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INTRODUCTION: When overwintering, most endoparasitoids are protected from the cold inside their hosts. However, some endoparasitoids, along with ectoparasitoids, fall into the category called outside-hosts-overwintering parasitoids (OHOP) at immature or adult stages. We compared the cold-hardiness capacity and strategy between adult OHOP and their hosts (HOST) by examining their supercooling points (SCP), with acclimation periods and acclimation temperatures, and their lower lethal temperatures at 50% mortality (LLT50). We hypothesized that OHOP are more cold-hardy than their HOST, with lower SCP and LLT50. MATERIALS AND METHODS: Throughout the summers of 2020, 2021, and 2022, adult cabbage seedpod weevils (HOST) were sampled with a sweep net at the canola pod stage, and thousands of pods were collected and placed in emergence boxes to retrieve the adult OHOP Trichomalus perfectus. Regarding SCP measures, OHOP and HOST were separated according to various treatments. Each treatment considered a target exposure temperature (5, 10, or 20 °C) or a target exposure period (5, 15 or 25 days) at 5 °C. Regarding LLT measures, OHOP and HOST were categorized into five treatments, each corresponding to a specific exposure temperature (-5, -10, -15, -20 or -25 °C). RESULTS AND CONCLUSION: Acclimations to a lower temperature (5 °C) and a longer period (25 days) led to a significantly lower SCP of OHOP than HOST. Regarding OHOP, the average SCP was -19.71 °C when the acclimation temperature was 20 °C and significantly decreased to -23.20 °C when it was 5 °C. The average SCP was -18.82 °C when the acclimation period was five days and significantly decreased to -23.20 °C when it was 25 days. Conversely, the average SCP for HOST was never below -20 °C. At 20 °C acclimation temperature, HOST exhibited a significantly higher SCP of -14.64 °C compared to acclimations at 5 °C (-19.19 °C) and 10 °C (-20.00 °C), but there were no significant differences between 5 and 10 °C nor between acclimation periods. Therefore, the adult OHOP is more cold-hardy than its HOST. OHOP also exhibited a lower LLT50 than HOST, with -19.20 °C versus -17.59 °C. Finally, OHOP and HOST employ the same freeze-avoidance strategy, as evidenced by their SCP values (-19.57 °C versus -16.80 °C) which closely align with their respective LLT50. Adult OHOP better survive winter than their HOST in cold environments.

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Hylurgus ligniperda invaded Shandong, China, through imported forest timber, posing a threat to China's forest health. Exotic insects with broad environmental tolerance, including low temperatures, may have a better chance of surviving the winters and becoming invasive. Understanding the cold-tolerance strategies of H. ligniperda may help to design sustainable pest management approaches. In this study, we aim to investigate the cold-tolerance ability and relevant physiological indicators in overwintering H. ligniperda adults to determine any possible overwintering strategies. Supercooling points (SCPs) for adults H. ligniperda differed significantly across months and reached the lowest level in the mid- and post-overwintering period, the minimum SCPs -6.45 ±â€…0.18 °C. As the cold exposure temperature decreased, the survival rate of adults gradually decreased, and no adult survived more than 1 day at -15 °C, and the LLT50 for 1 day was -7.1 °C. Since H. ligniperda adults can survive internal ice formation, they are freeze-tolerant insects. Throughout the overwintering period, the SCPs and the water, protein, sorbitol, and glycerol content in adults decreased initially and then increased. We reported significant correlations between total protein, sorbitol, trehalose, and glycerol content in the beetles and SCPs. Glycogen, lipid, protein, trehalose, and sorbitol content in adult beetles may directly affect their cold-tolerance capacity and survival during winter. This study provides a physiological and biochemical basis for further study of metabolism and cold-tolerance strategies in H. ligniperda adults, which may help predict population dynamics and distribution potential of pests.

