Fish mortality in the Amazonian drought of 2023: the role of experimental biology in our response to climate change.

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

Thermal Tolerance and Preferred Temperature in the Critical Endangered Montseny Brook Newt (Calotriton arnoldi).

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

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

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

Insect thermal limits in warm and perturbed habitats: Dragonflies and damselflies as study cases.

Disturbance (e.g. loss of plant cover) increases ambient temperature which can be lethal for ectotherm insects especially in hot places. We compared the thorax temperatures of 26 odonate species as a function of body size, habitat quality ("conserved" and cooler vs "perturbed" and warmer) and suborder (Anisoptera vs Zygoptera), as well as critical thermal maximum (CTmax) and as a function of habitat quality in Argia pulla (Zygoptera) and Orthemis ferruginea (Anisoptera). We expected thorax temperatures to differ between suborders based on their differences in body size and habitat quality status, and that populations in perturbed sites would have higher critical thermal maxima compared to those in conserved sites. This study was done in a tropical region with high ambient temperatures. Anisopterans had a higher body temperature than zygopterans, with no difference between habitats. Thoracic and air temperature were positively related, yet body temperatures were higher than the ambient temperature. A. pulla had higher CTmax in the perturbed sites, while O. ferruginea showed the opposite trend. Microenvironmental changes increase the ambient temperature, perhaps filtering insect species. The apparent resilience of odonates to disturbance should be examined more closely (using more species), especially in small species like the zygopterans which appear to be more strongly affected by ambient temperature.

Leaf-cutting ants' critical and voluntary thermal limits show complex responses to size, heating rates, hydration level, and humidity.

Thermal variation has complex effects on organisms and they respond to these effects through combined behavioral and physiological mechanisms. However, it is less clear how these traits combine in response to changes in body condition (e.g., size, hydration) and environmental factors that surround the heating process (e.g., relative humidity, start temperatures, heating rates). We tested whether these body conditions and environmental factors influence sequentially measured Voluntary Thermal Maxima (VTmax) and Critical Thermal Maxima, (CTmax) in leaf-cutting ants (Atta sexdens rubropilosa, Forel, 1908). VTmax and CTmax reacted differently to changes in body size and relative humidity, but exhibited similar responses to hydration level, start temperature, and heating rate. Strikingly, the VTmax of average-sized workers was closer to their CTmax than the VTmax of their smaller and bigger sisters, suggesting foragers maintain normal behavior at higher temperatures than sister ants that usually perform tasks within the colony. Previous experiments based on hot plate designs might overestimate ants' CTmax. VTmax and CTmax may respond concomitantly or not to temperature rises, depending on body condition and environmental factors.

Thermal Acclimation to the Highest Natural Ambient Temperature Compromises Physiological Performance in Tadpoles of a Stream-Breeding Savanna Tree Frog.

Amphibians may be more vulnerable to climate-driven habitat modification because of their complex life cycle dependence on land and water. Considering the current rate of global warming, it is critical to identify the vulnerability of a species by assessing its potential to acclimate to warming temperatures. In many species, thermal acclimation provides a reversible physiological adjustment in response to temperature changes, conferring resilience in a changing climate. Here, we investigate the effects of temperature acclimation on the physiological performance of tadpoles of a stream-breeding savanna tree frog (Bokermannohyla ibitiguara) in relation to the thermal conditions naturally experienced in their microhabitat (range: 18.8-24.6°C). We quantified performance measures such as routine and maximum metabolic rate at different test (15, 20, 25, 30, and 34°C) and acclimation temperatures (18 and 25°C). We also measured heart rate before and after autonomic blockade with atropine and sotalol at the respective acclimation temperatures. Further, we determined the critical thermal maximum and warming tolerance (critical thermal maximum minus maximum microhabitat temperature), which were not affected by acclimation. Mass-specific routine and mass-specific maximum metabolic rate, as well as heart rate, increased with increasing test temperatures; however, acclimation elevated mass-specific routine metabolic rate while not affecting mass-specific maximum metabolic rate. Heart rate before and after the pharmacological blockade was also unaffected by acclimation. Aerobic scope in animals acclimated to 25°C was substantially reduced, suggesting that physiological performance at the highest temperatures experienced in their natural habitat is compromised. In conclusion, the data suggest that the tadpoles of B. ibitiguara, living in a thermally stable environment, have a limited capacity to physiologically adjust to the highest temperatures found in their micro-habitat, making the species more vulnerable to future climate change.

