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Higher temperatures exacerbate drought conditions by increasing evaporation rates, reducing soil moisture and altering precipitation patterns. As global temperatures rise as a result of climate change, these effects intensify, leading to more frequent and severe droughts. This link between higher temperatures and drought is particularly evident in sensitive ecosystems like the Amazon rainforest, where reduced rainfall and higher evaporation rates result in significantly lower water levels, threatening biodiversity and human livelihoods. As an example, the serious drought experienced in the Amazon basin in 2023 resulted in a significant decline in fish populations. Elevated water temperatures, reaching up to 38°C, led to mass mortality events, because these temperatures surpass the thermal tolerance of many Amazonian fish species. We know this because our group has collected data on critical thermal maxima (CTmax) for various fish species over multiple years. Additionally, warmer waters can cause hypoxia, further exacerbating fish mortality. Thus, even Amazon fish species, which have relatively high thermal tolerance, are being impacted by climate change. The Amazon drought experienced in 2023 underscores the urgent need for climate action to mitigate the devastating effects on Amazonian biodiversity. The fact that we have been able to link fish mortality events to data on the thermal tolerance of fishes emphasizes the important role of experimental biology in elucidating the mechanisms behind these events, a link that we aim to highlight in this Perspective.

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Mangrove habitats can serve as nursery areas for sharks and rays. Such environments can be thermally dynamic and extreme; yet, the physiological and behavioural mechanisms sharks and rays use to exploit such habitats are understudied. This study aimed to define the thermal niche of juvenile mangrove whiprays, Urogymnus granulatus. First, temperature tolerance limits were determined via the critical thermal maximum (CTMax) and minimum (CTMin) of mangrove whiprays at summer acclimation temperatures (28 °C), which were 17.5 °C and 39.9 °C, respectively. Then, maximum and routine oxygen uptake rates (MO2max and MO2routine, respectively), post-exercise oxygen debt, and recovery were estimated at current (28 °C) and heatwave (32 °C) temperatures, revealing moderate temperature sensitivities (i.e., Q10) of 2.4 (MO2max) and 1.6 (MO2routine), but opposing effects on post-exercise oxygen uptake. Finally, body temperatures (Tb) of mangrove whiprays were recorded using external temperature loggers, and environmental temperatures (Te) were recorded using stationary temperature loggers moored in three habitat zones (mangrove, reef flat, and reef crest). As expected, environmental temperatures varied between sites depending on depth. Individual mangrove whiprays presented significantly lower Tb relative to Te during the hottest times of the day. Electivity analysis showed tagged individuals selected temperatures from 24.0 to 37.0 °C in habitats that ranged from 21.1 to 43.5 °C. These data demonstrate that mangrove whiprays employ thermotaxic behaviours and a thermally insensitive aerobic metabolism to thrive in thermally dynamic and extreme habitats. Tropical nursery areas may, therefore, offer important thermal refugia for young rays. However, these tropical nursery areas could become threatened by mangrove and coral habitat loss, and climate change.

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Climate change, driven by increased human greenhouse gas emissions since the beginning of the industrial revolution up to the present day, is considered one of the major threats to biodiversity in the twenty-first century. One of the most affected groups is the ectotherms due to their direct dependence on environmental temperatures. In recent years, several studies have analysed the effects of temperature and thermal tolerance on several species of ectotherms. However, there are species whose thermal tolerances are still unknown. Such is the case of the critically endangered species, the Montseny Brook Newt (Calotriton arnoldi), endemic to the Montseny massif in Spain and whose thermal biology is unknown. Its critical situation makes it essential to know its tolerance to cooling, warming and thermopreferendum in water environments where the newt lives. Three experimental procedures were conducted from the western and eastern subspecies of C. arnoldi, considering four classes separately (males, females, juveniles and larvae). The results obtained showed that the CTmax of the species exceeded 31 °C, with a significant difference between the two subspecies. We found that the species tolerates low temperatures (<1 °C) well because the genera Calotriton is adapted to live in cold waters with temperatures below 15 °C. Although the thermopreference of the species was expected to trend to cold temperatures, some individuals chose relatively high temperatures, obtaining a range of 11.7 °C to 21.6 °C. The results presented in this study are an advance in the knowledge of the thermal physiology of this species and support the importance of the temperature of the torrent on its survival. Knowing their thermal limits and their preferred temperature range will help to propose management measures that promote the conservation of streams and riparian forest cover to mitigate temperature increases due to climate change.