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The level of diversity and abundance of darkling beetles (Coleoptera, Tenebrionidae) is the main difference between the late Pleistocene and modern insect faunas of arid regions. In the Pleistocene assemblages they are extremely rare, whereas in the modern ones they predominate. It is assumed that the reason for their rarity in fossil entomological complexes is their lack of cold resistance. The supercooling points (SCP) and low lethal temperatures (LLT) of adults from five species of Altai darkling beetles that overwinter in the soil and larvae from one such species were measured in the laboratory. All beetles supercooled at negative temperatures but could not survive freezing, with the average SCP of the most cold-resistant species between -25.7 and -21.7 °C (Bioramix picipes, Anatolica dashidorzsi, and Penthicus altaicus). However, 50% of the individuals from different species in the experiment died after exposure during two days at temperatures ranging from -22 to -20 °C. The focal species are distributed in parts of Central Asia with an extreme continental climate, and the temperatures measured in the soil of these natural areas turned out to be lower than or close to the limit of cold resistance of the beetles. Overwintering of darkling beetles is therefore only possible in areas with deep snow: in hollows, under bushes, and under large cereals. Darkling beetles with poor cold resistance could not have existed in the colder climate of the late Pleistocene, which explains their absence from fossil fauna.

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A comparison was made of the supercooling points (SCPs) of questing Dermacentor variabilis adults from two populations located ca. 800 km apart on the Canadian prairies. This is also the first study to examine whether there are seasonal fluctuations in the SCP of questing D. variabilis adults. The SCPs of adult ticks from Lizard Lake Community Pasture, a recently established population in west-central Saskatchewan, varied over spring and summer, with the median SCP warming over time. In addition, the SCPs of ticks from Lizard Lake Community Pasture were significantly higher than those of adult ticks collected from Sandy Hook in Manitoba, a population that has been established for decades. The off-host survival of adults from Sandy Hook between summer and spring has been shown previously to be significantly greater than that of adults from Lizard Lake Community Pasture. The findings of the present study suggest that there may be geographical variation in the SCPs of D. variabilis adults which may be associated with differences in overwinter survival. The relatively low SCPs of questing D. variabilis adults, and the ability of some adults to survive off-host during winter, may be factors contributing to the range expansion of this tick species in Canada.

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The climatic variability hypothesis (CVH) predicts that organisms in more thermally variable environments have wider thermal breadths and higher thermal plasticity than those from more stable environments. However, due to evolutionary trade-offs, taxa with greater absolute thermal limits may have little plasticity of such limits (trade-off hypothesis). The CVH assumes that climatic variability is the ultimate driver of thermal tolerance variation across latitudinal and altitudinal gradients, but average temperature also varies along such gradients. We explored intraspecific variation of thermal tolerance in three typical Mediterranean saline water beetles (families Hydrophilidae and Dytiscidae). For each species, we compared two populations where the species coexist, with similar annual mean temperature but contrasting thermal variability (continental vs. coastal population). We estimated thermal limits of adults from each population, previously acclimated at 17, 20, or 25 °C. We found species-specific patterns but overall, our results agree with the CVH regarding thermal ranges, which were wider in the continental (more variable) population. In the two hydrophilid species, this came at the cost of losing plasticity of the upper thermal limit in this population, supporting the trade-off hypothesis, but not in the dytiscid one. Our results support the role of local adaptation to thermal variability and trade-offs between basal tolerance and physiological plasticity in shaping thermal tolerance in aquatic ectotherms, but also suggest that intraspecific variation of thermal tolerance does not fit a general pattern among aquatic insects. Overlooking such intraspecific variation could lead to inaccurate predictions of the vulnerability of aquatic insects to global warming.

