The characterization of ancient Methanococcales malate dehydrogenases reveals that strong thermal stability prevents unfolding under intense γ-irradiation.

Malate dehydrogenases (MalDH) (EC.1.1.1.37), which are involved in the conversion of oxaloacetate to pyruvate in the tricarboxylic acid cycle, are a relevant model for the study of enzyme evolution and adaptation. Likewise, a recent study showed that Methanococcales, a major lineage of Archaea, is a good model to study the molecular processes of proteome thermoadaptation in prokaryotes. Here, we use ancestral sequence reconstruction and paleoenzymology to characterize both ancient and extant MalDHs. We observe a good correlation between inferred optimal growth temperatures (OGTs) and experimental optimal temperatures for activity (A-Topt). In particular, we show that the MalDH present in the ancestor of Methanococcales was hyperthermostable and had an A-Topt of 80°C, consistent with a hyperthermophilic lifestyle. This ancestor gave rise to two lineages with different thermal constraints, one remaining hyperthermophilic while the other underwent several independent adaptations to colder environments. Surprisingly, the enzymes of the first lineage have retained a thermoresistant behavior (i.e., strong thermostability and high A-Topt), whereas the ancestor of the second lineage shows a strong thermostability, but a reduced A-Topt. Using mutants, we mimic the adaptation trajectory towards mesophily and show that it is possible to significantly reduce the A-Topt without altering the thermostability of the enzyme by introducing a few mutations. Finally, we reveal an unexpected link between thermostability and the ability to resist γ-irradiation-induced unfolding.

Experimental Evolution in a Warming World: The Omics Era.

A comprehensive understanding of the genetic mechanisms that shape species responses to thermal variation is essential for more accurate predictions of the impacts of climate change on biodiversity. Experimental evolution with high-throughput resequencing approaches (evolve and resequence) is a highly effective tool that has been increasingly employed to elucidate the genetic basis of adaptation. The number of thermal evolve and resequence studies is rising, yet there is a dearth of efforts to integrate this new wealth of knowledge. Here, we review this literature showing how these studies have contributed to increase our understanding on the genetic basis of thermal adaptation. We identify two major trends: highly polygenic basis of thermal adaptation and general lack of consistency in candidate targets of selection between studies. These findings indicate that the adaptive responses to specific environments are rather independent. A review of the literature reveals several gaps in the existing research. Firstly, there is a paucity of studies done with organisms of diverse taxa. Secondly, there is a need to apply more dynamic and ecologically relevant thermal environments. Thirdly, there is a lack of studies that integrate genomic changes with changes in life history and behavioral traits. Addressing these issues would allow a more in-depth understanding of the relationship between genotype and phenotype. We highlight key methodological aspects that can address some of the limitations and omissions identified. These include the need for greater standardization of methodologies and the utilization of new technologies focusing on the integration of genomic and phenotypic variation in the context of thermal adaptation.

Temperature-sensitive contacts in disordered loops tune enzyme I activity.

Homologous enzymes with identical folds often exhibit different thermal and kinetic behaviors. Understanding how an enzyme sequence encodes catalytic activity at functionally optimal temperatures is a fundamental problem in biophysics. Recently it was shown that the residues that tune catalytic activities of thermophilic/mesophilic variants of the C-terminal domain of bacterial enzyme I (EIC) are largely localized within disordered loops, offering a model system with which to investigate this phenomenon. In this work, we use molecular dynamics simulations and mutagenesis experiments to reveal a mechanism of sequence-dependent activity tuning of EIC homologs. We find that a network of contacts in the catalytic loops is particularly sensitive to changes in temperature, with some contacts exhibiting distinct linear or nonlinear temperature-dependent trends. Moreover, these trends define structurally clustered dynamical modes and can distinguish regions that tend toward order or disorder at higher temperatures. Assaying several thermophilic EIC mutants, we show that complementary mesophilic mutations to the most temperature-sensitive positions exhibit the most enhanced activity, while mutations to relatively temperature insensitive positions exhibit the least enhanced activities. These results provide a mechanistic explanation of sequence-dependent temperature tuning and offer a computational method for rational enzyme modification.