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Elevated water temperatures and low dissolved oxygen (hypoxia) are pervasive stressors in aquatic systems that can be exacerbated by climate change and anthropogenic activities, and there is growing interest in their interactive effects. To explore this interaction, we quantified the effects of acute and long-term hypoxia exposure on the critical thermal maximum (CTmax) of Redside Dace (Clinostomus elongatus), a small-bodied freshwater minnow with sparse populations in the Great Lakes Basin of Canada and designated as Endangered under Canada's Species at Risk Act. Fish were held at 18°C and acclimated to four levels of dissolved oxygen (>90%, 60%, 40%, and 20% air saturation). CTmax was measured after 2 and 10 weeks of acclimation and after 3.5 weeks of reoxygenation, and agitation behavior was quantified during CTmax trials. Aquatic surface respiration behavior was also quantified at 14 weeks of acclimation to oxygen treatments. Acute hypoxia exposure decreased CTmax in fish acclimated to normoxia (>90% air saturation), but acclimation to hypoxia reduced this effect. There was no effect of acclimation oxygen level on CTmax when measured in normoxia, and there was no effect of exposure time to hypoxia on CTmax. Residual effects of hypoxia acclimation on CTmax were not seen after reoxygenation. Agitation behavior varied greatly among individuals and was not affected by oxygen conditions. Fish performed aquatic surface respiration with low frequency, but performed it earlier when acclimated to higher levels of oxygen. Overall, this work sheds light on the vulnerability of fish experiencing acute hypoxia and heat waves concurrently.

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Understanding the intricate relationship between temperature and physiological processes in ectotherm vertebrates is crucial for predicting how these animals respond to environmental changes, including those associated with climate change. This is particularly relevant for the anurans, given their limited capacity for thermoregulation, particularly in larval stages. Herein, we investigated the capacity for thermal acclimatization in Thoropa taophora tadpoles, an endemic species in the Atlantic rainforest of Southeast Brazil, inhabiting distinct thermal environments. These semi-terrestrial tadpoles develop on rocky surfaces, with some populations inhabiting exposed regions near the marine coast where temperatures may reach up to 30°C in sunny conditions, while other populations occupy forested areas near waterfalls that maintain lower temperatures. We aimed to understand the effects of temperature on locomotor performance and on the activity of metabolic enzymes that support performance in tadpoles sampled in four different populations. Moreover, we measured several aspects of thermoregulation, including the critical thermal maximum (CTmax), the body temperature of activity (Tb), the preferred temperature (Tpref) and the effectiveness of thermoregulation (E). Despite differences in body size, tadpoles from warmer environments consistently demonstrated higher locomotor performance, with minimal or no acclimatization seen in other variables. Correlations between habitat temperature and biological endpoints underscore the significance of maximum locomotor performance in shaping physiological responses. Our results show how temperature can impact tadpole behavior and performance, without changes in many organismal measures of thermal acclimatization, providing insights into potential ecological implications, particularly in the context of climate change.

