Effects of thermophily-relevant temperature variation and sex on digestive performance in pythons.

Different physiological performances are often optimized at slightly varying temperatures, which can lead to ectotherms selecting higher body temperatures during certain physiological efforts (e.g., digestion, reproduction). Such thermophilic responses can lead to temperature-based tradeoffs between two physiological activities with differing optimal temperatures or between optimizing a physiological activity and water balance, as water loss is elevated at higher temperatures. For example, ectotherms will often select a higher body temperature after consuming a meal, but the extent to which body temperature is elevated after eating is affected by its hydric state. Despite this known hydration state-based suppression of thermophily associated with digestion, the impact of this reduced body temperature on digestion performance is unknown. Accordingly, we determined whether small, thermophily-relevant changes in body temperature impact digestive efficiency or passage time and whether sex influenced the extent of the effect. Eighteen (9 female and 9 male) Children's pythons (Antaresia childreni) each consumed a meal at three temperatures (29 °C, 30 °C, and 31 °C), and gut passage time and digestive efficiency were determined. We found that neither metric was affected by temperature over the range tested. However, digestive efficiency was significantly impacted by the interaction between sex and temperature with males having significantly lower digestive efficiency than females at 31 °C, but not 29 °C or 30 °C. Our results provide insight into the effects of temperature on digestive physiology across narrow temperature ranges as well as demonstrate a sex-based difference in digestive physiology.

Hydration state does not affect selected body temperature during gravidity or gravidity duration in pythons (Antaresia childreni).

The embryonic development of many ectothermic species are highly sensitive to temperature and typically have a higher thermal optima than do most other physiological processes. Thus, female ectotherms often maintain a higher and more carefully controlled body temperature when she is supporting developing embryos (early development in oviparous species, throughout development in viviparous species). Considering the positive correlation between body temperature and evaporative water loss, this response could potentially exacerbate female water imbalance in water-limited environments, suggesting that female water balance and egg development may be in conflict. Using Children's pythons (Antaresia childreni), we hypothesized that water deprivation reduces thermophily during gravidity. We split reproductive females into two thermal treatments: those provided with a continuously available thermal gradient of 25-45 °C and those kept at a constant 31 °C. We also had seven non-reproductive females that were provided a thermal gradient. Within each thermal treatment group, we alternatingly assigned females to either have or not have water throughout gravidity. We found that reproduction increased female body temperature, but this increase was not affected by water regime. Reproduction also increased plasma osmolality, and lack of water during gravidity exacerbated this effect. We also found that thermal treatment, but not water regime, significantly influenced gravidity duration, with females given a thermogradient having a shorter gravidity duration, likely as a result of having a higher average body temperature than did the females provided constant heat. Finally, we found that females provided water throughout gravidity had greater clutch masses than did females without water. Further research is needed to improve scientific understanding of the interactions among water balance, body temperature, and various physiological performances.

Gynecological surgery in adulthood imparts cognitive and brain changes in rats: A focus on hysterectomy at short-, moderate-, and long-term intervals after surgery.

Premenopausal hysterectomy is associated with a greater relative risk of dementia. We previously demonstrated cognitive impairments in adult rats six weeks after hysterectomy with ovarian conservation compared with intact sham-controls and other gynecological surgery variations. Here, we investigated whether hysterectomy-induced cognitive impairments are transient or persistent. Adult rats received sham-control, ovariectomy (Ovx), hysterectomy, or Ovx-hysterectomy surgery. Spatial working memory, reference memory, and anxiety-like behavior were tested either six-weeks post-surgery, in adulthood; seven-months post-surgery, in early middle-age; or twelve-months post-surgery, in late middle-age. Hysterectomy in adulthood yielded spatial working memory deficits at short-, moderate-, and long-term post-surgery intervals. Serum hormone levels did not differ between ovary-intact, but differed from Ovx, groups. Hysterectomy had no significant impact on healthy ovarian follicle or corpora lutea counts for any post-surgery timepoint compared with intact sham-controls. Frontal cortex, dorsal hippocampus, and entorhinal cortex were assessed for activity-dependent markers. In entorhinal cortex, there were alterations in FOSB and ΔFOSB expression during the early middle-age timepoint, and phosphorylated ERK1/2 levels at the adult timepoint. Collectively, results suggest a primary role for the uterus in regulating cognition, and that memory-related neural pathways may be modified following gynecological surgery. This is the first preclinical report of long-term effects of hysterectomy with and without ovarian conservation on cognition, endocrine, ovarian, and brain assessments, initiating a comprehensive framework of gynecological surgery effects. Translationally, findings underscore critical needs to decipher how gynecological surgeries, especially those involving the uterus, impact the brain and its functions, the ovaries, and overall aging from a systems perspective.

