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

Published online by Cambridge University Press:  11 July 2012

Patrícia Ribeiro Neves ,
Ricardo Pereira Ribeiro ,
Danilo Pedro Streit Jr ,
Maria Raquel M. Natali ,
Darci Carlos Fornari ,
Alexandra Inês Santos  and
Leandro C. Godoy
Patrícia Ribeiro Neves
Affiliation:
Departamento Zootecnia – Universidade Estadual de Maringá, Av. Colombo, 5790 – Campus Universitário, CEP: 87020-900 Maringá, Paraná, Brasil.
Ricardo Pereira Ribeiro
Affiliation:
Universidade Estadual de Maringá (UEM) – Depto. de Zootecnia e Biologia–Maringá, Paraná State, Brasil.
Danilo Pedro Streit Jr
Affiliation:
Universidade Federal do Rio Grande do Sul – UFRGS, Brasil.
Maria Raquel M. Natali
Affiliation:
Universidade Estadual de Maringá (UEM) – Depto. de Zootecnia e Biologia–Maringá, Paraná State, Brasil.
Darci Carlos Fornari*
Affiliation:
Departamento Zootecnia – Universidade Estadual de Maringá, Av. Colombo, 5790 – Campus Universitário, CEP: 87020-900 Maringá, Paraná, Brasil.
Alexandra Inês Santos
Affiliation:
Universidade Estadual de Maringá (UEM) – Grupo de Pesquisa PeixeGen, Brasil.
Leandro C. Godoy
Affiliation:
Universidade Federal do Rio Grande do Sul – UFRGS, Brasil.
*
All correspondence to: Darci Carlos Fornari. Departamento Zootecnia – Universidade Estadual de Maringá, Av. Colombo, 5790 – Campus Universitário, CEP: 87020-900 Maringá, Paraná, Brasil. Tel: +5544 3261 8969. e-mail: darci.peixegen@gmail.com
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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.

Keywords

CryoprotectantMethanolMorphologicalPacuReproductionSucrose
Type
Research Article
Information
Zygote , Volume 22 , Issue 1 , February 2014 , pp. 25 - 31
Copyright
Copyright © Cambridge University Press 2012 

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