Summary Chill tolerance of insects is defined as the ability of insects to tolerate low temperature under circumstances not involving freezing of intra- or extracellular fluids. For many insects chill tolerance is crucial for their ability to persist in cold environments and mounting evidence indicate that chill tolerance is associated with the ability to maintain ion- and water-homeostasis, thereby ensuring muscular function and preventing chill injury at low temperature. The present study describes the relationship between muscle and hemolymph ion-homeostasis and time to regain posture following cold shock (CS, 2h at -4°C) in the chill susceptible locust, Locusta migratoria. This relationship is examined in animals with and without a prior rapid cold hardening treatment (RCH, 2h at 0°C) to investigate the physiological underpinnings of RCH. Cold shock elicited a doubling of hemolymph [K+] and this disturbance was greater in locusts pre-exposed to RCH. Recovery of ion homeostasis was, however, markedly faster in RCH treated animals which correlated well with whole organism performance as hardened individuals regained posture more than 2 minutes faster than non-hardened individuals following CS. The present study indicates that loss and recovery of muscular function is associated with resting membrane potential of excitable membranes as attested from the changes in the equilibrium potential for K+ (EK) following CS. Both hardened and non-hardened animals recovered movement once K+ homeostasis was recovered to a fixed level (EK≈ -41 mV). RCH is therefore not associated with altered sensitivity to ion disturbance but instead a faster recovery of hemolymph [K+].

SUMMARYEctothermic animals inhabiting the subarctic and temperate regions have evolved strategies to deal with periods of continuous frost during winter. The earthworm Dendrobaena octaedra is freeze tolerant and accumulates large concentrations of glucose upon freezing. The present study investigates the roles of glucose accumulation for long-term freeze tolerance in worms kept frozen at -2°C for 47 days. During this period, worms were sampled periodically for determination of survival and for measurements of glucose, glycogen, lactate, alanine and succinate. In addition we performed calorimetric measurements to assess metabolic rate of frozen and unfrozen worms. Long-term freezing was associated with a gradual depletion of glucose and worms that succumbed during this period were always characterised by low glucose and glycogen levels. The anaerobic waste products lactate and alanine increased slightly whereas succinate levels remained constant. However, it is argued that other waste products (particularly propionate) could be the primary end product of a continued anaerobic metabolism. Calorimetric measures of the metabolic rate of frozen worms were in accord with values calculated from the reduction in glucose assuming that most (~90%) glucose was metabolised anaerobically. Both estimates of metabolic rate demonstrated a 10-fold metabolic depression associated with freezing. Thus, in addition to the suspected role of glucose as cryoprotectant, the present study demonstrates that glucose accumulation is vital to ensure substrate for long-term anaerobic metabolism in frozen worms. On the basis of the estimated metabolite levels, we calculate that the combined effect of metabolic depression and large glucose stores enables a projected 3 months survival of freezing at -2°C of the 'average' D. octaedra. Such conditions are very likely to occur in the northern distribution ranges of this stress-tolerant earthworm.