Zebrafish embryo neonicotinoid developmental neurotoxicity in the FET test and behavioral assays

Rebecca von Hellfeld* (Corresponding Author), Viktoriia Ovcharova, Samantha Bevan, Maria-Agapi Lazaridi, Caroline Bauch, Paul Walker, Susanne Hougaard Bennekou, Anna Forsby, Thomas Braunbeck* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)


The need for reliable, sensitive (developmental) neurotoxicity testing of chemicals has steadily increased. Given the limited capacities for routine testing according to accepted regulatory guidelines, there is potential risk to human health and the environment. Most toxicity studies are based on mammalian test systems, which have been questioned for low sensitivity, limited relevance for humans, and animal welfare considerations. This increased the need for alternative models, one of which is the zebrafish (Danio rerio) embryo. This study assessed selected neonicotinoids at sub-lethal concentrations for their effects on embryonic development and behavior. The fish embryo acute toxicity test (OECD TG 236) determined the lowest observable effective concentrations, which were used as the highest test concentrations in subsequent behavioral assays. In the FET test, no severe compound-induced sublethal effects were seen at < 100 µM. In the coiling assay, exposure to ≥ 1.25 µM nicotine (positive control) affected both the burst duration and burst count per minute, whereas ≥ 50 µM thiacloprid affected the mean burst duration. Exposure to ≥ 50 µM acetamiprid and imidacloprid induced significant alterations in both mean burst duration and burst count per minute. In the swimming assay, 100 µM acetamiprid induced alterations in the frequency and extent of movements, whilst nicotine exposure only induced non-significant changes. All behavioral changes could be correlated to findings in mammalian studies. Given the quest for alternative test methods of (developmental) neurotoxicity, zebrafish embryo behavior testing could be integrated into a future tiered testing scheme.
Original languageEnglish
Pages (from-to)367-387
Number of pages21
JournalALTEX - Alternatives to Animal Experiments
Issue number3
Early online date23 Feb 2022
Publication statusPublished - 20 Jul 2022


  • alternative test system
  • OECD TG 236
  • coiling assay
  • swimming assay
  • neonicotinoid toxicity


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