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The intensity and duration of heat waves, as well as average global temperatures, are expected to increase due to climate change. Heat waves can cause physiological stress and reduce fitness in animals. Species can reduce overheating risk through phenotypic plasticity, which allows them to raise their thermal tolerance limits over time. This mechanism could be important for ectotherms whose body temperatures are directly influenced by available environmental temperatures. Geckos are a large, diverse group of ectotherms that vary in their thermal habitats and times of daily activity, which could affect how they physiologically adjust to heat waves. Data on thermal physiology are scarce for reptiles, with only one study in geckos. Understanding thermal tolerance and plasticity, and their relationship, is essential for understanding how some species are able to adjust or adapt to changing temperatures. In this study, we estimated thermal tolerance and plasticity, and their interaction, in the crepuscular gecko, Eublepharis macularius, a species that is emerging as a model for reptile biology. After estimating basal thermal tolerance for 28 geckos, thermal tolerance was measured for each individual a second time at several timepoints (3, 6, or 24 h) to determine thermal tolerance plasticity. We found that thermal tolerance plasticity (1) does not depend on the basal thermal tolerance of the organism, (2) was highest after 6 h from initial heat shock, and (3) was negatively influenced by individual body mass. Our findings contribute to the increasing body of work focused on understanding the influence of biological and environmental factors on thermal tolerance plasticity in organisms and provide phenotypic data to further investigate the molecular basis of thermal tolerance plasticity in organisms.

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Winter climate is changing rapidly in northern latitudes, and these temperature events have effects on salmonid thermal biology. Stressors during winter egg incubation could reduce hatching success and physiological performance of fall-spawning fishes. Here we quantified the potential for ontogenic carryover effects from embryonic thermal stress in multiple wild and hatchery-origin populations of brook trout (Salvelinus fontinalis), a temperate ectotherm native to northeastern North America. Fertilized eggs from four populations were incubated over the winter in the laboratory in four differing thermal regimes: ambient stream-fed water, chronic warming (+2 °C), ambient with a mid-winter cold-shock, and short-term warming late during embryogenesis (to stimulate an early spring). We examined body size and upper thermal tolerance at the embryonic, fry (10 weeks post-hatch and 27-30 weeks post-hatch) and gravid adult (age 2+) life stages (overall N = 1482). In a separate experiment, we exposed developing embryos to acute seven-day heat stress events immediately following fertilization and at the eyed-egg stage, and then assessed upper thermal tolerance (CTmax) 37 weeks post-hatch. In all cases, fish were raised in common garden conditions after hatch (i.e., same temperatures). Our thermal treatments during incubation had effects that varied by life stage, with incubation temperature and life stage both affecting body size and thermal tolerance. Embryos incubated in warmer treatment groups had higher thermal tolerance; there was no effect of the mid-winter melt event on embryo CTmax. Ten weeks after hatch, fry from the ambient and cold-shock treatment groups had higher and less variable thermal tolerance than did the warmer treatment groups. At 27-30 post-hatch and beyond, differences in thermal tolerance among treatment groups were negligible. Collectively, our study suggests that brook trout only exhibit short-term carryover effects from thermal stressors during embryo incubation, with no lasting effects on phenotype beyond the first few months after hatch.

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Environmental variation experienced during early periods of development can lead to persistent phenotypic alteration, known as carryover effects. Such effects increase concern for threatened or endangered species such as the white sturgeon (Acipenser transmontanus), particularly considering expected thermal changes due to climate change. We evaluated how temperature during embryonic development affects physiological parameters such as larval and early juvenile growth and thermal tolerance. Nechako River white sturgeon embryos were incubated at different environmental temperatures (Te) of 12 °C (the natural spawning temperature of this population), 15 °C (the hatchery incubation temperature), and 18 °C (representing potential increases in river temperatures given global climate change). After hatch, fish were reared at a common 15 °C for 80 days post-hatch (dph). Individuals from each temperature treatment were tested for thermal tolerance using the critical thermal maximum method (CTmax), euthanized, and measured. Fish were examined at regular intervals from 13 to 80 dph, which bridged the time from the start of exogenous feeding through the transition into early juveniles. We found carryover effects of high embryonic Te in the short term for both thermal tolerance and growth. Fish that developed at 18 °C had the lowest thermal tolerance during the start of exogenous feeding. However, differences in thermal tolerance were small for early juveniles and were unlikely to be ecologically relevant in the longer term. Fish that developed at 18 °C were smallest over the observation period, indicating a possible cost for survival from increasing environmental temperatures during embryonic development. This research represents a window into a critical period of development during which fish are particularly vulnerable to climatic variation, and shows that cooler temperatures (12 °C) during incubation are optimal for this population. The results can inform environmental managers on the best strategies to help conserve current white sturgeon populations across their range.

