SPERM CRYOPRESERVATION OF GIANT GROUPER Epinephelus lanceolatus USING TUALANG HONEY AS A NATURAL ALTERNATIVE CRYOPROTECTIVE AGENT
DOI:
https://doi.org/10.46754/Keywords:
Sustainability, cryoprotective agent, cooling rate, antioxidant, post-thaw motilityAbstract
The current sperm cryopreservation protocol for Epinephelus lanceolatus uses Dimethyl Sulfoxide (DMSO) as a Cryoprotective Agent (CPA), which may exhibit toxicity at high concentrations. To address this limitation, this study investigated Tualang Honey (TH) as a natural, non-toxic alternative CPA. Specifically, this study aimed to determine the best concentration of TH and TH mixed with DMSO (THD), as well as the optimum cooling protocol when using TH as a CPA in the sperm cryopreservation protocol of E. lanceolatus. We used five concentrations (5%, 10%, 15%, 20%, and 25%) of TH and five combinations (5%, 10%, 15%, 20%, and 25%) of THD (1:1 TH:DMSO) as CPA. The cooling protocol using TH was optimised by manipulating the cooling heights (3, 5, 7, and 9 cm) during the cooling process. Sperm Post-Thaw Motility (PTM) and motility duration were used to evaluate sperm quality. All data were presented as mean ± Standard Error (SE) and analysed using one-way Analysis of Variance (ANOVA, p < 0.05) followed by Tukey’s post hoc test. The results revealed that all concentrations of TH and THD were not significantly different (p > 0.05) from the control (42.00% ± 10.0%). Among all treatments, 20% TH and 15% THD yielded the highest PTM values (29.33% ± 1.33% and 25.67% ± 5.17%, respectively). Furthermore, the highest PTM value was obtained when cooling at 7 cm above the Liquid Nitrogen (LN) surface (42.00% ± 1.00%). Overall, the results suggest that TH and THD can be used as alternatives to CPA for sperm cryopreservation of E. lanceolatus, with 20% TH providing the highest PTM values.
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