Heat tolerance, oxidative stress response tuning and robust gene activation in early-stage Drosophila melanogaster embryos.

In organisms with complex life cycles, life stages that are most susceptible to environmental stress may determine species persistence in the face of climate change. Early embryos of Drosophila melanogaster are particularly sensitive to acute heat stress, yet tropical embryos have higher heat tolerance than temperate embryos, suggesting adaptive variation in embryonic heat tolerance. We compared transcriptomic responses to heat stress among tropical and temperate embryos to elucidate the gene regulatory basis of divergence in embryonic heat tolerance. The transcriptomes of tropical and temperate embryos differed in both constitutive and heat-stress-induced responses of the expression of relatively few genes, including genes involved in oxidative stress. Most of the transcriptomic response to heat stress was shared among all embryos. Embryos shifted the expression of thousands of genes, including increases in the expression of heat shock genes, suggesting robust zygotic gene activation and demonstrating that, contrary to previous reports, early embryos are not transcriptionally silent. The involvement of oxidative stress genes corroborates recent reports on the critical role of redox homeostasis in coordinating developmental transitions. By characterizing adaptive variation in the transcriptomic basis of embryonic heat tolerance, this study is a novel contribution to the literature on developmental physiology and developmental genetics.

Thermal adaptation in plants: understanding the dynamics of translation factors and condensates.

Plants, as sessile organisms, face the crucial challenge of adjusting growth and development with ever-changing environmental conditions. Protein synthesis is the fundamental process that enables growth of all organisms. Since elevated temperature presents a substantial threat to protein stability and function, immediate adjustments of protein synthesis rates are necessary to circumvent accumulation of proteotoxic stress and to ensure survival. This review provides an overview of the mechanisms that control translation under high-temperature stress by the modification of components of the translation machinery in plants, and compares them to yeast and metazoa. Recent research also suggests an important role for cytoplasmic biomolecular condensates, named stress granules, in these processes. Current understanding of the role of stress granules in translational regulation and of the molecular processes associated with translation that might occur within stress granules is also discussed.

Variability in thermal tolerance of clutches from different mothers indicates adaptation potential to climate warming in sea turtles.

The current climate warming is a challenge to biodiversity that could surpass the adaptation capacity of some species. Hence, understanding the means by which populations undergo an increase in their thermal tolerance is critical to assess how they could adapt to climate warming. Specifically, sea turtle populations could respond to increasing temperatures by (1) colonizing new nesting areas, (2) nesting during cooler times of the year, and/or (3) by increasing their thermal tolerance. Differences in thermal tolerance of clutches laid by different females would indicate that populations have the potential to adapt by natural selection. Here, we used exhaustive information on nest temperatures and hatching success of leatherback turtle (Dermochelys coriacea) clutches over 14 years to assess the occurrence of individual variability in thermal tolerance among females. We found an effect of temperature, year, and the interaction between female identity and nest temperature on hatching success, indicating that clutches laid by different females exhibited different levels of vulnerability to high temperatures. If thermal tolerance is a heritable trait, individuals with higher thermal tolerances could have greater chances of passing their genes to following generations, increasing their frequency in the population. However, the high rate of failure of clutches at temperatures above 32°C suggests that leatherback turtles are already experiencing extreme heat stress. A proper understanding of mechanisms of adaptation in populations to counteract changes in climate could greatly contribute to future conservation of endangered populations in a rapidly changing world.

Multidecadal changes in coastal benthic species composition and ecosystem functioning occur independently of temperature-driven community shifts.

