In situ investigation of ice fractions and water states during partial freezing of pork loins and shrimps.

Ice fractions and water states in partially frozen muscle foods greatly affect their quality. In the study, a variable temperature nuclear magnetic resonance (VT-NMR) with a liquid nitrogen temperature control system was employed to in situ investigate the relationship between ice fractions and temperatures and changes in water states during partial freezing and thawing of pork and shrimp. Results indicated that changes in ice fractions ranging from -2 âˆ¼ -20 °C could be divided into 3 stages including slow increase, random leap and remarkable leap. More serious damages to the structures related to immobile water occurred in shrimp than in pork, and partial freezing also caused deterioration in muscle fibres related to free water. Additionally, -2 âˆ¼ -3 °C and - 3.5 °C were the appropriate partial freezing temperatures for pork and shrimp, respectively. Therefore, the VT-NMR method possessed great potential for fundamental studies and applications of partial freezing of muscle foods.

Supercooling and freezing as eco-physiological alternatives rather than mutually exclusive strategies: A case study in Pyrrhocoris apterus.

Overwintering insects are categorized either as freeze tolerant or freeze avoiding (supercooling) based on their ability or inability, respectively, to tolerate the formation of ice in their body. The freeze tolerant insects set their supercooling point (SCP) higher for winter to stimulate freezing at higher temperatures, while freeze avoiding insects survive winter in a supercooled state by depressing their SCP. Some supercooling insects, however, were found to survive in frozen state when freezing occurred through inoculation by external ice at mild subzero temperatures. Here, we assessed the potential relevance of inoculative freezing and freeze tolerance strategy in an insect that was so far considered as a classical example of a 'supercooler', the linden bug (Pyrrhocoris apterus). Microclimatic conditions of the overwintering microhabitat of P. apterus (leaf litter layer with buffered temperature fluctuations, mild sub-zero extremes, high humidity, and presence of ice) present a potentially high risk of inoculative freezing. We found that P. apterus is highly susceptible to inoculation by external ice. The temperature at which inoculative freezing occurred (above -3°C) was much higher compared to SCP (-16 °C to -20 °C in winter). The insects were inoculated through body openings and across cuticle and were able to survive after freezing. There was, however, a distinct critical ice fraction, corresponding to 38.7-42.8% of total body water, beyond which survival rapidly decreased to zero. We found that P. apterus adaptively reduces the actual ice fraction below critical ice fraction for winter season. Since many insect species overwinter in habitats similar to that of P. apterus, the ability to tolerate freezing after inoculation by external ice crystals could be much more common among 'supercooling' insects than it is currently appreciated.

Thermal analysis of ice and glass transitions in insects that do and do not survive freezing.

Some insects rely on the strategy of freeze tolerance for winter survival. During freezing, extracellular body water transitions from the liquid to the solid phase and cells undergo freeze-induced dehydration. Here, we present results of a thermal analysis (from differential scanning calorimetry) of ice fraction dynamics during gradual cooling after inoculative freezing in variously acclimated larvae of two drosophilid flies, Drosophila melanogaster and Chymomyza costata Although the species and variants ranged broadly between 0 and close to 100% survival of freezing, there were relatively small differences in ice fraction dynamics. For instance, the maximum ice fraction (IFmax) ranged between 67.9% and 77.7% total body water (TBW). Chymomyza costata larvae showed statistically significant phenotypic shifts in parameters of ice fraction dynamics (melting point and IFmax) upon entry into diapause, cold acclimation and feeding on a proline-augmented diet. These differences were mostly driven by colligative effects of accumulated proline (ranging between 6 and 487 mmol kg-1 TBW) and other metabolites. Our data suggest that these colligative effects per se do not represent a sufficient mechanistic explanation for high freeze tolerance observed in diapausing, cold-acclimated C. costata larvae. Instead, we hypothesize that accumulated proline exerts its protective role via a combination of mechanisms. Specifically, we found a tight association between proline-induced stimulation of glass transition in partially frozen body liquids (vitrification) and survival of cryopreservation in liquid nitrogen.