Coral reefs are experiencing unprecedented degradation due to human activities, and protecting specific reef habitats may not stop this decline, because the most serious threats are global (i.e., climate change), not local. However, ex situ preservation practices can provide safeguards for coral reef conservation. Specifically, modern advances in cryobiology and genome banking could secure existing species and genetic diversity until genotypes can be introduced into rehabilitated habitats. We assessed the feasibility of recovering viable sperm and embryonic cells post-thaw from two coral species, Acropora palmata and Fungia scutaria that have diffferent evolutionary histories, ecological niches and reproductive strategies. In vitro fertilization (IVF) of conspecific eggs using fresh (control) spermatozoa revealed high levels of fertilization (>90% in A. palmata; >84% in F. scutaria; P>0.05) that were unaffected by tested sperm concentrations. A solution of 10% dimethyl sulfoxide (DMSO) at cooling rates of 20 to 30°C/min most successfully cryopreserved both A. palmata and F. scutaria spermatozoa and allowed producing developing larvae in vitro. IVF success under these conditions was 65% in A. palmata and 53% in F. scutaria on particular nights; however, on subsequent nights, the same process resulted in little or no IVF success. Thus, the window for optimal freezing of high quality spermatozoa was short (∼5 h for one night each spawning cycle). Additionally, cryopreserved F. scutaria embryonic cells had∼50% post-thaw viability as measured by intact membranes. Thus, despite some differences between species, coral spermatozoa and embryonic cells are viable after low temperature (−196°C) storage, preservation and thawing. Based on these results, we have begun systematically banking coral spermatozoa and embryonic cells on a large-scale as a support approach for preserving existing bio- and genetic diversity found in reef systems.
Abstract. Bleaching profoundly impacts coral reproduction, often for years after an event. However, detailed reproductive characteristics of coral after bleaching have not been broadly described, especially as they relate to cryopreservation. Therefore, in the present study we measured several reproductive characteristics in coral in Kaneohe Bay, Hawaii, for two species, namely Fungia scutaria and Montipora capitata, during the bleaching period of 2014 and 2015. We examined spawning periods, egg morphometry, sperm concentration, fresh and cryopreserved sperm motility exposed to different concentrations of dimethyl sulfoxide, time of first cleavage, larval survival with fresh and cryopreserved spermatozoa, infection success and settlement success. Many of these reproductive parameters were reduced in 2015, especially sperm motility. Once the reduced-motility spermatozoa from 2015 post-bleach were cryopreserved, there was a steep decline in post-thaw viability and this would prevent any substantive further use of these samples in reproduction for conservation benefit. Worldwide, as bleaching events become more frequent, the ability to bank and conserve coral ex situ may be significantly reduced. Thus, it is imperative that while genetic diversity is still high in these populations, intensive efforts are made to bank coral species during non-bleaching periods.
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