Body mass, temperature, and pathogen intensity differentially affect critical thermal maxima and their population-level variation in a solitary bee.

Climate change presents a major threat to species distribution and persistence. Understanding what abiotic or biotic factors influence the thermal tolerances of natural populations is critical to assessing their vulnerability under rapidly changing thermal regimes. This study evaluates how body mass, local climate, and pathogen intensity influence heat tolerance and its population-level variation (SD) among individuals of the solitary bee Xenoglossa pruinosa. We assess the sex-specific relationships between these factors and heat tolerance given the differences in size between sexes and the ground-nesting behavior of the females. We collected X. pruinosa individuals from 14 sites across Pennsylvania, USA, that varied in mean temperature, precipitation, and soil texture. We measured the critical thermal maxima (CTmax) of X. pruinosa individuals as our proxy for heat tolerance and used quantitative PCR to determine relative intensities of three parasite groups-trypanosomes, Spiroplasma apis (mollicute bacteria), and Vairimorpha apis (microsporidian). While there was no difference in CTmax between the sexes, we found that CTmax increased significantly with body mass and that this relationship was stronger for males than for females. Air temperature, precipitation, and soil texture did not predict mean CTmax for either sex. However, population-level variation in CTmax was strongly and negatively correlated with air temperature, which suggests that temperature is acting as an environmental filter. Of the parasites screened, only trypanosome intensity correlated with heat tolerance. Specifically, trypanosome intensity negatively correlated with the CTmax of female X. pruinosa but not males. Our results highlight the importance of considering size, sex, and infection status when evaluating thermal tolerance traits. Importantly, this study reveals the need to evaluate trends in the variation of heat tolerance within and between populations and consider implications of reduced variation in heat tolerance for the persistence of ectotherms in future climate conditions.

Heat stress survival and thermal tolerance of Australian stingless bees.

Stingless bees (Meliponini) are important pollinators throughout the world's tropical and subtropical regions. Understanding their thermal tolerance is key to predicting their resilience to changing climates and increasingly frequent extreme heat events. We examined critical thermal maxima (CTmax), survival during 1-8 h heat periods, chill coma recovery and thermal preference for Australian meliponine species that occupy different climates across their ranges: Tetragonula carbonaria (tropical to temperate regions), T. hockingsi (tropical and subtropical regions only) and Austroplebeia australis (widely distributed including arid regions). We found interspecific differences in thermal tolerance consistent with differences in the climate variability observed in each species' range. Foragers of A. australis had a faster chill coma recovery (288 s) than foragers of T. hockingsi (1059 s) and T. carbonaria (872 s). Austroplebeia australis also had the highest CTmax of 44.5 °C, while the CTmax of the two Tetragonula species was ∼43.1 °C. After a 1-h heat exposure, T. carbonaria foragers experienced 95% mortality at 42 °C, and 100% at 45 °C. Surprisingly, larvae and pupae of both Tetragonula species were more resistant to heat exposure than foragers. Within an enclosed temperature gradient apparatus (17-38 °C), no clear preference was found for foragers; however, they were most frequently observed at ∼18 °C. Results indicate that in some regions of Australia, meliponines already experience periodic heat events exceeding their thermal maxima. Employing effective management strategies (such as nest site insulation and habitat preservation) may be crucial to colony survival under continued climate change.

Loss of mitochondrial performance at high temperatures is correlated with upper thermal tolerance among populations of an intertidal copepod.

Environmental temperatures have pervasive effects on the performance and tolerance of ectothermic organisms, and thermal tolerance limits likely play key roles underlying biogeographic ranges and responses to environmental change. Mitochondria are central to metabolic processes in eukaryotic cells, and these metabolic functions are thermally sensitive; however, potential relationships between mitochondrial function, thermal tolerance limits and local thermal adaptation in general remain unresolved. Loss of ATP synthesis capacity at high temperatures has recently been suggested as a mechanistic link between mitochondrial function and upper thermal tolerance limits. Here we use a common-garden experiment with seven locally adapted populations of intertidal copepods (Tigriopus californicus), spanning approximately 21.5° latitude, to assess genetically based variation in the thermal performance curves of maximal ATP synthesis rates in isolated mitochondria. These thermal performance curves displayed substantial variation among populations with higher ATP synthesis rates at lower temperatures (20-25 °C) in northern populations than in southern populations. In contrast, mitochondria from southern populations maintained ATP synthesis rates at higher temperatures than the temperatures that caused loss of ATP synthesis capacity in mitochondria from northern populations. Additionally, there was a tight correlation between the thermal limits of ATP synthesis and previously determined variation in upper thermal tolerance limits among populations. This suggests that mitochondria may play an important role in latitudinal thermal adaptation in T. californicus, and supports the hypothesis that loss of mitochondrial performance at high temperatures is linked to whole-organism thermal tolerance limits in this ectotherm.

