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Abstract
Coral reefs are suffering on a global scale due to human impacts, thereby necessitating cryopreservation efforts. The objective of this study was to develop a suitable vitrification and laser warming protocol for larvae of the scleractinian coral Seriatopora caliendrum, which inherit their dinoflagellate algal symbionts vertically. Toxicity experiments were conducted with the cryoprotectants (CPAs) ethylene glycol (EG), propylene glycol (PG), dimethyl sulfoxide (DMSO), glycerol (GLY), and methanol (METH; listed in order from least to most toxic), and larvae were subjected to vitrification and laser warming using 2 M EG + 1 M PG and 2 M EG + 1 M DMSO. Vitrification and laser warming (300 V, 10 ms pulse width, 2 mm beam diameter) using a vitrification solution of 2 M EG + 1 M PG, 40% w/v Ficoll, and 10% v/v gold nanobars (GNB) at a final concentration of 1.2 × 1018 GNB/mL and a characteristic wavelength of 535 nm resulted in larvae with vitality and settlement percentages of 55 and 9%, respectively. This represents the first successful instance of cryopreservation of coral larvae that proceeded to settle upon warming, and suggests that the vitrification and ultra-fast laser warming approach may be applicable to other threatened marine species.
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1 Department of Marine Biotechnology and Resources, National Sun Yai-sen University, Kaohsiung, Taiwan; National Museum of Marine Biology & Aquarium, Pingtung, Taiwan
2 Department of Marine Biotechnology and Resources, National Sun Yai-sen University, Kaohsiung, Taiwan
3 He Wei Precision Company Limited, Hsinchu, Taiwan
4 Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan
5 National Museum of Marine Biology & Aquarium, Pingtung, Taiwan; Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan
6 Department of Marine Biotechnology and Resources, National Sun Yai-sen University, Kaohsiung, Taiwan; National Museum of Marine Biology & Aquarium, Pingtung, Taiwan; Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan
7 Smithsonian Conservation Biology Institute, Washington, USA; Hawaii Institute of Marine Biology, Hawaii, USA
8 Department of Post Modern Agriculture, Mingdao University, Chang Hua, Taiwan