Low Temperature Exposure Research Articles

Lethal and Sublethal Implications of Low Temperature Exposure for Three Intertidal Predators

Lethal and Sublethal Implications of Low Temperature Exposure for Three Intertidal Predators

Low Temperature Effect on Different Varieties of Corchorus capsularis and Corchorus olitorius at Seedling Stage

To address the demand for natural fibers, developing new varieties that are resistant to abiotic stress is necessary. The present study was designed to investigate the physiological and biochemical traits of three varieties of C. capularis (Y49, Y38, and Y1) and four varieties C. olitorius (T8, W57, M33, M18) under low temperature to identify the cold-tolerant varieties and elucidate the mechanisms involved in enhancing cold tolerance. Research findings revealed that the varieties Y49 and M33 exhibited the highest chlorophyll and carotenoid content. Biochemical profiles revealed that varieties Y49 and M33 were found to be able to withstand low-temperature stress by accumulating different enzymatic and non-enzymatic antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APx), glutathione (GSH), and phenolics, which participated in reducing the content of malondialdehyde (MDA) and hydrogen peroxide (H2O2) caused by low temperature. Osmolytes compounds, such as total soluble sugar, significantly increased in Y49 and M33; and proline content decreased in all varieties except Y49 and M33 after low-temperature exposure. The rise in these osmolytes molecules can be a defense mechanism for the jute’s osmotic readjustment to reduce the oxidative damage induced by low temperature. Furthermore, PCA and hierarchical cluster analysis distinguished the seven varieties into three separate groups. Results confirmed that group I (Y49 and M33 varieties) were low-temperature tolerant, group II (M18, W57) were intermediate, whereas III groups (Y38, T8, and Y1) were low temperature susceptible. PCA also explained 88.36% of the variance of raw data and clearly distinguished three groups that are similar to the cluster heat map. The study thus confirmed the tolerance of selected varieties that might be an efficient adaptation strategy and utilized them for establishing breeding programs for cold tolerance.

Acclimations to Cold and Warm Conditions Differently Affect the Energy Metabolism of Diapausing Larvae of the European Corn Borer Ostrinia nubilalis (Hbn.).

The European corn borer Ostrinia nubilalis is a pest species, whose fifth instar larvae gradually develop cold hardiness during diapause. The physiological changes underlying diapause progression and cold hardiness development are still insufficiently understood in insects. Here, we follow a complex of changes related to energy metabolism during cold acclimation (5°C) of diapausing larvae and compare this to warm-acclimated (22°C) and non-diapause controls. Capillary electrophoresis of nucleotides and coenzymes has shown that in gradually cold-acclimated groups concentrations of ATP/ADP and, consequently, energy charge slowly decrease during diapause, while the concentration of AMP increases, especially in the first months of diapause. Also, the activity of cytochrome c oxidase (COX), as well as the concentrations of NAD+ and GMP, decline in cold-acclimated groups, until the latter part of diapause, when they recover. Relative expression of NADH dehydrogenase (nd1), coenzyme Q-cytochrome c reductase (uqcr), COX, ATP synthase (atp), ADP/ATP translocase (ant), and prohibitin 2 (phb2) is supressed in cold-acclimated larvae during the first months of diapause and gradually increases toward the termination of diapause. Contrary to this, NADP+ and UMP levels significantly increased in the first few months of diapause, after gradual cold acclimation, which is in connection with the biosynthesis of cryoprotective molecules, as well as regeneration of small antioxidants. Our findings evidence the existence of a cold-induced energy-saving program that facilitates long-term maintenance of larval diapause, as well as gradual development of cold hardiness. In contrast, warm acclimation induced faster depletion of ATP, ADP, UMP, NAD+, and NADP+, as well as higher activity of COX and generally higher expression of all energy-related genes in comparison to cold-acclimated larvae. Moreover, such unusually high metabolic activity, driven by high temperatures, lead to premature mortality in the warm-acclimated group after 2 months of diapause. Thus, our findings strongly support the importance of low temperature exposure in early diapause for gradual cold hardiness acquisition, successful maintenance of the resting state and return to active development. Moreover, they demonstrate potentially adverse effects of global climate changes and subsequent increase in winter temperatures on cold-adapted terrestrial organisms in temperate and subpolar regions.