Use of complex physiological traits as ecotoxicological biomarkers in tropical freshwater fishes.

We review the use of complex physiological traits, of tolerance and performance, as biomarkers of the toxicological effects of contaminants in subtropical and tropical freshwater fishes. Such traits are growing in relevance due to climate change, as exposure to contaminants may influence the capacity of fishes to tolerate and perform in an increasingly stressful environment. We review the evidence that the critical oxygen level, a measure of hypoxia tolerance, provides a valuable biomarker of impacts of diverse classes of contaminants. When coupled with measures of cardiorespiratory variables, it can provide insight into mechanisms of toxicity. The critical thermal maximum, a simple measure of tolerance of acute warming, also provides a valuable biomarker despite a lack of understanding of its mechanistic basis. Its relative ease of application renders it useful in the rapid evaluation of multiple species, and in understanding how the severity of contaminant impacts depends upon prevailing environmental temperature. The critical swimming speed is a measure of exercise performance that is widely used as a biomarker in temperate species but very few studies have been performed on subtropical or tropical fishes. Overall, the review serves to highlight a critical lack of knowledge for subtropical and tropical freshwater fishes. There is a real need to expand the knowledge base and to use physiological biomarkers in support of decision making to manage tropical freshwater fish populations and their habitats, which sustain rich biodiversity but are under relentless anthropogenic pressure.

Combined effect of pCO2 and temperature levels on the thermal niche in the early benthic ontogeny of a keystone species.

We evaluated the effects of projected, near future ocean acidification (OA) and extreme events of temperature (warming or cooling) on the thermal tolerance of Concholepas concholepas, a coastal benthic keystone species. Three separate trials of an experiment were conducted by exposing juvenile C. concholepas for 1 month to one of two contrasting pCO2 levels (~500 and ~1200 µatm). In addition, each pCO2 level was combined with one of four temperature treatments. The control was 15 °C, whilst the other temperatures were 10 °C (Trial 1), 20 °C (Trial 2) and 25 °C (Trial 3). At the end of each trial, we assessed Critical Thermal maximum (CTmax) and minimum (CTmin) via self-righting success, calculated partial thermal tolerance polygons, measured somatic growth, determined transcription of Heat Shock Proteins 70 (HSP70) and measured oxygen consumption rates. Regardless of pCO2 level, HSP70 transcript levels were significantly higher in juveniles after exposure to extreme temperatures (10 °C and 25 °C) indicating physiological stress. Oxygen consumption rates increased with increasing temperature from 10 °C to 20 °C though showed a decrease at 25 °C. This rate was not affected by pCO2 or the interaction between temperature and pCO2. Juveniles exposed to present-day and near future pCO2 levels at 20 °C showed similar thermal tolerance polygonal areas; whilst changes in both CTmin and CTmax at 25 °C and 10 °C caused narrower and broader areas, respectively. Temperature affected growth, oxygen consumption and HSP70 transcription in small juvenile C. concholepas. Exposure to elevated pCO2 did not affect thermal tolerance, growth or oxygen consumption at temperatures within the thermal range normally experienced by this species in northern Chile (15-20 °C). At elevated pCO2 conditions, however, exposure to warmer (25 °C) or colder (10 °C) temperatures reduced or increased the thermal area, respectively. This study demonstrates the importance of examining the thermal-tolerance edges to better understand how OA and temperature will combine to physiologically challenge inter-tidal organisms.

Ocean warming and acidification pose synergistic limits to the thermal niche of an economically important echinoderm.