Rising global temperatures are often identified as the key driver impacting ecosystems and the services they provide by affecting biodiversity structure and function. A disproportionate amount of our understanding of biodiversity and function is from short-term experimental studies and static values of biodiversity indices, lacking the ability to monitor long-term trends and capture community dynamics. Here, we analyse a biennial dataset spanning 32 years of macroinvertebrate benthic communities and their functional response to increasing temperatures. We monitored changes in species' thermal affinities to examine warming-related shifts by selecting their mid-point global temperature distribution range and linking them to species' traits. We employed a novel weighted metric using Biological Trait Analysis (BTA) to gain better insights into the ecological potential of each species by incorporating species abundance and body size and selecting a subset of traits that represent five ecosystem functions: bioturbation activity, sediment stability, nutrient recycling and higher and lower trophic production. Using biodiversity indices (richness, Simpson's diversity and vulnerability) and functional indices (richness, Rao's Q and redundancy), the community structure showed no significant change over time with a narrow range of variation. However, we show shifts in species composition with warming and increases in the abundance of individuals, which altered ecosystem functioning positively and/or non-linearly. Yet, when higher taxonomic groupings than species were excluded from the analysis, there was only a weak increase in the measured change in community-weighted average thermal affinities, suggesting changes in ecosystem functions over time occur independently of temperature increase-related shifts in community composition. Other environmental factors driving species composition and abundance may be more important in these subtidal macrobenthic communities. This challenges the prevailing emphasis on temperature as the primary driver of ecological response to climate change and emphasises the necessity for a comprehensive understanding of the temporal dynamics of complex systems.

Genetic background and thermal regime influence adaptation to novel environment in the seed beetle, Callosobruchus maculatus.

Climate change is associated with the increase in both the mean and variability of thermal conditions. Therefore, the use of more realistic fluctuating thermal regimes is the most appropriate laboratory method for predicting population responses to thermal heterogeneity. However, the long- and short-term implications of evolving under such conditions are not well understood. Here, we examined differences in key life-history traits among populations of seed beetles (Callosobruchus maculatus) that evolved under either constant control conditions or in an environment with fluctuating daily temperatures. Specifically, individuals from two distinct genetic backgrounds were kept for 19 generations at one of two temperatures, a constant temperature (T = 29 °C) or a fluctuating daily cycle (Tmean = 33 °C, Tmax = 40 °C, and Tmin = 26 °C), and were assayed either in their evolved environment or in the other environment. We found that beetles that evolved in fluctuating environments but were then switched to constant 29 °C conditions had far greater lifetime reproductive success compared with beetles that were kept in their evolved environments. This increase in reproductive success suggests that beetles raised in fluctuating environments may have evolved greater thermal breadth than control condition beetles. In addition, the degree of sexual dimorphism in body size and development varied as a function of genetic background, evolved thermal environment, and current temperature conditions. These results not only highlight the value of incorporating diel fluctuations into climate research but also suggest that populations that experience variability in temperature may be better able to respond to both short- and long-term changes in environmental conditions.

Acclimation to constant and fluctuating temperatures promote distinct metabolic responses in Arctic char (Salvelinus alpinus).

The Arctic is warming three times faster than the global average, imposing challenges to cold-adapted fish, like Arctic char (Salvelinus alpinus). We evaluated stress and metabolic responses of Arctic char to different thermal acclimation scenarios to determine if responses to thermal variation differed from those to stable exposures. Fish were exposed for 7 days to one of 4 treatments: (1) control (12 °C), (2) mean (16 °C), corresponding to the mean temperature of the diel thermal cycle, (3) constant high temperature (20 °C), and (4) diel thermal cycling (12 to 20 °C every 24 h). Exposure to 20 °C causes increases plasma lactate and glucose, an imbalance in antioxidant systems, and oxidative stress in the liver. The 20 °C treatment also elevated fractional rates of protein synthesis and caused oxidative stress in the heart. Stress responses were more pronounced in diel thermal cycling than in mean (16 °C) fish, indicating that peak exposure temperatures or variation are physiologically important. Cortisol was highest in diel thermal cycling fish and oxidative stress was noted in the liver. Gill Na+/K+-ATPase activity was also significantly reduced in diel thermal cycling fish, suggesting gill remodeling in response to an osmoregulatory stress. Exposure to a constant 20 °C, was more challenging than a diel thermal cycle, demonstrating the importance of daily cooling to recovery. Arctic char inhabit a thermally variable environment and understanding how this impacts their physiology will be critical for informing conservation strategies in the context of a rapidly warming Arctic.

