Calling behaviour under climate change: geographical and seasonal variation of calling temperatures in ectotherms

Calling behaviour is strongly temperature‐dependent and critical for sexual selection and reproduction in a variety of ectothermic taxa, including anuran amphibians, which are the most globally threatened vertebrates. However, few studies have explored how species respond to distinct thermal environments at time of displaying calling behaviour, and thus it is still unknown whether ongoing climate change might compromise the performance of calling activity in ectotherms. Here, we used new audio‐trapping techniques (automated sound recording and detection systems) between 2006 and 2009 to examine annual calling temperatures of five temperate anurans and their patterns of geographical and seasonal variation at the thermal extremes of species ranges, providing insights into the thermal breadths of calling activity of species, and the mechanisms that enable ectotherms to adjust to changing thermal environments. All species showed wide thermal breadths during calling behaviour (above 15 °C) and increases in calling temperatures in extremely warm populations and seasons. Thereby, calling temperatures differed both geographically and seasonally, both in terrestrial and aquatic species, and were 8–22 °C below the specific upper critical thermal limits (CT_max) and strongly associated with the potential temperatures of each thermal environment (operative temperatures during the potential period of breeding). This suggests that calling behaviour in ectotherms may take place at population‐specific thermal ranges, diverging when species are subjected to distinct thermal environments, and might imply plasticity of thermal adjustment mechanisms (seasonal and developmental acclimation) that supply species with means of coping with climate change. Furthermore, the thermal thresholds of calling at the onset of the breeding season were dissimilar between conspecific populations, suggesting that other factors besides temperature are needed to trigger the onset of reproduction. Our findings imply that global warming would not directly inhibit calling behaviour in the study species, although might affect other temperature‐dependent features of their acoustic communication system.     

Thermal dependence of behavioural performance of anuran amphibians

The thermal dependence of performance capacity was assessed in two anuran amphibians: Bufo boreas (western toad) and Rana pipiens (leopard frog). Quantitative measurements of performance showed that Bufo could sustain slow rates of walking for 10 min and cover greater distances than Rana, which initially jumped more vigorously but fatigued within 5 min. Changes in performance with changes in body temperature were virtually instantaneous, and performance exhibited no acclimation over 7 days. Within the range of temperatures studied, performance capacity increased with increasing body temperature and reached a maximum at 28 C in Bufo and 20 to 29 C in Rana. Performance capacity and the underlying metabolic processes had a similar thermal dependence within a species. The behavioural capacity for activity is apparently maximal for both species at body temperatures normally encountered in the field. Anuran behaviours requiring sustained activity (migration to breeding sites, mating, foraging) must therefore be markedly temperature-sensitive.     

Population‐specific effects of developmental temperature on body condition and jumping performance of a widespread European frog

All physiological processes of ectotherms depend on environmental temperature. Thus, adaptation of physiological mechanisms to the thermal environments is important for achieving optimal performance and fitness. The European Common Frog, Rana temporaria, is widely distributed across different thermal habitats. This makes it an exceptional model for studying the adaptations to different thermal conditions. We raised tadpoles from Germany and Croatia at two constant temperature treatments (15°C, 20°C), and under natural temperature fluctuations (in outdoor treatments), and tested how different developmental temperatures affected developmental traits, that is, length of larval development, morphometrics, and body condition, as well as jumping performance of metamorphs. Our results revealed population‐specific differences in developmental time, body condition, and jumping performance. Croatian frogs developed faster in all treatments, were heavier, in better body condition, and had longer hind limbs and better jumping abilities than German metamorphs. The populations further differed in thermal sensitivity of jumping performance. While metamorphs from Croatia increased their jumping performance with higher temperatures, German metamorphs reached their performance maximum at lower temperatures. These population‐specific differences in common environments indicate local genetic adaptation, with southern populations being better adapted to higher temperatures than those from north of the Alps.     

