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Effect of cryoprotectants on the survival of cascudo preto (Rhinelepis aspera) embryos stored at –8°C

Published online by Cambridge University Press:  28 August 2011

Darci Carlos Fornari ,
Ricardo Pereira Ribeiro ,
Danilo Streit Jr ,
Leandro Cesar Godoy ,
Patrícia Ribeiro Neves ,
Diego de Oliveira  and
Rodolfo Nardez Sirol
Darci Carlos Fornari*
Affiliation:
Departamento de Zootecnia, Universidade Estadual de Maringá, Av. Colombo 5790, CEP 87020–900, Maringá-PR, Brazil.
Ricardo Pereira Ribeiro
Affiliation:
PeixeGen Research Group, Maringa State University, Department of Animal Science, Maringa, Brazil.
Danilo Streit Jr
Affiliation:
Aquam Research Group, Federal University of Rio Grande do Sul, Department of Animal Science, Porto Alegre, Brazil.
Leandro Cesar Godoy
Affiliation:
Aquam Research Group, Federal University of Rio Grande do Sul, Department of Animal Science, Porto Alegre, Brazil.
Patrícia Ribeiro Neves
Affiliation:
Ponta Grossa State University, Department of Animal Science, Ponta Grossa, Brazil.
Diego de Oliveira
Affiliation:
Aquam Research Group, Federal University of Rio Grande do Sul, Department of Animal Science, Porto Alegre, Brazil.
Rodolfo Nardez Sirol
Affiliation:
Environment manager, CPFL Energy, São Paulo, Brazil.
*
All correspondence to: Darci Carlos Fornari. Departamento de Zootecnia, Universidade Estadual de Maringá, Av. Colombo 5790, CEP 87020–900, Maringá-PR, Brazil. Tel: +55 44 3261 8969. Fax: +55 44 3261 4729. E-mail: darci.peixegen@gmail.com
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Summary

Cryopreservation of germplasm provides a promising method to preserve fish genetic material, which is of great importance in preservation of species diversity, aquaculture, and management of fish models used in biomedical research. In the present study, cryopreservation of Rhinelepis aspera embryos, a Brazilian endangered species, was studied for the first time using a short-term cooling protocol. Embryos at blastoporous closing stage were selected, placed in 6-ml glass vials and stored at −8°C for 6 h in 10 different cryoprotectant solutions: S1 (17.1% sucrose + 9% methanol); S2 (17.1% sucrose + 9% DMSO); S3 (8.5% sucrose + 8.5% glucose + 9% methanol); S4 (8.5% sucrose + 8.5% glucose + 9% DMSO); S5 (17.1% sucrose + 9% ethylene glycol); S6 (8.5% sucrose + 8.5% glucose + 9% ethylene glycol); S7 (17.1% sucrose + 4.5% methanol + 4.5% DMSO); S8 (17.1% sucrose + 4.5% methanol + 4.5% ethylene glycol); S9 (17.1% sucrose + 4.5% DMSO + 4.5% ethylene glycol); and S10 (100% water). Embryo viability was assessed by hatching rate, counting live larvae and number of failed eggs under a stereomicroscope. The results showed that only the cryoprotectant solutions that contained methanol associated to sucrose (S1, S7 and S8) provided partial protection of Rhinelepis aspera embryos from cold damage (over 50% hatching rate in S1), while the use of DMSO and ethylene glycol, isolated or in combination, resulted in no hatching rate. Further studies are needed in order to extend the storage time and to improve the hatching rate for the species.

Keywords

ConservationCryobiologyFreshwater fishGermplasm bankingReproduction management
Type
Research Article
Information
Zygote , Volume 22 , Issue 1 , February 2014 , pp. 58 - 63
Copyright
Copyright © Cambridge University Press 2011 

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