From laboratory to the field: Validating molecular markers of effect in Folsomia candida exposed to a fungicide-based formulation

Tiago Simões*, Sara C. Novais, Tiago Natal-da-Luz, Mathieu Renaud, Sara Leston, Fernando Ramos, Jörg Römbke, Dick Roelofs, Nico M. van Straalen, José P. Sousa, Marco F.L. Lemos

*Corresponding author for this work

    Research output: Contribution to JournalArticleAcademicpeer-review


    Under controlled laboratory conditions, toxicity data tend to be less variable than in more realistic in-field studies and responses may thus differ from those in the natural environment, creating uncertainty. The validation of data under environmental conditions is therefore a major asset in environmental risk assessment of chemicals. The present study aimed to validate the mode of action of a commercial fungicide formulation in the soil invertebrate F. candida, under more realistic exposure scenarios (in-field bioassay), by targeting specific molecular biomarkers retrieved from laboratory experiments. Organisms were exposed in soil cores under minimally controlled field conditions for 4 days to a chlorothalonil fungicide dosage causing 75% reduction of reproduction in a previous laboratory experiment (127 mg a.i. kg −1 ) and half this concentration (60 mg a.i. kg −1 ). After exposure, organisms were retrieved and RNA was extracted from each pool of organisms. According to previous laboratorial omics results with the same formulation, ten genes were selected for gene expression analysis by qRT-PCR, corresponding to key genes of affected biological pathways including glutathione metabolism, oxidation-reduction, body morphogenesis, and reproduction. Six of these genes presented a dose-response trend with higher up- or down-regulation with increasing pesticide concentrations. Highly significant correlations between their expression patterns in laboratory and in-field experiments were observed. This work shows that effects of toxicants can be clearly demonstrated in more realistic conditions using validated biomarkers. Our work outlines a set of genes that can be used to assess the early effects of pesticides in a realistic agricultural scenario.

    Original languageEnglish
    Pages (from-to)522-530
    Number of pages9
    JournalEnvironment International
    Early online date11 Apr 2019
    Publication statusPublished - 1 Jun 2019


    • Biomarkers
    • Chlorothalonil
    • Collembola
    • Fungicides
    • In situ validation
    • qPCR


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