To make robust projectios of the impacts of climate change, it is critical to understand how abiotic factors may interact to constrain the distribution and productivity of marine flora and fauna. We evaluated the effects of projected end of the century ocean acidification (OA) and warming (OW) on the thermal tolerance of an important living marine resource, the sea urchin Loxechinus albus, a benthic shallow water coastal herbivore inhabiting part of the Pacific coast of South America. After exposing young juveniles for a 1-month period to contrasting pCO2 (~500 and 1400 µatm) and temperature (~15 °C and 20 °C) levels, critical thermal maximum (CTmax) and minimum (CTmin) as well as thermal tolerance polygons were assessed based on self-righting success as an end point. Transcription of heat shock protein 70 (HSP70), a chaperone protecting cellular proteins from environmental stress, was also measured. Exposure to elevated pCO2 significantly reduced thermal tolerance by increasing CTmin at both experimental temperatures and decreasing CTmax at 20 °C. There was also a strong synergistic effect of OA × OW on HSP70 transcription levels which were 75 times higher than in control conditions. If this species is unable to adapt to elevated pCO2 in the future, the reduction in thermal tolerance and HSP response suggests that near-future warming and OA will disrupt their performance and reduce their distribution with ecological and economic consequences. Given the wider latitudinal range (6 to 56°S) and environmental tolerance of L. albus compared to other members of this region's benthic invertebrate community, OW and OA may cause substantial changes to the coastal fauna along this geographical range.

Waterborne copper is more toxic to the killifish Poecilia vivipara in elevated temperatures: Linking oxidative stress in the liver with reduced organismal thermal performance.

In this study, we measured the interactive effect of temperature (22 °C and 28 °C) and waterborne copper (Cu) contamination (9 µg/L and 20 µg/L) on the killifish Poecilia vivipara. Endpoints analyzed included parameters involved in Cu-accumulation, antioxidant capacity (antioxidant capacity against peroxyl radicals [ACAP] and total antioxidant capacity [TAC]), oxidative damage (lipid peroxidation [LPO]) and upper thermal tolerance (critical thermal maximum [CTMax]). Results show that Cu hepatic accumulation was elevated in 28 °C in comparison to 22 °C in both exposure groups. For gills, this was true only in 20 µg/L. Moreover, hepatic and brachial accumulation were concentration-dependent in both acclimation temperatures. Additionally, Hepatic ACAP and TAC were elevated in animals acclimated to 28 °C and only the animals kept at this temperature had reduced ACAP and TAC levels facing metal exposure (9 and 20 µg/L). Similarly, the combination of elevated temperature and Cu exposure raised hepatic LPO levels. Finally, animals acclimated to 28 °C had higher CTMax levels in comparison to fish acclimated to 22 °C both in control and exposed animals, however, CTMax of contaminated fish were only reduced in comparison to control in animals kept at 28 °C. Concluding, we show that the physiological mechanism besides the potentiating effect of elevated temperature in Cu toxicity is related to higher hepatic and branchial metal accumulation and elevated oxidative stress in the liver, outlined by reduced antioxidant capacity and elevated oxidative damage. We also show that these outcomes lead to compromised organismal performance, characterized by reduced CTMax. Finally, it is concluded that Cu exposure in warmer periods of the year or within global warming predictions may be more hazardous to fish populations.

Effect of salinity on locomotor performance and thermal extremes of metamorphic Andean Toads (Rhinella spinulosa) from Monte Desert, Argentina.

Rhinella spinulosa is distributed from Peru to Argentina (from 1200 to 5000 m elevation), inhabiting arid mountain valleys of the Andes, characterized by salty soils. The variations in soil salinity, caused by high evapotranspiration of water, can create an osmotic constraint and high thermal oscillations for metamorphsed Andean toad (R. spinulosa), affecting their thermoregulation and extreme thermal tolerances. We investigated the changes in thermal tolerance parameters (critical thermal maximum and crystallization temperature) of a population of metamorphosed R. spinulosa from the Monte Desert of San Juan, Argentina, under different substrate salinity conditions. Our results suggest that the locomotor performance of metamorphs of R. spinulosa is affected by increasing salinity concentrations in the environment where they develop. On the other hand, the thermal extremes of metamorphs of R. spinulosa also showed changes associated with different salinity conditions. According to other studies on different organisms, the increase of the osmolarity of the internal medium may increase the thermal tolerance of this species. More studies are needed to understand the thermo-osmolar adjustments of the metamorphs of toads to environmental variability.