Divergent physiological acclimation responses to warming between two co-occurring salamander species and implications for terrestrial survival.

Small differences in physiological responses are known to influence demographic rates such as survival. We tested for differences in the physiological acclimation responses of two closely-related salamander species that often co-occur, Ambystoma maculatum and A. opacum. Specifically, we measured changes in critical thermal maxima (CTmax), standard metabolic rates (SMRs), and respiratory surface area water loss (RSAWL) following exposure to three temperature treatments under laboratory conditions. While the magnitude of RSAWL and CTmax acclimation responses to warming did not differ between the study species, SMR was maintained across acclimation temperatures among A. maculatum, but declined among A. opacum acclimated to warmer temperatures. Metabolic compensation may facilitate maintained A. maculatum activity levels during warm periods following the relatively cool spring breeding season. In contrast, metabolic suppression may allow A. opacum to conserve energy when exposed to surface conditions during fall breeding and nest guarding. We simulated how these different SMR responses would likely alter post-metamorphic survival in our study species using previously collected data representing six weeks under relatively warm seminatural conditions. Our simulation indicated that, following warming and under identical study conditions, metabolic compensation may allow juvenile A. maculatum to maintain survival likelihoods, whereas metabolic depression may cause juvenile A. opacum to experience increased survivorship. These findings underscore that comparable physiological responses among ecologically similar, sympatric species cannot be assumed. Further, results of this study suggest that metabolic responses may play an important role in amphibian species persistence as temperatures increase due to habitat modification and climate change.

Bacterial Symbionts Confer Thermal Tolerance to Cereal Aphids Rhopalosiphum padi and Sitobion avenae.

High-temperature events are evidenced to exert significant influence on the population performance and thermal biology of insects, such as aphids. However, it is not yet clear whether the bacterial symbionts of insects mediate the thermal tolerance traits of their hosts. This study is intended to assess the putative association among the chronic and acute thermal tolerance of two cereal aphid species, Rhopalosiphum padi (L.) and Sitobion avenae (F.), and the abundance of their bacterial symbionts. The clones of aphids were collected randomly from different fields of wheat crops and were maintained under laboratory conditions. Basal and acclimated CTmax and chronic thermal tolerance indices were measured for 5-day-old apterous aphid individuals and the abundance (gene copy numbers) of aphid-specific and total (16S rRNA) bacterial symbionts were determined using real-time RT-qPCR. The results reveal that R. padi individuals were more temperature tolerant under chronic exposure to 31 °C and also exhibited about 1.0 °C higher acclimated and basal CTmax values than those of S. avenae. Moreover, a significantly higher bacterial symbionts' gene abundance was recorded in temperature-tolerant aphid individuals than the susceptible ones for both aphid species. Although total bacterial (16S rRNA) abundance per aphid was higher in S. avenae than R. padi, the gene abundance of aphid-specific bacterial symbionts was nearly alike for both of the aphid species. Nevertheless, basal and acclimated CTmax values were positively and significantly associated with the gene abundance of total symbiont density, Buchnera aphidicola, Serratia symbiotica, Hamilton defensa, Regiella insecticola and Spiroplasma spp. for R. padi, and with the total symbiont density, total bacteria (16S rRNA) and with all aphid-specific bacterial symbionts (except Spiroplasma spp.) for S. avenae. The overall study results corroborate the potential role of the bacterial symbionts of aphids in conferring thermal tolerance to their hosts.

Estimating the differences in critical thermal maximum and metabolic rate of Helicoverpa punctigera (Wallengren) (Lepidoptera: Noctuidae) across life stages.