Physiological basis for diurnal activity in dispersing juvenile Bufo granulosus in the Caatinga, a Brazilian semi-arid environment

Diurnal activity is characteristic of many toad species, including Bufo granulosus from the Brazilian semi-arid biome called the Caatinga. Because of their patterns of activity, juvenile toads are exposed to hot and dehydrating conditions. Our investigation focuses on temperature and water relationships, and is based on the prediction that anuran diurnal activity in a semi-arid environment must be associated with morphological, physiological and behavioral traits enhancing thermal tolerances, capacity for performance at high temperatures and water balance. To test specific hypothesis related with this prediction, we investigated postmetamorphic B. granulosus and collected data on thermal tolerances and preferences, thermal safety margins, thermal dependence of locomotor behavior, thermal and kinetic properties of citrate synthase (CS), and skin morphophysiology. This information was compared with additional data from adult conspecifics and adult toads from sympatric species or from species from more moderate environments. We found that juvenile B. granulosus exhibit the highest critical maximum temperature reported for toads (44.2 degrees C) and are well suited to move at high temperatures. However, and in contrast with juveniles of other Bufo species, they do not show thermal preferences in a gradient and appear to hydroregulate more than thermoregulate. The CS of adult and juvenile toads shows typical patterns of thermal sensibility, but the thermal stability of this enzyme is much higher in juveniles than in adult Bufo of any other species studied. The inguinal skin exhibits a complex folding pattern and seems highly specialized for capillary water uptake. Diurnal activity in juvenile B. granulosus is possible given high thermal tolerances, keen ability to detect and uptake water, and avoidance behaviors.     

Evolution of morphology and locomotor performance in anurans: relationships with microhabitat diversification

The relationships between morphology, performance, behavior and ecology provide evidence for multiple and complex phenotypic adaptations. The anuran body plan, for example, is evolutionarily conserved and shows clear specializations to jumping performance back at least to the early Jurassic. However, there are instances of more recent adaptation to habit diversity in the post‐cranial skeleton, including relative limb length. The present study tested adaptive models of morphological evolution in anurans associated with the diversity of microhabitat use (semi‐aquatic arboreal, fossorial, torrent, and terrestrial) in species of anuran amphibians from Brazil and Australia. We use phylogenetic comparative methods to determine which evolutionary models, including Brownian motion (BM) and Ornstein‐Uhlenbeck (OU) are consistent with morphological variation observed across anuran species. Furthermore, this study investigated the relationship of maximum distance jumped as a function of components of morphological variables and microhabitat use. We found there are multiple optima of limb lengths associated to different microhabitats with a trend of increasing hindlimbs in torrent, arboreal, semi‐aquatic whereas fossorial and terrestrial species evolve toward optima with shorter hindlimbs. Moreover, arboreal, semi‐aquatic and torrent anurans have higher jumping performance and longer hindlimbs, when compared to terrestrial and fossorial species. We corroborate the hypothesis that evolutionary modifications of overall limb morphology have been important in the diversification of locomotor performance along the anuran phylogeny. Such evolutionary changes converged in different phylogenetic groups adapted to similar microhabitat use in two different zoogeographical regions.     

Behavioral responses to immune-system activation in an anuran (the cane toad, Bufo marinus): field and laboratory studies

The challenges posed by parasites and pathogens evoke behavioral as well as physiological responses. Such behavioral responses are poorly understood for most ectothermic species, including anuran amphibians. We quantified effects of simulated infection (via injection of bacterial lipopolysaccharide [LPS]) on feeding, activity, and thermoregulation of cane toads Bufo marinus within their invasive range in tropical Australia. LPS injection reduced feeding rates in laboratory trials. For toads in outdoor enclosures, LPS injection reduced activity and shifted body temperature profiles. Although previous research has attributed such thermal shifts to behavioral fever (elevated body temperatures may help fight infection), our laboratory studies suggest instead that LPS-injected toads stopped moving. In a thermal gradient, LPS-injected toads thus stayed close to whichever end of the gradient (hot or cold) they were first introduced; the introduction site (rather than behavioral thermoregulation) thus determined body temperature regimes. Shifts in thermal profiles of LPS-injected toads in outdoor enclosures also were a secondary consequence of inactivity. Thus, the primary behavioral effects of an immune response in cane toads are reduced rates of activity and feeding. Thermoregulatory modifications also occur but only as a secondary consequence of inactivity.     

