pdf35Ribogospod. nauka Ukr., 2017; 2(40): 99-113
DOI: https://doi.org/10.15407/fsu2017.02.099 
UDC 621.59:[597-146.31:639.371.2]

Development of cryoprotective media for low-temperature freezing of sterlet (Acipenser ruthenus) sperm

I. Kononenko, This email address is being protected from spambots. You need JavaScript enabled to view it. , National University of Life and Environmental Sciences of Ukraine, Kyiv

Purpose. To develop the optimum composition of a cryoprotective solution for freezing sterlet sperm with the use of new and standard constituents and compare the results of the fertilization of eggs with sperm frozen by different ways and in different media. To analyze the results of keeping sterlet males in simulated conditions of natural spawning and to assess the quality of sexual products obtained during early spawning periods in comparison with natural ones.

Methodology. The tasks set for the optimization of cryoprotective media for freezing sterlet sperm were solved according to generally accepted cryobiology methods and in accordance with Kopyeyka Ye. F. recommendations. The works with brооd sterlet were conducted in accordance with the generally accepted methods of sturgeon breeding and recommendations for natural spawning modeling.

Findings. The studies demonstrated a positive effect of creatine, the introduction of which into the solution allowed increasing the cryoprotection of sperm from damaging factors of cryopreservation. During low-temperature freezing of sperm by different ways, the best results were obtained when freezing sperm in samples of smallest volume — granules that was evidenced by the number of live thawed spermatozoa, fertilization rates of eggs and the number of developing embryos. The simulation of the conditions of natural spawning allowed obtaining high-quality sexual products from sterlet males earlier in comparison with natural spawning periods, freezing them and fertilizing eggs after several weeks of storage.

Originality. Among a variety of substances used in cryobiology for the optimization of the composition of protective solutions, positive results were tested and obtained with the use of creatine. Optimization of the cryopreservation solution composition was carried out using sterlet sperm obtained at early spawning terms compared to the natural ones.

Practical value. The obtained results can be used in the conditions of hatcheries for obtaining sterlet offspring both for market and for the restoration of their populations in natural water bodies. In addition, the obtained results are the basis for further studies on the development of optimum composition of cryoprotective media for freezing sterlet sperm.

Keywords: sterlet (Acipenser ruthenus), simulated conditions of natural spawning, pre-spawning group, spermatozoa, methanol, creatine.

