Field Mark-Recapture of Calcein-Stained Larval Oysters (Crassostrea virginica) in a Freshwater-Dominated Estuary

  • Haley N. GancelEmail author
  • Ruth H. Carmichael
  • Kyeong Park
  • Jeffrey W. Krause
  • Scott Rikard


Knowledge of larval transport is important for restoration and management efforts; yet, there are no established methods to determine larval transport in situ. Calcein staining of oyster larvae may help fill this void, and a two-part study was conducted to determine its effectiveness at tracking larval oyster transport in the field. First, it was tested whether oysters could be successfully stained, survive, and grow at estuarine salinities (15, 20, 26), and at sufficiently large numbers (millions of oysters) to support field mark-recapture studies. Second, the field-based application was tested by releasing 22 million stained larvae twice (high and low salinity) into a major estuary, and two methods (fluorescent microscopy and FlowCam) were used to detect recaptured larvae. Results were compared with expected larval movement patterns simulated by an existing larval transport model. Calcein concentrations (100 mg L−1) did not affect larval growth or survival, but handling conditions (water salinity manipulations and tank size) did affect growth and survival during the post-staining period. Microscopy had double the detection capacity, but FlowCam was more practical and time efficient for the large-volume, high particulate load field samples. Larvae (n = 2) were recaptured during the second, higher salinity release, and model comparison showed a 1–2-day time-lag between field recapture and model predictions, suggesting need for model refinement. Calcein has potential to be a useful marker to track larval movement at large scales needed for field-based studies, providing critical information to aid in selection of restoration sites and management of commercially important shellfish species.


Larval transport model FlowCam Salinity Growth Survival 



We thank the Dauphin Island Sea Lab-Food and Drug Administration Fellowship for refitting our FlowCam with laser capabilities. We thank the Auburn University Shellfish Laboratory for letting us use their hatchery facilities and Elizabeth Hieb, Casey Fulford, Audrey Palombo, Neil Berglund, Heather Patterson, Chris Williams, and Max Han for laboratory and field help. A special thanks to Sydney Acton for FlowCam logistics help and advice. We also thank two anonymous reviewers who greatly improved this manuscript.

Funding Information

This work was funded by the Mississippi-Alabama Sea Grant Consortium (project number #R/SFA-03).

Supplementary material

12237_2019_582_Fig5_ESM.png (354 kb)
Fig. S1

FlowCam images of stained oysters recaptured following the second release (July 28, 2014) at site 4 on day 2 (190 μm, anterior to posterior orientation; left panel) and day 5 (220 μm, posterior orientation; right panel) (PNG 353 kb)

12237_2019_582_MOESM1_ESM.tiff (818 kb)
High Resolution Image (TIFF 817 kb)
12237_2019_582_Fig6_ESM.png (1.2 mb)
Fig. S2

The forcing conditions for freshwater discharge and wind used for the model simulations for the first (May 19, 2014) and the second (July 28, 2014) releases (PNG 1235 kb)

12237_2019_582_MOESM2_ESM.tif (14.9 mb)
High Resolution Image (TIF 15294 kb)
12237_2019_582_MOESM3_ESM.docx (19 kb)
ESM 1 (DOCX 19 kb)


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Copyright information

© Coastal and Estuarine Research Federation 2019

Authors and Affiliations

  1. 1.University of South AlabamaMobileUSA
  2. 2.Dauphin Island Sea LabDauphin IslandUSA
  3. 3.Texas A&M University at GalvestonGalvestonUSA
  4. 4.Auburn University Shellfish LaboratoryDauphin IslandUSA

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