Veterinary Research Forum

Abstract This study aimed to determine whether the sperm acrosome-associated 1 (SPACA1) gene exhibits differential expression between X- and Y-bearing spermatozoa of Bali bulls and assess its potential as an RNA biomarker for sperm sexing. Sexed frozen semen (X or Y fractions) from Bali bulls was obtained from the Singosari National Artificial Insemination Center, Malang, Indonesia. Post-thaw semen quality was evaluated for motility, viability, abnormality, concentration, and plasma membrane integrity. Total RNA was extracted separately from X and Y fractions using easy-BLUE reagent, and SPACA1 expression was quantified by RT-qPCR using GAPDH as an internal control. Relative expression was calculated by the 2^−ΔΔCt method, and group differences were analyzed with the Mann-Whitney U test. The SPACA1 expression was numerically higher in X-bearing than Y-bearing spermatozoa (1.00 versus 0.51), but the difference was not significant. Thus, SPACA1 transcripts do not provide a reliable molecular marker for sperm sex differentiation. Future work should focus on protein-level characterization of SPACA1 and exploration of sex-biased surface proteins for immunological sexing. This research supports Sustainable Development Goal 2: Zero Hunger, by contributing to reproductive efficiency in livestock, and Sustainable Development Goal 12: Responsible Consumption and Production, through development of cost-effective, sustainable sperm sexing technologies.

Abstract In vitro maturation is a critical step in in vitro fertilization, significantly impacting oocyte quality and subsequent embryonic development. This study examines how insulin-like growth factor (IGF)-1 supplementation in the maturation medium affects mitogen-activated protein kinase and maturation promoting factor expressions, as well as the amount of mitochondrial DNA copies in Kacang goat oocytes. Oocytes were collected from Kacang goat ovaries and matured in vitro with varying IGF-1 concentrations, including 0.00 (control), 50.00, 100, and 150 ng mL^-1. Immuno-cytochemistry was used to assess mitogen-activated protein kinase and maturation promoting factor expressions, while quantitative polymerase chain reaction quantified mitochondrial DNA copy number. Results showed that the 100 ng mL^-1 dose of IGF-1 group had substantially increased mitogen-activated protein kinase and maturation promoting factor expressions and also mitochondrial DNA copy numbers compared to the other groups. These findings suggest that IGF-1 supplementation with 100 ng mL^-1 dose optimally enhances oocyte maturation by activating key signaling pathways and promoting mitochondrial replication. In conclusion, IGF-1 supplementation at 100 ng mL^-1 is recommended to improve oocyte quality in Kacang goats, potentially enhancing in vitro fertilization outcomes.