Experimental studies on cold survival of enchytraeid cocoons

The effect of different culture methods, cold acclimation and desiccation on the supercooling point (SCP), the melting point (MP) of fluids and the quantity of water freezing (osmotically active water) was investigated in individual cocoons of Enchytraeus crypticus and an undescribed Enchytraeus species using differential scanning calorimetry (DSC) techniques. Both species can be easily cultured in the laboratory in agar where the development and hatching of the worms can be observed. Culture methods (agar with nettle leaves or oats as food and wet filter paper without food supply) had a significant effect on fresh weight and SCP of E. crypticus cocoons. The water content (as a proportion of fresh weight) was slightly lower in the cocoons from the wet filter paper cultures. Acclimation at —3 °C did not affect the supercooling ability of E. crypticus cocoons, whereas the SCP of E. sp.1 cocoons was lowered from a mean of —8.7 to —12.4 °C. The supercooling ability of cocoons of E. crypticus was only slightly increased by desiccation and cold acclimation, but the quantity of frozen water was significantly reduced with acclimation and desiccation (2 and 3 min) at room temperature. The MP values of the cocoon fluids reflected these changes in both species. No cocoons of E. crypticus and E. sp.1 survived desiccation and freezing in our experiments and no worms of Enchytraeus albidus, E. crypticus and Henlea perpusilla survived freezing to —10 or —20 °C.