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Penthaleus major (Dugés) is a significant agricultural pest that attacks various pasture, vegetable, and crop plants. Temperature plays a critical role in the life history of P. major. However, there is limited understanding of its life table at different temperatures and cold tolerance. This study aimed to elucidate the performance of P. major by constructing life tables at 6, 9, 12, 15, 18, 21, and 24 °C. The results showed that P. major successfully developed at 9‒21 °C. However, no adults emerged at 6°C, and no eggs hatched at 24 °C. The highest intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), and gross reproductive rate (GRR) were observed at 12 °C. The supercooling point (SCP) exhibited significant variations at different developmental stages. The highest SCP (‒9.75 °C) was recorded in 10-day-old female adults, while the lowest SCP (‒24.37 °C) was observed in larvae. For female adult mites of 2, 6, and 10 days old, the low lethal temperatures (LLT50) were ‒14.63, ‒12.03, and ‒11.08 °C, respectively. This study provided valuable insights for modeling and predicting the population dynamics of P. major in the field and offered implications for developing successful management strategies.

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A major component of mosquito's climate change response is their heat tolerance, and any ability to rapidly adjust to extreme environmental conditions through phenotypic plasticity. The excessive use of insecticides for the control of major mosquito species leads to resistant populations, however it is largely unclear if this concurrently impacts thermal stress resistance and their potential to adjust tolerance via phenotypic plasticity. Culex pipiens pipiens, Culex pipiens molestus and Aedes albopictus populations obtained from the same region were subjected for 12 generations to selection trials to larvicides Diflubenzuron (DFB) and Bacillus thuringiensis subsp. israelensis (Bti) to develop insecticide resistance. Adults emerging from the selected populations were acclimated at different temperatures and the upper and lower critical thermal limits (CTmax and CTmin) were estimated using dynamic thermal assays. In addition, the supercooling points (SCPs) of non-acclimated adults of resistant and control populations were determined. Our results revealed marked differences in thermal response among the three species, the different acclimation regimes and sexes. Aedes albopictus was more resistant in high than low temperatures compared to both Culex pipiens biotypes. Culex forms responded similarly to heat but differently to cold stress. In both forms, females responded better than males to all thermal stressors. Acclimation at higher and lower temperatures improves CTmax and CTmin values, respectively in both insecticide resistant and control populations of all three species. Overall, selection to insecticides did not affect the thermal performance of adults. Hence, insecticide-resistant mosquito populations perform similarly to untreated ones and are capable of readily adapting to new environmental changes rising concerns regarding their geographic range expansion and disease transmission globally.

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The chalcid wasp Tetrastichus septentrionalis Yang (Hymenoptera: Eulophidae) is one of the dominant pupal parasitoids of Hyphantria cunea (Drury) (Lepidoptera: Erebidae). In this study, the photoperiod's effect on diapause induction in T. septentrionalis using the alternative host Tenebrio molitor was measured, revealing that T. septentrionalis is of the long-day type. The critical photoperiods for diapause induction in T. septentrionalis were estimated to be between photoperiods of 13:11 and 14:10 (L:D) h at 18 °C, and between photoperiods of 12:12 and 13:11 (L:D) h at 21 °C and 24 °C. We also found that T. septentrionalis diapausing larvae were grey-brown, while normally developed (non-diapausing) individuals were light yellow. The diapause-sensitive insect state was the larval stage, and the short light exposure treatment had a significant cumulative effect on diapause induction. The least squares method was used to calculate a lower developmental threshold of 13.34 ± 0.50 °C and an effective cumulative temperature of 184.46 ± 11.46 d·°C for post-diapause development. The average supercooling point of diapausing mature larvae was significantly lower than that of non-diapausing ones. Our research on T. septentrionalis provides a reference for the biological control of H. cunea and other leaf-eating pests.

