Freezing and anoxia tolerance of slugs: a metabolic perspective

Freezing survival was assessed in three species of terrestrial slugs, a holarctic but native North American species, Deroceras laeve, and two species introduced from Europe, D. reticulatum and Arion circumscriptus. The introduced species showed very poor freezing survival. Supercooling points of the introduced species were quite high (similar to-3 degrees C) and their freezing survival was very poor, limited to short-term freezing at -1.2 to -1.5 degrees C and low ice contents (23-44%). D. laeve showed a significant elevation of supercooling point between slugs collected in the autumn (-4.8 degrees C +/- 0.5) and those collected early in the spring(-3.1 degrees C +/- 0.4). This species also showed substantial freezing survival which was greater for spring-collected slugs (100% survival of 1 h freezing at -2 degrees C with an ice content of 65%) than for autumn animals (100% survival for 1 h at -1 degrees C with similar to 40% ice). Carbohydrate and amino acid responses to freezing and anoxia exposures were compared in the two Deroceras species. D. laeve showed a strong hyperglycemic response to freezing, a 100-fold increase in glucose levels that suggested that glucose may have a cryoprotective function in this species. D. reticulatum did not accumulate glucose and neither species produced glycerol or lactate. Both species showed typical responses to anoxia (aspartate and glutamate catabolism, alanine and succinate accumulation) and D. laeve also showed this pattern during freezing, suggesting a natural switch to anaerobiosis to support freezing survival.