Disentangling physiological and physical explanations for body size-dependent thermal tolerance.

The effects of climate change are often body size dependent. One contributing factor could be size-dependent thermal tolerance (SDTT), the propensity for heat and cold tolerance to vary with body size among species and among individuals within species. SDTT is hypothesized to be caused by size differences in the temperature dependence of underlying physiological processes that operate at the cellular and organ/system level (physiological SDTT). However, temperature-dependent physiology need not change with body size for SDTT to be observed. SDTT can also arise because of physical differences that affect the relative body temperature dynamics of large and small organisms (physical SDTT). In this Commentary, I outline how physical SDTT occurs, its mechanistic differences from physiological SDTT, and how physical and physiological SDTT make different predictions about organismal responses to thermal variation. I then describe how physical SDTT can influence the outcome of thermal tolerance experiments, present an experimental framework for disentangling physical and physiological SDTT, and provide examples of tests for physiological SDTT that control for physical effects using data from Anolis lizards. Finally, I discuss how physical SDTT can affect organisms in natural environments and influence their vulnerability to anthropogenic warming. Differentiating between physiological and physical SDTT is important because it has implications for how we design and interpret thermal tolerance experiments and our fundamental understanding of thermal ecology and thermal adaptation.

Early life exposure to high temperature enhances locomotor performance without alteration in thermal ecology in different populations of Thoropa taophora tadpoles (Anura, Cycloramphidae).

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.

Projecting kelp (Ecklonia radiata) gametophyte thermal adaptation and persistence under climate change.

BACKGROUND AND AIMS: Kelp forests underpin temperate marine ecosystems but are declining due to ocean warming, causing loss of associated ecosystem services. Projections suggest significant future decline but often only consider the persistence of adult sporophytes. Kelps have a biphasic life cycle, and the haploid gametophyte can be more thermally tolerant than the sporophyte. Therefore, projections may be altered when considering the thermal tolerance of gametophytes. METHODS: We undertook thermal tolerance experiments to quantify the effect of temperature on gametophyte survival, relative growth rate (RGR) and sex ratio for three genetically distinct populations of Ecklonia radiata gametophytes from comparatively high, mid- and low latitudes (43°, 33° and 30°S). We then used these data to project the likely consequences of climate-induced thermal change on gametophyte persistence and performance across its eastern Australian range, using generalized additive and linear models. KEY RESULTS: All populations were adapted to local temperatures and their thermal maximum was 2-3 °C above current maximum in situ temperatures. The lowest latitude population was most thermally tolerant (~70 % survival up to 27 °C), while survival and RGR decreased beyond 25.5 and 20.5 °C for the mid- and low-latitude populations, respectively. Sex ratios were skewed towards females with increased temperature in the low- and high-latitude populations. Spatially explicit model projections under future ocean warming (2050-centred) revealed a minimal decline in survival (0-30 %) across populations, relative to present-day predictions. RGRs were also projected to decline minimally (0-2 % d-1). CONCLUSIONS: Our results contrast with projections for the sporophyte stage of E. radiata, which suggest a 257-km range contraction concurrent with loss of the low-latitude population by 2100. Thermal adaptation in E. radiata gametophytes suggests this life stage is likely resilient to future ocean warming and is unlikely to be a bottleneck for the future persistence of kelp.

Maintaining Breed Integrity: Successful Introgression of the SLICK1 Allele into the Holstein Breed.

