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Biogeosciences An interactive open-access journal of the European Geosciences Union
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https://doi.org/10.5194/bg-2020-199
© Author(s) 2020. This work is distributed under
the Creative Commons Attribution 4.0 License.
https://doi.org/10.5194/bg-2020-199
© Author(s) 2020. This work is distributed under
the Creative Commons Attribution 4.0 License.

Submitted as: research article 04 Jun 2020

Submitted as: research article | 04 Jun 2020

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This preprint is currently under review for the journal BG.

Sample preservation and pre-treatment in stable isotope analysis: Implications for the study of aquatic food webs

Marc Jürgen Silberberger, Katarzyna Koziorowska-Makuch, Karol Kuliński, and Monika Kędra Marc Jürgen Silberberger et al.
  • Institute of Oceanology Polish Academy of Sciences, Powstańców Warszawy 55, 81-712 Sopot, Poland

Abstract. Stable isotope analysis has become one of the most widely used techniques in ecology. However, uncertainties about the effects of sample preservation and pre-treatment on the ecological interpretation of stable isotope data and especially on Bayesian stable isotope mixing models remain. Here, Bayesian mixing models were used to study how three different preservation methods (drying, freezing, formalin) and two pre-treatments (acidification, lipid removal) affect the estimation of diet composition for two benthic invertebrate species (Limecola balthica, Crangon crangon). Furthermore, commonly used mathematical lipid normalization and formalin correction were applied to check if they improve the model results. Preservation effects were strong on model outcomes for frozen as well as formalin preserved L. balthica samples, but not for C. crangon. Pre-treatment effects varied with species and preservation method and neither lipid normalization nor mathematical formalin correction consistently resulted in improved model outcomes. Our analysis highlights that particularly small changes in δ15N introduced by different preservation and pre-treatments display a so far unrecognized source of error in stable isotope studies. We conclude that mathematical correction of stable isotopes data should be avoided for Bayesian mixing models and that previously unaddressed effects of sample preservation (especially those arising from preservation by freezing) have potentially biased our understanding of the utilization of organic matter in aquatic food webs.

Marc Jürgen Silberberger et al.

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Marc Jürgen Silberberger et al.

Marc Jürgen Silberberger et al.

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Short summary
The use of stable isotope ratios to study food webs, requires multiple decisions about sample preservation and pre-treatments. In this study we demonstrate how different preservation and pre-treatment methods affect the interpretation of stable isotope data and highlight that today's guidelines are not applicable when data are used in Bayesian mixing models. Particularly the identified effects of freezing demonstrate that our understanding of the utilization of organic matter might be biased.
The use of stable isotope ratios to study food webs, requires multiple decisions about sample...
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