Effects of Sucrose and Different Cryoprotectants on Sperm Quality, DNA Damage, and Fertilization Rate During Cryopreservation of Mesopotamian Catfish (Silurus triostegus) Sperm
DOI:
https://doi.org/10.24925/turjaf.v14i3.677-685.8394Keywords:
Silurus triostegus, cryopreservation, sucrose, sperm motility, DNA damage, fertilizationAbstract
This study aimed to investigate the effects of sucrose added to a glucose-based extender in combination with different cryoprotectants such as dimethyl sulfoxide (DMSO), methanol, and methylglycol on post-thaw sperm motility, DNA damage, and fertilization rates in the cryopreservation of Mesopotamian catfish (Silurus triostegus) semen. Semen samples obtained by abdominal massage from individuals (4-5 years old) from Atatürk Dam Lake were used. Samples were diluted at a 1:3 ratio with glucose extenders (0.3 M glucose + 20% egg yolk) containing 5% or 10% DMSO, methanol, or methylglycol with or without 50 mM sucrose. Diluted samples were frozen in liquid nitrogen vapor and stored at -196°C. Post-thaw sperm motility rate and duration, DNA damage (8-OHdG ELISA and Comet Assay), apoptosis (ELISA), and fertilization rates were evaluated. Data were analyzed using ANOVA and Duncan's test. The highest post-thaw sperm motility rate (30.00 ± 3.51%) and motility duration (45.00 ± 6.50 s) were detected in the 5% DMSO group (P<0.05). The control group containing only sucrose showed the lowest motility (10.00 ± 4.50%). According to the 8-OHdG test, the methanol+sucrose group showed the highest DNA damage (123.86 ± 4.08 ng/ml) (P<0.05), while the DMSO+sucrose group caused low-level damage similar to the control groups. In the Comet Assay, the DMSO+sucrose group had significantly lower DNA damage compared to the DMSO control group. The highest fertilization rates were obtained in the DMSO (24.66 ± 3.51%), methylglycol (20.66 ± 1.15%), and DMSO+sucrose (20.33 ± 1.52%) groups (P<0.05). No fertilization was observed in the group containing only sucrose (0.0 ± 0.00%). Study findings showed that 5% DMSO and 50 mM sucrose containing glucose extender was the most suitable combination for cryopreservation of S. triostegus sperm because it provided high sperm motility, low DNA damage and satisfactory fertilization rates after thawing.
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