Thermal Biology Research Articles

Variability in the summer movements, habitat use and thermal biology of two fish species in a temperate river

The ability of fish to cope with warm water temperatures in summer depends on factors including their thermal traits and the ability of individuals to access cool-water refugia. Knowledge is highly limited on the in situ responses of many fishes to elevated summer temperatures, including whether they express behavioural thermoregulation. The responses of two riverine species to summer water temperatures were tested here using the movement metrics, spatial habitat use and body temperatures of individual European barbel Barbus barbus (‘barbel’) and common bream Abramis brama (‘bream’) versus river temperatures. Acoustic biotelemetry was applied in the lower River Severn basin, western Britain, in summer 2021 (barbel) and 2022 (bream), where individuals could move across > 150 km of river, including a tributary of cooler water. Across all individuals, bream occupied 37 km of river length (mainstem only), with low inter-individual variability in their spatial habitat use, movements and body temperatures. In contrast, barbel occupied 62 km of river (main river/tributary), with relatively high inter-individual variability in spatial habitat use, movements and body temperatures, with higher variation in body temperatures as river temperatures increased (maximum mean daily temperature difference between individuals on the same day: 4.2 °C). Although warmer individuals generally moved more, their activity was greatest at relatively low temperatures and higher flows, and neither species revealed any evidence of behavioural thermoregulation during elevated temperatures. Enabling phenotypically diverse fish populations to express their natural behaviours and thermal preferences in summer water temperatures thus requires maintaining their free-ranging in thermally heterogenous habitats.

Fitness surfaces and local thermal adaptation in Drosophila along a latitudinal gradient.

Local adaptation is commonly cited to explain species distribution, but how fitness varies along continuous geographical gradients is not well understood. Here, we combine thermal biology and life-history theory to demonstrate that Drosophila populations along a 2500 km latitudinal cline are adapted to local conditions. We measured how heat tolerance and viability rate across eight populations varied with temperature in the laboratory and then simulated their expected cumulative Darwinian fitness employing high-resolution temperature data from their eight collection sites. Simulations indicate a trade-off between annual survival and cumulative viability, as both mortality and the recruitment of new flies are predicted to increase in warmer regions. Importantly, populations are locally adapted and exhibit the optimal combination of both traits to maximize fitness where they live. In conclusion, our method is able to reconstruct fitness surfaces employing empirical life-history estimates and reconstructs peaks representing locally adapted populations, allowing us to study geographic adaptation in silico.

Temperature impacts on dengue incidence are nonlinear and mediated by climatic and socioeconomic factors: A meta-analysis

Temperature can influence mosquito-borne diseases like dengue. These effects are expected to vary geographically and over time in both magnitude and direction and may interact with other environmental variables, making it difficult to anticipate changes in response to climate change. Here, we investigate global variation in temperature–dengue relationship by analyzing published correlations between temperature and dengue and matching them with remotely sensed climatic and socioeconomic data. We found that the correlation between temperature and dengue was most positive at intermediate (near 24°C) temperatures, as predicted from an independent mechanistic model. Positive temperature–dengue associations were strongest when temperature variation and population density were high and decreased with infection burden and rainfall mean and variation, suggesting alternative limiting factors on transmission. Our results show that while climate effects on diseases are context-dependent they are also predictable from the thermal biology of transmission and its environmental and social mediators.

Thermal Relationship in Tropical Anurans from Two Contrasting Habitats Along an Elevation Gradient in Colombia.

Anurans are ectothermic organisms highly susceptible to variations in the environmental temperature that changes with elevation and between habitats in tropical mountains. The aim of this study was to evaluate the variation of body temperature (BT) of nocturnal anurans from two contrasting habitats (open and forest habitats) along an elevation gradient in Colombia. We measured the environmental temperatures (substrate and air) and BT of 135 adult frogs of 11 species from open and forest habitats at three elevational zones of an Andean Mountain. The BT had a positive and significant relationship with environmental temperatures and showed a higher thermal dependence for substrate than air temperature, which suggests that anurans are thermoconformers and potentially tigmotherms. Additionally, BT of anurans from both habitats decreased with the elevation, but species from open habitats had a higher BT than forest species. Therefore, the impact of environmental temperatures on anurans that live at a similar altitude level is not the same, as the type of habitat has a strong influence on their BT. This information is important to a better understanding of anuran thermal biology, refine conservation strategies, and to improve the predictive power of environmental data in forecasting the effects of climate change on small ectotherms such as amphibians.