REFERENCES

  1. Alkhimov, Ie. M., & Shevchenko, V. Iu. (2017). Suchasnyi stan vyroshchuvannia remontnykh tsoholitok osetropodibnykh ryb (Acipenseriformes) v umovakh pivdnia Ukrainy. Rybohospodarska nauka Ukrainy, 1(39), 52-63. https://doi.org/10.15407/fsu2017.01.052 
  2. Chebanov, M. S., & Galich, E. V. (2013). Rukovodstvo po iskusstvennomu vosproizvodstvu osetrovyh ryb: Tehnicheskij doklad FAO po rybnomu hozjajstvu. Ankara.
  3. Ponomareva, E. N., Sorokina, M. N., Grigor'ev, V. A., Kovaleva, A. V., & Korchunov, A. A. (2010). Rezul'taty razrabotki metodov formirovanija matochnyh stad sterljadi v uslovijah zamknutogo vodoobespechenija. Vestnik AGTU, 1, 86-91.
  4. Veprincev, B. N., & Piliev, S. A. (1989). Sohranit' genofond ryb i vodnyh bespozvonochnyh. Rybnoe hozjajstvo, 6, 29-32.
  5. Karnauhov, V. N. (1994). Problemy i perspektivy sozdanija geneticheskih kriobankov dlja celej sohranenija bioraznoobrazija. Biofizika zhivoj kletki, 6, 1-6.
  6. Ponomareva, E. N., Tihomirov, A. M., Bogatyreva, M. M., & Krasil'nikova, A. A. (2012). Kriokonservacija reproduktivnogo materialy ryb: razrabotki Juzhnogo nauchnogo centra Rossijskoj akademii nauk. Sovremennye rybohozjajstvennye i jekologicheskie problemy Azovo-Chernomorskogo regiona: VІІ Mezhdunarodnaja konferencija. Kerch', 55-58.
  7. Anan'ev, V. I., Gahova, Je. N., Katasonov, V. Ja., Rott, N. N., & Cvetkova, L. I. (2002). Koncepcija sohranenija i ustojchivogo ispol'zovanija bioraznoobrazija s primineniem metodov kriokonservacii genomov gidrobiontov. Izbrannye trudy VNIIPRH. Dmitrov: Sever Podmoskov'ja, 385-399.
  8. Belous, A. M., Gordienko, E. A., & Rozanov, L. F. (1987). Biohimija membran. Zamorazhivane i krioprotekcija. Moskva: Vysshaja shkola.
  9. Vojnova, N. V., Mirzojan, A. V., Ivanov, G. Ju., & Fedchenko, V. M. (2003). Ispol'zovanie novyh tehnologij v akvakul'ture. Holodnovodnaja akvakul'tura: start v HHІ vek: mezhdunarodnyj simpozium. Sankt-Peterburg, 78-80.
  10. Cvetkova, L. I., Dokina, O. B., Pronina, N. D., & Milenko, V. A. (2001). Tehnologija kriokonservacii i hranenija v nizkotemperaturnom banke spermy ryb. Sbornik nauchno-tehnicheskoj i metodicheskoj dokumentacii po akvakul'ture. Moskva: VNIRO.
  11. Glogowski, J., Kolman, R., Szczepkowski, M., Horvath, A., Urbanyi, B., Sieczynski, P., Rzemieniecki, A., Domagala, J., Demianowicz, W., Kowalski, R., & Ciereszko, A. (2002). Fertilization rate of Siberian sturgeon (Acipenser baeri Brandt) milt cryopreserved with methanol. Aquaculture, 211, 367-373. https://doi.org/10.1016/S0044-8486(02)00003-0
  12. Ponomareva, E. N., Metallov, G. F., Korchunov, A. A., & Kovaleva, A. V. (2012). Osobennosti razvitija reproduktivnoj sistemy donskoj sterljadi v zaregulirovannyh uslovijah vodnoj sredy. Izvestija Samarskogo nauchnogo centra RAN, 5, 191-194.
  13. Ponomareva, E. N., Grigor'ev, V. A., Sorokina, M. N., Kovaleva, A. V., & Korchunov, A. A. (2011). Osobennosti gametogeneza sterljadi v zaregulirovannyh uslovijah vodnoj sredy. Vestnik AGTU, 2, 112-117.
  14. Matishov, G. G., Ponomarev, S. V., & Bakaneva, Ju. M. et. al. (2013). Spravochnik rybovoda. Innovacionnye tehnologii akvakul'tury Juga Rossii. Rostov-na-Donu: JuNC RAN.
  15. Korchunov, A. A. (2011). Metody uskorennogo formirovanija matochnogo stada sterljadi v uslovijah UZV. Aktual'nye problemy obespechenija prodovol'stvennoj bezopasnosti Juga Rossii: innovacionnye tehnologii dlja sohranenija bioresursov, plodorodija pochv, melioracii i vodoobespechenija: Mezhdunarodnaja nauchnaja konf. Rostov-na-Donu, 24-25.
  16. Ponomareva, E. N., Sorokina, M. N., Grigor'ev, V. A., Kovaleva, A. V., & Korchunov, A. A. (2010). Rezul'taty razrabotki metodov formirovanija matochnyh stad sterljadi v uslovijah zamknutogo vodosnabzhenija. Vestnik AGTU, 1, 86-91.
  17. Sudakova, N. V. (2006). Tehnologii i normativy po tovarnomu osetrovodstvu v VI rybovodnoj zone. Moskva: VNIRO.
  18. Chebanov, M. S., Galich, E. V., & Chmyr', Ju. M. (2004). Rukovodstvo po razvedeniju i vyrashhivaniju osetrovyh ryb. Moskva: Rosinformagrotekh.
