Helix aspersa hibernates in Brittany (western France), where it may experience subzero temperatures. Studies on cold hardiness, although scarce in land snails, have shown a seasonal variation in supercooling ability, associated with high temperatures of crystallization (Tc). In the present work, two key environmental factors, temperature and photoperiod, were studied to elucidate, how they may affect the enhancement of supercooling ability in the snails from the end of summer to winter. Nine groups of adult snails were acclimated to different combinations of photoperiod (LD-16:8, LD-12:12, and LD-8:16 h) and temperature (15, 10, and 5 degreesC). Temperature of crystallization, hemolymph osmolality, and water content were measured. The results demonstrate a significant effect of the photoperiod on Tc, i.e., shorter photoperiods induce lower Tc (LD-16:8 h, mean Tc = -3.0 degreesC, SD = 2.0; LD-12:12 h, mean Tc = -4.3 degreesC, SD = 1.9; LD-8:16 h, mean Tc = -5.2 degreesC, SD = 1.9; n = 90), whereas the acclimation temperature had no effect. Measurements of hemolymph osmolality and water content showed that osmolality is negatively correlated with water content. Mechanisms such as dehydration are involved in the decrease of Tc. A declining photoperiod triggers a lower Tc, long before the onset of winter conditions. This response may have an adaptive component, allowing individuals to cope with the mild winters typically observed in oceanic regions. (C) 2001 Elsevier Science.