Seasonal changes in physiology and development of cold hardiness in the hatchling painted turtle Chrysemys picta

Hatchling painted turtles (Chrysemys picta) commonly hibernate in shallow, natal nests where winter temperatures may fall below −10 degrees C. Although hatchlings are moderately freeze-tolerant, they apparently rely on supercooling to survive exposure to severe cold. We investigated seasonal changes in physiology and in the development of supercooling capacity and resistance to inoculative freezing in hatchling Chrysemys picta exposed in the laboratory to temperatures that decreased from 22 to 4 degrees C over a 5.5 month period. For comparison, we also studied hatchling snapping turtles (Chelydra serpentina), a less cold-hardy species that usually overwinters under water. Although Chrysemys picta and Chelydra serpentina differed in some physiological responses, both species lost dry mass, catabolized lipid and tended to gain body water during the acclimation regimen. Recently hatched, 22 degrees C-acclimated Chrysemys picta supercooled only modestly (mean temperature of crystallization −6.3+/−0.2 degrees C; N=6) and were susceptible to inoculation by ice nuclei in a frozen substratum (mean temperature of crystallization −1.1+/−0.1 degrees C; N=6) (means +/− s.e.m.). In contrast, cold-acclimated turtles exhibited pronounced capacities for supercooling and resistance to inoculative freezing. The development of cold hardiness reflected the elimination or deactivation of potent endogenous ice nuclei and an elevation of blood osmolality that was due primarily to the retention of urea, but was not associated with accumulation of the polyols, sugars or amino acids commonly found in the cryoprotection systems of other animals. Also, Chrysemys picta (and Chelydra serpentina) lacked both antifreeze proteins and ice-nucleating proteins, which are used by some animals to promote supercooling and to initiate freezing at the high temperatures conducive to freezing survival, respectively.