Unexpected behavioural adaptation of yellow fever mosquitoes in response to high temperatures

Temperature is a major ecological driver of mosquito-borne diseases as it influences the life-history of both the mosquito and the pathogen harboured within it. Understanding the mosquitoes’ thermal biology is essential to inform risk prediction models of such diseases. Mosquitoes can respond to temperatures by microhabitat selection through thermal preference. However, it has not yet been considered that mosquitoes are likely to adapt to changing temperatures, for example during climate change, and alter their preference over evolutionary time. We investigated this by rearing six cohorts of the yellow fever mosquito Aedes aegypti at two temperatures (24 °C, 30 °C) for 20 generations and used these cohorts to explicitly separate the effects of long-term evolution and within-generation acclimation on their thermal preferences in a thermal gradient of 20–35 °C. We found that warm-evolved mosquitoes spent 31.5% less time at high temperatures, which affects their efficiency as a vector. This study reveals the complex interplay of experimental evolution, rearing temperatures, and thermal preference in Ae. aegypti mosquitoes. It highlights the significance of incorporating mosquito microhabitat selection in disease transmission models, especially in the context of climate change.

Tadpoles Develop Elevated Heat Tolerance in Urban Heat Islands Regardless of Sex

The ability of wildlife to endure the effects of high temperatures is increasingly important for biodiversity conservation under climate change and spreading urbanization. Organisms living in urban heat islands can have elevated heat tolerance via phenotypic or transgenerational plasticity or microevolution. However, the prevalence and mechanisms of such thermal adaptations are barely known in aquatic organisms. Furthermore, males and females can differ in heat tolerance, which may lead to sex-biased mortality, yet it is unknown how sex differences in thermal biology influence urban phenotypic divergence. To address these knowledge gaps, we measured critical thermal maxima (CTmax) in male and female agile frog (Rana dalmatina) tadpoles captured from warm urban ponds and cool woodland ponds, and in a common-garden experiment where embryos collected from both habitat types were raised in the laboratory. We found higher CTmax in urban-dwelling tadpoles compared to their counterparts living in woodland ponds. This difference was reversed in the common-garden experiment: tadpoles originating from urban ponds had lower CTmax than tadpoles originating from woodland ponds. We found no effect of sex on CTmax or its difference between habitats. These results demonstrate that aquatic amphibian larvae can respond to the urban heat island effect with increased heat tolerance similarly to other, mostly terrestrial taxa studied so far, and that phenotypic plasticity may be the main driver of this response. Our findings also suggest that heat-induced mortality may be independent of sex in tadpoles, but research is needed in many more taxa to explore potentially sex-dependent urban thermal responses.

Thermal heterogeneity in a Mediterranean pond: perspectives for the thermal biology of amphibian larvae

Abstract In Mediterranean climates, amphibian larvae often occupy temporary ponds where they must avoid deleterious temperatures and develop before these ponds dry out. We measured thermal heterogeneity in one of these ponds and discussed its relationship with development time and thermal tolerance in three observed species. We found large daily thermoclines that may reach 14°C and persist into the early evening. To our knowledge, this is the first time that such a large degree of thermal heterogeneity has been reported in temporary ponds. Thus, we raise hypotheses linking thermoregulation, habitat use, and growth that Mediterranean amphibian larvae, or other aquatic larvae inhabiting water bodies with strong thermoclines, might exhibit. We argue that studies on thermoregulation of larvae are badly needed to help us understand the responses of Mediterranean amphibians to global warming.

Cross-species gut microbiota transplantation predictably affects host heat tolerance.