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Dermacentor variabilis (Say) is expanding its range northwards in Saskatchewan and Manitoba (Canada). Knowledge of the relative ability of different life cycle stages to survive off-host over winter is important for assessing the potential for further range expansion and risk of pathogen transmission. In the present study, the off-host survival of D. variabilis adults was determined in outdoor terraria within a field plot close to Lizard Lake Community Pasture (LLCP) situated near the northern distributional limit in Saskatchewan. Only a small proportion (7%) of adult ticks in this relatively recently established population survived from mid-summer to late winter. Off-host survival had declined further to <4% by mid-spring. A significantly greater proportion of female ticks (6%) survived from mid-summer to mid-spring than did male ticks (1%). The supercooling points (SCPs), the temperature below 0 °C when body fluids spontaneously freeze, of the 17 ticks that survived through to mid-spring did not differ significantly from the SCPs of questing ticks in the previous summer, suggesting adult D. variabilis do not undergo cold hardening during winter diapause. The off-host survival of D. variabilis adults near LLCP was also much lower than that previously reported for adults from Sandy Hook in Manitoba, a population that has been established for decades. This finding suggests that there is geographical variation in the ability of D. variabilis in southern Canada to survive from mid-summer to the subsequent spring. However, the ecological and genetic factors influencing the survival of northern populations of D. variabilis requires further investigation.

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The saffron mite, Rhizoglyphus robini Claparède (Acari, Astigmata: Acaridae), is one of the most important pests of saffron-producing regions in Iran. It causes yellowing and decreases saffron growth, and finally it destroys the bulbs. In this research, the cold tolerance and supercooling point (SCP) of the saffron mite were measured in three populations and two temperature regimes. Our results showed that the mean SCP of the saffron mite was approximately -14.6 °C without significant difference among the populations. On the contrary, acclimation of the mites significantly decreased their SCP to a mean of approximately -16.5 °C. Exposure of the mites for 24 h to 0 and -2.5 °C had no significant effect on the survival of the mites but when the mites were exposed to -5.0 °C for 24 h, survival of the three populations reached the lowest level of roughly 60%. By 24-h exposure to -7.5 °C, survival of the mites was almost negligible. As a large proportion of mortality was observed above the SCP, and LT50 > SCP, it can be inferred that the saffron mite is likely a chill-susceptible species. This suggests that the saffron mite lacks the ability to withstand extracellular ice formation. Overall, the results of the current study suggest no significant physiological differences between populations of the saffron mite.

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Broad-spectrum insecticides are the main control measure of the invasive and economically damaging soybean aphid (Aphis glycines) in North America, although biological control by resident natural enemies can also greatly diminish population levels. One such natural enemy is the accidentally introduced Eurasian parasitoid Aphelinus certus (Hymenoptera: Aphelinidae), though its impact appears to be limited by low rates of parasitism early in the growing season. We tested the hypothesis that A. certus might experience high overwintering mortality. In the laboratory, we used thermocouple thermometry to measure the supercooling points of diapausing parasitoids and assessed parasitoid survival after exposure to ecologically relevant durations of low temperature. We found A. certus to be freeze-intolerant with a median supercooling point of -28°C. When exposed to temperatures of 0°C for up to 7 months, adults emerged only after exposures of at least 60 days and survival decreased with durations beyond 150 days. We also conducted in-field studies at sites from northern Minnesota to southern Iowa to determine if diapausing A. certus could overwinter above and below the snowpack. Survival was negatively correlated with increasing latitude and was greater for parasitoids placed on the ground than 1 meter off the ground, likely due to the warmer and stabler temperatures of the subnivean microclimate. Our results suggest that A. certus is capable of overwintering in the region inhabited by soybean aphid but may experience substantial mortality even under ideal conditions. Climate change is predicted to bring warmer, drier winters to the North American Midwest, with decreased depth and duration of snow cover, which may further reduce overwintering survival.