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Despite the generally negative impact of urbanization on insect biodiversity, some insect species persist in urban habitats. Understanding the mechanisms underpinning the ability of insects to tolerate urban habitats is critical given the contribution of land-use change to the global insect decline. Compensatory mechanisms such as phenotypic plasticity and evolutionary change in thermal physiological traits could allow urban populations to persist under the altered thermal regimes of urban habitats. It is important to understand the contributions of plasticity and evolution to trait change along urbanization gradients as the two mechanisms operate under different constraints and timescales. Here, we examine the plastic and evolutionary responses of heat and cold tolerance (critical thermal maximum [CTmax] and critical thermal minimum [CTmin]) to warming among populations of the cabbage white butterfly, Pieris rapae, from urban and non-urban (rural) habitats using a two-temperature common garden experiment. Although we expected populations experiencing urban warming to exhibit greater CTmax and diminished CTmin through plastic and evolutionary mechanisms, our study revealed evidence only for plasticity in the expected direction of both thermal tolerance traits. We found no evidence of evolutionary divergence in either heat or cold tolerance, despite each trait showing evolutionary potential. Our results suggest that thermal tolerance plasticity contributes to urban persistence in this system. However, as the magnitude of the plastic response was low and comparable to other insect species, other compensatory mechanisms likely further underpin this species' success in urban habitats.

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Wild ectotherms are exposed to multiple stressors, including parasites, that can affect their responses to environmental change. Simultaneously, unprecedented warm temperatures are being recorded worldwide, increasing both the average and maximum temperatures experienced in nature. Understanding how ectotherms, such as fishes, will react to the combined stress of parasites and higher average temperatures can help predict the impact of extreme events such as heat waves on populations. The critical thermal method (CTM), which assesses upper (CTmax) and lower (CTmin) thermal tolerance, is often used in acclimated ectotherms to help predict their tolerance to various temperature scenarios. Despite the widespread use of the CTM across taxa, few studies have characterized the response of naturally infected fish to extreme temperature events or how acute thermal stress affects subsequent survival. We acclimated naturally infected pumpkinseed sunfish (Lepomis gibbosus) to four ecologically relevant temperatures (10, 15, 20 and 25°C) and one future warming scenario (30°C) for 3 weeks before measuring CTmax and CTmin. We also assessed individual survival the week following CTmax. Parasites were counted and identified following trials to relate infection intensity to thermal tolerance and survival. Interestingly, trematode parasites causing black spot disease were negatively related to CTmax, suggesting that heavily infected fish are less tolerant to acute warming. Moreover, fish infected with yellow grub parasites showed decreased survival in the days following CTmax implying that the infection load has negative survival consequences on sunfish during extreme warming events. Our findings indicate that, when combined, parasite infection and high prolonged average temperatures can affect fish thermal tolerance and survival, emphasizing the need to better understand the concomitant effects of stressors on health outcomes in wild populations. This is especially true given that some parasite species are expected to thrive in warming waters making host fish species especially at risk.

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Temperature in many natural aquatic environments follows a diel cycle, but to date, we know little on how diel thermal cycles affect fish biology. The current study investigates the growth, development and physiological performance of wild Atlantic salmon collected from the Miramichi and Restigouche rivers (NB, Canada). Fish were collected as parr and acclimated to either 16-21 or 19-24°C diel thermal cycles throughout the parr and smolt life stages. Both Miramichi and Restigouche Atlantic salmon parr grew at similar rates during 16-21 or 19-24°C acclimations. However, as smolts, the growth rates of the Miramichi (-8% body mass day-1) and Restigouche (-38% body mass day-1) fish were significantly slower at 19-24°C, and were in fact negative, indicating loss of mass in this group. Acclimation to 19-24°C also increased Atlantic salmon CTmax. Our findings suggest that both life stage and river origin impact Atlantic salmon growth and performance in the thermal range used herein. These findings provide evidence for local adaptation of Atlantic salmon, increased vulnerability to warming temperatures, and highlight the differential impacts of these ecologically relevant diel thermal cycles on the juvenile life stages in this species.