The gut microbiome is known to influence and have regulatory effects in diverse physiological functions of host animals, but only recently has the relationship between host thermal biology and gut microbiota been explored. Here, we examined how early-life manipulations of the gut microbiota in larval amphibians influenced their critical thermal maximum (CTmax) at different acclimation temperatures. We stripped the resident microbiome from egg masses of wild-caught wood frogs (Lithobates sylvaticus) via an antibiotic wash, and then inoculated the eggs with pond water (control), no inoculation, or the intestinal microbiota of another species that has a wider thermal tolerance - green frogs (Lithobates clamitans). We predicted that this cross-species transplant would increase the CTmax of the recipient wood frog larvae relative to the other treatments. In line with this prediction, green frog microbiome-recipient larvae had the highest CTmax while those with no inoculum had the lowest CTmax. Both the microbiome treatment and acclimation temperature significantly influenced the larval gut microbiota communities and α-diversity indices. Green frog microbiome-inoculated larvae were enriched in Rikenellaceae relative to the other treatments, which produce short-chain fatty acids and could contribute to greater energy availability and enhanced heat tolerance. Larvae that received no inoculation had a higher relative abundance of potentially pathogenic Aeromonas spp., which negatively affects host health and performance. Our results are the first to show that cross-species gut microbiota transplants alter heat tolerance in a predictable manner. This finding has repercussions for the conservation of species that are threatened by climate change and demonstrates a need to further explore the mechanisms by which the gut microbiota modulate host thermal tolerance.

ECR Spotlight – Jason Dallas

ECR Spotlight is a series of interviews with early-career authors from a selection of papers published in Journal of Experimental Biology and aims to promote not only the diversity of early-career researchers (ECRs) working in experimental biology but also the huge variety of animals and physiological systems that are essential for the ‘comparative’ approach. Jason Dallas is an author on ‘ Cross-species gut microbiota transplantation predictably affects host heat tolerance’, published in JEB. Jason conducted the research described in this article while a PhD student in Dr Robin Warne's lab at Southern Illinois University, USA. He is now a postdoctoral research assistant in the lab of Dr Donald Walker at Middle Tennessee State University, USA, investigating herpetofauna ecophysiology including thermal biology, host microbiome and responses to human activity.

Seasonal Variation in Thermal Biology and Water Balance in a Year-Round Active Neotropical Treefrog, Scinax fuscovarius

Seasonal Variation in Thermal Biology and Water Balance in a Year-Round Active Neotropical Treefrog, Scinax fuscovarius

Термобиологические характеристики ушастых круглоголовок (Phrynocephalus mystaceus) (Reptilia, Agamidae) на бархане Сарыкум (Дагестан, Россия), полученные с помощью логгеров температуры

The paper compares two variants of thermobiological studies of toad-headed agamas (Phrynocephalus mystaceus) – the traditional one, based on route trapping and measurements of body temperature in captured lizards, and the new one, using temperature loggers that register body temperature once a minute and implemented interperitoneally. Both work options are not alternative. The main thing is the correct interpretation of the results: the traditional method provides data obtained from many individuals characterizing the temperature parameters of the activity of the studied reptiles. The new technique allows, although on a limited number of individuals, but with a huge number of measurements, to obtain the calculated characters of the reptiles' thermoregulation. Both of them successfully complement each other and, with subsequent refinement, could represent some single whole, allowing for a more complete and accurate understanding and description of the thermal biology of different reptile species.

Time of day-dependent effects of heat treatment on thermal tolerance and sex differentiation in Nile tilapia (Oreochromis niloticus)

The present research aimed to determine the influence of the time of day at which heat treatment (HT) is applied to sexual differentiation and thermal tolerance mechanisms in Nile tilapia (Oreochromis niloticus). To this end, fish larvae were subjected to HT (36 °C) for 12 days at different times of the day during the thermosensitive period (i.e. 11–23 days post-fertilization, dpf). The heat treatment was applied during the light phase (L-HT); during the dark phase (D-HT), or during both light-dark phases (LD-HT). A control group maintained at constant rearing temperature (28 °C, CTRL) was also used. At 25 dpf, thermal tolerance was determined (survival) and whole larvae samples were collected for the mRNA expression analysis (qPCR). There after, all the groups remained at constant temperature until 270 dpf, when some fish were sacrificed and each individual's gonadal lobules were collected for histological and qPCR analyses. At the same time, each experimental group was subjected to acute heat shock (HS; 36 °C for 2 h) in either the mid-light (ML) or the mid-dark (MD) phase. When HS exposure finished, adults' brains were extracted to determine the HS response by qPCR. The expression of the genes involved in female (cyp19a1a, foxl2, era) and male (amh, ara, sox9a, dmrt1) sexual differentiation and thermal tolerance (hsp70, hsp90a, hsp27) in the 25 dpf larvae and adults was studied. The results revealed that all the HT-treated larvae showed an up-regulation of testicular differentiation gene expression and inhibition of ovarian differentiation gene expression, which resulted in a higher percentage of males. The time of day of HT during development influenced thermotolerance in the larval and adult stages, with the lowest survival and highest cellular stress (higher HSPs expression) observed in the larvae of the D-HT and LD-HT groups. These findings reveal the need to further investigate the link between sexual differentiation and thermotolerance daily rhythms in thermal biology research. These results may be useful for optimizing masculinization protocols in the tilapia industry, while ensuring animal welfare and minimizing negative HT effects on fish physiology.

