Many fishes exploit the terrestrial environment. Some of these amphibious species use it as an ecological release, others for feeding, while others use land to escape aquatic stressors. Another hypothesis is that amphibious fishes leave water to thermoregulate, but there is little corroborating data. One of the most intriguing fish that emerges onto land is the mangrove rivulus (Kryptolebias marmoratus), which can survive on land for up to 75 days. The mangrove rivulus emerges to escape from warm water and initially benefits from enhanced evaporative cooling on land. However, emergence imposes an immediate hypoxaemia due to reduced gas exchange over the gills and an accumulation of CO2 that impairs haemoglobin-O2 binding. In other ectothermic species, hypoxia is known to decrease thermal preference. Before physiological acclimation restores respiratory function, it is therefore plausible that fish that have emerged onto land will seek cooler temperatures beyond what is achieved by simple evaporative cooling. To test this idea, two strains of laboratory-raised mangrove rivulus were placed in a thermal gradient that provided the option of land and water to determine how their preferred temperature was informed by choice of the surrounding environment. We hypothesized that selecting land would be associated with anapyrexia, or a purposeful selection of lower temperatures. In trials where thermal gradients were provided in aquatic, terrestrial or combined settings, mangrove rivulus selected cooler temperatures only when on land. These findings support the premise that terrestrial emergence serves an active thermoregulatory strategy.

Chagas disease, caused by the parasite Trypanosoma cruzi, is a major neglected tropical disease affecting 6-7 million people worldwide. Rhodnius prolixus, one of the most important vectors of Chagas disease in Latin America, is known to be highly sensitive to environmental temperature, which influences both its biology and parasite development. However, few studies have investigated how this vector behaviorally modulates the effects of temperature by shifting their thermopreference, particularly in response to infection. We investigated how T. cruzi infection of R. prolixus fourth-instar nymphs influences thermopreference along a temperature gradient, while examining differences across times of day and time since blood feeding. Additionally, parasite load and infection maintenance were compared between free-moving nymphs and nymphs kept at a constant 26°C. Infected nymphs exhibited a preference for temperatures approximately 1°C cooler than uninfected controls. This cold-seeking behavior emerged around 15 days post-infection and persisted until shortly after molting. Importantly, infected insects allowed to thermoregulate showed significantly lower intestinal parasite loads and a higher rate of infection clearance compared to those kept at a constant 26°C. A diurnal cycle in temperature preference was also observed, with higher preferred temperatures shortly after the beginning of the photophase, followed by a gradual decline over the day and night. These results suggest the existence of an infection-induced behavioral anapyrexia response in R. prolixus that limits T. cruzi development. This potential form of adaptive thermoregulation has important implications for the ecology of Chagas disease transmission and the development of behaviorally informed vector control strategies.

Hypoxia-induced anapyrexia is thought to be a regulated decrease in body core temperature (Tcore), but the underlying mechanism remains unclear. Recent evidence suggests that lactate, a glycolysis product, could modulate neuronal excitability through the G protein-coupled receptor 81 (GPR81). The present study aims to elucidate the role of central lactate and GPR81 in a rat model of hypoxia-induced anapyrexia. The findings revealed that hypoxia (11.1% O2, 2 h) led to an increase in lactate in cerebrospinal fluid (CSF) and a decrease in Tcore. Injection of dichloroacetate (DCA, 5 mg/kg, 1 μL), a lactate production inhibitor, to the third ventricle (3 V), alleviated the increase in CSF lactate and the decrease in Tcore under hypoxia. Immunofluorescence staining showed GPR81 was expressed in the preoptic area of hypothalamus (PO/AH), the physiological thermoregulation integration center. Under normoxia, injection of GPR81 agonist 3-chloro-5-hydroxybenzoic acid (CHBA, 0.05 mg/kg, 1 μL) to the 3 V, reduced Tcore significantly. In addition, hypoxia led to a dramatic increase in tail skin temperature and a decrease in interscapular brown adipose tissue skin temperature. The number of c-Fos[+] cells in the PO/AH increased after exposure to 11.1% O2 for 2 h, but administration of DCA to the 3 V blunted this response. Injection of CHBA to the 3 V also increased the number of c-Fos[+] cells in the PO/AH under normoxia. In light of these, our research has uncovered the pivotal role of central lactate-GPR81 signaling in anapyrexia, thereby providing novel insights into the mechanism of hypoxia-induced anapyrexia.

