Effects of fixation on freshwater invertebrate carbon and nitrogen isotope composition and its arithmetic correction

Retrospective investigations using carbon and nitrogen stable isotope composition of archived material have a great potential for describing past effects of anthropogenic ecosystem alterations or natural shifts in ecosystems. In this study, we examined the effects of two commonly used preservation substances of freshwater invertebrates, ethanol and lugol, on δ¹³C and δ¹⁵N of various planktonic and benthic taxa. For both isotopes, the average effect of fixation in ethanol was stronger than in lugol, and the effects on δ¹³C were stronger than on δ¹⁵N (average ± SD: 1.18 ± 0.94 and −0.47 ± 0.99 for δ¹³C ethanol and lugol fixed samples, respectively, and 0.39 ± 0.68 and 0.17 ± 0.77 for δ¹⁵N, respectively). The changes in the isotopic composition were not dependent on the initial isotopic composition of each taxon, but were related with concomitant changes in the carbon or nitrogen content. Application of a mass balance correction equation to the fixed samples resulted in a significantly lower average effect of fixation in ethanol (0.01 ± 0.59 and 0.44 ± 0.65 for δ¹⁵N and δ¹³C, respectively), while corrections had little effect for lugol fixed samples (0.24 ± 0.53 and −0.39 ± 0.85, respectively). For both isotopes and fixatives, corrections resulted in linear relationships between fixed vs. control samples, with slopes and intercepts not significantly different from 1 and 0, respectively. Therefore, mass balance correction of stable isotopes in fixed invertebrates is recommended for minimising the effects of fixation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Handling editor: M. Power