Temperature is a crucial driver of insect activity and physiological processes throughout their life-history, and heat stress may impact life stages (larvae, pupae and adult) in different ways. Using thermolimit respirometry, we assessed the critical thermal maxima (CTmax-temperature at which an organism loses neuromuscular control), CO2 emission rate (V́CO2) and Q10 (a measure of V́CO2 temperature sensitivity) of three different life stages of Helicoverpa punctigera (Wallengren) by increasing their temperature exposure from 25 °C to 55 °C at a rate of 0.25 °C min-1 . We found that the CTmax of larvae (49.1 °C ± 0.3 °C) was higher than pupae (47.4 °C ± 0.2 °C) and adults (46.9 °C ± 0.2 °C). The mean mass-specific CO2 emission rate (ml V́CO2 h-1) of larvae (0.26 ± 0.03 ml V́CO2 h-1) was also higher than adults (0.24 ± 0.04 ml V́CO2 h-1) and pupae (0.06 ± 0.02 ml V́CO2 h-1). The Q10: 25-35 °C for adults (2.01 ± 0.22) was significantly higher compared to larvae (1.40 ± 0.06) and Q10: 35-45 °C for adults (3.42 ± 0.24) was significantly higher compared to larvae (1.95 ± 0.08) and pupae (1.42 ± 0.98) respectively. We have established the upper thermal tolerance of H. punctigera, which will lead to a better understanding of the thermal physiology of this species both in its native range, and as a pest species in agricultural systems.

Sub-lethal temperature thresholds indicate acclimation and physiological limits in brook trout Salvelinus fontinalis.

The upper thermal tolerance of brook trout Salvelinus fontinalis was estimated using critical thermal maxima (CTmax ) experiments on fish acclimated to temperatures that span the species' thermal range (5-25°C). The CTmax increased with acclimation temperature but plateaued in fish acclimated to 20, 23 and 25°C. Plasma lactate was highest, and the hepato-somatic index (IH ) was lowest at 23 and 25°C, which suggests additional metabolic costs at those acclimation temperatures. The results suggest that there is a sub-lethal threshold between 20 and 23°C, beyond which the fish experience reduced physiological performance.

Nitrite-induced reductions in heat tolerance are independent of aerobic scope in a freshwater teleost.

Nitrite is a widespread form of pollution that directly lowers the blood oxygen carrying capacity of aquatically respiring species. It is unknown if this impairment of oxygen transport translates into an increased susceptibility to elevated temperatures. We hypothesised that nitrite exposure would lower blood oxygen carrying capacity and decrease both aerobic scope (maximum-standard metabolic rate) and heat tolerance. To test these hypotheses, juvenile European carp (Cyprinus carpio) were exposed to two levels of nitrite (0 mmol l-1 or 1 mmol l-1) for 7 days and haematological parameters, critical thermal maxima (CTmax) and aerobic scope were assessed. Nitrite exposure reduced total haemoglobin by 32.9%. Aerobic scope remained unchanged in fish exposed to nitrite; however, marked declines in CTmax (1.2°C reduction) were observed in nitrite-exposed fish. These findings demonstrate that nitrite exposure can significantly impair heat tolerance, even when aerobic capacity is maintained.

Are model organisms representative for climate change research? Testing thermal tolerance in wild and laboratory zebrafish populations.

Model organisms can be useful for studying climate change impacts, but it is unclear whether domestication to laboratory conditions has altered their thermal tolerance and therefore how representative of wild populations they are. Zebrafish in the wild live in fluctuating thermal environments that potentially reach harmful temperatures. In the laboratory, zebrafish have gone through four decades of domestication and adaptation to stable optimal temperatures with few thermal extremes. If maintaining thermal tolerance is costly or if genetic traits promoting laboratory fitness at optimal temperature differ from genetic traits for high thermal tolerance, the thermal tolerance of laboratory zebrafish could be hypothesized to be lower than that of wild zebrafish. Furthermore, very little is known about the thermal environment of wild zebrafish and how close to their thermal limits they live. Here, we compared the acute upper thermal tolerance (critical thermal maxima; CTmax) of wild zebrafish measured on-site in West Bengal, India, to zebrafish at three laboratory acclimation/domestication levels: wild-caught, F1 generation wild-caught and domesticated laboratory AB-WT line. We found that in the wild, CTmax increased with increasing site temperature. Yet at the warmest site, zebrafish lived very close to their thermal limit, suggesting that they may currently encounter lethal temperatures. In the laboratory, acclimation temperature appeared to have a stronger effect on CTmax than it did in the wild. The fish in the wild also had a 0.85-1.01°C lower CTmax compared to all laboratory populations. This difference between laboratory-held and wild populations shows that environmental conditions can affect zebrafish's thermal tolerance. However, there was no difference in CTmax between the laboratory-held populations regardless of the domestication duration. This suggests that thermal tolerance is maintained during domestication and highlights that experiments using domesticated laboratory-reared model species can be appropriate for addressing certain questions on thermal tolerance and global warming impacts.

