Skip to main content Accessibility help
×
Home

Injuries in pacu embryos (Piaractus mesopotamicus) after freezing and thawing

  • Patrícia Ribeiro Neves (a1), Ricardo Pereira Ribeiro (a2), Danilo Pedro Streit (a3), Maria Raquel M. Natali (a2), Darci Carlos Fornari (a4), Alexandra Inês Santos (a5) and Leandro C. Godoy (a3)...

Summary

Although the sperm cryopreservation of freshwater and marine teleosts has been feasible for years, the cryopreservation of some fish embryos still remains elusive. Thus, the objective of this experiment was to analyze the embryo morphology after freezing and thawing 40 embryos of Piaractus mesopotamicus immersed into methanol and ethylene glycol, both at 7, 10 and 13% plus 0.1 M sucrose for 10 min. Soon after thawing, three embryos were treated with historesin, stained with hematoxylin–eosin and analyzed under an optical microscope. From every treatment, one palette containing embryos was thawed and incubated, but none of the eggs hatched. Samples containing two embryos were immersed into 10% methanol or 10% ethylene glycol both in association with sucrose, and embryos immersed into only water or sucrose solution were frozen, processed and analyzed using scanning electron microscopy (SEM). In both cases, the control group was immersed into only water. Although the embryos had the chorion, vitello, yolk syncytial layer and blastoderm, all of them were found altered under the optical microscope and by SEM. The chorion was irregular and injured; there was no individuality in the yolk granules; the yolk syncytial layer had an irregular shape, thickness and size; the blastoderm showed injuries in the nucleus shape and sometimes was absent; the blastoderm was located in atypical areas and absent in some embryos. In conclusion, no treatment was effective in preserving the embryos, and none of the embryos avoided injury from intracellular ice formation. These morphological injuries during the freezing process made the P. mesopotamicus embryos unfeasible for hatching.

Copyright

Corresponding author

All correspondence to: Darci Carlos Fornari. Departamento ZootecniaUniversidade Estadual de Maringá, Av. Colombo, 5790 – Campus Universitário, CEP: 87020-900 Maringá, Paraná, Brasil. Tel: +5544 3261 8969. e-mail: darci.peixegen@gmail.com

References

Hide All
Ballou, J.D. (1992). Potential contribution of cryopreserved germ plasm to the preservation of genetic diversity and conservation of endangered species in captivity. Cryobiology 29, 1925.
Castagnolli, N. & Zuim, S.M.F. (1985). Consolidação do conhecimento adquirido sobre o pacu, Colossoma mitrei–Berg, 1895. Boletim Técnico do CEPTA 5, 12.
Denniston, R., Michelet, S. & Godke, R.A. (2000). Principles of cryopreservation. In: Tiersch, T.R. & Mazik, P.M.Cryopreservation in Aquatic Species. World Aquaculture Society, pp. 5974.
Dobrinsky, J.R. (1996). Cellular approach to cryopreservation of embryos. Theriogenology 45, 1726.
Fornari, D.C., Ribeiro, R.P., Streit, D.P. Jr., Vargas, L. & Moraes, V.G. (2010). Freezing injuries in the embryos of Piaractus mesopotamicus. Zygote 18, 16.
Hagedorn, M., Hsu, E., Kleinhans, F.W. & Wildt, D.E. (1997a). New approaches for studying the permeability of fish embryos: toward successful cryopreservation. Cryobiology 34, 335347.
Hagedorn, M., Kleinhans, F.W., Weidt, D.E. & Rall, W.E. (1997b). Chill sensitivity and cryoprotectant permeability of dechorionated zebrafish embryos, Brachydanio rerio. Cryobiology, 34, 251–63.
Hagedorn, M., Kleinhans, F.W., Artmov, D., Artmov, D. & Pilatus, U. (1998). Characterization of a major permeability barrier in the zebrafish embryo. Biol. Reprod. 59, 1240–50.
Hagedorn, M., Peterson, A., Mazur, P. & Kleinhans, F.W. (2004). High ice nucleation temperature of zebrafish embryos: slow-freezing is not an option. Cryobiology 49, 181–9.
Harvey, B. (1983). Cooling of embryonic cells, isolated blastoderms, and intact embryos of the zebrafish Brachydanio rerio to –196 Celsius. Cryobiology 20, 440–7.
IBAMA. (2007). Estatística da pesca 2005: Brasil, grandes regiões e unidades da federação. Brasília: IBAMA, 2007. 137 pp.
Janik, M., Kleinhans, F.W. & Hagedorn, M. (2000). Overcoming a permeability by microinjecting cryoprotectants into zebrafish embryos (Brachidanio rerio) Cryobiology 41, 2534.
Kasai, M. (1996). Simple and efficient methods for vitrification of mammalian embryos. Anim. Reprod. Sci. 42, 6775.
Mazur, P. (1984). Freezing of living cells: mechanisms and implications. Am. J. Physiol. 247, 125142.
Ninhaus-Silveira, A., Foresti, F. & Azevedo, A. (2006). Structural and ultrastructural analysis of embryonic development of Prochilodus lineatus (Valenciennes, 1836) (Characiforme; Prochilodontidae). Zygote 14, 217–29.
Ninhaus-Silveira, A., Foresti, F., Azevedo, A. & Augustine, C.A. (2007). Structural and ultrastructural characteristics of the yolk syncytial layer in Prochilodus lineatus (Valenciennes, 1836) (Teleostei, Prochilodontidae). Zygote 15, 267–71.
Robles, V., Cabrita, E., Real, M., Alvarez, R. & Herraez, M.P. (2003). Vitrification of turbot embryos: preliminary assays. Cryobiology 47, 30–9.
Streit, D.P. Jr, Digmayer, M., Ribeiro, R.P., Sirol, R.N., Moraes, G.V. & Cock, J.M. (2007). Embriões de pacu submetidos a diferentes protocolos de resfriamento. Pesquisa Agropecuária Brasileira 42, 1199–202.
Vajta, G. (2000). Vitrification of the oocytes and embryos of domestic animals. Anim. Reprod. Sci. 60–61, 357–64.
Wildt, D.E., Seal, U.S. & Rall, W.F. (1993). Genetic resource banks and reproductive technology for wildlife conservation. In: Genetic Conservation of Salmonid Fishes. (Cloud, J.G. and Thorgaard, G.H., eds) pp. 159–73.
Zhang, T. & Rawson, D.M. (1995). Studies on chilling sensitivity of zebrafish (Brachydanio rerio) embryos. Cryobiology 32, 239–46.
Zhang, T. & Rawson, D.M. (1996). Studies on vitrification of intact zebrafish (Brachydanio rerio) embryos. Cryobiology 33, 113.
Zhang, T., Rawson, D.M. & Morris, G.J. (1993). Cryopreservation of pre-hatch embryos of zebrafish (Brachydanio rerio) embryos. Aquat. Living Resour. 6, 145–53.

Keywords

Injuries in pacu embryos (Piaractus mesopotamicus) after freezing and thawing

  • Patrícia Ribeiro Neves (a1), Ricardo Pereira Ribeiro (a2), Danilo Pedro Streit (a3), Maria Raquel M. Natali (a2), Darci Carlos Fornari (a4), Alexandra Inês Santos (a5) and Leandro C. Godoy (a3)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed