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Direct Setting of Eastern Oyster (Crassostrea virginica) Larvae Confirmed with Calcein, a Fluorochrome Dye

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Abstract

Ongoing efforts to restore eastern oyster (Crassostrea virginica) populations in regions of low natural recruitment rely on stocking of juveniles (spat on shell) to rebuild populations. Often, remote setting is used, entailing releasing hatchery-produced larvae into recirculating tanks filled with oyster shells. Other restoration methods that rely on releasing C. virginica larvae directly onto the shell in situ, called “direct setting,” have shown promise but were unable to definitively prove larval origin without the use of enclosures. The objective of this study was to determine if tagging C. virginica with calcein, a fluorochrome dye, could be a viable method for confirming larval origin in studies of direct setting in Chesapeake Bay. To do so, C. virginica larvae conditioned in water from adult C. virginica were marked with calcein and released by divers directly onto three 3.6-m2 research sites constructed of oyster shell bags during July 2019 and September 2019 and recovered after 7 days. All shell bags were moved to flow-through tanks on land and spat on a subsample of valves were counted in each bag 8–12 days after deployment. Spat on the remaining valves were counted 42 to 46 days post deployment. A total of 119,020 spat were found on 84 shell bags from the two deployments during the initial settlement counts conducted just after shell bags were recovered. All recovered juveniles that were viewed under blue light excitation (n = 84) contained the calcein tag, indicating that these spat were derived from larvae released over the reefs. Initial settlement efficiencies on the sites ranged from 0.1 to 3.4% in July and September, respectively. The salinities experienced in July were below average and may have contributed to reduced larval survival compared to that in September. Shell bags contained zero to 90 spat per shell. Spat settlement was greatest closest to where the larvae were released (87% of spat were found in 12% of bags; the high-count bags were clustered around the larval release locations). Overall, 6 shell bags out of 190 deployed had spat per shell estimates similar to remote larval setting (hatchery) targets (10–20 spat per shell) and 6 had spat per shell values higher than hatchery targets. The presence of the calcein mark in recovered spat confirmed larval origin, and together with the observed setting efficiencies suggests there is promise for developing remote larval setting as a stock enhancement technique. However, more work is needed to understand the limitations of the technique, including its efficaciousness at a larger scale.

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Data Availability

The data that support the findings of this study are available from the corresponding author [JES].

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Acknowledgements

This work would not have been possible without the help and support of the following organizations: NOAA Cooperative Oxford Laboratory, Maryland Department of Natural Resources Piney Point Aquaculture Center, University of Maryland Center for Environmental Science Horn Point Laboratory Oyster Hatchery, NOAA Chesapeake Bay Office, Chesapeake Research Consortium, US Food and Drug Administration, US Fish and Wildlife Administration and the Fryling Fund of the Mid-Shore Community Foundation. We thank, Grace Reising, Prian Vidal, Ava Ellett, Amy Freitag, Andrew Mason, Bart Merrick, David Bruce, Elise Trelegan, Eric Daniels, Gretchen Messick, Janine Harris, Jay Lazar, Jay Lewis, John Jacobs, Krysta Hougen, Deidre Gibson, Jennifer Kraus, Melanie Gange, Patrick Hurley, Steve Thur, Amanda Ault, Brian Richardson, Gary Culver, Dave Williams, Jacob Shaner, John Gallagher, Kevin Rosemary, Matt Baldwin, Carol McCollough, Shawn McLaughlin, John Jacobs, Suzanne Skelley, and A.K. Leight for providing invaluable laboratory, field, and logistical support. We appreciate Howard Townsend, Eric Weissberger, Michael Wilberg, Matt Gray, Shannon Hood, and Stephanie Alexander for the valuable advice and comments. Additionally, we thank the NOAA internal reviewers for the careful and insightful review of our manuscript.

Funding

This work was funded by the National Oceanic and Atmospheric Administration National Centers for Coastal Ocean Science.

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Authors and Affiliations

  1. Horn Point Laboratory, University of Maryland Center for Environmental Science, 2020 Horns Point Road, Cambridge, MD, 21613, USA

    Jason E. Spires & Elizabeth W. North

  2. Cooperative Oxford Laboratory, National Centers for Coastal Ocean Science, National Ocean Service, National Oceanic and Atmospheric Administration, 904 S. Morris St., Oxford, MD, 21654, USA

    Jason E. Spires

  3. Restoration Center, National Oceanic and Atmospheric Administration, 200 Harry S. Truman Parkway, Annapolis, MD, 21401, USA

    Stephanie R. Westby

  4. Oceanography Department, United States Naval Academy, 572C Holloway Rd, Annapolis, MD, 21402, USA

    Cecily N. Steppe

  5. Maryland Department of Natural Resources, 17996 Piney Point Rd, Piney Point, MD, 20674, USA

    James M. Dumhart II

  6. Hampton University, 100 E Queen St, Hampton, VA, 23669, USA

    Sierra S. Hildebrandt

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  1. Jason E. Spires
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  2. Elizabeth W. North
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Correspondence to Jason E. Spires.

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Communicated by Eric N. Powell.

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Spires, J.E., North, E.W., Westby, S.R. et al. Direct Setting of Eastern Oyster (Crassostrea virginica) Larvae Confirmed with Calcein, a Fluorochrome Dye. Estuaries and Coasts 46, 723–739 (2023). https://doi.org/10.1007/s12237-023-01178-x

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