cryopreservation straws. Asian J. Androl. 18, 851–857. |
29. Vásquez, F., Soler, C., Camps, P., Valverde, A., and García-Molina, A. (2016). Spermiogram and sperm head morphometry assessed by multivariate cluster analysis results during adolescence (12–18 years) and the effect of varicocele. Asian J. Androl.18, 824–830. |
30. Vladic, T. V., and Järvi, T. (2001). Sperm quality in alternative reproductive tactics of Atlantic salmon: the importance of the loaded raffle. Proc. Biol. Sci.268, 2375–2381. doi:10.1098/RSPB.2001.1768 |