Larval Hatchery Crasn 16s Amplicon Sequencing

Matthew W. Gray, Stephanie Alexander, Brian Beal, Tom Bliss, Colleen A. Burge, Jacob Cram, Michael De Luca, James Dumhart, Patricia M. Glibert, Michael Gonsior, Andrew Heyes, Klaus Huebert, Vyacheslav Lyubchich, Matt Parker, Louis Plough, Eric Schott, Lisa Wainger, Ami Wilbur. 2021. Hatchery crashes among shellfish research hatcheries along the Atlantic coast of the United States: a case study at Horn Point Laboratory oyster hatchery. Aquaculture (Manuscript#: AQUACULTURE-D-21-01351R1)In brief, larvae were collected from experiments adjacent to the HPL larval hatchery. In both cases a wild type strain and non wild type strain (NEH in the crash experiment, AQUA/LOLA in the case of the non-crash experiment) of larvae were either starved, or not starved, in triplicate tanks. In one experiment in , all larvae died, concurrently with an ongoing crash in the hatchery. In the other, no larvae died.Larvae were 7 (crash) and 5 (non-crash) days old at the "pre" time point. During the Starved and Fed time-points in the crash group they were 12 days old. During the Fed and Starved non-crash time-points they were 8 days old.Samples were extracted with SDS, heating and bead-beeting, followed by protinase K incubation. Samples were washed with phenol-chlorophorm three times, precipitated with isopropanol and ammonium acitate, rinsed with ethanol and re-suspended in a Tris-EDTA solution. Amplicon library prep and sequencing was carried out by the Institute for Marine and Enviornmental Technologies' Bioanalytical Services Laboratory (IMET-BASLab). The Crashed samples, run after a laboratory shutdown following the 2020 pandemic, were initially of low quaity and had to be re-run three additional times. The data presented here are from the best, fourth, run.