Phylogenetic Analysis of the Plant U2 snRNP Auxiliary Factor Large Subunit A Gene Family in Response to Developmental Cues and Environmental Stimuli.

In all organisms, splicing occurs through the formation of spliceosome complexes, and splicing auxiliary factors are essential during splicing. U2AF65 is a crucial splicing cofactor, and the two typical RNA-recognition motifs at its center recognize and bind the polypyrimidine sequence located between the intron branch site and the 3′-splice site. U2AF65A is a member of the U2AF65 gene family, with pivotal roles in diseases in mammals, specifically humans; however, few studies have investigated plant U2AF65A, and its specific functions are poorly understood. Therefore, in the present study, we systematically identified U2AF65A in plant species from algae to angiosperms. Based on 113 putative U2AF65A sequences from 33 plant species, phylogenetic analyses were performed, followed by basic bioinformatics, including the comparisons of gene structure, protein domains, promoter motifs, and gene expression levels. In addition, using rice as the model crop, we demonstrated that the OsU2AF65A protein is localized to the nucleus and cytoplasm, and it is involved in responses to various stresses, such as drought, high salinity, low temperature, and heavy metal exposure (e.g., cadmium). Using Arabidopsis thaliana and rice mutants, we demonstrated that U2AF65A is involved in the accumulation of plant biomass, growth of hypocotyl upon thermal stimulation, and reduction of tolerance of high temperature stress. These findings offer an overview of the U2AF65 gene family and its stress response functions, serving as the reference for further comprehensive functional studies of the essential specific splicing cofactor U2AF65A in the plant kingdom.

Study of low-temperature exposure on biotissue using an elongated cryoapplicator

The article presents the results of a study of low-temperature exposure on animal biological tissue using the novel prototype of a liquid nitrogen cryoapplicator. The data obtained are compared with the cryoapplicator characteristics cooled by nitrogen dioxide that are currently used for the atrial fibrillation treatment. Data analysis confirmed the liquid nitrogen cryoapplicators effectiveness and made it possible to highlight their advantages.

Orientation of Tribolium castaneum (Coleoptera: Tenebrionidae) adults to 4,8-dimethyldecanal, kairomone and botanical oils following ambient, low, or high temperature exposure

The aggregation pheromone 4,8-dimethyldecanal (4,8 DMD) used alone or with kairomone-baited traps generally is used for monitoringTribolium castaneum (Herbst) but low efficiency is reported. Furthermore, insect orientation to pheromones and kairomones following low or high temperature exposure is not yet understood. This research evaluated the orientation of T. castaneum adults to 4,8 DMD and common kairomones following exposure to ambient, low or high temperatures. Fifty adults were introduced to the middle of rectangular glass chamber, and movement to the treatment or control was determined after 1 h. In experiment 1, insects reared at 30°C were used. Experiments 2 and 3 used insects exposed to 10°C for 4 days (cold temperature) and 42°C for 12 h (brief high temperature), respectively at 2–8 h following cold or heat exposure. The highest trapping occurred when 4,8DMD was combined with neem oil whereas the lowest was in coconut oil alone. Neem oil alone demonstrated repellent action. Prior exposure to cold or heat initially reduced taxis of T. castaneum adults to traps. The attraction for adults exposed to cold and heat was restored after 6–8 h when traps contained pheromone + neem or coconut oil. This study marks the first to experimentally test how exposure to high or low temperatures, two key IPM tactics in the post-harvest supply chain, affects the behavioral response of an important stored-product species to pheromone- and kairomones-baited traps.

Perirenal Adipose Tissue from Healthy Donor: Characteristics and Promise as Potential Therapeutic Cell Source.

