Use of Formalin-Preserved Collections to Infer Trophic Indicators of Marine Zooplankton from Stable Isotopes

Formalin preservation affects the stable isotope composition of zooplankton samples, thus limiting the analysis of valuable collections covering large time intervals. Here, we compare different procedures for correcting the bias caused by formalin in δ13C and δ15N of zooplankton community samples. Zooplankton samples representative of seasonal variations in the period 2000–2009 were collected off A Coruña (NW Spain). Part of the sample was immediately dried and analysed for δ13C, δ15N, and elemental composition within 3 years of collection. These values were used as the unpreserved reference. The remaining sample was preserved in 4% formaldehyde and aliquots obtained after a period ranging from 3 years to more than 10 years of storage were analysed as the originally dried samples. Additionally, the copepod fraction of total biomass was determined in the preserved samples. Corrections of formalin effects based on ordinary least squares regression had large uncertainties, while mass balance corrections based on the change in C:N ratio (only possible for δ13C) overestimated reference values. However, either simple corrections based on the mean difference between values in dry and preserved samples or more complex generalised additive models considering seasonality, copepod biomass, and time of sample storage, produced estimations with relatively low uncertainty. Our results highlight the importance of determining specific correction solutions for each preserved collection before reconstructing stable isotope time series. Furthermore, the uncertainties associated with the estimates can be used in sensitivity analysis to assess their potential impact on the interpretation of the series.

This research was funded by projects QLOCKS (PID2020-115620RB-100, MCIN/AEI/10.13039/501100011033) and RADIALES (Instituto Español de Oceanografía, Spain). C.M. was supported by a FPI fellowship from the Instituto Español de Oceanografía (Spain).

Peer reviewed