Utilization of Vitamin C as an Antioxidant in the Cryopreservation of Astyanax lacustris Semen

• Grant number: 24/10393-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2024
• End date: August 31, 2025
• Field of knowledge: Biological Sciences - Zoology - Applied Zoology
• Principal Investigator: Alexandre Ninhaus Silveira
• Grantee: Nathaly Godoy da Silva
• Host Institution: Faculdade de Engenharia (FEIS). Universidade Estadual Paulista (UNESP). Campus de Ilha Solteira. Ilha Solteira , SP, Brazil
• Associated scholarship(s): 25/00149-1 - Technical study of the characteristics of sperm kinetics in freshwater fish using a computerized seminal analysis system and cryopreservation of fish semen., BE.EP.IC

Abstract

Astyanax lacustris is a species of fish native to the Paraná River, with great importance for the aquatic ecosystem of this river and also for the country's aquaculture production, related to its use as live bait in sport fishing, for human consumption and, in recent years, as a model species for scientific experiments. Therefore, the development of a genetic and/or a germplasm bank for the species is essential for projects that work to maintain the native biodiversity, as it can also be used to optimize commercial production by inserting improved genes into the breeding stock. In this sense, this project aims to evaluate the effect of vitamin C supplementation, as an antioxidant substance, in the cryoprotective solution composition in the cryopreservation of A. lacustris semen with a view to improving the quality of cryopreserved semen. To this end, adult and mature males will be used, and seminal characteristics will be evaluated using CASA System (Computer Assisted Sperm Analysis). For the cryopreservation experiment, seven pools (15 animals per pool) per treatment will be carried out, the semen will be diluted in a 1:9 ratio (semen:solution), using diluents composed of 10% dimethylformamide + 0.8% NaCl, supplemented with vitamin C, at test dosages of 0, 1, 2 and 4 mM. Packaged in 0.25mL straws frozen in nitrogen vapor in a Styrofoam box, positioned 2 cm from the surface of the nitrogen for 10 mins, in a cryogenic cylinder (-195ºC). The cryopreserved semen will be thawed, subjected to sperm motility and morphology using CASA System, membrane integrity and fertilization analyses. All data will be subjected to statistical analysis for conclusions.