  19. Kopejka, E. F. (1986). Instrukcija po nizkotemperaturnoj konservacii spermy karpa. Moskva: VNIIPRH.
  20. Zalepuhin, V. V. (2009). Individual'naja rabochaja plodovitost' kak harakteristika jendogennoj raznokachestvennosti ryb. Vestnik AGTU. Rybnoe hozjajstvo, 1, 73-78.
  21. Stein, H. (1979). Zum Einflus von Schutzsubstanz und Konfektionierung auf das Befruchtungsergebnis mit tiefgefrorenem Bachforellensperma (Salmo trutta morpha fario L.). Berl. Munch. Tierarztliche Wochenschrift, 92, 420–421.
  22. Cvetkova, L. I., Titareva, L. N., Kochetov, A. A. Voronina, V. P., Milenko, V. A., & Pronina, N. D. (2002). O nekotoryh faktorah, vlijajushhih na krioustojchivost' spermy karpa Cyprinus carpio. Izbrannye trudy VNIIPRH. Dmitrov: Sever Podmoskov'ja, 377-384.
  23. Krasil'nikova, A. A., & Tihomirov, A. M. (2014). Ob’em zamorazhivaemogo obraztsa kak odin iz faktorov vyzhivaemosti spermatozoidov osetrovyh vidov ryb pri kriokonservacii. Estestvennye nauki. Biotehnologija, 2 (47), 62-69.
  24. Linhart, O., Mims, S. D., Gomelsky, B., Cvetkova, L. I., Cosson, J., Rodina, M., Horvath, A., & Urbanyi, B. (2006). Effects of cryoprotectants and male on motility parameters and fertilizabilization rate in paddlefish (Polyodon spathula) frozen-thawed spermatozoa. J. Appl. Ichthyol., 22, 384-388. https://doi.org/10.1111/j.1439-0426.2007.00992.x 
  25. Cvetkova, L. I., Dokina, O. B., Pronina, N. D., Milenko, V. A., & Tansykbaev, A. N. (2004). Akvakul'tura osetrovyh ryb: dostizhenija i perspektivy razvitija. Astrahan'.
  26. Psenicka, M., Dietrich, G. J., Wojtczak, M., Nynca, J., Rodina, M., Linhart, O., Cosson, J., & Ciereszko, A. (2008). Acrosome staining and motility characteristics of sterlet spermatozoa after cryopreservation with use of methanol and DMSO. Cryobiology, 56, 251-253. https://doi.org/10.1016/j.cryobiol.2008.03.006 
  27. Horvath, A. Wayman, W. R., Urbanyi, B., Ware, K. M., Dean, J. C., & Tiersch T. R. (2005). The relationship of the cryoprotectants methanol and dimethylsulfoxide and hyperosmotic extenders on sperm cryopreservation of two North-American sturgeon species. Aquaculture, 247, 243-251. https://doi.org/10.1016/j.aquaculture.2005.02.007 
  28. Chao, N.-H., & Liao, I. C. (2001). Cryopreservation of finfish and shellfish gametes end embryos. Aquaculture, 197, 161-189. https://doi.org/10.1016/S0044-8486(01)00586-5 
  29. Nahiduzzaman, Md., Hassan, Md. M., Roy, P. K., Hossain, Md. A., Hossain, M. Ali R., & Tierach, T. R. (2012). Sperm cryopreservation of the Indian major carp, Labeo calbasu: Effects of cryoprotectants, cooling rates and thawing rates on egg fertilization. Animal Reproduction Science, 136, 133-138. https://doi.org/10.1016/j.anireprosci.2012.10.023 
  30. Mounib, M. S. (1978). Cryogenic preservation of fish and mammalian spermatozoa. J. Reprod. Fert., 1 (53), 13-18. https://doi.org/10.1530/jrf.0.0530013 
  31. Mounib, M. S., Hwang, P. C., & Idler, R. R. (1968). Cryogenic preservation of Atlantic Cod (Gadus morhua) sperm. J. Fish. Res. Board. Can., 12(25), 2623-2632. https://doi.org/10.1139/f68-232
  32. Chernomashencev, A. I., & Mil'shtejn, V. V. (1983) Rybovodstvo. Moskva: Legkaja promyshlennost'.  
  33. Ivanov, A. P. (1988). Rybovodstvo v estestvennyh vodoemah. Moskva: Agropromizdat.
  34. Horváth, Á., Wayman, W. R., & Urbányi B. (2005). The relationship of the cryoprotectants methanol and dimethylsulfoxide and hyperosmotic extenders on sperm cryopreservation of two North-American sturgeon species. Aquaculture, 247, 243-251. https://doi.org/10.1016/j.aquaculture.2005.02.007 
  35. Lahnsteiner, F., Berger, B., Horvath, A., & Urbanyi, B. (2004). Studies on the sperm biology and sperm cryopreservation in the sterlet, Acipenser ruthenus L. Aquaculture Research, 35, 519-528. https://doi.org/10.1111/j.1365-2109.2004.01034.x  
  36. Lahnsteiner, F., Weismann, T., & Patzner R. A. (1997). Methanol as cryoprotectant and the suitability of 1.2 ml and 5 ml straws for cryopreservation of sperm from salmonid fishes. Aquaculture Research, 28, 471-479. https://doi.org/10.1111/j.1365-2109.2004.01034.x