The climate crisis necessitates predicting how organisms respond to changing environments, but this requires understanding the mechanisms underlying thermal tolerance. The Hierarchical Mechanisms of Thermal Limitation (HMTL) hypothesis proposes that respiratory capacity and marginal stability of proteins and membranes interact hierarchically to determine thermal performance and limits. An untested prediction of the HMTL hypothesis is that behavioral anapyrexia (i.e., reduced body temperature in hypoxia) is exacerbated when metabolic demand is high. We tested this prediction by manipulating western fence lizards' (Sceloporus occidentalis) metabolic demand and oxygen environment, then measuring selected body temperatures. Lizards with elevated metabolic demand selected lower body temperatures at higher oxygen concentrations than resting lizards, but this occurred only at oxygen concentrations <12% O2 , suggesting thermal limits are unaffected by naturally-occurring oxygen variation. Given our results and the ubiquity of behavioral anapyrexia, the HMTL hypothesis may generally explain how oxygen and temperature interactively affect reptile performance.

The costs associated with immune and thermal responses may exceed the benefits to the host during severe inflammation. In this case, regulated hypothermia instead of fever can occur in rodents as a beneficial strategy to conserve energy for vital functions with consequent tissue protection and hypoxia prevention. We tested the hypothesis that this phenomenon is not exclusive to mammals, but extends to the other endothermic group, birds. A decrease in metabolic rate without any failure in mitochondrial respiration, nor oxygen delivery, is the main evidence supporting the regulated nature of endotoxin-induced hypothermia in chicks. Thermolytic mechanisms such as tachypnea and cutaneous vasodilatation can also be recruited to facilitate body temperature decrease under lipopolysaccharide treatment, especially in the cold. Our findings bring a new perspective for evolutionary medicine studies on energy trade-off in host defence because regulated hypothermia may be a phenomenon spread among vertebrates facing a severe immune challenge.

Behavioural flexibility allows ectotherms to exploit the environment to govern their metabolic physiology, including in response to environmental stress. Hydrogen sulfide (H2S) is a widespread environmental toxin that can lethally inhibit metabolism. However, H2S can also alter behaviour and physiology, including a hypothesized induction of hibernation-like states characterized by downward shifts of the innate thermal set point (anapyrexia). Support for this hypothesis has proved controversial because it is difficult to isolate active and passive components of thermoregulation, especially in animals with high resting metabolic heat production. Here, we directly test this hypothesis by leveraging the natural behavioural thermoregulatory drive of fish to move between environments of different temperatures in accordance with their current physiological state and thermal preference. We observed a decrease in adult zebrafish (Danio rerio) preferred body temperature with exposure to 0.02% H2S, which we interpret as a shift in the thermal set point. Individuals exhibited consistent differences in shuttling behaviour and preferred temperatures, which were reduced by a constant temperature magnitude during H2S exposure. Seeking lower temperatures alleviated H2S-induced metabolic stress, as measured by reduced rates of aquatic surface respiration. Our findings highlight the interactions between individual variation and sublethal impacts of environmental toxins on behaviour.

The anteroventral preoptic region (AVPO) of the hypothalamus is involved in both temperature and breathing regulation. This area densely express cannabinoid receptors type 1 (CB1) that modulate both excitatory and inhibitory synaptic transmission. However, it is still unknown if the endocannabinoid system located in the AVPO participates in breathing control and thermoregulation. Therefore, we tested the participation of CB1 in the AVPO in the modulation of ventilation and thermal control during normoxia and hypoxia. To this end, body temperature (Tb) of Wistar rats was monitored by datallogers and ventilation (VE) by whole body plethysmography before and after intra-AVPO microinjection of AM-251 (CB1 antagonist, 50 and 100 pmol) followed by 60 min of hypoxia exposure (7% O2). Intra-AVPO microinjection of the higher dose of AM-251 increased VE but did not change Tb under resting conditions. Exposure of rats to 7% of inspired oxygen evoked typical hypoxia-induced anapyrexia and hyperventilation after vehicle microinjection. The higher dose of the cannabinoid antagonist increased the hypoxia-induced hyperventilation, in the same magnitude as observed under normoxic condition, whereas the drop in Tb elicited by hypoxia was attenuated. Therefore, the present results demonstrate that the endocannabinoid system acting on CB1 receptors in the AVPO exerts a tonic inhibitory modulation on breathing but seem not be involved in thermoregulation during resting conditions. In addition, activation of CB1 receptors in the AVPO stimulate thermal response during hypoxia, reducing energetically expensive responses, such as the hypoxic hyperventilation.

