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Impact of test chamber design on spontaneous behavioral responses of model crustacean zooplankton Artemia franciscana

Abstract

The use of small aquatic model organisms to investigate the behavioral effects of chemical exposure is becoming an integral component of aquatic ecotoxicology research and neuroactive drug discovery. Despite the increasing use of invertebrates for behavioral phenotyping in toxicological studies and chemical risk assessments, little is known regarding the potential for environmental factors—such as geometry, size, opacity and depth of test chambers—to modulate common behavioral responses. In this work, we demonstrate that test chamber geometry, size, opacity and depth can affect spontaneous, unstimulated behavioral responses of euryhaline crustacean Artemia franciscana first instar larval stages. We found that in the absence of any obvious directional cues, A. franciscana exhibited a strong innate wall preference behavior. Using different test chamber sizes and geometries, we found both increased wall preference and lowered overall distance traveled by the test shrimp in a smaller chamber with sharper-angled vertices. It was also determined through quantifiable changes in the chambers’ color that the A. franciscana early larval stages can perceive, differentiate and react to differences in color or perhaps rather to light transmittance of the test chambers. The interaction between innate edge preference and positive phototaxis could be consistently altered with a novel photic stimulus system. We also observed a strong initial preference for depth in A. franciscana first instar larval stages, which diminished through the acclimatization. We postulate that the impact of test chamber designs on neurobehavioral baseline responses warrants further investigation, in particular considering the increased interest in behavioral eco-neurotoxicology applications.

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Fig. 1: Impact of test chamber size and geometry on behavioral endpoints.
Fig. 2: The color of test chamber sidewalls induces directional alterations in spontaneous behaviors.
Fig. 3: A crosstalk between positive phototaxis and wall-preference behaviors.
Fig. 4: Influence of test chamber depth on innate behavioral preferences.
Fig. 5: Influence of test chamber depth on innate behavioral preferences with removal of edge preference through horizontal filming.

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Data from all experiments are available upon request.

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Authors and Affiliations

Authors

Contributions

J.H. and D.Wlodkowic developed and designed the experimental concepts. J.H., D.Williams, Y.B. and A.L. performed the experiments and collected and analyzed the data. J.H. wrote the manuscript. Y.B., A.F. and D.Wlodkowic reviewed and edited the manuscript.

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Correspondence to Donald Wlodkowic.

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The authors declare no competing interests.

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Lab Animal thanks Maria Violetta Brundo, Jake Martin and Joseph Covi for their contribution to the peer review of this work.

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Supplementary Table 1. Detailed statistical results from a GLMM.

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Henry, J., Bai, Y., Williams, D. et al. Impact of test chamber design on spontaneous behavioral responses of model crustacean zooplankton Artemia franciscana. Lab Anim 51, 81–88 (2022). https://doi.org/10.1038/s41684-021-00908-7

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