Stable Isotope Analysis of Ozark Hellbender (Cryptobranchus alleganiensis bishopi) Living and Preserved Museum Tissue Reveals a Shift in Their Generalist Diet Composition

Abstract

Ozark hellbenders (Cryptobranchus alleganiensis bishopi) have undergone marked population declines across their entire distribution. A variety of ecological life history research has been conducted to determine the cause(s) of the declines. Historically, hellbender diet studies used stomach content examination methods; however, alternative approaches such as less intrusive stable isotope analyses are now options for researchers. The goals of our study were to conduct stable isotope analysis on live and formalin-preserved museum specimen Ozark hellbender tissues to identify diet composition in the Eleven Point and Spring rivers, Arkansas. Also, we used stable isotope analysis to investigate if Spring River hellbender diets have changed over time. We sampled fish, live hellbenders (non-destructively), and formalin-preserved hellbender tissues from museum collections for stable isotope analysis. We sampled crayfish for assemblage composition and stable isotope analysis. The results of our stable isotope study revealed three main findings: (1) there were no statistically significant differences between hellbender δ13C and δ15N values among sites and hellbender stable C and N isotopes were correlated with body length; (2) traditional δ13C versus δ15N bi-plots and trophic discrimination values did not provide complete discernment in hellbender diets; however, Bayesian MixSIAR models revealed hellbenders to be generalists, and (3) the use of δ13C and δ15N values adjusted historic formalin-fixed and ethanol preserved hellbenders matched well with current crayfish and fish stable isotope values based on Bayesian MixSIAR models. These findings provide important diet information and a possible tool to examine dietary patterns from preserved specimens that may be used for hellbender conservation and management.