Effects of Tissue Preservation on Carbon and Nitrogen Stable Isotope Signatures in Syngnathid Fishes and Prey

SIMPLE SUMMARY: Stable isotope analysis (SIA) was used to assess the influence of various preservation methods (freezing, ethanol and formaldehyde) on syngnathid (seahorses and pipefishes) fins, seahorse newborns (seahorses), and prey (copepods and Artemia). The first available conversion models for syngnathids are provided, enabling their application to isotopic studies in the field and in the laboratory. ABSTRACT: Isotopic stable analysis (SIA) is a powerful tool in the assessment of different types of ecological and physiological studies. For that, different preservation methods for sampled materials are commonly used prior to isotopic analysis. The effects of various preservation methods (freezing, ethanol and formaldehyde) were analyzed for C:N, and δ(13)C and δ(15)N signals on a variety of tissues including dorsal fins (three seahorse and two pipefish species), seahorse newborns (three seahorses species), and prey (copepods and different stages of Artemia) commonly used to feed the fishes under rearing conditions. The aims of the study were: (i) to evaluate isotopic effects of chemical preservation methods across different types of organisms and tissues, using frozen samples as controls, and (ii) to construct the first conversion models available in syngnathid fishes. The chemical preservation in ethanol and, to a lesser extent, in formaldehyde significantly affected δ(13)C values, whereas the effects on δ(15)N signatures were negligible. Due to their low lipid content, the isotopic signals in fish fins was almost unaffected, supporting the suitability of dorsal fins as the most convenient material in isotopic studies on vulnerable species such as syngnathids. The regression equations provided resulted convenient for the successful conversion of δ(13)C between preservation treatments. Our results indicate that the normalization of δ(15)N signatures in preserved samples is unnecessary. The conversion models should be applicable in isotopic field studies, laboratory experiments, and specimens of historical collections.