Burrowing is a common trait among crayfish thought to help species deal with adverse environmental challenges. However, little is known about the microhabitat ecology of crayfish taxa in relation to their burrows. To fill this knowledge gap, we assessed the availability of oxygen inside the crayfish shelter by series of in-vivo and in-silico modelling experiments. Under modeled condition, we found that, except for the entrance region of the 200 mm, a flooded burrow microenvironment became anoxic within 8 h, on average. Multiple 12-hour day-night cycles, with burrows occupied by crayfish for 12 h and empty for 12 h, were not sufficient for refreshing the burrow microenvironment. We then examined the degree to which crayfish species with different propensities for burrowing are tolerant of self-created anoxia. From these experiments, primary and secondary burrowers showed best and most consistent tolerance-exhibiting ≥ 64% survival to anoxia and 25-91% survival of ≥ 9 h at anoxia, respectively. Tertiary burrowers exhibited little to no tolerance of anoxia with 0-50% survival to anoxia and only one species exhibiting survival (2%) of ≥ 9 h at anoxia. Results suggest that moderate to strongly burrowing crayfish can quickly draw down the dissolved oxygen in burrow water but appear to have conserved a legacy of strong tolerance of anoxia from their monophyletic ancestors-the lobsters-whereas tertiary burrowers have lost (or never evolved) this ability.

Center for Bottomland Hardwoods Research Southern Research Station USDA Forest Service Oxford MS United States of Ameirca

Central Fisheries Research Institute Yoichi Hokkaido Japan

Coastal and Marine Research Centre Griffith University Gold Coast Queensland Australia

Crayfish Research Centre Institute for Advanced Environmental Research West University of Timisoara Timisoara Romania

Department of Biochemistry and Pharmacology Faculty of Medicine Victor Babeș University of Medicine and Pharmacy Timisoara Timisoara Romania

Department of Biology Chemistry Faculty of Chemistry Biology Geography West University of Timisoara Timisoara Romania

Faculty of Physics West University of Timisoara Timisoara Romania

Laboratório de Carcinologia Programa de Pós Graduação em Biodiversidade Animal Universidade Federal de Santa Maria Santa Maria RS Brazil

School of Fisheries Aquaculture and Aquatic Sciences Auburn University Auburn AL United States of America

South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Faculty of Fisheries and Protection of Waters University of South Bohemia in České Budějovice Vodňany Czech Republic

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Some Like It Cold: Long-Term Assessment of a Near-Global Invader

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