Thermal biology of Lanthanotus borneensis (Lanthanotidae) in Sarawak, Borneo

Abstract Lanthanotus borneensis, the Bornean earless monitor, is a monotypic member of the family Lanthanotidae, and restricted to the island of Borneo. Little has been published on its field ecology. This study investigated aspects of its thermal biology through an analysis of surface body temperatures of free ranging individuals against corresponding environmental temperatures, in order to explore aspects of microhabitat utilisation in relation to thermoregulation. A generalised linear mixed model shows significant effect of air and substrate temperatures, but not of water temperature. Further, the fixed effects of substrate temperature (coefficient estimate 0.396; ) versus ambient temperature (0.264; ), is suggestive of thigmothermy as the primary mode of thermoregulation. The species does not appear to utilise specific microhabitat structures to thermoregulate, the results of these observations suggesting that it is a thermoconformer.

How seasonality influences the thermal biology of lizards with different thermoregulatory strategies: a meta-analysis.

Ectotherms that maintain thermal balance in the face of varying climates should be able to colonise a wide range of habitats. In lizards, thermoregulation usually appears as a variety of behaviours that buffer external influences over physiology. Basking species rely on solar radiation to raise body temperatures and usually show high thermoregulatory precision. By contrast, species that do not bask are often constrained by climatic conditions in their habitats, thus having lower thermoregulatory precision. While much focus has been given to the effects of mean habitat temperatures, relatively less is known about how seasonality affects the thermal biology of lizards on a macroecological scale. Considering the current climate crisis, assessing how lizards cope with temporal variations in environmental temperature is essential to understand better how these organisms will fare under climate change. Activity body temperatures (Tb ) represent the internal temperature of an animal measured in nature during its active period (i.e. realised thermal niche), and preferred body temperatures (Tpref ) are those selected by an animal in a laboratory thermal gradient that lacks thermoregulatory costs (i.e. fundamental thermal niche). Both traits form the bulk of thermal ecology research and are often studied in the context of seasonality. In this study, we used a meta-analysis to test how environmental temperature seasonality influences the seasonal variation in the Tb and Tpref of lizards that differ in thermoregulatory strategy (basking versus non-basking). Based on 333 effect sizes from 137 species, we found that Tb varied over a greater magnitude than Tpref across seasons. Variations in Tb were not influenced by environmental temperature seasonality; however, body size and thermoregulatory strategy mediated Tb responses. Specifically, larger species were subjected to greater seasonal variations in Tb , and basking species endured greater seasonal variations in Tb compared to non-basking species. On the other hand, the seasonal variation in Tpref increased with environmental temperature seasonality regardless of body size. Thermoregulatory strategy also influenced Tpref , suggesting that behaviour has an important role in mediating Tpref responses to seasonal variations in the thermal landscape. After controlling for phylogenetic effects, we showed that Tb and Tpref varied significantly across lizard families. Taken together, our results support the notion that the relationship between thermal biology responses and climatic parameters can be taxon and trait dependent. Our results also showcase the importance of considering ecological and behavioural aspects in macroecological studies. We further highlight current systematic, geographical, and knowledge gaps in thermal ecology research. Our work should benefit those who aim to understand more fully how seasonality shapes thermal biology in lizards, ultimately contributing to the goal of elucidating the evolution of temperature-sensitive traits in ectotherms.

