Revival of cryopreserved larvae from the important aquaculture species Pacific White Shrimp (Litopenaeus vannamei) using vitrification and ultra-rapid laser warming

Abstract

Cryopreservation of aquatic embryos or larvae is needed to help safeguard genetics from important wild and captive species, increase aquaculture output, and meet the global demand for protein. To this end, the development of a cryopreservation protocol for nauplius larvae of the commercially important aquaculture species Litopenaeus vannamei, or Pacific White Shrimp, was pursued. Toxicity screening was performed using multiple cryoprotective agents (CPA), and a multi-constituent CPA cocktail was developed to achieve reliable vitrification of shrimp larvae encapsulated in 1.0-μL droplets containing gold nanoparticles. Vitrification and ultra-rapid laser warming were used to cryopreserve and revive nauplius-V stage larvae. Laser warming parameters were optimized to protect the pigmented eye spot from laser-induced ablation, and ice recrystallization inhibitors (IRIs) were tested to induce long-term survival. Approximately 54 % of revived larvae resumed active swimming, but all failed to molt to the zoea-I stage of development or live beyond 15 h post warming.