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Thermal tolerance greatly influences the geographic distribution, seasonality, and feeding habits of mosquitoes; this study aimed to examine the impacts of species, sex, and diet on thermal tolerance in mosquitoes. We found that Culex quinquefasciatus was inherently significantly more cold tolerant than Aedes aegypti, while Ae. aegypti had improved heat tolerance compared to Cx. quinquefasciatus. There were no differences in thermal tolerance between sexes within either species. We observed similar levels of cold tolerance between all diets tested, but observed decreased heat tolerance in mannitol-fed mosquitoes. Our results suggest that although dietary factors such as sugar alcohols and sugars may play a role in thermal tolerance in mosquitoes, there are likely physiological and genetic factors that can have a greater influence on the limits of thermal tolerance within a species.

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Winter low-temperature confines species distribution. Intertidal gastropods are distributed from tropical to polar zones, facing variable intensities and durations of low temperatures. They usually set their supercooling points (SCPs) at high subzero temperatures to stimulate freezing. However, the variations in SCP in intertidal gastropods at intraspecific and interspecific levels remain poorly understood. Here, we measured the body size, cooling rate, and SCP of nine intertidal gastropod species in China. These species were distributed in high or middle intertidal zone with different geographic distributions. The average SCPs (-4.27~-7.10 °C) and the coefficients of variation of SCP (22.6%~45.9%) were high in all species. At the intraspecific level, the supercooling capacity was positively correlated with the cooling rate. Interspecifically, the supercooling capacity was closely related to the cooling rate, and also to the species' geographical distribution. Northern high-shore species showed lower SCPs, while southern high-shore species had higher SCPs. There was no difference in SCP between widespread high- and mid-shore species. Our results indicated that the supercooling capability is potentially an adaptative response to the local winter temperatures, and the cooling rate is a factor in determining the difference in SCP at the intraspecific and interspecific levels.

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The variegated ladybug, Hippodamia variegata is one of the most effective predators of various pests that hibernate as adult beetles. During the overwintering period from April 2021 to March 2022, we examined the supercooling point (SCP), cold tolerance, and physiological adaptations of beetles in Kerman, Iran. The beetles exhibited their greatest cold tolerance (63.4% after 24 h at -5°C) when their SCP was lowest (-23.2°C). Conversely, from April to October 2021, the SCP reached its peak (approximately -13.0°C), while cold tolerance was at its lowest level (6.7% after 24 h at -5°C). Cryoprotectant content (trehalose, glycerol, and glucose) was at its highest level in September (11.15, 10.82, and 6.31 mg/g, respectively). The critical thermal minimum (CTmin) reached its lowest point of -2.2°C in January and February. The lowest point of the lower lethal temperature (LLT) coincided with the lowest level of the SCP and the highest level of cold tolerance (in February, LT50 = -5.3°C, SCP = -23.2°C, and survival = 77.78% at -4°C/24 h). Chill-coma recovery time (CCRT) was examined at five different temperatures and two different exposure durations. The CCRT increased with a decrease in exposure temperature and time (68.0 s at -2°C after 2 h and 102.0 s at -2°C after 4 h). As the majority of the overwintering beetle's mortality occurred at temperatures significantly higher than SCP, the adults of H. variegata are chill-susceptible insects that primarily rely on a depressed supercooling point to cope with unfavorable conditions during the overwintering period.

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The swallowtail butterfly, Sericinus montelus Gray, is endemic to East Asia, has high ornamental value but faces an increased risk of extinction. To understand the overwintering strategies of this species, the dynamic changes in supercooling point (SCP) and water and biochemical contents of diapause-destined and non-diapause S. montelus pupae were investigated. The SCP of laboratory-reared diapause pupae was as low as -26°C compared to -24°C in diapause pupae in the field. Although there was no significant difference in total water content between diapause-destined and non-diapause pupae, the free water of diapause-destined pupae was significantly lower, and the bound water was significantly higher, than that of non-diapause pupae. Lipid, glycogen, and protein contents of diapause-destined pupae showed a downward trend, whereas the total sugar content showed the opposite trend after pupation. The glycogen content decreased rapidly during the initial stage of pupation, whereas the lipid content decreased significantly after 30 days of pupation, suggesting that diapause-destined pupae deplete glycogen stores during the pre-diapause period and then switch to using lipids during the diapause maintenance phase. Trehalose levels in diapause-destined pupae increased significantly and remained high after pupation. Meanwhile, the trehalose content of overwintering pupae during the diapause maintenance period was significantly higher than that of diapause termination pupae in the field. These results suggest that trehalose is the main cryoprotectant for overwintering pupae. Thus, diapausing S. montelus pupae appear to be freeze avoidant, accumulate trehalose as a cryoprotectant, and reduce the free water content to decrease the SCP, enhancing their cold tolerance.