Perirenal adipose tissue, one of the fat masses surrounding the kidneys, can be obtained from healthy donors during a kidney transplant. Perirenal adipose tissue has only ever been known as a connective tissue to protect the kidneys and renal blood vessels from external physical stimulation. Yet, recently, as adipose tissue has begun to be considered an endocrine organ, and perirenal adipose tissue is now regarded to have a direct effect on metabolic diseases. The characteristics of perirenal adipose tissue from a healthy donor are that: (1) There are a large number of brown adipose cells (70–80% of the total), (2) Most of the brown adipose cells are inactive in the resting cell cycle, (3) Activating factors are constant low-temperature exposure, hormones, metastasis factors, and environmental factors, (4) Anatomically, a large number of brown adipose cells are distributed close to the adrenal glands, (5) Beige cells, produced by converting white adipocytes to brown-like adipocytes, are highly active, (6) Activated cells secrete BATokines, and (7) Energy consumption efficiency is high. Despite these advantages, all of the perirenal adipose tissue from a healthy donor is incinerated as medical waste. With a view to its use, this review discusses the brown adipocytes and beige cells in perirenal adipose tissue from a healthy donor, and proposes opportunities for their clinical application.

Cardiac and respiratory metabolic analysis of low-temperature tolerance in two geographic genotypes of Haliotis diversicolor and the hybrid

Evident survival heterosis exists in the hybrid (HdDY) of two Haliotis diversicolor geographic genotypes (maternal genotype HdTW and paternal genotype HdJP). To explore the physiological basis underlying the heterosis, we compared the survival rate, cardiac performance, oxygen consumption rate, and ammonia excretion rate of the three H. diversicolor genotypes at the low temperature. After one year of cultivation in the abalone farm, the survival rates of HdTW, HdJP, and HdDY were 27.76%, 86.90%, and 88.57%, respectively. The 50% lethal low temperature (LLT50) of HdTW was higher than HdJP and HdDY. During low-temperature exposure, the abalone’s heartbeat variation pattern appeared as three stages: the heartbeat decreasing stage, arrhythmia stage, and cardiac recovery stage. Four cardiac indices were developed to compare the three genotypes’ cardiac differences: the low-temperature heartbeat decreasing rate (LDR), cardiac arrhythmia temperature (CAT), low-temperature heartbeat recovering rate (LRR), and cardiac recovery time (CRT). HdTW had lower LDR and higher CAT than HdJP and HdDY, while HdJP had lower LRR and higher CRT than HdTW and HdDY, suggesting the hybrid’s cardiac function is parent-like in low-temperature tolerance and mother-like in low-temperature recovery. Meanwhile, the heart rates of HdJP and HdDY were always higher than HdTW at both normal temperature (20 °C) and low temperature (12 °C). The oxygen consumption rate and ammonia excretion rate of HdJP and HdDY were also higher than HdTW after low-temperature exposure. The results indicate the hybrid’s advantageous respiratory and metabolic functions at low temperatures, which could be a vital physiological mechanism of the hybrid’s survival heterosis. This study elucidates the cardiac and respiratory metabolic differences among different abalone genotypes at low temperatures, which may help us understand the physiological basis of mollusks’ low-temperature tolerance. The evaluation protocol and indices developed in this study could be helpful in mollusks’ physiological researches, such as the low-temperature tolerance heterosis, low-temperature adaption, and habitat differentiation.

Differences in lipid homeostasis and membrane lipid unsaturation confer differential tolerance to low temperatures in two Cycas species

BackgroundC. panzhihuaensis is more tolerant to freezing than C. bifida but the mechanisms underlying the different freezing tolerance are unclear. Photosynthesis is one of the most temperature-sensitive processes. Lipids play important roles in membrane structure, signal transduction and energy storage, which are closely related to the stress responses of plants. In this study, the chlorophyll fluorescence parameters and lipid profiles of the two species were characterized to explore the changes in photosynthetic activity and lipid metabolism following low-temperature exposure and subsequent recovery.ResultsPhotosynthetic activity significantly decreased in C. bifida with the decrease of temperatures and reached zero after recovery. Photosynthetic activity, however, was little affected in C. panzhihuaensis. The lipid composition of C. bifida was more affected by cold and freezing treatments than C. panzhihuaensis. Compared with the control, the proportions of all the lipid categories recovered to the original level in C. panzhihuaensis, but the proportions of most lipid categories changed significantly in C. bifida after 3 d of recovery. In particular, the glycerophospholipids and prenol lipids degraded severely during the recovery period of C. bifida. Changes in acyl chain length and double bond index (DBI) occurred in more lipid classes immediately after low-temperature exposure in C. panzhihuaensis compare with those in C. bifida. DBI of the total main membrane lipids of C. panzhihuaensis was significantly higher than that of C. bifida following all temperature treatments.ConclusionsThe results of chlorophyll fluorescence parameters confirmed that the freezing tolerance of C. panzhihuaensis was greater than that of C. bifida. The lipid metabolism of the two species had differential responses to low temperatures. The homeostasis and plastic adjustment of lipid metabolism and the higher level of DBI of the main membrane lipids may contribute to the greater tolerance of C. panzhihuaensis to low temperatures.

