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Cryopreservation Protocol Optimization for Penaeus monodon Sperm: Reagent Screening and Parameter Refinement
Dewei Kong1, Song Jiang1,2, Jianzhi Shi1
1South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, Guangzhou 510300, China.
Biology
|April 26, 2025
Summary
This study optimized cryopreservation for black tiger shrimp (Penaeus monodon) sperm using natural seawater and 10% dimethyl sulfoxide (DMSO). The developed protocol achieved a 53.33% survival rate after 15 days of liquid nitrogen storage.
Area of Science:
- Aquaculture and Marine Biology
- Reproductive Biology and Cryopreservation
Background:
- Penaeus monodon (black tiger shrimp) is a vital species in global aquaculture.
- Effective sperm cryopreservation is crucial for genetic resource conservation and sustainable aquaculture development.
Purpose of the Study:
- To establish an optimal cryopreservation protocol for Penaeus monodon sperm.
- To evaluate the effects of different diluents, cryoprotectants, equilibration times, cooling rates, and thawing temperatures.
Main Methods:
- Screening of diluents (natural seawater) and cryoprotectants (dimethyl sulfoxide - DMSO).
- Optimization of equilibration (4°C for 30 min), controlled-rate freezing (-5°C/min to -20°C, -10°C/min to -80°C, -20°C/min to -180°C), and thawing (37°C water bath).
- Assessment of sperm motility, survival rates, structural integrity (SEM, TEM), and enzyme activity (AKP, ACP, MDA, CAT, T-SOD) post-cryopreservation.
Main Results:
- The optimal protocol utilized natural seawater with 10% DMSO, 1:1 (v/v) ratio, 30 min equilibration at 4°C, followed by programmable cooling and 37°C thawing.
- Sperm survival rate reached 53.33 ± 9.18% after 15 days of liquid nitrogen storage.
- Cryopreservation induced some structural damage (acrosomal spikes, membrane) but enzyme activity analysis revealed changes in AKP, ACP, MDA, CAT, and T-SOD over storage time.
Conclusions:
- A robust cryopreservation protocol for Penaeus monodon sperm has been successfully developed.
- This protocol, involving specific diluents, cryoprotectants, and freezing/thawing parameters, significantly improves sperm cryosurvival.
- Findings offer critical insights for optimizing cryopreservation techniques to enhance sperm viability and support Penaeus monodon aquaculture.
