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Cryoprotectant agents and cooling effect on embryos of Macrobrachium amazonicum

Published online by Cambridge University Press:  15 April 2018

Caroline Costa Lucas ,
Luana Rolim Melo ,
Míriam Luzia Nogueira Martins de Sousa ,
Glayciane Bezerra de Morais ,
Moisés Fernandes Martins ,
Francisco Antônio Félix Xavier Junior ,
Janaina Serra Azul Monteiro Evangelista  and
Célia Maria de Souza Sampaio
Caroline Costa Lucas*
Affiliation:
Department of Biology, State University of Ceará, UECE, Fortaleza, CE, 60740-000, Brazil.
Luana Rolim Melo
Affiliation:
Postgraduate Program in Veterinary Science, UECE, Fortaleza, CE, Brazil.
Míriam Luzia Nogueira Martins de Sousa
Affiliation:
Postgraduate Program in Veterinary Science, UECE, Fortaleza, CE, Brazil.
Glayciane Bezerra de Morais
Affiliation:
Postgraduate Program in Veterinary Science, UECE, Fortaleza, CE, Brazil.
Moisés Fernandes Martins
Affiliation:
Department of Biology, State University of Ceará, UECE, Fortaleza, CE, Brazil.
Francisco Antônio Félix Xavier Junior
Affiliation:
Department of Medicine Veterinary, State University of Ceará, UECE, Fortaleza, CE, Brazil.
Janaina Serra Azul Monteiro Evangelista
Affiliation:
Department of Medicine Veterinary, State University of Ceará, UECE, Fortaleza, CE, Brazil.
Célia Maria de Souza Sampaio
Affiliation:
Department of Biology, State University of Ceará, UECE, Fortaleza, CE, Brazil.
*
All correspondence to: Caroline Costa Lucas. Department of Biology, State University of Ceará, UECE, Fortaleza, CE, 60740-000, Brazil. Tel:/Fax: +85 3101 9927. E-mail: [email protected]
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Summary

There are few reports of cryopreservation and injuries in Macrobrachium amazonicum embryos. Thus, the aim of this study was to analyze the effects of cryoprotectants agents and cooling on stage VIII of this species. Fertilized eggs from ovigerous females were removed from the incubation chamber, then placed in 10 ml Falcon tubes with a cryoprotectant solution and saline-free calcium solution. Thus, the embryos underwent a cooling curve of 1°C per min until reaching 5°C, and then were stored for 2 h. The tubes containing the embryos were washed to remove the cryoprotectant, acclimated for 5 min and then transferred to 50 ml incubators. At the end of the 24-h period, living embryos from each tube were counted and tabulated. A pool of embryos was fixed with 4% formaldehyde and then subjected to histology using 3-mm thick sections and stained with haematoxylin/eosin. Another pool was used for biometric analysis in which length, width and volume were analyzed. The cryoprotectants agents used were: dimethylsulfoxide (DMSO), methyl alcohol, ethylene glycol at 1, 5 and 10% and sucrose (0.5 M). Variance analysis was performed followed by Tukey's honest significant difference (HSD) test at 5% significance level. DMSO cryoprotectant affected embryo survival the least with rates of 71.8, 36.2 and 0% for concentrations of 1, 5 and 10%, respectively. Ethylene glycol caused 100% mortality at all the concentrations used. It was not possible to observe the interference of cooling and cryoprotectants on embryonic structures in this study.

Keywords

Amazon river prawnChillingCooling protectorsFertilized eggsMorphology
Type
Research Article
Information
Zygote , Volume 26 , Issue 2 , April 2018 , pp. 111 - 118
Copyright
Copyright © Cambridge University Press 2018 

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