Ecological constraints on the evolutionary association between field and preferred temperatures in Tropidurinae lizards

Ectothermic body temperatures affect organismal performances and presumably fitness, and are strongly influenced by the thermal environment. Therefore, the processes of colonization of novel thermal habitats by lizards might involve changes in thermal preferences, performance curves (reaction norms) and field activity temperatures. According to theory based on optimality analysis, diverse aspects of the thermal biology of vertebrate ectotherms should co-evolve as to maximize performance at the temperature range more often experienced by animals in the field. One corollary of this premise is that derived lizard clades that experienced a significant shift in thermal ecology, in comparison with the ancestral condition, should prefer and select temperatures in a thermal gradient similar to those experienced in nature. Here we report an analysis of the premise stated before. Specifically, we verify whether or not Tropidurinae species from three major Brazilian habitats (the Rainforests, the semi-arid Caatingas and the Cerrados, a Savannah-like biome) differ in thermal ecology and thermoregulatory behavior. The Caatinga is believed to be the ancestral habitat of this sub-family, and differences are expected because species from semi-arid habitats usually exhibit high body temperatures for lizards, whereas forest specialists might be thermoconformers and active at low temperatures. We also compared selected temperatures in the laboratory by species from the two open habitats (Caatingas and Cerrados). Data were analyzed using both conventional and phylogenetic analysis tools. Although species from Caatingas exhibited higher activity temperatures in nature than those from Cerrados, mean selected temperatures were similar between ecological groups. Phylogenetic analyses confirmed these findings and evidenced large␣evolutionary divergence in field activity temperatures between sister species from different␣open habitats without coupled divergence in selected temperatures. Therefore, thermoregulatory behavior and ecological parameters did not evolve similarly during the colonization of contrasting open habitats by Tropidurinae.     

Variation in thermal sensitivity of performance among colour morphs of a pygmy grasshopper

Populations of pygmy grasshoppers, Tetrix subulata, display genetically coded discrete variation in colour pattern and there are differences among morphs in the capacity to achieve body heating. To determine whether colour morphs differ in thermal physiology, I assessed reaction distance and jumping performance of individuals belonging to different morphs at two different temperatures. Individuals allowed a potential predator to approach less closely and jumped longer distances at high than at low temperature. My analyses also uncovered variation among morphs in average reaction distance and jumping capacity, as well as in thermal sensitivity of these two traits. Matrix correlation analysis further revealed that pair-wise differences between morphs in thermal sensitivity of jumping performance (but not reaction distance) could be accurately predicted by differences in body temperatures preferred in a laboratory thermal gradient. These results support the view that morphology, behaviour and thermal physiology of ectotherms may evolve in concert. The relationship between reaction distance and jumping performance varied among colour morphs at high temperature, and the common within-morph relationship between these two traits deviated from the corresponding among-morph relationship. This suggests that the variation among morphs has partially arisen through active divergence, with selection having influenced both traits and modifications having occurred to different degrees in different morphs. My data further suggest that pale colour morphs, with a limited capacity to attain high body temperatures, may not necessarily be at a selective disadvantage, because their physiology may be adapted to lower body temperatures.     

Abiotic noise,call frequency and stream-breeding anuran assemblages

Environmental noise can be an important selective force modulating signal evolution in species with acoustic communication. Many anuran species breed alongside streams; hence, the sound produced by the flowing water is an important source of noise for acoustic communication. Since calling is physiologically very expensive in anurans, and communication is essential for reproduction, we expected adaptations that reduce environmental masking effects and allow acoustic communication in streamside breeders. This basic assumption of the acoustic adaptation hypothesis has not been yet evaluated at a large phylogenetic scale. We combined ahistorical and phylogenetic methods to test whether anuran species that breed alongside streams call at higher frequencies than species that breed away from streams. We compiled primary and secondary data on body size, breeding habitat, and the dominant frequency of the advertisement call for 110 species; 40 of them breed alongside streams and 70 away from streams. Call frequency was slightly higher and body size was significantly smaller in streamside breeding species. After controlling for the effects of body size and phylogenetic signal, only differences in body size persisted between species breeding at both kinds of habitats. Our data suggest that habitat filtering rather than acoustic adaptation explains the high call frequency of stream breeders. Species with large body size, pleiotropically constrained to utter low-frequency calls, would have succeeded less often in establishing viable populations alongside streams, due to the masking effect of low-frequency noise. Thus, small species calling at relatively high frequencies would be more common there. Although our data do not preclude adaptations to noisy habitats in some anuran species, they do not provide support for the acoustic adaptation hypothesis at a wider phylogenetic scale.     