Rate dynamics of ectotherm responses to thermal stress.

Critical thermal limits (CTLs) show much variation associated with the experimental rate of temperature change used in their estimation. Understanding the full range of variation in rate effects on CTLs and their underlying basis is thus essential if methodological noise is not to overwhelm or bias the ecological signal. We consider the effects of rate variation from multiple intraspecific assessments and provide a comprehensive empirical analysis of the rate effects on both the critical thermal maximum (CTmax) and critical thermal minimum (CTmin) for 47 species of ectotherms, exploring which of the available theoretical models best explains this variation. We find substantial interspecific variation in rate effects, which takes four different forms (increase, decline, no change, mixed), with phylogenetic signal in effects on CTmax, but not CTmin. Exponential and zero exponential failure rate models best explain the rate effects on CTmax. The majority of the empirical rate variation in CTmin could not be explained by the failure rate models. Our work demonstrates that rate effects cannot be ignored in comparative analyses, and suggests that incorporation of the failure rate models into such analyses is a useful further avenue for exploration of the fundamental basis and implications of such variation.

The effect of salinity and photoperiod on thermal tolerance of Atlantic and coho salmon reared from smolt to adult in recirculating aquaculture systems.

Land-based, closed containment salmon aquaculture involves rearing salmon from smolt to adult in recirculating aquaculture systems (RAS). Unlike in open-net pen aquaculture, rearing conditions can be specified in RAS in order to optimize growth and physiological stress tolerance. The environmental conditions that yield optimal stress tolerance in salmon are, however, unknown. To address this knowledge gap, we reared Atlantic (Salmo salar) and coho (Oncorhynchus kisutch) salmon in 7 separate RASs for 400 days post-smoltification under 2 photoperiods (24:0 or 12:12, light:dark) and 4 salinities (2.5, 5, 10 or 30 ppt.) and assessed the effects of these conditions on thermal tolerance. We found that over the first 120 days post-smoltification, rearing coho under a 24:0 photoperiod resulted in a ~2 °C lower critical thermal maxima (CTmax) than in coho reared under a 12:12 photoperiod. This photoperiod effect did not persist at 200 and 400 days, which was coincident with an overall decrease in CTmax in coho. Finally, Atlantic salmon had a higher CTmax (~28 °C) compared to coho (~26 °C) at 400 days post-smoltification. Overall, these findings are important for the future implications of RAS and for the aquaculture industry to help identify physiologically sensitive time stages.

Intraspecific variation in lizard heat tolerance alters estimates of climate impact.

Research addressing the effects of global warming on the distribution and persistence of species generally assumes that population variation in thermal tolerance is spatially constant or overridden by interspecific variation. Typically, this rationale is implicit in sourcing one critical thermal maximum (CTmax ) population estimate per species to model spatiotemporal cross-taxa variation in heat tolerance. Theory suggests that such an approach could result in biased or imprecise estimates and forecasts of impact from climate warming, but limited empirical evidence in support of those expectations exists. We experimentally quantify the magnitude of intraspecific variation in CTmax among lizard populations, and the extent to which incorporating such variability can alter estimates of climate impact through a biophysical model. To do so, we measured CTmax from 59 populations of 15 Iberian lizard species (304 individuals). The overall median CTmax across all individuals from all species was 42.8°C and ranged from 40.5 to 48.3°C, with species medians decreasing through xeric, climate-generalist and mesic taxa. We found strong statistical support for intraspecific differentiation in CTmax by up to a median of 3°C among populations. We show that annual restricted activity (operative temperature > CTmax ) over the Iberian distribution of our study species differs by a median of >80 hr per 25-km2 grid cell based on different population-level CTmax estimates. This discrepancy leads to predictions of spatial variation in annual restricted activity to change by more than 20 days for six of the study species. Considering that during restriction periods, reptiles should be unable to feed and reproduce, current projections of climate-change impacts on the fitness of ectotherm fauna could be under- or over-estimated depending on which population is chosen to represent the physiological spectra of the species in question. Mapping heat tolerance over the full geographical ranges of single species is thus critical to address cross-taxa patterns and drivers of heat tolerance in a biologically comprehensive way.

