Abstract
Sample fixation and preservation are crucial steps for biological research. This is especially true for resolving ecological changes across various spatial and temporal scales based on sediment and/or macrozoobenthos samples, recognized to be efficient indicators of habitat quality and ecosystem health in aquatic environments. Thus, it is very important to find a convenient method for long-term sample storage. In this study, we compared effectiveness of five different solutions (glyoxal, 3.6% formaldehyde, NBF, 70% ethanol, 99% ethanol) for the preservation of macroinvertebrate taxa commonly found in stream habitats (i.e. families of aquatic insects, Chironomidae, Baetidae, Ephemeridae, Heptageniidae), with the aim to find a new “gold standard” fixative that could replace formalin, recognized to have significantly adverse health effects if exposed to it. To estimate the effectiveness of these solutions on all macrozoobenthos samples/taxa, color contrast and tissue preservation within hematoxylin and eosin stained slides were assessed as well as the preservation of body parts and tissues of collected specimens. Our results show that different preservatives/fixatives yield different effects on the tissue preservation of different organisms. None of the tested preservation liquids prove to be unique formalin successor that could serve as a new “gold standard” for preserving macroinvertebrate taxa commonly found in morphological and stream ecology studies. The efficiency of different solutions in preserving macroinvertebrate tissue integrity depends on the chemical properties and concentrations of the solutions for sample storage, tissue type as well as on the dimensions and structure (e.g., the extent of chitin coverage) of the studied organisms.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
This study was funded by Croatian Science Foundation (grant IP-CORONA-2020-12-3798), and partial financial support (tested solutions) was received from company BioGnost.
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Conceptualization: [Valerija Begić, Ivan Marchiotti, Ines Radanović, Petra Korać]; Data curation: [Valerija Begić, Mirela Sertić Perić, Tea Gabud]; Formal analysis: [Valerija Begić, Mirela Sertić Perić, Suzana Hančić, Petra Korać]; Funding acquisition: [Ivan Marchiotti, Petra Korać, Ines Radanović]; Investigation: [Valerija Begić, Mirela Sertić Perić, Suzana Hančić, Ines Radanović, Petra Korać]; Methodology: [Mirela Sertić Perić, Suzana Hančić, Ivan Marchiotti, Tea Gabud, Ivana Šestak Panižić, Ines Radanović, Petra Korać; Resources, Ivan Marchiotti]; Supervision: [Petra Korać; Validation, Valerija Begić, Mirela Sertić Perić, Petra Korać]; Visualization: [Valerija Begić, Mirela Sertić Perić, Petra Korać]; Writing – original draft: [Valerija Begić, Mirela Sertić Perić, Petra Korać]; Writing – review and editing: [Valerija Begić, Mirela Sertić Perić, Suzana Hančić, Ivan Marchiotti, Tea Gabud, Ivana Šestak Panižić, Ines Radanović, Petra Korać].
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Authors IŠP and TG work for the company BioGnost that produces and sells tested solutions. Author IM is director, co-founder and co-owner of BioGnost Ltd. Other authors have no relevant financial or non-financial interests to disclose.
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Begić, V., Sertić Perić, M., Hančić, S. et al. Effectiveness of five different solutions for preserving aquatic insects commonly used in morphological and stream ecology studies. Biologia 78, 1011–1026 (2023). https://doi.org/10.1007/s11756-022-01298-y
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DOI: https://doi.org/10.1007/s11756-022-01298-y