Heat exchange processes between the body and the environment are introduced. The definition of the thermoneutral zone as the ambient temperature range within which body temperature (Tb) regulation is achieved only by nonevaporative processes is explained. Thermoreceptors, thermoregulatory effectors (both physiologic and behavioral), and neural pathways and Tb signals that connect receptors and effectors into a thermoregulation system are reviewed. A classification of thermoeffectors is proposed. A consensus concept is presented, according to which the thermoregulation system is organized as a dynamic federation of independent thermoeffector loops. While the activity of each effector is driven by a unique combination of deep (core) and superficial (shell) Tbs, the regulated variable of the system can be viewed as a spatially distributed Tb with a heavily represented core and a lightly represented shell. Core Tb is the main feedback; it is always negative. Shell Tbs (mostly of the hairy skin) represent the auxiliary feedback, which can be negative or positive, and which decreases the system's response time and load error. Signals from the glabrous (nonhairy) skin about the temperature of objects in the environment serve as feedforward signals for various behaviors. Physiologic effectors do not use feedforward signals. The system interacts with other homeostatic systems by "meshing" with their loops. Coordination between different thermoeffectors is achieved through the common controlled variable, Tb. The term balance point (not set point) is used for a regulated level of Tb. The term interthreshold zone is used for a Tb range in which no effectors are activated. Thermoregulatory states are classified, based on whether: Tb is increased (hyperthermia) or decreased (hypothermia); the interthreshold zone is narrow (homeothermic type of regulation) or wide (poikilothermic type); and the balance point is increased (fever) or decreased (anapyrexia). During fever, thermoregulation can be either homeothermic or poikilothermic; anapyrexia is always a poikilothermic state. The biologic significance of poikilothermic states is discussed. As an example of practical applications of the concept presented, thermopharmacology is reviewed. Thermopharmacology uses drugs to modulate specific temperature signals at the level of a thermoreceptor (transient receptor potential channel).

Temperature is a crucial factor determining biology and ecology of poikilothermic animals. It often constitutes an important barrier for invasive species originating from different climate zones but, on the other hand, may facilitate the invasion process of animals with wide thermal preferences and high resistance to extreme temperatures. In our experimental study, we investigated the thermal behaviour of two Ponto-Caspian amphipod crustaceans-Dikerogammarus villosus and Dikerogammarus haemobaphes. Both species are known to live under a wide range of thermal conditions which may promote their invasion. Moreover, both these amphipods are hosts for microsporidian parasites which co-evolved with them within the Ponto-Caspian region and spread in European waters. As the presence of a parasite may influence the thermal preferences of its host, we expected to observe behavioural changes in infected individuals of the studied amphipods leading to (1) behavioural fever (selecting a warmer habitat) or (2) anapyrexia (selecting a colder habitat). The experiment (N = 20) was carried out for 30 min in a 100 cm. 20 cm from boths sides were not avaliable for amphipods long thermal gradient (0-40 °C), using 30 randomly selected adult amphipod individuals of one species. At the end of each trial, we checked the position of amphipods along the gradient and determined their sex and infection status (uninfected or infected by one of microsporidium species). D. villosus was infected with Cucumispora dikerogammari whereas D. haemobaphes was a host for C. dikerogammari, Dictyocoela muelleri or D. berillonum. Thermal preferences of amphipods depended on their species and sex. Females of D. villosus preferred warmer microhabitats (often much above 30 °C) than conspecific males and females of D. haemobaphes, whereas no significant differences were found among males of both species and both sexes of D. haemobaphes. Moreover, infected males of D. villosus stayed in warmer water more often than uninfected males of this species, selecting temperatures higher than 30 °C, which may be explained either as a behavioural fever constituting a defence mechanism of a host against the infection, or as a parasite manipulation of the host behaviour increasing the parasite fitness. On the other hand, none of the parasite species affected the thermal preferences of D. haemobaphes, including also C. dikerogammari, changing the behaviour of D. villosus. Our research presents the complexity of the thermal behaviour of studied amphipods and the evidence that microsporidia may trigger a change in temperature preferendum of their host species and those observations may be the result of different host-parasite coevolution time which may vary for the two host species (Poulin, 2010).