Modification of the effects of nitrogen dioxide and sulfur dioxide on congenital limb defects by meteorological conditions.

Can meteorological conditions modify the associations between NO2 and SO2 exposure and congenital limb defects (CLDs) during the first trimester of pregnancy? Increases in NO2 and SO2 exposure were consistently associated with higher risks of CLDs during the first trimester of pregnancy; both low- and high-temperature exposure and high air humidity act synergistically with the two air pollutants on CLDs. Animal studies have indicated air pollutants are associated with CLDs, but corresponding epidemiological studies are limited with equivocal conclusions. Meteorological conditions are closely connected to the generation, diffusion, distribution and even chemical toxicity of air pollutants. This case-control study included 972 cases of CLDs and 9720 controls in Changsha, China during 2015-2018. Cases from the hospital based monitoring system for birth defects (including polydactyly, syndactyly, limb shortening, and clubfoot) and healthy controls from the electronic medical records system were studied. Complete data on daily average NO2 and SO2 concentrations and meteorological variables were obtained from local monitoring stations to estimate monthly individual exposures during the first trimester of pregnancy, using the nearest monitoring station approach for NO2 and SO2 concentrations, and the city-wide average approach for temperature and relative humidity, respectively. The 25th and 75th percentiles of daily mean temperature, as well as the 50th percentile of daily mean relative humidity during the study period were used to classify high- and low-temperature exposure, and high humidity exposure based on existing evidence and local climate characteristics. Multivariate logistic regression models were used to estimate the independent effects per 10 μg/m3 increase in NO2 and SO2 on CLDs, and the attribute proportions of interaction (API) were used to quantify the additive joint effects of air pollutants with meteorological conditions after including a cross product interaction term in the regression models. NO2 and SO2 exposures during the first trimester of pregnancy were consistently and positively associated with overall CLDs and subtypes, with adjusted odd ratios (aORs) ranging from 1.13 to 1.27 for NO2, and from 1.37 to 2.49 for SO2. The effect estimates were generally observed to be the strongest in the first month and then attenuated in the second and third months of pregnancy. Synergistic effects of both low and high temperature in combination with NO2 (with APIs ranging from 0.07 to 0.38) and SO2 (with APIs ranging from 0.18 to 0.51) appeared in the first trimester of pregnancy. Several significant modifying effects by high humidity were also observed, especially for SO2 (with APIs ranging from 0.13 to 0.38). Neither NO2 nor SO2 showed an interactive effect with season of conception. The methods used to estimate individual exposure levels of air pollutants and meteorological factors may lead to the misclassification bias because of the lack of information on maternal activity patterns and residential mobility during pregnancy. Moreover, we were unable to consider several potentially confounding factors, including socioeconomic status, maternal nutrient levels, alcohol use and smoking during early pregnancy due to unavailable data, although previous studies have suggested limited change to the results after when including these factors in the analysis. The findings are helpful for understanding the combined effects of air pollution and meteorological conditions on birth defects. Environmental policies and practices should be formulated and implemented to decrease air pollutant emissions and improve meteorological conditions to reduce their harmful effects on pregnancy. Additionally, pregnant women should be suggested to reduce outdoor time when the air quality is poor, especially when ambient temperature is higher or lower than what is comfortable, or when it is excessively humid. The study is funded by Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defects in Hunan Province (2019SK1012), Major Research and Development Projects in Hunan Province (2018SK2060) and Scientific and Technological Department Projects in Hunan Province (2017SK50802). There are no competing interests. N/A.