Native Chinook salmon Oncorhynchus tshawytscha and non-native brook trout Salvelinus fontinalis prefer similar water temperatures.

Preferred water temperatures and acute temperature tolerance limits of two salmonids in California were assessed: juvenile Chinook salmon Oncorhynchus tshawytscha, a native anadromous species, and sub-adult brook trout Salvelinus fontinalis, an introduced game species. These two species preferred similar temperatures across an 18 h temperature preference experiment and showed similar critical thermal tolerance limits, suggesting a substantial thermal habitat overlap in the wild.

Negligible differences in metabolism and thermal tolerance between diploid and triploid Atlantic salmon (Salmo salar).

The mechanisms that underlie thermal tolerance in aquatic ectotherms remain unresolved. Triploid fish have been reported to exhibit lower thermal tolerance than diploids, offering a potential model organism to better understand the physiological drivers of thermal tolerance. Here, we compared triploid and diploid juvenile Atlantic salmon (Salmo salar) in freshwater to investigate the proposed link between aerobic capacity and thermal tolerance. We measured specific growth rates (SGR) and resting (aerobic) metabolic rates (RMR) in freshwater at 3, 7 and 9 weeks of acclimation to 10, 14 and 18°C. Additionally, maximum metabolic rates (MMR) were measured at 3 and 7 weeks of acclimation, and critical thermal maxima (CTmax) were measured at 9 weeks. Mass, SGR and RMR differed between ploidies across all temperatures at the beginning of the acclimation period, but all three metrics were similar across ploidies by week 7. Aerobic scope (MMR-RMR) remained consistent across ploidies, acclimation temperatures and time. At 9 weeks, CTmax was independent of ploidy, but correlated positively with acclimation temperature despite the similar aerobic scope between acclimation groups. Our findings suggest that acute thermal tolerance is not modulated by aerobic scope, and the altered genome of triploid Atlantic salmon does not translate to reduced thermal tolerance of juvenile fish in freshwater.

Evolutionary and environmental determinants of freshwater fish thermal tolerance and plasticity.

Understanding the extent to which phylogenetic constraints and adaptive evolutionary forces help define the physiological sensitivity of species is critical for anticipating climate-related impacts in aquatic environments. Yet, whether upper thermal tolerance and plasticity are shaped by common evolutionary and environmental mechanisms remains to be tested. Based on a systematic literature review, we investigated this question in 82 freshwater fish species (27 families) representing 829 experiments for which data existed on upper thermal limits and it was possible to estimate plasticity using upper thermal tolerance reaction norms. Our findings indicated that there are strong phylogenetic signals in both thermal tolerances and acclimation capacity, although it is weaker in the latter. We found that upper thermal tolerances are correlated with the temperatures experienced by species across their range, likely because of spatially autocorrelated processes in which closely related species share similar selection pressures and limited dispersal from ancestral environments. No association with species thermal habitat was found for acclimation capacity. Instead, species with the lowest physiological plasticity also displayed the highest thermal tolerances, reflecting to some extent an evolutionary trade-off between these two traits. Although our study demonstrates that macroecological climatic niche features measured from species distributions are likely to provide a good approximation of freshwater fish sensitivity to climate change, disentangling the mechanisms underlying both acute and chronic heat tolerances may help to refine predictions regarding climate change-related range shifts and extinctions.

Self-heating by large insect larvae?

Do insect larvae ever self-heat significantly from their own metabolic activity and, if so, under what sets of environmental temperatures and across what ranges of body size? We examine these questions using larvae of the Japanese rhinoceros beetle (Trypoxylus dichotomus), chosen for their large size (>20g), simple body plan, and underground lifestyle. Using CO2 respirometry, we measured larval metabolic rates then converted measured rates of gas exchange into rates of heat production and developed a mathematical model to predict how much steady state body temperatures of underground insects would increase above ambient depending on body size. Collectively, our results suggest that large, extant larvae (20-30g body mass) can self-heat by at most 2°C, and under many common conditions (shallow depths, moister soils) would self-heat by less than 1°C. By extending the model to even larger (hypothetical) body sizes, we show that underground insects with masses >1kg could heat, in warm, dry soils, by 1.5-6°C or more. Additional experiments showed that larval critical thermal maxima (CTmax) were in excess of 43.5°C and that larvae could behaviorally thermoregulate on a thermal gradient bar. Together, these results suggest that large larvae living underground likely regulate their temperatures primarily using behavior; self-heating by metabolism likely contributes little to their heat budgets, at least in most common soil conditions.