Dehydrated snakes reduce postprandial thermophily.

Transient thermophily in ectothermic animals is a common response during substantial physiological events. For example, ectotherms often elevate body temperature after ingesting a meal. In particular, the increase in metabolism during the postprandial response of pythons - known as specific dynamic action - is supported by a concurrent increase in preferred temperature. The objective of this study was to determine whether hydration state influences digestion-related behavioral thermophily. Sixteen (8 male and 8 female) Children's pythons (Antaresia childreni) with surgically implanted temperature data loggers were housed individually and provided with a thermal gradient of 25-45°C. Body temperature was recorded hourly beginning 6 days prior to feeding and for 18 days post-feeding, thus covering pre-feeding, postprandial and post-absorptive stages. Each snake underwent this 24 day trial twice, once when hydrated and once when dehydrated. Our results revealed a significant interaction between temperature preference, digestive stage and hydration state. Under both hydrated and dehydrated conditions, snakes similarly increased their body temperature shortly after consuming a meal, but during the later days of the postprandial stage, snakes selected significantly lower (∼1.5°C) body temperature when they were dehydrated compared with when they were hydrated. Our results demonstrate a significant effect of hydration state on postprandial thermophily, but the impact of this dehydration-induced temperature reduction on digestive physiology (e.g. passage time, energy assimilation) is unknown and warrants further study.

Corticosterone and immune responses to dehydration in squamate reptiles.

Many environments present some degree of seasonal water limitations; organisms that live in such environments must be adapted to survive periods without permanent water access. Often this involves the ability to tolerate dehydration, which can have adverse physiological effects and is typically considered a physiological stressor. While having many functions, the hormone corticosterone (CORT) is often released in response to stressors, yet increasing plasma CORT while dehydrated could be considered maladaptive, especially for species that experience predictable bouts of dehydration and have related coping mechanisms. Elevating CORT could reduce immunocompetence and have other negative physiological effects. Thus, such species likely have CORT and immune responses adapted to experiencing seasonal droughts. We evaluated how dehydration affects CORT and immune function in eight squamate species that naturally experience varied water limitation. We tested whether hydric state affected plasma CORT concentrations and aspects of immunocompetence (lysis, agglutination, bacterial killing ability and white blood cell counts) differently among species based on how seasonally water limited they are and whether this is constrained by phylogeny. The species represented four familial pairs, with one species of each pair inhabiting environments with frequent access to water and one naturally experiencing extended periods (>30 days) with no access to standing water. The effects of dehydration on CORT and immunity varied among species. Increases in CORT were generally not associated with reduced immunocompetence, indicating CORT and immunity might be decoupled in some species. Interspecies variations in responses to dehydration were more clearly grouped by phylogeny than by habitat type.

The correlated evolution of foraging mode and reproductive effort in lizards.

Life-history theory suggests that the optimal reproductive effort of an organism is affected by factors such as energy acquisition and predation risk. The observation that some organisms actively search for their prey and others ambush them creates the expectation of different energy needs and predation risk associated with each foraging behaviour, the so-called 'foraging-mode paradigm'. Although this paradigm has been around for decades, the empirical evidence consists of conflicting results derived from competing models based on different mechanisms. For instance, models within the foraging-mode paradigm suggest that widely foraging females have evolved low reproductive effort, because a heavy reproductive load decreases their ability to escape from predators. By contrast, a long-standing prediction of evolutionary theory indicates that organisms subject to high extrinsic mortality, should invest more in reproduction. Here, we present the first partial evidence that widely foraging species have evolved greater reproductive effort than have sit-and-wait species, which we attribute to a larger body size and greater mortality among mobile foragers. According to our findings, we propose a theoretical model that could explain the observed pattern in lizards, suggesting ways for evolutionary ecologists to test mechanistic hypotheses at the intraspecific level.