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Evergestis extimalis (Scopoli) is a significant pest of spring oilseed rape in the Qinghai-Tibet Plateau. It has developed resistance to many commonly used insecticides. Therefore, biopesticides should be used to replace the chemical pesticides in pest control. In this study, the effects of ice-nucleation-active (INA) microbes (Pseudomonas syringae 1.7277, P. syringae 1.3200, and Erwinia pyrifoliae 1.3333) on E. extimalis were evaluated. The supercooling points (SCP) were markedly increased due to the INA bacteria application when they were compared to those of the untreated samples. Specifically, the SCP of E. extimalis after its exposure to a high concentration of INA bacteria in February were -10.72 °C, -13.73 °C, and -14.04 °C. Our findings have demonstrated that the trehalase (Tre) genes were up-regulated by the application of the INA bacteria, thereby resulting in an increased trehalase activity. Overall, the INA bacteria could act as effective heterogeneous ice nuclei which could lower the hardiness of E. extimalis to the cold and then freeze them to death in an extremely cold winter. Therefore, the control of insect pests with INA bacteria goes without doubt, in theory.

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Diapause and cold hardiness are essential components of winter survival for most insects in temperate zones. The fall webworm, Hyphantria cunea, overwinters in a pupal diapause. In this study, we investigated the energy consumption and cold hardiness of diapausing pupae. We found that lipid content decreased from October to November and stabilized from November to March. Glycogen content decreased by 61.3% and 52.2% for females and males, respectively, from October to November, and decreased slowly from November to March. We also observed a significant increase in trehalose concentrations as ambient temperatures decreased from October to November and a decrease in trehalose as temperatures increased again in March. We did not observe substantial changes in pupal supercooling points among the dates sampled. In addition, prolonged pupal development time reduced their survival rate and had no significant effect on post-diapause adult body mass and fecundity but reduced egg diameter in females. These results suggest that the energy consumption of H. cunea pupae during early diapause depends on lipid and glycogen, while it shifts to depend on glycogen or other energy stores in the mid- and late diapause stages. Our results also suggest that the prolonged development time of diapausing pupae had a negative effect on post-diapause fitness.

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Global changes in soil surface temperatures are altering the abundances and distribution ranges of invertebrate species worldwide, including effects on soil microarthropods such as springtails (Collembola), which are vital for maintaining soil health and providing ecosystem services. Studies of thermal tolerance limits in soil invertebrates have the potential to provide information on demographic responses to climate change and guide assessments of possible impacts on the structure and functioning of ecosystems. Here, we review the state of knowledge of thermal tolerance limits in Collembola. Thermal tolerance metrics have diversified over time, which should be taken into account when conducting large-scale comparative studies. A temporal trend shows that the estimation of 'Critical Thermal Limits' (CTL) is becoming more common than investigations of 'Supercooling Point' (SCP), despite the latter being the most widely used metric. Indeed, most studies (66%) in Collembola have focused on cold tolerance; fewer have assessed heat tolerance. The majority of thermal tolerance data are from temperate and polar regions, with fewer assessments from tropical and subtropical latitudes. While the hemiedaphic life form represents the majority of records at low latitudes, euedaphic and epedaphic groups remain largely unsampled in these regions compared to the situation in temperate and high latitude regions, where sampling records show a more balanced distribution among the different life forms. Most CTL data are obtained during the warmest period of the year, whereas SCP and 'Lethal Temperature' (LT) show more variation in terms of the season when the data were collected. We conclude that more attention should be given to understudied zoogeographical regions across the tropics, as well as certain less-studied clades such as the family Neanuridae, to identify the role of thermal tolerance limits in the redistribution of species under changing climates.