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Bees are essential pollinators and understanding their ability to cope with extreme temperature changes is crucial for predicting their resilience to climate change, but studies are limited. We measured the response of the critical thermal maximum (CTMax) to short-term acclimation in foragers of six bee species from the Greek island of Lesvos, which differ in body size, nesting habit, and level of sociality. We calculated the acclimation response ratio as a metric to assess acclimation capacity and tested whether bees' acclimation capacity was influenced by body size and/or CTMax. We also assessed whether CTMax increases following acute heat exposure simulating a heat wave. Average estimate of CTMax varied among species and increased with body size but did not significantly shift in response to acclimation treatment except in the sweat bee Lasioglossum malachurum. Acclimation capacity averaged 9% among species and it was not significantly associated with body size or CTMax. Similarly, the average CTMax did not increase following acute heat exposure. These results indicate that bees might have limited capacity to enhance heat tolerance via acclimation or in response to prior heat exposure, rendering them physiologically sensitive to rapid temperature changes during extreme weather events. These findings reinforce the idea that insects, like other ectotherms, generally express weak plasticity in CTMax, underscoring the critical role of behavioral thermoregulation for avoidance of extreme temperatures. Conserving and restoring native vegetation can provide bees temporary thermal refuges during extreme weather events.

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Rapid evolutionary adaptation could reduce the negative impacts of climate change if sufficient heritability of key traits exists under future climate conditions. Plastic responses to climate change could also reduce negative impacts. Understanding which populations are likely to respond via evolution or plasticity could therefore improve estimates of extinction risk. A large body of research suggests that the evolutionary and plastic potential of a population can be predicted by the degree of spatial and temporal climatic variation it experiences. However, we know little about the scale at which these relationships apply. Here, we test if spatial and temporal variation in temperature affects genetic variation and plasticity of fitness and a key thermal tolerance trait (critical thermal maximum; CTmax) at microgeographic scales using a metapopulation of Daphnia magna in freshwater rock pools. Specifically, we ask if (a) there is a microgeographic adaptation of CTmax and fitness to differences in temperature among the pools, (b) pools with greater temporal temperature variation have more genetic variation or plasticity in CTmax or fitness, and (c) increases in temperature affect the heritability of CTmax and fitness. Although we observed genetic variation and plasticity in CTmax and fitness, and differences in fitness among pools, we did not find support for the predicted relationships between temperature variation and genetic variation or plasticity. Furthermore, the genetic variation and plasticity we observed in CTmax are unlikely sufficient to reduce the impacts of climate change. CTmax plasticity was minimal and heritability was 72% lower when D. magna developed at the higher temperatures predicted under climate change. In contrast, the heritability of fitness increased by 53% under warmer temperatures, suggesting an increase in overall evolutionary potential unrelated to CTmax under climate change. More research is needed to understand the evolutionary and plastic potential under climate change and how that potential will be altered in future climates.

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AbstractTemperature and interspecific competition are fundamental drivers of community structure in natural systems and can interact to affect many measures of species performance. However, surprisingly little is known about the extent to which competition affects extinction temperatures during extreme warming. This information is important for evaluating future threats to species from extreme high-temperature events and heat waves, which are rising in frequency and severity around the world. Using experimental freshwater communities of rotifers and ciliates, this study shows that interspecific competition can lower the threshold temperature at which local extinction occurs, reducing time to extinction during periods of sustained warming by as much as 2 weeks. Competitors may lower extinction temperatures by altering biochemical characteristics of the natural environment that affect temperature tolerance (e.g., levels of dissolved oxygen, nutrients, and metabolic wastes) or by accelerating population decline through traditional effects of resource depletion on life history parameters that affect population growth rates. The results suggest that changes in community structure in space and time could drive variability in upper thermal limits.

