Cryogenic tolerance to freezing temperature and sperm motility characteristics of frozen-thawed spermatozoa of Philippine native pigs (Sus scrofa)
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Abstract
Philippine native pigs (PnPs) in rural farming communities still thrive due to their adaptability and resilience to adverse environment challenges, perceived disease resistance traits and consumer demand for their tastier meat. These qualities make them likely candidates for gametes conservation for Animal Genetic Resources (AnGRs) Cryobanking. PnPs spermatozoa were cryopreserved using Lactose egg yolk-based Cooling Extender (CE; 0.31M Lactose 80% v/v and 20% Egg yolk v/v) and Freezing Extender (FE) solutions (CE 92.5% v/v, Glycerol 6% v/v and Equex paste 1.5% v/v). With the use of a Computer Assisted Sperm Analyzer (CASA) the average ± s.e.m sperm motility (CASA-MOT) and progressively motile sperm (CASA PMOT) were determined before and after cryopreservation. In study 1, epididymal spermatozoa (EpS) had a pre-freeze CASA MOT values of 71.75 ± 2.32 % and CASA PMOT of 44.0 ±1.96%, that decreased significantly to 30.1± 5.0% and 12.0 ± 3.0% after freezing, respectively (Student’s T-test:p<0.05). In study 2, ejaculated semen of native Boar stud #1 and #2 registered a pre-freezing CASA MOT values of 53.1±3.6% and 52.3± 3.0% with sperm post thaw motility values of 29.1 ±4.1% and 31.0± 4.9%, respectively.The CASA PMOT values for Boar Stud #1 and #2 before cryopreservation was 45.9 ±3.6% and 46.4.±3.3%, which manifested a significant decline to 25.0 ± 3.9% and 26.6 ± 4.6, respectively. Fluorescence microscopy using Live/Dead® Sperm Viability Kit revealed sperm head with green fluorescence confirming live sperm and viability indicative of intact plasma membrane integrity post thawing. These results demonstrated the cryogenic tolerance to freezing temperature and survivability of Philippine native pig spermatozoa for future research on in vitro fertilization and eventual AnGR cryo-conservation/cryobanking.
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