Embryotoxicity and Cardiotoxicity Assessment of Chemical Compounds Using Eggs of Lymnaea stagnalis

There is a paucity of invertebrate models for the assessment of general and organ-specific toxicity caused by chemicals, such as chemotherapeutics and environmental pollutants. To address this, we have established the embryo of Lymnaea stagnalis (great pond snail) as a simple and rapid model to assay toxicity in a medium-throughput manner. For measuring general toxicity, we have successfully repurposed Alamar Blue dye to assay physiological activity of the embryos. Due to the transparent nature of the L. stagnalis eggs, we have employed a microscope-based phenotypic assay to determine cardiotoxicity. Furthermore, we have previously shown that this model can be used to test cardioprotective strategies to overcome toxicity indued by cancer chemotherapeutics. Due to similarities of the L. stagnalis heart with that of a vertebrate heart, combined with the advantages of easy handling, L. stagnalis eggs serve as an emerging model for embryotoxicity and cardiotoxicity screening of pharmaceutical agents and environmental pollutants. © 2026 Wiley Periodicals LLC. Basic Protocol 1: Isolation of clusters and separation of individual eggs of L. stagnalis Basic Protocol 2: Assessment of cardiotoxicity and evaluation of cardioprotective strategies Basic Protocol 3: Alamar Blue dye-based embryotoxicity test.