CFD-Based Safety Assessment of a Novel Concept of a CNG Ship

The GASVESSEL project aims to prove the techno-economic feasibility of a new CNG (Compressed Natural Gas) transport concept, enabled by a novel patented Pressure Vessel manufacturing technology and a new conceptual ship design including safe on- and off-loading solution. This article describes the CFD models that have been defined for a preliminary safety assessment on the ship. Analytical models and numerical simulations were used in order to have a better understanding of the problem and give to the partners of the project the indications necessary to identify the proper solutions to improve safety on the ship. First, a possible gas leakage in the hold is analyzed, considering all side effects caused by the low temperature exposure, and a CFD study on the flow through a small crack with a high pressure differential is studied. Then the focus is moved to the compressor room, where different ventilation solutions are analyzed to reduce explosion risks in case of gas dispersion and to allow partners to understand which one best suits the requirements. Finally, an emergency vent mast system, that is activated in case of critical gas dispersion to evacuate the gas from the hold, is analyzed. In particular, the aim is to determine if the gas cloud will impact the exhaust chimneys at the ship's stern or the accommodations at the ship's bow.

Trans-seasonal activation of the neuroendocrine reproductive axis: Low-temperature winter dormancy modulates gonadotropin-releasing hormone neurons in garter snakes.

All animals use external cues from the environment to accurately time life-history events. How the brain decodes environmental stimuli to effect changes in physiology and behavior, however, is poorly understood, particularly with regard to supplementary environmental cues such as temperature. We asked if low-temperature dormancy alters the synthesis and/or release of gonadotropin-releasing hormone (GnRH). We used the well-studied red-sided garter snake (Thamnophis sirtalis) for this study, as low-temperature exposure is both necessary and sufficient to induce reproduction in northern populations of this species. Snakes were collected from the field and hibernated at 4°C or 10°C in complete darkness for up to 16 weeks. In males, increasing duration of low-temperature dormancy significantly increased GnRH-immunoreactive cell number and GnRH soma size (a proxy for relative cell activity) in the forebrain. These changes mirrored those in male reproductive behavior (reported previously) and plasma androgen concentrations. The changes in GnRH cell area observed in males were specific to the neuroendocrine population of cells in the medial preoptic area; soma size in the rostral GnRH cells did not change. Finally, temperature-induced changes in GnRH were sexually dimorphic: neither hibernation temperature nor the duration of winter dormancy significantly modulated GnRH cell number or soma size in females, despite the fact that plasma estradiol and corticosterone increased significantly in response to both. These data demonstrate that the neuroendocrine GnRH system is sensitive to environmental temperature and suggest that GnRH neurons play a conserved but trans-seasonal role in mediating changes in reproductive physiology and behavior in dissociated breeders.

QTL mapping of seedling tolerance to exposure to low temperature in the maize IBM RIL population.

Maize is a cold sensitive crop that exhibits severe retardation of growth and development when exposed to cold spells during and right after germination, including the slowdown in development of new leaves and in formation of the photosynthetic apparatus. Improving cold tolerance in maize would allow early sowing to improve crop yield by prolonging a growing season and by decreasing the negative effects of summer drought, diseases, and pests. Two maize inbreds widely incorporated into American maize germplasm, B73 and Mo17, exhibit different levels of tolerance to low temperature exposure at seedling stage. In addition, thirty seven diverse inbred maize lines showed large variation for seedling response to low temperature exposure with lines with extremely low tolerance to seedling exposure to low temperatures falling into stiff stalk, non-stiff stalk, and tropical clades. We employed the maize intermated B73×Mo17 (IBM) recombinant inbred line population (IBM Syn4 RIL) to investigate the genetic architecture of cold stress tolerance at a young seedling stage and to identify quantitative trait loci (QTLs) controlling this variation. A panel of 97 recombinant inbred lines of IBM Syn4 were used to measure, and score based on several traits related to chlorophyll concentration, leaf color, and tissue damage. Our analysis resulted in detection of two QTLs with high additive impact, one on chromosome 1 (bin 1.02) and second on chromosome 5 (bin 5.05). Further investigation of the QTL regions using gene expression data provided a list of the candidate genes likely contributing to the variation in cold stress response. Among the genes located within QTL regions identified in this study and differentially expressed in response to low temperature exposure are the genes with putative functions related to auxin and gibberellin response, as well as general abiotic stress response, and genes coding for proteins with broad regulatory functions.

