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Be Wary of the Edge: Well Position Influences Larval Zebrafish Behavior (CSETAC)
Citation:
Hill, B., D. Hunter, K. Jarema, J. Olin, M. Lowery, B. Knapp, AND S. Padilla. Be Wary of the Edge: Well Position Influences Larval Zebrafish Behavior (CSETAC). Carolinas Society for Environmental Toxicology and Chemistry (CSETAC) Regional Meeting, Charleston, SC, April 19 - 21, 2023. https://doi.org/10.23645/epacomptox.22798685
Impact/Purpose:
Poster presented to the Carolinas SETAC Annual Meeting and ToxExpo April 2023. Analysis of larval zebrafish behavior, recognized as a higher-throughput testing strategy to identify developmentally neurotoxic chemicals, is being utilized as a new approach method at the U.S. Environmental Protection Agency. Aiming to obtain more consistent data across laboratories, we have been investigating confounders that may contribute to variability in results. To our knowledge, edge effects have not been previously explored for larval zebrafish behavior tested in a 96-well plate. Here we compare the locomotor activity of larvae tested under two different light intensities in the outside wells (rows A and H; columns 1 and 12) versus the inside wells. Larvae in the outside wells exhibited higher locomotor activity compared to those in the inside wells, and this effect was even greater when tested with the higher light intensity (about a 14% difference between outside and inside wells when tested with 3,500 lux). These results highlight the necessity of randomization of treatment groups across the entire 96-well plate to avoid having this edge-effect confounding the interpretation of behavioral results.
Description:
Analysis of larval zebrafish behavior, recognized as a higher-throughput testing strategy to identify developmentally neurotoxic chemicals, is being utilized as a new approach method at the U.S. Environmental Protection Agency. Aiming to obtain more consistent data across laboratories, we have been investigating confounders that may contribute to variability in results. To our knowledge, edge effects have not been previously explored for larval zebrafish behavior tested in a 96-well plate. Here we compare the locomotor activity of larvae tested under two different light intensities in the outside wells (rows A and H; columns 1 and 12) versus the inside wells. Larval zebrafish (6 dpf) reared and tested in square 96-well microtiter plates were tested under a light/dark transition locomotor assay consisting of 40 minutes of light (either 335 or 3,500 lux) followed by 40 minutes of dark (12 lux) using a Viewpoint Zebrabox. Results showed no difference in the distance moved with either light intensity. There was, however, a main effect of outside versus inside wells regardless of light intensity. Larvae in the outside wells exhibited higher locomotor activity compared to those in the inside wells, and this effect was even greater when tested with the higher light intensity (about a 14% difference between outside and inside wells when tested with 3,500 lux). These results highlight the necessity of randomization of treatment groups across the entire 96-well plate to avoid having this edge-effect confounding the interpretation of behavioral results. This abstract does not reflect US EPA policy.
URLs/Downloads:
DOI: Be Wary of the Edge: Well Position Influences Larval Zebrafish Behavior (CSETAC)
B HILL CSETAC 2023 POSTER FINAL_04.12.23.PDF (PDF, NA pp, 1120.879 KB, about PDF)