Water temperature affects aggressive interactions in a Neotropical cichlid fish

Changes in water temperature may affect the aggressive behavior of aquatic organisms, such as fish, either by changing some physiological mechanisms or by increasing the probability of encounters between individuals as a result of variation in their swimming activity. In our study, we evaluated the influence of increasing and decreasing temperature on the aggressive behavior of the Neotropical cichlid fish Cichlasoma paranaense. Firstly, we tested the critical thermal maximum (CTMax) tolerated by this species. Then, we tested the effect of decreasing or increasing the water temperature in 6o C (starting at 27° C) on the aggressive interactions of fish under isolation or housed in groups. We found a CTMax value of 39° C for C. paranaense. We also observe that a 6° C decrease in water temperature lowers swimming activity and aggressive interactions in both isolated and group-housed fish, as expected. On the other hand, the increase in temperature had no effect on the fish's aggressive behavior, neither for isolated nor for grouped fish. We concluded that C. paranaense shows high tolerance to elevated temperatures and, in turn, it does not affect aggressive behavior. Nevertheless, we cannot dismiss possible effects of elevated temperatures on aggressive interactions over longer periods.(AU)

Mudanças na temperatura da água podem afetar o comportamento agressivo de organismos aquáticos, como peixes, seja alterando alguns mecanismos fisiológicos ou aumentando a probabilidade de encontros entre os indivíduos como um resultado da variação na sua atividade natatória. Em nosso estudo, nós avaliamos a influência do aumento e da diminuição da temperatura sobre o comportamento agressivo do peixe ciclídeo neotropical Cichlasoma paranaense. Primeiramente, nós testamos a temperatura crítica máxima (CTMax) tolerada por essa espécie. Depois disso, nós testamos o efeito de reduzir ou aumentar a temperatura da água em 6° C (a partir de 27° C) sobre as interações agressivas de peixes sob isolamento ou alojados em grupos. Nós encontramos um valor de CTMax de 39° C para C. paranaense. Nós também observamos que uma redução de 6° C na temperatura da água diminui a atividade natatória e as interações agressivas tanto em peixes isolados quanto agrupados, como esperado. Por outro lado, o aumento na temperatura não teve efeito no comportamento agressivo dos peixes nem isolados, nem agrupados. Nós concluímos que C. paranaense apresenta alta tolerância a temperaturas elevadas, e que isso, por sua vez, não afeta o comportamento agressivo. No entanto, nós não podemos descartar possíveis efeitos de temperaturas elevadas sobre interações agressivas durante longos períodos.(AU)

Water temperature affects aggressive interactions in a Neotropical cichlid fish

Changes in water temperature may affect the aggressive behavior of aquatic organisms, such as fish, either by changing some physiological mechanisms or by increasing the probability of encounters between individuals as a result of variation in their swimming activity. In our study, we evaluated the influence of increasing and decreasing temperature on the aggressive behavior of the Neotropical cichlid fish Cichlasoma paranaense. Firstly, we tested the critical thermal maximum (CTMax) tolerated by this species. Then, we tested the effect of decreasing or increasing the water temperature in 6o C (starting at 27° C) on the aggressive interactions of fish under isolation or housed in groups. We found a CTMax value of 39° C for C. paranaense. We also observe that a 6° C decrease in water temperature lowers swimming activity and aggressive interactions in both isolated and group-housed fish, as expected. On the other hand, the increase in temperature had no effect on the fish's aggressive behavior, neither for isolated nor for grouped fish. We concluded that C. paranaense shows high tolerance to elevated temperatures and, in turn, it does not affect aggressive behavior. Nevertheless, we cannot dismiss possible effects of elevated temperatures on aggressive interactions over longer periods.(AU)

