Comparison of Development and Antioxidative Ability in Fertilized Crossbred (Yorkshire × Landrace × Duroc) Oocytes Using Duroc and Landrace Sperm

Pig production through crossbreeding methods is a pillar of the swine industry; however, research on the fertilization ability of male pigs in crossbreeds is lacking. Therefore, this study investigated the effects of Duroc sperm (DS) and Landrace sperm (LS) on fertility in Yorkshire × Landrace × Duroc (YLD) oocytes. Sperm were collected from the Duroc and Landrace species, and sperm characteristics, viability, and acrosome reactions were analyzed using flow cytometry. Oocytes were collected from YLD ovaries, and the fertility of DS and LS was determined using in vitro fertilization (IVF). Reactive oxygen species (ROS) and antioxidative abilities were analyzed using H₂DCFDA and a Cell Tracker Red assay. Pluripotency (<i>OCT4</i>, <i>SOX2</i>, and <i>NANOG)</i>, antioxidative (<i>SOD1</i>, <i>SOD2</i>, <i>CAT</i>, and <i>GPx1</i>), apoptotic (<i>Bax</i> and <i>Bcl-2</i>), and cell cycle-related (<i>Cdc2</i> and <i>CCNB1</i>) genes were detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in oocytes fertilized with sperm. The results showed no significant difference in viability or acrosome reaction between DS and LS. ROS levels were significantly lower in the LS group than in the DS group, whereas glutathione (GSH) levels in the embryo did not significantly differ between the DS and LS groups. The <i>OCT4</i>, <i>GPx1</i>, and <i>Cdc2</i> mRNA expression levels were significantly higher in the LS than DS groups. Blastocyst formation was significantly higher in the LS than DS groups. ROS levels were reduced, and blastocyte formation was increased in LS-obtained embryos. In conclusion, these results provide a fundamental understanding of using Landrace semen in the three-way crossbreeding of YLD pigs.