This study evaluated the effectiveness of genetic introgression of the SLICK1 allele derived from Senepol cattle into the Holstein breed to enhance thermotolerance. The SLICK1 allele, located in PRLR gene, confers a short and sleek coat that is inherited as a simple dominant phenotype. Approximately 40 years ago, the University of Florida initiated efforts to introgress this allele into the Holstein population. Here we tracked the introgression of the SLICK1 allele using a medium-density genotyping array and a reference population of both breeds (50 Holstein, 46 Senepol). Among the 31 SLICK1+ Holsteins, there was 15.25% ± 11.11% (mean ± SD) Senepol ancestry on BTA20. Holsteins at the University of Florida descended from slick matings that did not inherit the SLICK1 allele (n=9) exhibited no Senepol ancestry. A secondary introgression of Senepol genetics in SLICK1+ animals was found on BTA4, spanning 54 markers and 15 genes, with 26.67% Senepol ancestry. This region, previously linked to heat stress adaptation, suggests that the introgression extends beyond the SLICK1 allele to incorporate additional beneficial genetics for thermal stress adaptation. These findings indicate that deliberate introgression of the SLICK1 allele enhances specific traits and potentially introduces other adaptive genetic variations. The study demonstrates the successful use of genetic interventions to improve livestock resilience against environmental challenges without significantly disrupting the recipient breed's genetic structure. The introgression of the SLICK1 allele serves as a model for breeding programs aimed at optimizing animal welfare and productivity in the face of global climate change while maintaining breed integrity.

Molecular basis and functional development of membrane-based microbial metabolism.

My research interest has so far been focused on metabolisms related to the "membrane" of microorganisms, such as the respiratory chain, membrane proteins, sugar uptake, membrane stress and cell lysis, and fermentation. These basic metabolisms are important for the growth and survival of cell, and their knowledge can be used for efficient production of useful materials. Notable achievements in research on metabolisms are elucidation of the structure and function of membrane-bound glucose dehydrogenase as a primary enzyme in the respiratory chain, elucidation of ingenious expression regulation of several operons or by divergent promoters, elucidation of stress-induced programed-cell lysis and its requirement for survival during a long-term stationary phase, elucidation of molecular mechanism of survival at a critical high temperature, elucidation of thermal adaptation and its limit, isolation of thermotolerant fermenting yeast strains, and development of high-temperature fermentation and green energy production technologies. These achievements are described together in this review.

Thermal Adaptations in Animals: Genes, Development, and Evolution.

Thermal adaptation to environmental temperature is a driving force in animal evolution. This chapter presents thermal adaptation in ectotherms and endotherms from the perspective of developmental biology. In ectotherms, there are known examples of temperature influencing morphological characteristics, such as seasonal color change, melanization, and sex determination. Furthermore, the timing of embryonic development also varies with environmental temperature. This review will introduce the cellular and molecular mechanisms underlying temperature-dependent embryogenesis. The evolution of thermal adaptation in endotherms is also important for survival in cold climates. Recent genome-wide studies have revealed adaptive mutations in the genomes of extant humans as well as extinct species such as woolly mammoths and Neanderthals. These studies have shown that single-nucleotide polymorphisms in physiologically related genes (e.g., CPT1A, LRP5, THATA, PRKG1, and FADS1-3) allow humans to live in cold climates. At the end of this chapter, we present the remaining questions in terms of genetic assimilation, heat shock protein Hsp90, and embryonic development.

Comparison of thermal comfort between different heating systems and adaptation to different indoor climates in winter.

