Optimization of Seminal Cryopreservation in Swine and Identification of Molecular Biomarkers of Cryotolerance

• Grant number: 25/04918-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2025
• End date: October 31, 2027
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: André Furugen Cesar de Andrade
• Grantee: Henricco Zapparoli e Silva
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 24/09891-0 - Small molecules and miRNAs as cryotolerance and fertility biomarkers in boar semen, AP.R

Abstract

Swine semen cryopreservation represents a challenge due to the high variability in freezing tolerance among ejaculates and boars. This study aims to investigate the potential of micro-RNAs ssc-miR-503, ssc-miR-130a, and ssc-miR-9 as biomarkers of sperm cryotolerance, allowing the prior identification of ejaculates with lower freezing capacity. The study is divided into two experiments. In the first, ejaculates from boars of different commercial lineages will be collected and analyzed for motility, plasma and acrosomal membrane integrity, followed by micro-RNA extraction and quantification via RT-qPCR. In the second experiment, semen will be cryopreserved using different extenders and sperm concentrations in the straws to evaluate the impact of these variables on post-thaw viability. Computer-assisted sperm analysis (CASA) and flow cytometry will be used to assess sperm quality. The expected results include confirming the relationship between the selected miRNAs and lower cryotolerance and optimizing freezing protocols to enhance the practical application of cryopreservation in swine reproduction.