Thermal biology of aquatic insects in alpine lakes: Insights from diving beetles

Abstract High mountain areas are especially vulnerable to global warming, as they experience faster temperature changes than lowlands in a climate change context. Notably, increased temperatures and frequency of extreme flooding and droughts, and the consequent decrease in ice cover and water availability fluctuations, will induce important physical changes in alpine freshwater systems. Thus, assessing thermal limits and exploring overwintering strategies of aquatic alpine insects is pivotal to understanding how aquatic communities of high‐mountain fresh waters will respond to climate change. However, knowledge on these topics is still scarce for aquatic alpine insects. Here, the thermal biology of adults of five diving beetle species from alpine lakes located in the Sierra Nevada mountain range (southern Iberia) was studied. Cold tolerance was measured estimating the supercooling point (SCP), lower lethal temperature (LLT), tolerance to ice enclosure and to submersion, whereas heat tolerance was assessed from the heat coma temperature and upper lethal temperature. All of the species survived ice enclosure for 3 h. Furthermore, three of the studied species had SCPs higher than their LLTs, suggesting that they could be freeze‐tolerant. All species except Agabus nevadensis also were tolerant to submersion, which could be a key adaptation for overwintering underwater below the ice cover as adults, reducing risk from freezing conditions in the air. The species did not differ significantly in their upper thermal limits, which were similar to those of other dytiscids from lower altitudes. Overall, our results suggest that increasing temperatures is not expected to be the most important threat for the water beetle populations in Sierra Nevada, but rather the colonisation of alpine lakes by lowland dytiscids in a warmer climate scenario.

Effect of thermal acclimation on the tolerance of the peach fruit fly (Bactrocera zonata: Tephritidae) to heat and cold stress

Understanding the thermal biology of insects is of increasing importance for predicting their geographic distribution, particularly in light of current and future global temperature increases. Within the limits set by genetic makeup, thermal tolerance is affected by the physiological conditioning of individuals (e.g., through acclimation). Considering this phenotypic plasticity may add to accurately estimating changes to the distribution of insects under a changing climate.We studied the effect of thermal acclimation on cold and heat tolerance of the peach fruit fly (Bactrocera zonata) – an invasive, polyphagous pest that is currently expanding through Africa and the Middle East. Females and males were acclimated at 20, 25 and 30 °C for up to 19 days following adult emergence. The critical thermal minimum (CTmin) and maximum (CTmax) were subsequently recorded as well adult survival following acute exposure to chilling (0 or −3 °C for 2 h). Additionally, we determined the survival of pupae subjected for 2 h to temperatures ranging from −12 °C to 5 °C.We demonstrate that acclimation at 30 °C resulted in significantly higher CTmax and CTmin values (higher heat resistance and lower cold resistance, respectively). Additionally, adult recovery following exposure to −3 °C was significantly reduced following acclimation at 30 °C, and this effect was significantly higher for females. Pupal mortality increased with the decrease in temperature, reaching LT50 and LT95 values following exposure to -0.32 °C and −6.88 °C, respectively. Finally, we found that the survival of pupae subjected to 0 and 2 °C steadily increased with pupal age.Our findings substantiate a physiological foundation for understanding the current geographic range of B. zonata. We assume that acclimation at 30 °C affected the thermal tolerance of the flies partly through modulating feeding and metabolism. Tolerance to chilling during the pupal stage probably changed according to temperature-sensitive processes occurring during metamorphosis, rendering younger pupae more sensitive to chilling.

Living in the mountains: Thermal ecology and freezing tolerance of the lizard Abronia taeniata (Squamata: Anguidae)

The impact of daily and seasonal variation in environmental temperature on lizards is important, since their physiological processes are body temperature dependent. Lizards that occupy mountainous areas must have been favoured to colonize such habitats through selection on thermal biology traits to thermoregulate effectively. Moreover, mountain lizards may be able to maintain their activity near their minimum critical temperature and even have antifreeze mechanisms. Tolerance of freezing is related to the biosynthesis of cryoprotective molecules, such as glucose, whose concentration may increase after freezing. The aims of the present work were: (1) study the thermoregulation of the viviparous lizard Abronia taeniata, and (2) determine its survival and/or tolerance to freezing. This species occurs in pine forests, pine-oak forests, and mountain mesophilic forests in areas that reach freezing temperatures. In the field, we recorded air, substrate, and body temperatures at capture time of the lizards, and registered operative temperatures at the study area. In the laboratory, we determined thermal preferences, crystallization point, and blood glucose levels of individuals before and after freezing. We found out that A. taeniata sustains activity in a wide range of temperatures, actively avoids thermally favourable microhabitats in spring, and is a moderate thermoregulator during autumn and winter. In A. taeniata, the body temperatures are tightly linked to air and substrate temperatures. Seasonality had an effect over body temperature, preferred temperatures and thermoregulatory effectiveness indices. When exposed to temperatures below zero, A. taeniata showed an increase in blood glucose levels, which aided them in surviving freezing. Taken together, our results suggest that A. taeniata may sustain activity at low environmental temperatures, due to an effective behavioural thermoregulation, and in case temperatures of its habitat go below zero, is also capable of tolerate freezing.

