Preprints
https://doi.org/10.5194/bg-2020-199
https://doi.org/10.5194/bg-2020-199
04 Jun 2020
 | 04 Jun 2020
Status: this preprint was under review for the journal BG but the revision was not accepted.

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

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, Katarzyna Koziorowska-Makuch, Karol Kuliński, and Monika Kędra
 
Status: closed
Status: closed
AC: Author comment | RC: Referee comment | SC: Short comment | EC: Editor comment
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Status: closed
Status: closed
AC: Author comment | RC: Referee comment | SC: Short comment | EC: Editor comment
Printer-friendly Version - Printer-friendly version Supplement - Supplement
Marc Jürgen Silberberger, Katarzyna Koziorowska-Makuch, Karol Kuliński, and Monika Kędra
Marc Jürgen Silberberger, Katarzyna Koziorowska-Makuch, Karol Kuliński, and Monika Kędra

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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.
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