Duration of Low Temperature Exposure Affects Egg Hatching of the Colorado Potato Beetle and Emergence of Overwintering Adults.

Simple SummaryThe Colorado potato beetle is a pest of Solanaceae in China, more than 30 species of Solanaceae as host plants of the Colorado potato beetle, and this beetle can cause yield losses to potato farms. Temperature is one of the main factors that affect the growth and survival of insects. In recent years, due to global climate change, sudden drops in short-term temperatures are also frequent, which will affect insects. This paper studies the influence of short-term low temperature on the Colorado potato beetle. In laboratory experiments, we evaluated the effect of short-term low temperature on the Colorado potato beetle eggs, and the emerged date, the number of Colorado potato beetles, were analyzed in combination with the low temperature conditions. Our results show that low temperature had adverse effects on the development of Colorado potato beetle eggs and the emerged of adults Colorado potato beetles, and the longer the low temperature treatment time, the greater the impact.The Colorado potato beetle is a serious pest of Solanaceae in China. In early summer, cold spells in later spring may occur for brief periods in the field environmental conditions, and temperatures often deviate far below the normal temperature for short periods, such as sudden short-term low temperature, may affect the development of Colorado potato beetle eggs. This paper studies the effects of low temperature stress at 8 °C for 0 d, 1 d, 3 d, 5 d, 7 d, and 10 d on the development of Colorado potato beetle eggs. Our results show that egg survival is significantly affected by short-term low temperature exposure. The percentage of eggs hatched is significantly affected by different treatment times (p = 0.000)—the percentage of eggs hatched decreases with increased treatment time, and Colorado potato beetles will extend the wintering time of their soil to resist the effects of lower temperatures. Thus, exposure of Colorado potato beetles to a short-term low temperature affects their emergence and population growth; this study could provide information for the occurrence, monitoring, and early warning of Colorado potato beetle during short-term temperature.

Intermittent high temperature reduces leaf sugar content and inhibits inflorescence initiation in Phalaenopsis hybrid

Phalaenopsis plants require several weeks of prolonged low-temperature periods for inflorescence initiation. We conducted a series of experiments to determine the inhibitory effects of intermittent high-temperature treatment on inflorescence initiation and the involvement of carbohydrate metabolism in the flowering inhibition of Phalaenopsis. In Expt. 1, Forcing temperatures from zero to ten weeks with two weeks of interval with different cultivars (‘Mantefon’, ‘Hong Seol’ and ‘Cutie’) or temperatures (constant 20 and 23 °C). After each treatment period, the plants were moved to 28 °C high temperature to prevent further low-temperature accumulation. In Expt. 2 and 3, under vegetative (continuous high temperature at 28/28 °C), forcing (continuous low temperature at 25/20 °C), and intermittent high-temperature conditions, growth and inflorescence initiation characteristics were observed, and transcriptomes and carbohydrate contents in leaves were also analyzed. Six to eight weeks of low-temperature periods were required for inflorescence initiation irrespective of cultivars, and different temperatures did not induce significant difference. The disruption of low-temperature exposure decreased the percentage of flowering-induced plants and delayed inflorescence emergence in all cultivars used in this study despite a sufficiently low-temperature period. Flowering-inhibited plants showed a significant increase in new leaf numbers, implying that vegetative growth was maintained by intermittent treatment. In transcriptome analysis, relatively low expression of differentially expressed genes (DEGs) of enzymes related to carbohydrate hydrolysis and gluconeogenesis was observed in vegetative and intermittent conditions. Moreover, the soluble sugar contents in leaves were significantly reduced under the intermittent condition compared with the forcing condition. These results indicated that discontinuous low-temperature exposure by intermittent high-temperature treatment prevented inflorescence initiation of Phalaenopsis and that the flowering inhibition could be attributed to the reduction of carbohydrate contents in leaves by decreasing sugar accumulation.

Autumn larval cold tolerance does not predict the northern range limit of a widespread butterfly species.

