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Methyl glycol, methanol and DMSO effects on post-thaw motility, velocities, membrane integrity and mitochondrial function of Brycon orbignyanus and Prochilodus lineatus (Characiformes) sperm

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Abstract

The aim of this study was to use more accurate techniques to investigate the effects of cryoprotectants (CPAs) and extenders on post-thaw sperm quality of Brycon orbignyanus and Prochilodus lineatus. Six freezing media comprising the combination of three CPAs (DMSO, methanol and methyl glycol) and two extenders (BTS and glucose) were used. Sperm was diluted in each medium, loaded into 0.5-mL straws, frozen in a nitrogen vapor vessel (dry-shipper), and stored in liquid nitrogen at −196 °C. Post-thaw sperm motility rate and velocities (curvilinear = VCL; straight line = VSL; average path = VAP) were evaluated using a computer-assisted sperm analyzer. Membrane integrity and mitochondrial function were determined using fluorochromes. Post-thaw quality was considered high when samples presented the following minimum values: 60 % motile sperm, 140 µm/s of VCL, 50 % intact sperm membrane and 50 % mitochondrial function integrity. High post-thaw quality was observed in B. orbignyanus sperm frozen in BTS–methyl glycol and in P. lineatus sperm frozen in BTS–methyl glycol, glucose–methyl glycol and glucose–methanol. All samples frozen in DMSO yielded low quality. The presence of ions in the BTS extender affected post-thaw sperm quality positively in B. orbignyanus and negatively in P. lineatus. Methyl glycol was the most suitable CPA for both fish species, leading to a good protection of cell membrane, mitochondrial function and motility apparatus during the cryopreservation process. For an improved protection, B. orbignyanus sperm should be frozen in an ionic freezing medium.

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Acknowledgments

This study received funding from the Brazilian fostering agencies CNPq (PQ 300994/2008-7; PQ 302434/2011-9; Grant 552471/2010-0), ANEEL P&D Furnas (017965) and FAPEMIG (Grant CVZ BPD 00216-11; PPM CVZ 00129-11), and from Czech programs CENAKVA CZ.1.05/2.1.00/01.0024, Strengthening of excellence scientific teams in USB FFPW CZ.1.07/2.3.00/20.0024, GACR P502/12/1973 and LO1205. This is part of A.F. Nascimento’s PhD project. The authors thank the undergraduate student M. Hamaue, MSc student T.R. Taffarel (UFLA), and the biologists D.M. Ribeiro and M.B. Goulart (Furnas) for assistance during the experiments.

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Authors and Affiliations

  1. Department of Animal Science (DZO), Federal University of Lavras (UFLA), Lavras, MG, 37200-000, Brazil

    Ana T. M. Viveiros, Ariane F. Nascimento, Marcelo C. Leal & Antônio C. S. Gonçalves

  2. Department of Agronomy, José do Rosário Vellano University (UNIFENAS), Alfenas, MG, 37130-000, Brazil

    Laura H. Orfão

  3. Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, University of South Bohemia in Ceske Budejovice, 38925, Vodnany, Czech Republic

    Jacky Cosson

Authors
  1. Ana T. M. Viveiros
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  2. Ariane F. Nascimento
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  3. Marcelo C. Leal
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  4. Antônio C. S. Gonçalves
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  5. Laura H. Orfão
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  6. Jacky Cosson
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Correspondence to Ana T. M. Viveiros.

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Viveiros, A.T.M., Nascimento, A.F., Leal, M.C. et al. Methyl glycol, methanol and DMSO effects on post-thaw motility, velocities, membrane integrity and mitochondrial function of Brycon orbignyanus and Prochilodus lineatus (Characiformes) sperm. Fish Physiol Biochem 41, 193–201 (2015). https://doi.org/10.1007/s10695-014-0016-7

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  • DOI: https://doi.org/10.1007/s10695-014-0016-7

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