Oocyte cryopreservation of Prochilodus lineatus (Valenciennes, 1836)

• Grant number: 13/10807-9
• Support Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2013
• End date: February 28, 2015
• Field of knowledge: Agronomical Sciences - Fishery Resources and Fishery Engineering - Aquaculture
• Principal Investigator: Alexandre Ninhaus Silveira
• Grantee: Douglas de Castro Ribeiro
• Host Institution: Faculdade de Engenharia (FEIS). Universidade Estadual Paulista (UNESP). Campus de Ilha Solteira. Ilha Solteira , SP, Brazil

Abstract

This project aims to develop a protocol for oocyte freezing Prochilodus lineatus. To this solution will be evaluated three extensor (Ringer's solution, Hank's solution, and phosphate buffer) and three cryoprotectant substances (Dimethyl sulfoxide, propylene glycol and methanol) at a concentration of 0.2M to 4M linked glucose. The oocytes are packaged in straws of 0.5 ml and exposed to a cooling curve 2°C/min to -12.5°C (for 5 minutes), 0.3°C/min to -40°C, 10°C min to -80°C and 50°C/min to -160°C, and then immersed in liquid nitrogen (LN2) and stored for 24 hours. The samples are defrosted in a "bath" to 27°C for 10s, and rehydrated by soaking in solutions of decreasing osmolarity of cryoprotectants (2M, 1M, 0.5M). The subsequent membrane integrity is assessed by using trypan blue dye, and the viability of the oocytes fertilization. The experiment will include samples consisting of 100 oocytes and 5 replicates in each treatment and their data were assessed for normality (Shapiro-Wilk) and homogeneity (Bartlett) and means were compared using an analysis of variance (ANOVA) and Tukey (± = 0 , 05). Morphologically, the embryos will undergo optical microscopy (stained with PAS, HE and toluidine blue), scanning electron microscopy and transmission electron microscopy. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

RIBEIRO, D. C.; CHAGAS, J. M. A.; BASHIYO-SILVA, C.; COSTA, R. S.; VERISSIMO-SILVEIRA, R.; NINHAUS-SILVEIRA, A.. Oocyte viability and cortical activation under different salt solutions in Prochilodus lineatus (Teleostei: Prochilodontidae). REPRODUCTION IN DOMESTIC ANIMALS, v. 52, n. 6, p. 932-938, DEC 2017. (13/10807-9)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

RIBEIRO, Douglas de Castro. Criopreservação de oócitos de Prochilodus lineatus (Valenciennes, 1836) (Teleostei, Prochilodontidae). 2015. Master's Dissertation - Universidade Estadual Paulista (Unesp). Faculdade de Engenharia. Ilha Solteira Ilha Solteira.