Direct writing of microheater for studying plant thermal biology

Microheaters have drawn extensive attention for their substantial applications in thermotherapy, gas sensors, thin film preparation, biological research, etc. In plant physiology, uncovering the mechanisms by which plants sense and respond to environmental temperature fluctuations will help us better understand the impact of climate change on crop yield and ecosystem resilience. Currently, microheaters with long-term heating capability have rarely been applied to investigate plant thermal responses. In this study, we applied a direct writing technique to fabricate microheaters suitable for studying plant thermal biology with silver conductive ink. The optimal printing conditions and the heating performance (e.g., stability, durability, reusability) of the printed heaters were thoroughly characterized. The printed microheaters can provide stable and constant heating to plant organs for over four days. When placed near plant leaves to create localized heating, the microheater could successfully activate the expression of a thermoresponsive marker gene in plants. These results demonstrate the potential of applying printed microheaters to study plant thermal biology at the organ and tissue level.

Differences between thermal preference and thermal performance in a wintry spider Mecicobothrium thorelli: are the spiders under evolutionary pressures on their seasonal activity?

Thermal preference and thermal performance are used to describe the thermal biology of an ectothermic organism through parameters, i.e., estimating locomotor performance by maximum running speed. In this study, we assessed the thermal preference and locomotor performance of the spider Mecicobothrium thorelli Holmberg, 1882, a wintry mygalomorph spider endemic to the native mountainous grasslands of central Argentina and Uruguay. The preferred temperatures of the 72.4% of the individuals were in the range of 10–20 °C. The highest frequencies of preferred temperatures were 10–15 °C in males and 15–20 °C in females. The sprint speed showed significant differences between all the temperatures evaluated and showed the highest speeds at 25 °C and the lowest at 3 °C. The optimal temperature was 26.09 °C, which was significantly higher than the preferred temperature in both males and females. We concluded that M. thorelli selects a wide range of temperatures and prefers to stay in medium and low temperatures, which are correlated with winter activity in the wild. However, the species showed maximum speed at higher temperatures, which implies that spiders would perform even better in nature and maximize their locomotion by being active during a warmer period.

Negative effects of the spatial clumping of thermal resources on lizard thermoregulation in a fragmented habitat

In ecosystems threatened by the expansion of croplands, habitat fragmentation and climate change, two of the main extinction drivers, may have thermoregulation-mediated interacting effects on demographic trends of terrestrial ectotherms. We studied the thermal biology of a metapopulation of the widespread Mediterranean lacertid Psammodromus algirus in ten fragments of evergreen or deciduous oak forests interspersed among cereal fields. We obtained thermoregulation statistics (selected temperature range, body and operative temperatures, thermal quality of the habitat, and precision, accuracy, and effectiveness of thermoregulation) that could be compared among fragments and with conspecific populations living in unfragmented habitat. We also measured the selection (use vs. availability) and spatial distribution of sunlit and shaded patches used for behavioral thermoregulation in fragments, and we estimated operative temperatures and thermal habitat quality in the agricultural matrix surrounding the fragments. Variation of the thermal environment was much larger within fragments than among them, and thermoregulation was accurate, precise, and efficient throughout the fragmented landscape; its effectiveness was similar to that of previously studied unfragmented populations. The average distance between sunlit and shaded patches was shorter in deciduous than in evergreen fragments, producing a more clumped distribution of the mosaic of thermal resources. Consequently, in evergreen habitat the cost of thermoregulation was higher, because lizards were more selective in their choice of sunlit sites (i.e. they used sunlit patches closer to shade and refuge than expected at random, and the extent of such selection was larger than at deciduous habitat). Temperatures available in croplands were too high to allow lizard dispersal, at least in the post-breeding season. This result confirms the role of croplands as a thermal barrier that promotes inbreeding and associated fitness losses in isolated fragments, and it forecasts a dark future for populations of forest lizards in agricultural landscapes under the combined effects of habitat fragmentation and global warming.