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The thermally dynamic nearshore Beaufort Sea, Alaska, is experiencing climate change-driven temperature increases. Measuring thermal tolerance of broad whitefish (Coregonus nasus) and saffron cod (Eleginus gracilis), both important species in the Arctic ecosystem, will enhance understanding of species-specific thermal tolerances. The objectives of this study were to determine the extent that acclimating broad whitefish and saffron cod to 5°C and 15°C changed their critical thermal maximum (CTmax) and HSP70 protein and mRNA expression in brain, muscle and liver tissues. After acclimation to 5°C and 15°C, the species were exposed to a thermal ramping rate of 3.4°C · h-1 before quantifying the CTmax and HSP70 protein and transcript concentrations. Broad whitefish and saffron cod acclimated to 15°C had a significantly higher mean CTmax (27.3°C and 25.9°C, respectively) than 5°C-acclimated fish (23.7°C and 23.2°C, respectively), which is consistent with trends in CTmax between higher and lower acclimation temperatures. There were species-specific differences in thermal tolerance with 15°C-acclimated broad whitefish having higher CTmax and HSP70 protein concentrations in liver and muscle tissues than saffron cod at both acclimation temperatures. Tissue-specific differences were quantified, with brain and muscle tissues having the highest and lowest HSP70 protein concentrations, respectively, for both species and acclimation temperatures. The differences in broad whitefish CTmax between the two acclimation temperatures could be explained with brain and liver tissues from 15°C acclimation having higher HSP70a-201 and HSP70b-201 transcript concentrations than control fish that remained in lab-acclimation conditions of 8°C. The shift in CTmax and HSP70 protein and paralogous transcripts demonstrate the physiological plasticity that both species possess in responding to two different acclimation temperatures. This response is imperative to understand as aquatic temperatures continue to elevate.

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Critical thermal maximum (CTmax ) is widely used to measure upper thermal tolerance in fish but is rarely examined in embryos. Upper thermal limits generally depend on an individual's thermal history, which molds plasticity. We examined how thermal acclimation affects thermal tolerance of brook trout (Salvelinus fontinalis) embryos using a novel method to assess CTmax in embryos incubated under three thermal regimes. Warm acclimation was associated with an increase in embryonic upper thermal tolerance. However, CTmax variability was markedly higher than is typical for juvenile or adult salmonids.

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Critical thermal maximum (CTmax) describes the upper thermal tolerance of an animal where biological functions start to fail. A period of acclimation can enhance CTmax through plasticity, potentially buffering animals from extreme temperatures caused by climate change. Basal and acclimated CTmax vary within and between species and may be explained by traits related to thermal physiology, such as body size and sex. Differences in CTmax have not been established among species of tsetse fly (Glossina spp.), vectors of animal and human African trypanosomiasis. Here, we investigated basal CTmax and its plasticity for five tsetse species following adult acclimation at constant 25 or 30 °C for five days. We then set our findings in context using a meta-analysis on 33 species of Diptera. We find that, of the five tsetse species considered, only Glossina palpalis gambiensis and Glossina brevipalpis exhibited plasticity of CTmax, with an increase of 0.12 °C and 0.10 °C per 1 °C acclimation respectively. Within some species, higher basal CTmax values were associated with larger body size and being female, while variation in plasticity (i.e., response to the acclimation temperature) could not be explained by sex or size. Our broader meta-analysis across Diptera revealed overall CTmax plasticity of 0.06 °C per 1 °C acclimation, versus a similar 0.05 °C mean increase in tsetse. In contrast, there was greater CTmax plasticity in males compared to females in Diptera. Our study highlights that CTmax and its plasticity varies even among closely related species. Broader patterns across groups are not always reflected at a finer resolution; we thus emphasise the need for detailed experimental studies across a wide range of insect species to capture their capacity to cope with rapidly warming temperatures.