Thermal tolerance and survival responses to scenarios of experimental climatic change: changing thermal variability reduces the heat and cold tolerance in a fly.

Climate change poses one of the greatest threats to biodiversity. Most analyses of the impacts have focused on changes in mean temperature, but increasing variance will also impact organisms and populations. We assessed the combined effects of the mean and the variance of temperature on thermal tolerances-i.e., critical thermal maxima, critical thermal minima, scope of thermal tolerance, and survival in Drosophila melanogaster. Our six experimental climatic scenarios were: constant mean with zero variance or constant variance or increasing variance; changing mean with zero variance or constant variance or increasing variance. Our key result was that environments with changing thermal variance reduce the scope of thermal tolerance and survival. Heat tolerance seems to be conserved, but cold tolerance decreases significantly with mean low as well as changing environmental temperatures. Flies acclimated to scenarios of changing variance-with either constant or changing mean temperatures-exhibited significantly lower survival rate. Our results imply that changing and constant variances would be just as important in future scenarios of climate change under greenhouse warming as increases in mean annual temperature. To develop more realistic predictions about the biological impacts of climate change, such interactions between the mean and variance of environmental temperature should be considered.

Behavioral thermoregulation and critical thermal limits of giant keyhole limpet Megathura crenulata(Sowerby 1825) (Mollusca; Vetigastropoda).

The thermoregulatory behavior of the giant keyhole limpet Megathura crenulata was determined in a horizontal thermal gradient during the day at 18.9 °C and 18.3 °C for the night. The final preferendum determined for giant keyhole limpets was of 18.6±1.2 °C. Limpets' displacement velocity was 10.0±3.9 cm h(-1) during the light phase and 8.4±1.6 cm h(-1) during the dark phase. The thermotolerance (measured as CTMax at 50%) was determined in a keyhole limpet in three acclimation temperatures 17, 20, and 23 °C. Limpets were subjected to water increasing temperatures at a rate of 1 °C every 30 min, until they detached from the substrate. The critical thermal maximum at 50% was 27.2, 27.9 and 28.3 °C respectively.

Climate change, multiple stressors, and the decline of ectotherms.

Climate change is believed to be causing declines of ectothermic vertebrates, but there is little evidence that climatic conditions associated with declines have exceeded critical (i.e., acutely lethal) maxima or minima, and most relevant studies are correlative, anecdotal, or short-term (hours). We conducted an 11-week factorial experiment to examine the effects of temperature (22 °C or 27 °C), moisture (wet or dry), and atrazine (an herbicide; 0, 4, 40, 400 µg/L exposure as embryos and larvae) on the survival, growth, behavior, and foraging rates of postmetamorphic streamside salamanders (Ambystoma barbouri), a species of conservation concern. The tested climatic conditions were between the critical maxima and minima of streamside salamanders; thus, this experiment quantified the long-term effects of climate change within the noncritical range of this species. Despite a suite of behavioral adaptations to warm and dry conditions (e.g., burrowing, refuge use, huddling with conspecifics, and a reduction in activity), streamside salamanders exhibited significant loss of mass and significant mortality in all but the cool and moist conditions, which were closest to the climatic conditions in which they are most active in nature. A temperature of 27 °C represented a greater mortality risk than dry conditions; death occurred rapidly at this temperature and more gradually under cool and dry conditions. Foraging decreased under dry conditions, which suggests there were opportunity costs to water conservation. Exposure to the herbicide atrazine additively decreased water-conserving behaviors, foraging efficiency, mass, and time to death. Hence, the hypothesis that moderate climate change can cause population declines is even more plausible under scenarios with multiple stressors. These results suggest that climate change within the noncritical range of species and pollution may reduce individual performance by altering metabolic demands, hydration, and foraging effort and may facilitate population declines of amphibians and perhaps other ectothermic vertebrates.