Reconstitution of fish species in danger of extinction: obtaining androgenetic Brycon orbignyanus

• Grant number: 14/16699-6
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: February 01, 2015
• End date: May 31, 2018
• Field of knowledge: Agronomical Sciences - Fishery Resources and Fishery Engineering - Aquaculture
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Pedro Manoel Galetti Junior
• Grantee: Francine Faustino Theodoro Costa
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Piracanjuba Brycon orbignyanus is a Brycon species in danger of extinction. For its economic and ecological importance, it's necessary and even imperative that there are efforts in preserving it, as the consolidation of ex situ gene banks. An alternative for maintenance of this species can be reconstitution by androgenesis, which generates progeny with only a paternal genome. This process comprises two main steps: inactivation of the oocyte genome, activation of the embryonic development from haploid gametes and, finally, recovery of diploid embryos from suppression of the first mitotic cleavage. The purpose of this study is to create a protocol for reconstitution of B. orbignyanus using androgenesis technique. The first step was partially developed during the PhD candidate in the previous experiments, with the matrinxã B. amazonicus, the oocyte donor species, the optimal solution has been defined for the preservation of these oocytes (UV irradiation medium). For the completion of this first stage will be followed the protocol found, being tested doses of UV irradiation for complete inactivation of the oocyte genome. Subsequently, the second step is executed, activating the embryonic development of fresh semen B. orbignyanus and finally the third stage of the technique represented by the recovery of the androgenetic embryo diploidy by deleting the first mitotic cleavage by shock heat. To confirm the ploidy of androgenetic embryos is essential to analysis by flow cytometry, and to prove the paternity of these androgenetic embryos is indispensable to perform paternity testing using molecular markers. If successful technique, this work can be considered the pioneer in involving androgenesis native species and provide an effective tool for use in ex situ gene banks. (AU)