Individual heating systems, such as the air-source heat pump (ASHP) air-conditioner or floor heating (FH), are usually used by people living in the hot summer and cold winter (HSCW) zone of China to heat indoor climates in the winter. However, little research has been conducted in the HSCW zone on the thermal comfort difference between indoor climates heated by ASHP air-conditioners and those heated by floor heating, as well as how occupants adapt to different indoor climates. We conducted a comparative field experiment in ASHP-heated and FH-heated apartments in Nanjing to investigate how different types of heating systems influence the thermal sensation of occupants, and we conducted a comparative field experiment in ASHP-heated office buildings and naturally ventilated teaching buildings in Shanghai to investigate how occupants adapt to different indoor thermal environments. Indoor environmental parameters and body surface temperatures were measured using instruments, and occupants' thermal sensation, activity level, and clothing were evaluated using the questionnaire. The results show that floor heating improves thermal comfort by raising foot temperature compared to the ASHP air-conditioner, and that occupants become acclimatized to different indoor climates by adjusting neutral operative temperature. According to the findings, there is no need to overheat the indoor environment in the HSCW zone because occupants can adapt to their experienced thermal environment and it is critical to maintain warm foot temperature in the cool/cold indoor environment.

Five weeks of intermittent hand exposures to alternating cold and hot stimuli do not modify finger cold-induced vasodilatation response.

We tested the hypothesis that prolonged intermittent hand exposures to transient contrast thermal stimuli would enhance the finger cold-induced vasodilatation (CIVD) response during localized cooling. Eight healthy men participated in a 5-week regimen, during which they immersed, thrice per week, the non-dominant (EXP) hand in 8° and 43 °C water, sequentially and at 3-min intervals, for a total period of 60 min. The contralateral (i.e., dominant) hand served as the control (CON) hand. Before and after the regimen, subjects conducted two 30-min hand cold (8 °C water) provocation trials, one with the EXP hand and the other with the CON hand. In addition, a flow-mediated dilatation test was performed in the brachial artery of the EXP arm. Regardless of the hand tested, the average finger skin temperature [CON hand: pre-trial = 10.5 (1.2)°C, post-trial = 10.8 (1.3)°C and EXP hand: pre-trial = 10.7 (1.1)°C, post-trial 10.9 (1.1)°C; p = 0.79], and the incidence of CIVD events [CON hand: pre-trial = 1.1 (1.2) events, post-trial = 1.2 (1.1) events and EXP hand: pre-trial = 1.1 (0.8) events, post-trial = 1.1 (0.8) events; p = 0.88] were not affected by the 5-week regimen. The sensation of cold-induced pain was transiently alleviated following the regimen (p = 0.02). The flow-mediated dilatation response of the EXP brachial artery remained unaltered [pre-trial = 5.4 (3.2)%, post-trial = 4.7 (3.6)%; p = 0.51]. Therefore, five weeks of intermittent hand exposures to alternating cold and hot stimuli do not improve finger temperature responsiveness to sustained localized cold.

Skin transcriptome analysis in Brangus cattle under heat stress.

Heat stress is a major factor that negatively affects animal welfare and production systems. Livestock should adapt to tropical and subtropical areas and to meet this, composite breeds have been developed. This work aimed to evaluate gene expression profiles in the skin of Brangus cattle under heat stress using a case-control design, and to correlate this with skin histological characteristics. Two groups of bulls were set using rectal temperature as a criterion to define stress conditions: stressed (N = 5) and non-stressed (N = 5) groups. Skin transcriptomics was performed and correlations between breed composition, phenotypic and skin histological traits were evaluated. Results showed 4309 differentially expressed genes (P < 0.01), 2113 downregulated and 2196 upregulated. Enrichment and ontology analyses revealed 132 GO terms and 67 pathways (P < 0.01), including thermogenesis, glycolysis, gluconeogenesis, mitochondrial activity, antioxidant and immune response, and apoptosis. The identity of the terms and pathways indicated the diversity of mechanisms directed to relieve the animals' suffering, acting from simple passive mechanisms (conduction, convection and radiation) to more complex active ones (behavioural changes, evaporation, vasodilation and wheezing). Furthermore, significant differences between phenotypic and skin histological traits and correlations between pairs of traits suggested a direction towards heat dissipation processes. In this sense, number of vessels was positively correlated with number of sweat glands (P < 0.001) and both were positively correlated with zebuine genetic content (P < 0.05 and P < 0.01, respectively), gland size was positively correlated with epidermal thickness and negatively with hair length (P < 0.05), and epidermal thickness was negatively correlated with gland-epidermis distance (P < 0.0005). These results support the notion that response to heat stress is physiologically complex, producing significant changes in the expression of genes involved in several biological pathways, while the animal's ability to face it depends greatly on their skin features.