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Bactrocera tau (Walker) (Diptera: Tephritidae) is a serious, economically important invasive pest that has spread and been established in many regions worldwide. Temperature is a crucial abiotic factor governing insect activity, fitness, and geographical distribution. Yet, surprisingly, the tolerance of B. tau to extreme cold and heat stress remains unclear. Here, we measured the supercooling point (SCP) of different life stages of B. tau. Further, several life stages of B. tau (egg, 1st, 2nd, and 3rd instar larvae, 1-day-old pupae, and 3-day-old adult) were subjected to six low temperatures (-9, -7, -5, -3, -1, and 0 °C) and six high temperatures (39, 40, 41, 42, 43, and 44 °C) for various durations (0.5, 1.0, 2.0, and 4.0 h), and three-way survival-time-temperature relationships were investigated. We found that the SCPs differed significantly among different life stages of B. tau, being the lowest for SCP of eggs, at -25.82 ± 0.51 °C. There was no significant effect of sex on the mean SCPs of B. tau adults, except for 45- to 50-day-old flies. In addition, an interaction effect was uncovered between tested temperatures and exposure duration upon B. tau mortality at different life stages. Eggs exhibited the strongest cold tolerance, yet the weakest heat tolerance. The 3rd instar larvae were the most heat- and cold tolerant among larval stages, followed by the 2nd and 1st instar larvae. The upper limit of the chill injury zone (ULCIZ) for 3-day-old adult and 1-day-old pupae was -2.51 °C and -2.50 °C, respectively, while their corresponding lower limit of thermal injury zone (LLTIZ) was 39.39 °C and 38.29 °C. This paper presents valuable data to provide an integrated knowledge for understanding the cold and heat tolerance potential of B. tau and ensure the proper implementation of post-harvest phytosanitary protocols for this pest's disinfestation.

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The honey locusts (genus Gleditsia) are a genus of high-value trees in Asia. Seed beetle, Megabruchidius dorsalis (Fåhraeus) (Col.: Chrysomelidae: Bruchinae), is a Gleditsia oligophagous pest that causes severe yield reduction. To understand the cold tolerance of M. dorsalis adults, this study investigated its cold tolerance strategy and the influence of low temperatures on its physiology and biochemistry. The low-temperature treatments were divided into three groups: long-term temperature acclimation (Group 1; 15°C, or 20°C, or 25°C, or 28°C [control check, CK] for 10 days), short-term low-temperature exposure (Group 2; 0°C or 4°C for 2 h), and long-term low-temperature induction (Group 3; 0°C or 4°C for 1, 3, or 5 d). The supercooling point (SCP; temperature at which spontaneous nucleation and ice lattice growth begin), freezing point (FP; temperature at which insect fluids freeze), low lethal temperature (LLT; temperature at which all individuals are killed), water, lipid, glycerol, and total sugars contents were measured under different temperature stresses. The results showed that M. dorsalis adults were a freeze-avoidant species. The SCP and LLT at 28°C were -10.62°C and -19.48°C, respectively. The SCP and FP of long-term temperature acclimation (15°C, or 20°C, or 25°C) were significantly lower than that of the control group (28°C). The water content of the long-term low temperature induction (0°C) group was significantly lower than that of the control group. The lipid and glycerol content in the acclimated group at 20°C and 25°C were significantly higher than in the control group. M. dorsalis adults may maintain their biofluids in a supercooled state via cryoprotectant accumulation and cryoprotective dehydration to prevent ice nucleation. This study provides a theoretical basis for future research on overwintering and potential distribution and related prediction of M. dorsalis adults.