title: Coral larvae conservation: physiology and reproduction.
abstract: Coral species throughout the world's oceans are facing severe environmental pressures. We are interested in conserving coral larvae by means of cryopreservation, but little is known about their cellular physiology or cryobiology. These experiments examined cryoprotectant toxicity, dry weight, water and cryoprotectant permeability using cold and radiolabeled glycerol, spontaneous ice nucleation temperatures, chilling sensitivity, and settlement of coral larvae. Our two test species of coral larvae, Pocillopora damicornis (lace coral), and Fungia scutaria (mushroom coral) demonstrated a wide tolerance to cryoprotectants. Computer-aided morphometry determined that F. scutaria larvae were smaller than P. damicornis larvae. The average dry weight for P. damicornis was 24.5%, while that for F. scutaria was 17%, yielding osmotically inactive volumes (V(b)) of 0.22 and 0.15, respectively. The larvae from both species demonstrated radiolabeled glycerol uptake over time, suggesting they were permeable to the glycerol. Parameter fitting of the F. scutaria larvae data yielded a water permeability 2 microm/min/atm and a cryoprotectant permeability = 2.3 x 10(-4) cm/min while modeling indicated that glycerol reached 90% of final concentration in the larvae within 25 min. The spontaneous ice nucleation temperature for F. scutaria larvae in filtered seawater was -37.8+/-1.4 degrees C. However, when F. scutaria larvae were chilled from room temperature to -11 degrees C at various rates, they exhibited 100% mortality. When instantly cooled from room temperature to test temperatures, they showed damage below 10 degrees C. These data suggest that they are sensitive to both the rate of chilling and the absolute temperature, and indicate that vitrification may be the only means to successfully cryopreserve these organisms. Without prior cryopreservation, both species of coral settled under laboratory conditions.
authors.affiliation: Department of Reproductive Sciences, Smithsonian National Zoological Park, Washington, DC 20008, USA. hagedornm@si.edu
authors.forename: M
authors.initials: M
authors.lastname: Hagedorn
authors.forename: R
authors.initials: R
authors.lastname: Pan
authors.forename: E F
authors.initials: EF
authors.lastname: Cox
authors.forename: L
authors.initials: L
authors.lastname: Hollingsworth
authors.forename: D
authors.initials: D
authors.lastname: Krupp
authors.forename: T D
authors.initials: TD
authors.lastname: Lewis
authors.forename: J C
authors.initials: JC
authors.lastname: Leong
authors.forename: P
authors.initials: P
authors.lastname: Mazur
authors.forename: W F
authors.initials: WF
authors.lastname: Rall
authors.forename: D R
authors.initials: DR
authors.lastname: MacFarlane
authors.forename: G
authors.initials: G
authors.lastname: Fahy
authors.forename: F W
authors.initials: FW
authors.lastname: Kleinhans
mesh_terms: D000818:Animals; D037421:Anthozoa / Q000254:growth & development / Q000502:physiology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D005615:Freezing; D005990:Glycerol / Q000378:metabolism / Q000494:pharmacology; D007053:Ice; D007814:Larva / Q000254:growth & development* / Q000502:physiology; D012098:Reproduction / Q000502:physiology*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013696:Temperature
chemical_list: D003451:Cryoprotective Agents; D007053:Ice; D005990:Glycerol