The Validity of Brine Shrimp (Artemia Sp.) Toxicity Assays to Assess the Ecological Function of Marine Natural Products

Many organisms employ toxic compounds for protection against predators. To understand the effectiveness of such compounds, chemoecological studies often use brine shrimp (Artemia spp.) as a model organism instead of more ecologically relevant species. This is mostly because brine shrimp assays are simple and quick, but also due to the ethical implications associated with inducing harm to vertebrate predators in toxicity assays. In this study, we examined whether brine shrimp assays produce similar results to ichthyological toxicity assays with the aim of validating the use of brine shrimp as a preliminary screening tool. We extracted compounds from eight nudibranch molluscs including six species that we consider to signal their chemical defenses via warning coloration to visually hunting vertebrate predators. We tested the relative toxicity of these compounds against brine shrimp and a vertebrate potential predator, the blue-green damselfish (Chromis viridis). We found that extracts toxic to brine shrimp were also toxic to damselfish; however, extracts non-toxic to brine shrimp may still be toxic to damselfish. We also produced and tested mantle vs whole-body extracts for some nudibranch species, which exhibited similar toxicities in both assays except for the whole-body extract of Goniobranchus splendidus which was harmless to shrimp but toxic to fish, while the mantle extract was toxic to both. Overall, we argue that the brine shrimp assay can reasonably indicate the potential toxicity of a compound to fish, but additional experiments with more ecologically relevant predators are required if a no dose-response is observed against brine shrimp.

Keywords: Brine shrimp; Chemical defenses; Nudibranch; Toxicity assay.

© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

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References

    1. Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18:265–267. https://doi.org/10.1093/jee/18.2.265a - DOI
    1. Aguado F, Marin A (2007) Warning coloration associated with nematocyst-based defences in aeolidiodean nudibranchs. J Molluscan Stud 73:23–28. https://doi.org/10.1093/mollus/eyl026 - DOI
    1. Akhtar Y, Isman MB (2003) Binary mixtures of feeding deterrents mitigate the decrease in feeding deterrent response to antifeedants following prolonged exposure in the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae). Chemoecology 13:177–182. https://doi.org/10.1007/s00049-003-0246-0 - DOI
    1. Ankley GT, Johnson RD (2004) Small fish models for identifying and assessing the effects of endocrine-disrupting chemicals. ILAR J 45:469–483. https://doi.org/10.1093/ilar.45.4.469 - DOI - PubMed
    1. Avila C, Paul VJ (1997) Chemical ecology of the nudibranch Glossodoris pallida: is the location of diet-derived metabolites important for defense? Mar Ecol Prog Ser 150:171–180 - DOI