Effect of repeated freeze-thaw cycles on geographically different populations of the freeze-tolerant worm Enchytraeus albidus (Oligochaeta).

Freeze-tolerant organisms survive internal ice formation; however, the adaptations to repeated freeze-thaw cycles are often not well investigated. Here we report how three geographically different populations of Enchytraeus albidus (Germany, Iceland and Svalbard) respond to three temperature treatments - constant thawed (0°C), constant freezing (-5°C) and fluctuating temperature (0 to -5°C) - over a period of 42 days. Survival varied between treatments and populations such that enchytraeids from arctic locations had a higher survival following prolonged freeze periods compared with temperate populations. However, enchytraeids from temperate locations had the same survival rate as arctic populations when exposed to repeated freeze-thaw events. Across all populations, metabolic rate decreased markedly in frozen animals (-5°C) compared with thawed controls (0°C). This decrease is likely due to the lower temperature of frozen animals, but also to the transition to the frozen state per se. Animals exposed to repeated freeze-thaw events had an intermediate metabolic rate and freeze-thaw events were not associated with pronounced excess energetic costs. Overwintering under either condition was not associated with a decrease in lipid content; however, during exposure to constant freezing and repeated freeze-thaw events there was a noticeable decrease in carbohydrate stores over time. Thus, animals exposed to constant freezing showed a decrease in glycogen stores, while both glucose and glycogen content decreased over time when the organisms were exposed to repeated freezing. The results therefore suggest that carbohydrate resources are important as a fuel for E. albidus during freezing whereas lipid resources are of marginal importance.