Linking energetics and overwintering in temperate insects.

Overwintering insects cannot feed, and energy they take into winter must therefore fuel energy demands during autumn, overwintering, warm periods prior to resumption of development in spring, and subsequent activity. Insects primarily consume lipids during winter, but may also use carbohydrate and proteins as fuel. Because they are ectotherms, the metabolic rate of insects is temperature-dependent, and the curvilinear nature of the metabolic rate-temperature relationship means that warm temperatures are disproportionately important to overwinter energy use. This energy use may be reduced physiologically, by reducing the slope or elevation of the metabolic rate-temperature relationship, or because of threshold changes, such as metabolic suppression upon freezing. Insects may also choose microhabitats or life history stages that reduce the impact of overwinter energy drain. There is considerable capacity for overwinter energy drain to affect insect survival and performance both directly (via starvation) or indirectly (for example, through a trade-off with cryoprotection), but this has not been well-explored. Likewise, the impact of overwinter energy drain on growing-season performance is not well understood. I conclude that overwinter energetics provides a useful lens through which to link physiology and ecology and winter and summer in studies of insect responses to their environment

The overwintering biology of the acorn weevil, Curculio glandium in southwestern Ontario.

The acorn weevil, Curculio glandium, is a widespread predator of acorns in eastern North America that overwinters in the soil as a larva. It is possible that low temperatures limit its northern geographic range, so we determined the cold tolerance strategy, seasonal variation in cold tolerance, and explored the physiological plasticity of overwintering larvae. Weevil larvae were collected from acorns of red and bur oak from Pelee Island, southwestern Ontario in fall 2010 and 2011. C. glandium larvae are freeze avoidant and larvae collected from bur oak acorns had lower supercooling points (SCPs: -7.6±0.36°C, LT50: -7.2°C) than those collected from red oak acorns (SCPs: -6.1±0.40°C, LT50: -6.1°C). In the winter of 2010-2011, SCPs and water content decreased, however these changes did not occur in 2011-2012, when winter soil temperatures fluctuated greatly in the absence of the buffering effect of snow. To examine whether larvae utilize cryoprotective dehydration, larvae from red oak acorns were exposed to -5°C in the presence of ice for seven days. These conditions decreased the SCP without affecting water content, suggesting that SCP and water content are not directly coupled. Finally, long-term acclimation at 0°C for six weeks slightly increased cold tolerance but also did not affect water content. Thus, although larval diet affects cold tolerance, there is limited plasticity after other treatments. The soil temperatures we observed were not close to lethal limits, although we speculate that soil temperatures in northerly habitats, or in years of reduced snow cover, has the potential to cause mortality in the field

Mechanisms underlying insect freeze tolerance.

Freeze tolerance - the ability to survive internal ice formation - has evolved repeatedly in insects, facilitating survival in environments with low temperatures and/or high risk of freezing. Surviving internal ice formation poses several challenges because freezing can cause cellular dehydration and mechanical damage, and restricts the opportunity to metabolise and respond to environmental challenges. While freeze-tolerant insects accumulate many potentially protective molecules, there is no apparent \u27magic bullet\u27 - a molecule or class of molecules that appears to be necessary or sufficient to support this cold-tolerance strategy. In addition, the mechanisms underlying freeze tolerance have been minimally explored. Herein, we frame freeze tolerance as the ability to survive a process: freeze-tolerant insects must withstand the challenges associated with cooling (low temperatures), freezing (internal ice formation), and thawing. To do so, we hypothesise that freeze-tolerant insects control the quality and quantity of ice, prevent or repair damage to cells and macromolecules, manage biochemical processes while frozen/thawing, and restore physiological processes post-thaw. Many of the molecules that can facilitate freeze tolerance are also accumulated by other cold- and desiccation-tolerant insects. We suggest that, when freezing offered a physiological advantage, freeze tolerance evolved in insects that were already adapted to low temperatures or desiccation, or in insects that could withstand small amounts of internal ice formation. Although freeze tolerance is a complex cold-tolerance strategy that has evolved multiple times, we suggest that a process-focused approach (in combination with appropriate techniques and model organisms) will facilitate hypothesis-driven research to understand better how insects survive internal ice formation

Insect Immunity Varies Idiosyncratically During Overwintering.

Overwintering insects face multiple stressors, including pathogen and parasite pressures that shift with seasons. However, we know little of how the insect immune system fluctuates with season, particularly in the overwintering period. To understand how immune activity changes across autumn, winter, and spring, we tracked immune activity of three temperate insects that overwinter as larvae: a weevil (Curculio sp., Coleoptera), gallfly (Eurosta solidaginis, Diptera), and larvae of the lepidopteran Pyrrharctia isabella. We measured baseline circulating hemocyte numbers, phenoloxidase activity, and humoral antimicrobial activity, as well as survival of fungal infection and melanization response at 12°C and 25°C to capture any potential plasticity in thermal performance. In Curculio sp. and E. solidaginis, hemocyte concentrations remained unchanged across seasons and antimicrobial activity against Gram-positive bacteria was lowest in autumn; however, Curculio sp. were less likely to survive fungal infection in autumn, whereas E. solidaginis were less likely to survive infection during the winter. Furthermore, hemocyte concentrations and antimicrobial activity decreased in P. isabella overwintering beneath snow cover. Overall, seasonal changes in activity were largely species dependent, thus it may be difficult to create generalizable predictions about the effects of a changing climate on seasonal immune activity in insects. However, we suggest that the relationship between the response to multiple stressors (e.g., cold and pathogens) drives changes in immune activity, and that understanding the physiology underlying these relationships will inform our predictions of the effects of environmental change on insect overwintering success