Climate change is driving range shifts, and a lack of cold tolerance is hypothesized to constrain insect range expansion at poleward latitudes. However, few, if any, studies have tested this hypothesis during autumn when organisms are subjected to sporadic low‐temperature exposure but may not have become cold‐tolerant yet. In this study, we integrated organismal thermal tolerance measures into species distribution models for larvae of the Giant Swallowtail butterfly, Papilio cresphontes (Lepidoptera: Papilionidae), living at the northern edge of its actively expanding range. Cold hardiness of field‐collected larvae was determined using three common metrics of cold‐induced physiological thresholds: the supercooling point, critical thermal minimum, and survival following cold exposure. P. cresphontes larvae were determined to be tolerant of chilling but generally die at temperatures below their SCP, suggesting they are chill‐tolerant or modestly freeze‐avoidant. Using this information, we examined the importance of low temperatures at a broad scale, by comparing species distribution models of P. cresphontes based only on environmental data derived from other sources to models that also included the cold tolerance parameters generated experimentally. Our modeling revealed that growing degree‐days and precipitation best predicted the distribution of P. cresphontes, while the cold tolerance variables did not explain much variation in habitat suitability. As such, the modeling results were consistent with our experimental results: Low temperatures in autumn are unlikely to limit the distribution of P. cresphontes. Understanding the factors that limit species distributions is key to predicting how climate change will drive species range shifts.

Evidence of Cold Induced Cytoskeletal Arrest in Hepatocytes of the Western Painted Turtle

The western painted turtle (Chrysemys picta belli) overwinters in ice-covered water bodies and can survive without oxygen (anoxia) for months, during which time metabolic rate decreases so that ATP demand does not exceed anoxic ATP supply. In order to characterize the contribution of the cytoskeleton to overwintering metabolic depression in non-excitable turtle cells, we have optimized a hepatocyte cell culture system. Actin and tubulin cycling are large consumers of cellular ATP pools, and we hypothesized that their cycling activity, and consequently capacity for structural rearrangement, would be downregulated by overwintering conditions. Turtles were acclimated to either 22°C or 4°C for at least 20 days and isolated hepatocytes were plated on either fibronectin or polylysine at varying temperatures, or in cyanide containing media, which acted as a chemical proxy for anoxia. Low incubation temperature, but not cyanide, prevented hepatocytes from forming stable attachment to fibronectin, suggesting inhibition of cell spreading and arrest of actin structural rearrangement by low temperature. Attachment to polylysine, a predominately passive and electrostatic phenomenon, was not inhibited by low temperature or cyanide. Hepatocytes isolated from 4°C acclimated turtles were 30% smaller in 2D area than those of 22°C acclimated turtles. Three-hour cyanide exposure of hepatocytes from 22°C acclimated turtles also resulted in a 30% shrinkage of 2D area, indicating that temperature depression and anoxia may have overlapping inhibitory effects on cell spreading and underlying cytoskeletal turnover. Previous research has shown that cyanide treatment causes reduction of hepatocyte mitochondrial membrane potential. Ongoing work is demonstrating that, regardless of whole animal temperature acclimation, low temperature exposure results in decreased mitochondrial membrane potential as well as increased cytosolic calcium, signaling which may contribute to inhibition of energetically costly actin rearrangement.

Cold tolerance of laboratory-reared Asian longhorned beetles

Low winter temperatures in temperate climates can limit the success of non-native species. The Asian longhorned beetle, Anoplophora glabripennis, is an invasive wood-boring pest of hardwood trees in North America and Europe. Native A. glabripennis populations are spread across several climate zones in China and the Korean Peninsula and are likely to encounter low temperatures in at least some of this range. Understanding the lethal limits of the overwintering life stages of A. glabripennis is essential for accurately modeling the risk that invasive populations pose to non-native environments. In this study, we provide the first systematic characterization of the cold tolerance strategy and lower lethal limits of A. glabripennis eggs, larvae, and pupae. In diapausing larvae, the most common overwintering stage in this species, we measure hemolymph glycerol and osmolality and identify the effects of prolonged low temperature exposure. In developing pupae, we identify sublethal effects caused by low temperature exposure before freezing. Eggs and larvae were the most cold-tolerant life stages; eggs were freeze-avoidant with an average supercooling point of −25.8 °C and larvae were freeze tolerant with an LT90 of −25 °C. Hemolymph osmolality of freeze-tolerant larvae, on average, increased to 811 mOsm during chilling. This increase was primarily driven by a concurrent, average increase of 232 mM hemolymph glycerol. Pupae died upon exposure to freezing temperatures, but accumulate strong sublethal effects prior to freezing, indicating that they are chill susceptible. Taken together, these data will be useful to inform species distribution modeling in A. glabripennis.