Trading heat and hops for water: Dehydration effects on locomotor performance,thermal limits,and thermoregulatory behavior of a terrestrial toad

Due to their highly permeable skin and ectothermy, terrestrial amphibians are challenged by compromises between water balance and body temperature regulation. The way in which such compromises are accommodated, under a range of temperatures and dehydration levels, impacts importantly the behavior and ecology of amphibians. Thus, using the terrestrial toad Rhinella schneideri as a model organism, the goals of this study were twofold. First, we determined how the thermal sensitivity of a centrally relevant trait—locomotion—was affected by dehydration. Secondly, we examined the effects of the same levels of dehydration on thermal preference and thermal tolerance. As dehydration becomes more severe, the optimal temperature for locomotor performance was lowered and performance breadth narrower. Similarly, dehydration was accompanied by a decrease in the thermal tolerance range. Such a decrease was caused by both an increase in the critical minimal temperature and a decrease in the thermal maximal temperature, with the latter changing more markedly. In general, our results show that the negative effects of dehydration on behavioral performance and thermal tolerance are, at least partially, counteracted by concurrent adjustments in thermal preference. We discuss some of the potential implications of this observation for the conservation of anuran amphibians.     

Seasonal metabolic changes in a year-round reproductively active subtropical tree-frog (Hypsiboas prasinus)

Although seasonal metabolic variation in ectothermic tetrapods has been investigated primarily in the context of species showing some level of metabolic depression during winter, but several species of anurans maintain their activity patterns throughout the year in tropical and subtropical areas. The tree-frog Hypsiboas prasinus occurs in the subtropical Atlantic Forest and remains reproductively active during winter, at temperatures below 10 degrees C. We compared males calling in summer and winter, and found that males of H. prasinus exhibit seasonal adjustments in metabolic and morphometric variables. Individuals calling during winter were larger and showed higher resting metabolic rates than those calling during summer. Calling rates were not affected by season. Winter animals showed lower liver and heart activity level of citrate synthase (CS), partially compensated by larger liver mass. Winter individuals also showed higher activity of pyruvate kinase (PK) and lower activity of CS in trunk muscles, and higher activity of CS in leg muscles. Winter metabolic adjustments seem to be achieved by both compensatory mechanisms to the lower environmental temperature and a seasonally oriented aerobic depression of several organs. The impact of seasonal metabolic changes on calling performance and the capacity of subtropical anurans for metabolic thermal acclimatization are also discussed.     

A metabolic view of amphibian local community structure: the role of activation energy

In the context of the metabolic theory of ecology (MTE), the activation energy (E) reflects the temperature dependence of metabolism and organism performance in different activities, such as calling behavior. In this contribution we test the role of temperature in affecting local amphibian community structure, particularly the number of species engaged in calling behavior across a temperature gradient. Toward this aim, we compiled phenological calling activity for 52 Neotropical anuran communities. For each community we estimated the activation energy of calling behavior (E), finding values significantly higher than previous reports. A wide range of methodological issues with the potential to produce overestimated E‐values were shown to have no significant effect on reported E‐values, supporting a biological interpretation of their high values and of geographic trends. Further, a path analysis related variation in E among communities with communities’ phylogenetic structure, local environmental conditions, richness, and seasonality. The decrease of activation energy at higher latitudes and less productive environments suggests that amphibians’ activity could become more dependent of internal individuals’ resources once external sources are reduced. The increase in phylogenetic attraction with latitude points to a rise in the role of niche conservatism and community filtering operating over conserved traits. Finally, flexibility in activation energy related to amphibians’ calling could be an important and poorly recognized determinant of their thermal dependence. The temporal structuring of amphians’ communities was related here with the interplay between ecological and evolutionary processes operating at different scales. Our results support the view of activation energy as an important parameter of biodiversity organization, which unravels the effects of ecological and evolutionary processes on biodiversity structure and function.     

