Optimization of Curimba (Prochilodus lineatus) Sperm Cryopreservation: Efficacy of Trehalose as a Cryoprotectant
DOI:
https://doi.org/10.1590/1983-21252026v3915118rcKeywords:
Curimba. Freshwater fish. Sugars. Flow cytometry. Sperm kinetics. Cryopreservation.Abstract
Semen cryopreservation is essential for the conservation and maintenance of germplasm stocks from several species, each of which has specific requirements for successful cryopreservation. The objective of this study was to develop a sperm preservation protocol for Prochilodus lineatus (curimba) using the sugars trehalose, sucrose, lactose, and raffinose. Because synergy among cryoprotectants is not always beneficial, each cryoprotectant was tested separately at three concentrations (50, 100, and 150 mM), and dimethyl sulfoxide (DMSO) was used as the control. After thawing, sperm kinetic parameters were analyzed using computer-assisted sperm analysis (CASA), and cellular functional parameters were assessed by flow cytometry. In the sperm kinetic analyses, only the amplitude of lateral head displacement did not differ statistically from the control (p > 0.05), whereas the other parameters were lower than those of the control. In the motility-period analysis, 100 mM trehalose was superior to the control (p < 0.05), supporting its use in the protocol. However, the results for the different parameters analyzed showed that the other sugars tested were not superior to the control (10% DMSO), indicating that they were not effective for cryopreservation. Trehalose at 100 mM showed the most favorable response among the tested sugars for P. lineatus sperm cryopreservation.
References
ALVES, J. P. et al. The role of amides in seminal cryopreservation of wild silverside, Odontesthes bonariensis. Cryobiology, 73: 83-87, 2016.
ASTURIANO, J. F.; CABRITA, E.; HORVÁTH, A. Progress, challenges and perspectives on fish gamete cryopreservation: A mini‐review. General and Comparative Endocrinology, 245: 69-76, 2017.
CARVALHO, A. F. S. et al. Canonical correlation analysis to identify the semen characteristics used to forecast the freeze survival of curimba (Prochilodus lineatus) spermatozoa. CryoLetters, 38: 263-268, 2017.
DZIEWULSKA, K. et al. Post-thawed motility and fertility from Atlantic salmon (Salmo salar L.) sperm frozen with four cryodiluents in straws or pellets. Theriogenology, 76: 300-311, 2011.
FADHILLAH, M. A. B. P. et al. The Effects of Brown Sugar as a Natural Cryoprotectant on Tor Soro (Valenciennes 1842) Spermatozoa Quality. Journal of Hunan University Natural Sciences, 49: 1-13, 2022.
FELIZARDO, V. et al. Effect of cryopreservant combinations on the motility and morphology of curimba (Prochilodus lineatus) sperm. Animal Reproduction Science, 122: 259-263, 2010.
FERNANDES, M. D. O. et al. Cryopreservation of sperm in annual fish Austrolebias minuano (Cyprinodontiformes; Rivulidae). Aquaculture Research, 51: 147-154, 2020.
FERNANDEZ, M. A. et al. Species-specific responses to cryoprotectants in fish semen cryopreservation. Aquaculture, 528: 735557, 2020.
GALLEGO, V.; ASTURIANO, J. F. Fish sperm motility assessment as a tool for aquaculture research: a historical approach. Reviews in Aquaculture, 10: 697-724, 2018.
JESUS PAULA, D. A. et al. Effects of cooling rates on the quality of Prochilodus lineatus (Valenciennes, 1836) sperm. Reproduction Domestic Animals, 54: 1034-1043, 2019.
LEE, Y. H. et al. Effects of cryoprotective agents and treatment methods on sperm cryopreservation of stone flounder, Kareius bicoloratus. Aquaculture, 531: 735969, 2021.
LIU, Q. et al. Effect of the addition of six antioxidants on sperm motility, membrane integrity and mitochondrial function in red seabream (Pagrus major) sperm cryopreservation. Fish Physiology and Biochemistry, 41: 413-422, 2015.
MAGNOTTI, C. et al. Cryopreservation and vitrification of fish semen: A review with special emphasis on marine species. Reviews in Aquaculture, 10: 15-25, 2018.
MARTÍNEZ-PÁRAMO, S. et al. Cryobanking of aquatic species. Aquaculture, 472: 156-177, 2017.
MILIORINI, A. B. et al. A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation. Aquaculture Research, 42: 177-187, 2011.
MURGAS, L. D. S. et al. Cryopreservation of curimba (Prochilodus lineatus) semen after addition of different diluters, activators and cryoprotectants. Revista Brasileira de Zootecnia, 36: 526-531, 2007.
NAVARRO, R. D. et al. Semen quality of curimba (Prochilodus lineatus) cryopreserved with vitamins. Acta Scientiarum. Technology, 36: 55-60, 2014.
PAN, J. M. et al. Cryopreservation of black seabream (Acanthopagrus schlegelii) sperm. Theriogenology, 210: 182-191, 2023.
PIRES, D. M. et al. Association between DMSO and sugars in the sperm cryopreservation of pacu. CryoLetters, 39: 121-130, 2018.
SANDOVAL-VARGAS, L. et al. Cryopreservation of coho salmon sperm (Oncorhynchus kisutch): Effect on sperm function, oxidative stress and fertilizing capacity. Aquaculture, 533: 736151, 2021.
SARVI, K. et al. Cryopreservation of semen from the endangered Caspian brown trout (Salmo trutta caspius). Aquaculture, 256: 564-569, 2006.
SCARABOTTI, P. A. et al. Long-term trends of fishery landings and target fish populations in the lower La Plata basin. Neotropical Ichthyology, 19: e210013, 2021.
SOTNIKOV, A. et al. Optimizing aquaculture-scale common carp artificial reproduction: a novel approach to sperm cryopreservation using large-volume containers and elevated thawing temperatures. Frontiers in Marine Science, 11: 1342483, 2024.
SOUZA, F. P. D. et al. Genetic variability of Prochilodus lineatus in artificial and semi-natural reproduction. Italian Journal of Animal Science, 17, 321-325, 2018.
VARELA JUNIOR, A. S. et al. Methods of cryopreservation of tambaqui semen, Colossoma macropomum. Animal Reproduction Science, 157: 71-77, 2015.
VARELA JUNIOR, A. S. et al. Trehalose in extenders for cryopreservation of tambaqui (Colossoma macropomum) sperm. CryoLetters, 43: 264-268, 2022.
VASCONCELOS, A. C. N. et al. Cryopreservation of Prochilodus lineatus semen: effect of cryoprotectant combinations on motility and morphology. Boletim do Instituto de Pesca, 41: 409-417, 2015.
WANG, Y. et al. A comprehensive review on genetically modified fish: key techniques, applications and future prospects. Reviews in Aquaculture, 13: 1635-1660, 2021.
WYLIE, M. J. et al. Fish germ cell cryobanking and transplanting for conservation. Molecular Ecology Resources, 25: e13868, 2025.
YANG, S. et al. Evaluation of different sugar extenders and large volume cryotube for sperm cryopreservation of brown‐marbled grouper (Epinephelus fuscoguttatus). Aquaculture Research, 53: 4069-4075, 2022.
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