Mudanças na temperatura da água podem afetar o comportamento agressivo de organismos aquáticos, como peixes, seja alterando alguns mecanismos fisiológicos ou aumentando a probabilidade de encontros entre os indivíduos como um resultado da variação na sua atividade natatória. Em nosso estudo, nós avaliamos a influência do aumento e da diminuição da temperatura sobre o comportamento agressivo do peixe ciclídeo neotropical Cichlasoma paranaense. Primeiramente, nós testamos a temperatura crítica máxima (CTMax) tolerada por essa espécie. Depois disso, nós testamos o efeito de reduzir ou aumentar a temperatura da água em 6° C (a partir de 27° C) sobre as interações agressivas de peixes sob isolamento ou alojados em grupos. Nós encontramos um valor de CTMax de 39° C para C. paranaense. Nós também observamos que uma redução de 6° C na temperatura da água diminui a atividade natatória e as interações agressivas tanto em peixes isolados quanto agrupados, como esperado. Por outro lado, o aumento na temperatura não teve efeito no comportamento agressivo dos peixes nem isolados, nem agrupados. Nós concluímos que C. paranaense apresenta alta tolerância a temperaturas elevadas, e que isso, por sua vez, não afeta o comportamento agressivo. No entanto, nós não podemos descartar possíveis efeitos de temperaturas elevadas sobre interações agressivas durante longos períodos.(AU)

Thermal ecology of the post-metamorphic Andean toad (Rhinella spinulosa) at elevation in the monte desert, Argentina.

Rhinella spinulosa is an anuran toad species distributed latitudinal and altitudinal (1200-5000m) from Peru to Argentina, inhabiting mountain valleys in the Andes. Considering the broad range of habitats where they live, it is important to understand the thermal physiological mechanisms, thermal tolerances and physiological adaptations for surviving in rigorous environments. We investigated the thermal parameters (field body temperature, selected body temperature, locomotor performance in field and laboratory conditions, and thermal extremes) during diurnal activity for a population of juvenile, post-metamorphosed toads (Rhinella spinulosa) from the Monte Desert of San Juan, Argentina. Post-metamorphic toads are active from approximately 1100-1900 (in contrast to nocturnal adult toads). Our findings show that these toads have a wide thermal tolerance range, ranging from a critical thermal maximum of 36.9°C to crystallization temperatures below 0°C. During their active period, toads always showed suboptimal thermal conditions for locomotion. Despite the suboptimal condition for the locomotion, diurnal activity is likely to confer thermal advantages, allowing them to search for food and increase digestion and growth rates. We also found that the toads are capable of super-cooling, which prevents mortality from freezing when the environmental temperatures drop below 0°C. The environmental temperatures are below zero at night, when toads are inactive and take refuge under rocks. In summary, this toad population demonstrates high thermal plasticity, as shown by a relatively high level of activity sustained over a wide range of ambient temperature (~35°C). These thermal adaptations allow this species of juvenile toads to inhabit a wide range of altitudes and latitudes.

Thermal physiology, disease, and amphibian declines on the eastern slopes of the Andes.

Rising temperatures, a widespread consequence of climate change, have been implicated in enigmatic amphibian declines from habitats with little apparent human impact. The pathogenic fungus Batrachochytrium dendrobatidis (Bd), now widespread in Neotropical mountains, may act in synergy with climate change causing collapse in thermally stressed hosts. We measured the thermal tolerance of frogs along a wide elevational gradient in the Tropical Andes, where frog populations have collapsed. We used the difference between critical thermal maximum and the temperature a frog experiences in nature as a measure of tolerance to high temperatures. Temperature tolerance increased as elevation increased, suggesting that frogs at higher elevations may be less sensitive to rising temperatures. We tested the alternative pathogen optimal growth hypothesis that prevalence of the pathogen should decrease as temperatures fall outside the optimal range of pathogen growth. Our infection-prevalence data supported the pathogen optimal growth hypothesis because we found that prevalence of Bd increased when host temperatures matched its optimal growth range. These findings suggest that rising temperatures may not be the driver of amphibian declines in the eastern slopes of the Andes. Zoonotic outbreaks of Bd are the most parsimonious hypothesis to explain the collapse of montane amphibian faunas; but our results also reveal that lowland tropical amphibians, despite being shielded from Bd by higher temperatures, are vulnerable to climate-warming stress.