Effects of formalin preservation on invertebrate stable isotope values over decadal time scales

Abstract

Stable isotope values derived from chemically preserved organisms are a valuable resource for documenting long-term ecosystem changes. However, isotopic correction factors of preservation effects applied to samples stored for decades are frequently based on studies lasting only months, assuming that the effects of preservation stabilize within a short time frame. Very few studies test this critical assumption. We validated this assumption for formalin-preserved invertebrate tissues, finding no significant difference between mean isotopic δ13C and δ15N values of material stored 1–15 years across taxa. Preservation effects were evaluated for Amphipoda, Chironomidae, Dreissenidae, Ephemeroptera, Gastropoda, Isopoda, Sphaeridae, Oligochaeta, and Trichoptera. On average, freshwater benthos δ13C was lower by approximately 2‰ after formalin fixation, whereas δ15N values were not different from control samples. Fixation effects were similar among taxa, but were more pronounced in Gastropoda and Sphaeridae for δ13C and in Trichoptera for δ15N. We reviewed the literature to show that preserved freshwater zooplankton δ13C were slightly but significantly lower relative to control samples (–0.2‰) and higher in δ15N (+0.25‰). The mean decline among marine invertebrate δ13C was greater than for freshwater invertebrates after 1+ years of formalin preservation, but effects on δ15N were not different between marine and freshwater invertebrates.