Assessing the effects of non-optimal temperature on risk of gestational diabetes mellitus in a cohort of pregnant women in Guangzhou, China

Previous observational studies have shown that exposure to ambient temperature and air pollution were associated with the incidence of gestational diabetes mellitus (GDM). However, the susceptible time window of non-optimal temperature on GDM is still unknown, and the interaction with air pollution has not been examined. We conducted a prospective cohort study in Guangzhou, China to investigate the windows of susceptibility of temperature extremes and variability on the risk of GDM and to explore any interaction effect with air pollution. Daily maximum (Tmax), minimum temperature (Tmin) and diurnal temperature range (DTR) were obtained from Guangdong Meteorological Service. Distributed lag non-linear models with a logistic regression were applied to assess the effect of temperature extremes and DTR in different weeks of gestation on GDM. To examine the interaction effect, relative excess risk due to interaction index, attributable proportion and synergy index were calculated. There were 5,165 pregnant women enrolled, of which 604 were diagnosed with GDM (11.7%). Compared with a reference temperature (50th percentile of Tmax), we found that extreme high temperature (99th percentile of Tmax) exposure during 21st and 22nd gestational weeks was associated with an increased risk of GDM. Extreme low temperature (1st percentile of Tmax) exposure during 14th to 17th weeks increased the risk of GDM. We observed that per 1 °C increment of DTR during 21st to 24th weeks was associated with an elevated GDM risk. No interaction effect of temperature extremes or variability with air pollution on GDM were observed. Our results suggested that non-optimal temperature is an independent risk factor of GDM. The time window of susceptibility for extreme temperatures and DTR exposure on the risk of GDM generally occurred in second trimester of pregnancy. In the context of climate change, our study has important implications for reproductive health and justifies more research in different climate zones.

Changes of Growth and Inflorescence Initiation by Exogenous Gibberellic Acid3 and 6-Benzylaminopurine Application in Phalaenopsis Orchids

This study was conducted to observe the effects of exogenous gibberellic acid3 (GA3) and 6-benzylaminopurine (BAP) treatments on the growth and inflorescence initiation of Phalaenopsis and to determine whether hormonal applications can substitute for low-temperature exposure for floral transition. Phalaenopsis Queen Beer ‘Mantefon’ clones were treated with no hormones (control), GA3 100 mg/L, GA3 200 mg/L, BAP 100 mg/L, and GA3 100 mg/L + BAP 100 mg/L by foliar spray. The treatments were carried at 28 °C for vegetative growth and 20 °C for forcing, respectively. At 28 °C of vegetative temperature, all exogenous hormonal treatments did not induce inflorescence initiation, but lateral shoots were observed in BAP-treated plants even though this plant is a monopodial orchid. GA3 significantly increased leaf length and decreased leaf width, and consequently increased length:width (L:W) ratio compared with the control and BAP alone. The trend grew as GA3 concentration increased. Also, the GA3 increased stem length and decreased stem diameter. At 20 °C of forcing temperature, L:W ratio responded similarly to 28 °C in GA3 treatments, but leaf size was smaller than for the control or BAP alone. BAP accelerated inflorescence emergence and significantly increased inflorescence numbers, whereas GA3 and GA3 + BAP slightly delayed inflorescence emergence. GA3 significantly promoted new leaf development at 20 °C of forcing condition. These results indicated that cytokinin was associated with the break of axillary vegetative and inflorescence meristems and exogenous GA3 spray did not improve inflorescence initiation in Phalaenopsis. Although exogenous hormonal application did not substitute for low-temperature exposure, it showed a possibility in promoting the growth and inflorescence initiation.