Is cold hardiness size-constrained? A comparative approach in land snails

Body water is a major element of the cold-hardiness strategies observed in ectothermic animals, in particular in freezing avoidant species for which body ice formation is lethal. Here, we investigate the relationships, in terrestrial snails, between the temperature of crystallisation (Tc) and body water (water mass and water content), shell shape, geographic and climatic distribution, taking into account phylogenetic inertia. Phylogenetic relationships among 31 species from 13 different families of terrestrial Gastropods were studied using 28S rRNA nuclear and COI mitochondrial sequence data, together with species-specific traits. Our results provide evidence for clear relationships between Tc and absolute/relative body water: smaller species with lower water content tended to be characterized by colder temperatures of crystallisation, although some exceptions were noticeable. Environmental conditions do not appear to affect Tc significantly, as well as shell shape which is however correlated with water content. This study confirmed that supercooling ability in land snails is size-constrained, with consequences on cold-hardiness strategies.