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Insecticides and climate change are among the multiple stressors that bees face, but little is known about their synergistic effects, especially for non-Apis bee species. In laboratory experiments, we tested whether the stingless bee Tetragonula hockingsi avoids insecticide in sucrose solutions and how T. hockingsi responds to insecticide and heat stress combined. We found that T. hockingsi neither preferred nor avoided sucrose solutions with either low (2.5 × 10-4 ng µl-1 imidacloprid or 1.0 × 10-4 ng µl-1 fipronil) or high (2.5 × 10-3 ng µl-1 imidacloprid or 1.0 × 10-3 ng µl-1 fipronil) insecticide concentrations when offered alongside sucrose without insecticide. In our combined stress experiment, the smallest dose of imidacloprid (7.5 × 10-4 ng) did not significantly affect thermal tolerance (CTmax). However, CTmax significantly reduced by 0.8°C (±0.16 SE) and by 0.5°C (±0.16 SE) when bees were fed as little as 7.5 × 10-3 ng of imidacloprid or 3.0 × 10-4 ng of fipronil, respectively, and as much as 1.5°C (±0.16 SE) and 1.2°C (±0.16 SE) when bees were fed 7.5 × 10-2 ng of imidacloprid or 3.0 × 10-2 ng of fipronil, respectively. Predictions of temperature increase, and increased insecticide use in the tropics suggest that T. hockingsi will be at increased risk of the effects of both stressors in the future.

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AbstractThermal physiology helps us understand how ectotherms respond to novel environments and how they persist when introduced to new locations. Researchers generally measure thermal physiology traits immediately after animal collection or after a short acclimation period. Because many of these traits are plastic, the conclusions drawn from such research can vary depending on the duration of the acclimation period. In this study, we measured the rate of change and extent to which cold tolerance (critical thermal minimum [CTmin]) of nonnative Italian wall lizards (Podarcis siculus) from Hempstead, New York, changed during a cold acclimation treatment. We also examined how cold acclimation affected heat tolerance (critical thermal maximum [CTmax]), thermal preference (Tpref), evaporative water loss (EWL), resting metabolic rate (RMR), and respiratory exchange ratio (RER). We predicted that CTmin, CTmax, and Tpref would decrease with cold acclimation but that EWL and RMR would increase with cold acclimation. We found that CTmin decreased within 2 wk and that it remained low during the cold acclimation treatment; we suspect that this cold tolerance plasticity reduces risk of exposure to lethal temperatures during winter for lizards that have not yet found suitable refugia. CTmax and Tpref also decreased after cold acclimation, while EWL, RMR, and RER increased after cold acclimation, suggesting trade-offs with cold acclimation in the form of decreased heat tolerance and increased energy demands. Taken together, our findings suggest that cold tolerance plasticity aids the persistence of an established population of invasive lizards. More generally, our findings highlight the importance of accounting for the plasticity of physiological traits when investigating how invasive species respond to novel environments.

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Thermal tolerance and associated mechanisms are often tested via the critical thermal maximum (CTmax). The agitation temperature is a recently described thermal limit in fishes that has received little mechanistic evaluation. The present study used a temperate elasmobranch fish to test the hypothesis that this thermal tolerance trait is partially set by the onset of declining cardiorespiratory performance and the cellular stress response. Pacific spiny dogfish (Squalus suckleyi) were screened for cardiorespiratory and whole-organism thermal limits to test for associations between thermal performance and tolerance. Then, biochemical markers of secondary stress, aerobic and anaerobic enzyme activities, and molecular markers of cellular stress were determined for various tissues at the agitation temperature and secondary stress markers were determined at CTmax. In dogfish, the agitation temperature was characterised by increased turning activity within experimental chambers and was equal to the temperature at which dogfish exhibited maximum heart rate. Citrate synthase activity increased at the agitation temperature in white muscle relative to unmanipulated dogfish. Furthermore, lactate dehydrogenase activity and accumulated lactate in the plasma and muscle were not affected by acute warming. Cellular stress was apparent in hypothalamus, gill filament and ventricle, denoted by elevated transcript abundance of the stress response gene hsp70 but not the oxygen homeostasis gene hif1α. Conversely, CTmax was characterised by metabolic acidosis driven by anaerobic lactate production, signifying an increased reliance on anaerobic metabolism between the agitation temperature and CTmax. Together, these data provide partial support for our hypothesis, in that cellular stress, but not declining thermal performance, occurred at the agitation temperature.