Trace amine-associated receptors (TAARs) belong to the class A G-protein-coupled receptors (GPCR) and are evolutionary related to aminergic receptors. TAARs have been identified to mediate effects of trace amines. TAAR1 signaling is mainly mediated via activation of the Gs/adenylyl cyclase pathway. In addition to classical trace amines, TAAR1 can also be activated by the thyroid hormone derivative 3-iodothyronamine (3-T1AM). Pharmacological doses of 3-T1AM induced metabolic and anapyrexic effects, which might be centrally mediated in the hypothalamus in rodents. However, the observed anapyrexic effect of 3-T1AM persists in Taar1 knock-out mice which raises the question whether further GPCRs are potential targets for 3-T1AM and mediate the observed physiological effect. Anapyrexia has been observed to be related to action on aminergic receptors such as the serotonin receptor 1b (5-HT1b). This receptor primarily activates the Gi/o mediated pathway and PLC signaling through the Gβγ of Gi/o. Since the expression profiles of TAAR1 and 5-HT1b overlap, we questioned whether 3-T1AM may activate 5-HT1b. Finally, we also evaluated heteromerization between these two GPCRs and tested signaling under co-expressed conditions. In this study, we showed, that 3-T1AM can induce Gi/o signaling through 5-HT1b in a concentration of 10 μM. Strikingly, at 5-HT1b the ligand 3-T1AM only activates the Gi/o mediated reduction of cAMP accumulation, but not PLC activation. Co-stimulation of 5-HT1b by both ligands did not lead to additive or synergistic signaling effects. In addition, we confirmed the capacity for heteromerization between TAAR1 and 5-HT1b. Under co-expression of TAAR1 and HTR1b, 3-T1AM action is only mediated via TAAR1 and activation of 5-HT1b is abrogated. In conclusion, we found evidence for 5-HT1b as a new receptor target for 3-T1AM, albeit with a different signaling effect than the endogenous ligand. Altogether, this indicates a complex interrelation of signaling effects between the investigated GPCRs and respective ligands.

Most small rodents reduce energy demand in hypoxia via behavioural strategies. For example, animals may reduce their activity, and/or move to colder environments or alter huddling strategies to take advantage of anapyretical energy savings. Naked mole rats (NMRs) are among the most hypoxia tolerant mammals and are highly social; social interactions also have a significant impact on behaviour. Therefore, this species offers a fascinating model in which to study trade-offs between social interactions and energy conservation in hypoxia. We hypothesized that the need to conserve energy in hypoxia supersedes the impetus of sociality in this species and predicted that, in hypoxia, behaviour would not differ between individuals or groups of NMRs. To test this hypothesis, we placed awake, freely behaving NMRs, alone or in groups of 2 or 4, into a temperature-controlled apparatus and measured behavioural activity during 1 h each of normoxia (21% O2), acute hypoxia (7% O2), and normoxic recovery. We found that in normoxia, groups of 4 NMRs were significantly more active in all temperatures than were groups of 1-2 NMRs. When exposed to hypoxia, individual NMRs were ~50% less active and their speed was reduced relative to normoxic levels. Conversely, groups of 2 or 4 NMRs exhibited minor or insignificant decreases in time spent active and speed in hypoxia and huddling behaviour was not altered. Our findings suggest that social interactions influence behavioural strategies employed by NMRs in hypoxia.

Thermal tolerance is an important variable in predictive models about the effects of global climate change on species distributions, yet the physiological mechanisms responsible for reduced performance at high temperatures in air-breathing vertebrates are not clear. We conducted an experiment to examine how oxygen affects three variables exhibited by ectotherms as they heat-gaping threshold, panting threshold, and loss of righting response (the latter indicating the critical thermal maximum)-in two lizard species along an elevational (and therefore environmental oxygen partial pressure) gradient. Oxygen partial pressure did not impact these variables in either species. We also exposed lizards at each elevation to severely hypoxic gas to evaluate their responses to hypoxia. Severely low oxygen partial pressure treatments significantly reduced the gaping threshold, panting threshold, and critical thermal maximum. Further, under these extreme hypoxic conditions, these variables were strongly and positively related to partial pressure of oxygen. In an elevation where both species overlapped, the thermal tolerance of the high elevation species was less affected by hypoxia than that of the low elevation species, suggesting the high elevation species may be adapted to lower oxygen partial pressures. In the high elevation species, female lizards had higher thermal tolerance than males. Our data suggest that oxygen impacts the thermal tolerance of lizards, but only under severely hypoxic conditions, possibly as a result of hypoxia-induced anapyrexia.