A Multi-method Approach for Assessing the Distribution of a Rare, Burrowing North American Crayfish Species

Primary burrowing crayfishes face high extinction risk, but are challenging to study, manage, and conserve due to their difficult-to-sample habitat (, terrestrial burrows) and low population densities. We apply here a variety of methods to characterize the distribution, habitat associations, and conservation status of the Boston Mountains Crayfish (Reimer, 1966), an endemic burrowing crayfish found only in the Ozark Mountains of Arkansas, United States. We used species distribution modeling (SDM) on historic occurrence records to characterize the distribution and macro-scale habitat associations of this species. We then ground-truthed SDM predictions with conventional sampling, modeled fine-scale habitat associations with generalized linear models (GLM), and lastly developed and tested an environmental DNA (eDNA) assay for this species in comparison to conventional sampling. This represents, to our knowledge, the first successful eDNA assay for a terrestrial burrowing crayfish. Our MaxEnt-derived SDM found a strong effect of average annual precipitation on the historic distribution of , which occurred most frequently at locations with moderately high average annual precipitation (140-150 cm/yr) within our study region. was difficult to detect by conventional sampling in 2019 and 2020, found at only 9 of 51 sites (17.6%) sampled by searching for and manually excavating crayfish burrows. Surprisingly, habitat suitability predicted from our MaxEnt models was not associated with contemporary occurrences per GLMs. Instead, presence was negatively associated with both sandy soils and the presence of other burrowing crayfish species. Poor SDM performance in this instance was likely caused by the omission of high resolution fine-scale habitat data (, soils) and biotic interactions from MaxEnt models. Finally, our eDNA assay detected from six of 25 sites (24.0%) sampled in 2020, out-performing conventional surveys by burrow excavation for this species. Given the difficulty of studying primary burrowing crayfishes and their high conservation need, we propose that eDNA may become an increasingly important monitoring tool for and similar species.