Seasonal and between-population variation in heat tolerance and cooling efficiency in a Mediterranean songbird.

Discrete populations of widely distributed species may inhabit areas with marked differences in climatic conditions across geographic and seasonal scales, which could result in intraspecific variation in thermal physiology reflecting genetic adaptation, phenotypic plasticity, or both. However, few studies have evaluated inter-population variation in physiological responses to heat. We evaluated within- and inter-population seasonal variation in heat tolerance, cooling efficiency and other key thermoregulatory traits in two Mediterranean populations of Great tit Parus major experiencing contrasting thermal environments: a lowland population subject to hotter summers and a higher annual thermal amplitude than a montane population. Specifically, we measured heat tolerance limits (HTL), body temperature, resting metabolic rate, evaporative water loss, and evaporative cooling efficiency (the ratio between evaporative heat loss to metabolic heat production) within and above the thermoneutral zone during winter and summer. Heat tolerance during summer was greater in lowland than in montane birds; indeed, lowland birds seasonally increased this trait to a significant level, while montane ones did to a lesser extent. Besides, lowland birds showed greater evaporative cooling efficiency during summer (possibly due in part to reductions in total endogenous heat load), while surprisingly montane ones showed the opposite trend. Thus, lowland birds displayed greater seasonal flexibility in HTL, body temperature and resting metabolic rate above thermoneutrality, thus giving some support to the climatic variability hypothesis - that flexibility in thermoregulatory traits should increase with climatic variability. Our results partially support the idea that songbirds' adaptive thermoregulation in the heat is flexible, highlighting the importance of considering intraspecific variation in thermoregulatory traits when modelling the future distribution and persistence of species under different climate change scenarios.

Hot, dry, and salty: The present and future of an Extremophile model lizard from Argentina.

Global warming poses a threat to lizard populations by raising ambient temperatures above historical norms and reducing thermoregulation opportunities. Whereas the reptile fauna of desert systems is relatively well studied, the lizard fauna of saline environments has not received much attention and-to our knowledge-thermal ecology and the effects of global warming on lizards from saline environments have not been yet addressed. This pioneer study investigates the thermal ecology, locomotor performance and potential effects of climate warming on Liolaemus ditadai, a lizard endemic to one of the largest salt flats on Earth. We sampled L. ditadai using traps and active searches along its known distribution, as well as in other areas within Salinas Grandes and Salinas de Ambargasta, where the species had not been previously recorded. Using ensemble models (GAM, MARS, RandomForest), we modeled climatically suitable habitats for L. ditadai in the present and under a pessimistic future scenario (SSP585, 2070). L. ditadai emerges as an efficient thermoregulator, tolerating temperatures near its upper thermal limits. Our ecophysiological model suggests that available activity hours predict its distribution, and the projected temperature increase due to global climate change should minimally impact its persistence or may even have a positive effect on suitable thermal habitat. However, this theoretical increase in habitat could be linked to the distribution of halophilous scrub in the future. Our surveys reveal widespread distribution along the borders of Salinas Grandes and Salinas de Ambargasta, suggesting a potential presence along the entire border of both salt plains wherever halophytic vegetation exists. Optimistic model results, extended distribution, and no evidence of flood-related adverse effects offer insights into assessing the conservation status of L. ditadai, making it and the Salinas Grandes system suitable models for studying lizard ecophysiology in largely unknown saline environments.