Changes in breeding phenology of eastern Ontario frogs over four decades

Global climate change has been implicated in phenological shifts for a variety of taxa. Amphibian species in particular are sensitive to changes in their environment due to their biphasic life history and restricted reproductive requirements. Previous research has shown that not all temperate amphibian species respond similarly to the same suite of climatic or environmental cues, nor are individual species necessarily uniform in their responses across their range. We examined both the timing of spring emergence and calling phenology of eight anuran species in southeastern Ontario, Canada, using an approximately 40‐year dataset of historical records of amphibian activity. Rana pipiens was the only species out of eight considered to emerge significantly earlier, by an estimated 22 days over four decades. Both R. pipiens and Bufo americanus have advanced initiation of calling over a four‐decade span significantly earlier by an estimated 37.2 and 19.2 days, respectively. Rana sylvatica showed a trend toward earlier emergence by 19 days, whereas we did not detect changes in emergence phenology for the remaining five species. This significant shift in breeding behavior for two species correlates to significant regional increases in spring temperatures of an estimated 2.7–2.8°C overall over four decades. Our study suggests that local temperature increases have affected the timing of emergence and the onset of calling activity in some Ontario anuran species. Global decline or range shifts ultimately may be related to changes in reproductive behavior and timing mediated by shifting climate.     

A metabolic syndrome in terrestrial ectotherms with different elevational and distribution patterns

The metabolic performance of ectotherms is expected to be driven by the environment in which they live. Ecologically similar species with contrasting elevation distributions occurring in sympatry at mid‐elevations, provide good models for studying how physiological responses to temperature vary as a function of adaptation to different elevations. Under sympatry, at middle elevations, where divergent species ranges overlap, sympatric populations are expected to have similar thermal responses, suggesting similar local acclimation or adaptation, while observed differences would suggest adaptation to each species’ core range. We analysed the metabolic traits of sympatric species pairs from three ectotherm groups: reptiles (Reptilia: Lacertidae), amphibians (Amphibia: Salamandridae) and beetles (Coleoptera: Carabidae), living at different elevations, in order to test how adaptation to different elevations affects metabolic responses to temperature. We experimentally tested the thermal response of respiration rate (RR) and estimated potential metabolic activity (PMA) at three temperature regimes surrounding the groups’ optimal activity body temperatures. RR was relatively similar among groups and showed a positive response to increasing temperature, which was more pronounced in the high‐elevation species of reptiles and beetles. Relative to RR, PMA displayed a stronger and more consistent positive response to increased temperature in all three groups. For all three groups, the average biochemical capacity for metabolism (PMA) was higher in the range‐restricted, high‐elevation species, and this difference increased at higher temperatures in a consistent manner. These results, indicating consistent pattern in three independently evolved animal groups, suggest a ubiquitous adaptive syndrome and represent a novel understanding of the mechanisms shaping spatial biodiversity patterns. Our results also highlight the importance of geographic patterns for the mechanistic understanding of adaptations in physiological traits, including species’ potential to respond/adapt to global climate changes.     

Lifestyle-Based Approaches Provide Insights into Body Size Variation Across Environmental Gradients in Anurans

Body size of organisms as a fitness-related phenotype has evolved in response to local conditions, often through the size-dependent thermoregulatory mechanisms. The direction and degree of this response should depend on animals’ lifestyle in terms of the preference for terrestrial or aquatic conditions, especially so for adult anurans that differ in lifestyle among species but all must maintain certain body temperatures for metabolism. It may be expected that anuran species frequently exposed to terrestrial environments characterized by fluctuant thermal conditions are more plastic in body size along thermal gradients than those highly relaying on aquatic environments where thermal conditions are relatively stable. We test this prediction using both interspecific and intraspecific data. With anurans in China as the model organisms, we show that across terrestrial species but not aquatic species, body size decreases with increasing ambient temperature. From the published literature worldwide, we summarized that more terrestrial versus fewer aquatic species follow the predicted ecogeographical size patterns. In addition, both interspecific and intraspecific data reveal that arboreal anurans do not exhibit the size cline, probably because relatively warm climates experienced by these species impose weak selective pressures on heat conservation or adaptation to tree-climbing constrains the variation in body size. Our finding highlights the importance of taking lifestyle into account when assessing macroevolutionary trends in body size for anurans in particular and ectothermic taxa in general.     