Overwintering Red Velvet Mites Are Freeze Tolerant

Although many arthropods are freeze tolerant (able to withstand internal ice), small-bodied terrestrial arthropods such as mites are thought to be constrained to freeze avoidance. We field-collected active adult red velvet mites, Allothrombium sp. (Trombidiidae), in winter in Southwestern Ontario, Canada, where temperatures drop below −20°C. These mites froze between −3.6° and −9.2°C and survived internal ice formation. All late-winter mites survived being frozen for 24 h at −9°C, and 50% survived 1 wk. The lower lethal temperature (LLT50; low temperature that kills 50% of mites) was ca. −20°C in midwinter. Hemolymph osmolality and glycerol concentration increased in midwinter, accompanied by decreased water content. Thus, this species is freeze tolerant, demonstrating that there is neither phylogenetic nor size constraint to evolving this cold tolerance strategy

Thermal variability and plasticity drive the outcome of a host-pathogen interaction

Variable, changing, climates may affect each participant in a biotic interaction differently. We explored the effects of temperature and plasticity on the outcome of a host-pathogen interaction to try to predict the outcomes of infection under fluctuating temperatures. We infected Gryllus veletis crickets with the entomopathogenic fungus Metarhizium brunneum under constant (6 °C, 12 °C, 18 °C or 25 °C) or fluctuating temperatures (6 °C to 18 °C or 6 °C to 25 °C). We also acclimated crickets and fungi to constant or fluctuating conditions. Crickets acclimated to fluctuating conditions survived best under constant conditions if paired with warm-acclimated fungus. Overall, matches and mismatches in thermal performance, driven by acclimation, determined host survival. Mismatched performance also determined differences in survival under different fluctuating thermal regimes: crickets survived best when fluctuating temperatures favoured their performance (6 °C to 25 °C), compared to fluctuations that favoured fungus performance (6 °C to 18 °C). Thus, we could predict the outcome of infection under fluctuating temperatures by averaging relative host-pathogen performance under constant temperatures, suggesting that it may be possible to predict responses to fluctuating temperatures for at least some biotic interactions

Overwintering biology of the carob moth Apomyelois ceratoniae (Lepidoptera: Pyralidae)

The pomegranate fruit moth, Apomyelois ceratoniae (Zeller), is the most important pest of pomegranate orchards in Iran, where infestations lead to 20%–80% fruit loss. A. ceratoniae overwinters as larvae in several instars. The success in overwintering determines the fruit loss in the following season, thus overwintering physiology of A. ceratoniae could provide insights into population prediction. To this end, overwintering strategy and some seasonal physiological and biochemical changes were investigated in the field-collected larvae of A. ceratoniae. The lowest supercooling point was recorded in November (−14.6 ± 0.9 °C) and the highest in both October and March (−10.2 ± 0.9 °C). The median lethal temperature (LT50) of larvae was higher than supercooling point, suggesting that A. ceratoniae is chill-susceptible. Overwintering larvae had slightly higher concentrations of glycerol and sorbitol compared to summer larvae. There were no significant seasonal changes in body water content or hemolymph osmolality. Current winter temperatures in Iranian orchards are higher than the cold tolerance thresholds of A. ceratoniae, suggesting that overwintering mortality is not a key factor in determining A. ceratoniae populations

Ion and water balance in Gryllus crickets during the first twelve hours of cold exposure.

Insects lose ion and water balance during chilling, but the mechanisms underlying this phenomenon are based on patterns of ion and water balance observed in the later stages of cold exposure (12 or more hours). Here we quantified the distribution of ions and water in the hemolymph, muscle, and gut in adult Gryllus field crickets during the first 12h of cold exposure to test mechanistic hypotheses about why homeostasis is lost in the cold, and how chill-tolerant insects might maintain homeostasis to lower temperatures. Unlike in later chill coma, hemolymph [Na(+)] and Na(+) content in the first few hours of chilling actually increased. Patterns of Na(+) balance suggest that Na(+) migrates from the tissues to the gut lumen via the hemolymph. Imbalance of [K(+)] progressed gradually over 12h and could not explain chill coma onset (a finding consistent with recent studies), nor did it predict survival or injury following 48h of chilling. Gryllus veletis avoided shifts in muscle and hemolymph ion content better than Gryllus pennsylvanicus (which is less chill-tolerant), however neither species defended water, [Na(+)], or [K(+)] balance during the first 12h of chilling. Gryllus veletis better maintained balance of Na(+) content and may therefore have greater tissue resistance to ion leak during cold exposure, which could partially explain faster chill coma recovery for that species