Thermal parameters changes in males of Rhinella arenarum (Anura:Bufonidae) related to reproductive periods

The regulation of body temperature in ectotherms has a major impact in their physiological and behavioral processes. Observing changes in thermal parameters related with reproduction allows us to better understand how Rhinella arenarum optimizes a thermal resource. The aim of this study was to compare the thermal parameters of this species between breeding and non-breeding periods. In the field, we recorded the body temperature from captured animals, the air temperature, and the temperature of the substrate. In the laboratory, we measured the temperature R. arenarum selected on a thermal gradient and the critical extreme temperatures. The results of our study show variations in thermal parameters between the two situations studied. This species makes efficient use of thermal resources during the breeding period by basking to significantly increase body temperature. Because calling is energetically costly for males, this behavior results in increased efficiency to callers during the breeding period.     

Herpetological diversity along Andean elevational gradients: links with physiological ecology and evolutionary physiology

A well-defined macroecological pattern is the decline in biodiversity with altitude. However, this decline is taxa-specific. For example, amphibians are more diverse than squamates at extreme elevations in the tropical Andes, but this pattern is reversed at extreme elevations in the southern latitudes. Several ecophysiological and evolutionary factors may be related to this difference. At high-elevations in southern latitudes temperature differs dramatically among seasons and dry soils dominate, characteristics that appear to favor lizard physiological ecology. Tropical high altitudes, in contrast, are humid and offer abundant and diverse water resources. These characteristics allow for a richer anuran community but might complicate lizard egg development through temperature and oxygen constrains. Differences in strategies of thermal adaptation might also modulate diversity patterns. The thermal physiology of anurans is extremely labile so that behavioral and physiological performance is maintained despite an altitudinal decrease in field body temperature. Lizards, in contrast, exhibit a conservative thermal physiology and rely on behavioral thermoregulation to face cold and variable temperatures. Both, lizard behavioral strategies and anuran physiological adjustments seem equally efficient in allowing ecological success and diversification for both groups in the tropics up to approximately 3000 m. At higher elevations physiological thermal adaptation is required, and lizards are ecologically constrained, perhaps at various ontogenetic stages. Patterns of biodiversity along environmental clines can be better understood through a physiological approach, and can help to refine and propose hypotheses in evolutionary physiology.     

The evolution of jumping performance in anurans: morphological correlates and ecological implications

We investigated the evolution of anuran locomotor performance and its morphological correlates as a function of habitat use and lifestyles. We reanalysed a subset of the data reported by Zug (Smithson. Contrib. Zool. 1978; 276: 1–31) employing phylogenetically explicit statistical methods (n = 56 species), and assembled morphological data on the ratio between hind-limb length and snout-vent length (SVL) from the literature and museum specimens for a large subgroup of the species from the original paper (n = 43 species). Analyses using independent contrasts revealed that classifying anurans into terrestrial, semi-aquatic, and arboreal categories cannot distinguish between the effects of phylogeny and ecological diversification in anuran locomotor performance. However, a more refined classification subdividing terrestrial species into 'fossorials' and 'non-fossorials', and arboreal species into 'open canopy', 'low canopy' and 'high canopy', suggests that part of the variation in locomotor performance and in hind-limb morphology can be attributed to ecological diversification. In particular, fossorial species had significantly lower jumping performances and shorter hind limbs than other species after controlling for SVL, illustrating how the trade-off between burrowing efficiency and jumping performance has resulted in morphological specialization in this group.     

Non-climatic thermal adaptation: implications for species' responses to climate warming

There is considerable interest in understanding how ectothermic animals may physiologically and behaviourally buffer the effects of climate warming. Much less consideration is being given to how organisms might adapt to non-climatic heat sources in ways that could confound predictions for responses of species and communities to climate warming. Although adaptation to non-climatic heat sources (solar and geothermal) seems likely in some marine species, climate warming predictions for marine ectotherms are largely based on adaptation to climatically relevant heat sources (air or surface sea water temperature). Here, we show that non-climatic solar heating underlies thermal resistance adaptation in a rocky–eulittoral-fringe snail. Comparisons of the maximum temperatures of the air, the snail''s body and the rock substratum with solar irradiance and physiological performance show that the highest body temperature is primarily controlled by solar heating and re-radiation, and that the snail''s upper lethal temperature exceeds the highest climatically relevant regional air temperature by approximately 22°C. Non-climatic thermal adaptation probably features widely among marine and terrestrial ectotherms and because it could enable species to tolerate climatic rises in air temperature, it deserves more consideration in general and for inclusion into climate warming models.