Glucose tolerance of iguanas is affected by high-sugar diets in the lab and supplemental feeding by ecotourists in the wild.

There is great interspecific variation in the nutritional composition of natural diets, and the varied nutritional content is physiologically tolerated because of evolutionarily based balances between diet composition and processing ability. However, as a result of landscape change and human exposure, unnatural diets are becoming widespread among wildlife without the necessary time for evolutionary matching between the diet and its processing. We tested how a controlled, unnatural high glucose diet affects glucose tolerance using captive green iguanas, and we performed similar glucose tolerance tests on wild Northern Bahamian rock iguanas that are either frequently fed grapes by tourists or experience no such supplementation. We evaluated both short and longer-term blood glucose responses and corticosterone (CORT) concentrations as changes have been associated with altered diets. Experimental glucose supplementation in the laboratory and tourist feeding in the wild both significantly affected glucose metabolism. When iguanas received a glucose-rich diet, we found greater acute increases in blood glucose following a glucose challenge. Relative to unfed iguanas, tourist-fed iguanas had significantly lower baseline CORT, higher baseline blood glucose, and slower returns to baseline glucose levels following a glucose challenge. Therefore, unnatural consumption of high amounts of glucose alters glucose metabolism in laboratory iguanas with short-term glucose treatment and free-living iguanas exposed to long-term feeding by tourists. Based on these results and the increasing prevalence of anthropogenically altered wildlife diets, the consequences of dietary changes on glucose metabolism should be further investigated across species, as such changes in glucose metabolism have health consequences in humans (e.g. diabetes).

Fear-based aggression and its relationship to corticosterone responsiveness in three species of python.

It has long been known that even closely related species can vary in their antipredator behavior, and in the last two decades there has been mounting interest in how these differences might relate to the hormonal stress response. We tested the relationship between fear-based aggression, a form of antipredator behavior, and plasma corticosterone levels in three species of python [Children's Python (Antaresia childreni), Ball Python (Python regius), Bismarck Ring Python (Bothrochilus boa)]. We recorded the amount of striking in response to perturbation before and after a controlled, stressful confinement. We also measured plasma corticosterone levels prior to confinement, after confinement, and after confinement plus an adrenocorticotropin hormone (ACTH) injection, the later to induce a maximal corticosterone response. We performed among species analyses using two mixed models, and we determined between individual variance within each species to estimate repeatability. Bismarck Ring Pythons struck more than either Ball Pythons or Children's Pythons, and Ball Pythons had a suppressed corticosterone response compared to Children's and Bismarck Ring Pythons. Thus, mean species fear-based aggression correlated with species level differences in corticosterone profile. We also found evidence suggesting behaviors are repeatable within individuals. Our results point to a need for further exploration of aggression, anti-predator behavior, and corticosterone profile.

Reproductive state and water deprivation increase plasma corticosterone in a capital breeder.

Plasma corticosterone (CORT) concentrations fluctuate in response to homeostatic demands. CORT is widely recognized as an important hormone related to energy balance. However, far less attention has been given to the potential role of CORT in regulating salt and water balance or responding to osmotic imbalances. We examined the effects of reproductive and hydric states on CORT levels in breeding Children's pythons (Antaresia childreni), a species with substantial energetic and hydric costs associated with egg development. Using a 2 × 2 experimental design, we examined how reproduction and water deprivation, both separately and combined, impact CORT levels and how these changes correlate with hydration (plasma osmolality) and energy levels (blood glucose). We found that reproduction leads to increased CORT levels, as does dehydration induced by water deprivation. The combined impact of reproduction and water deprivation led to the largest increases in CORT levels. Additionally, we found significant positive relationships among CORT levels, plasma osmolality, and blood glucose. Our results provide evidence that both reproductive activity and increased plasma osmolality can lead to increased plasma CORT in an ectotherm, which could be explained by either CORT having a role as a mineralocorticoid or CORT being elevated as part of a stress response to resource imbalances.

Energetic investment associated with vitellogenesis induces an oxidative cost of reproduction.

Oxidative stress is a potential cost of reproduction, but conclusive evidence for this relationship is lacking. The goal of this study was to serially assess across a seasonal gradient the relationship between reproduction, circulating plasma energy metabolites and oxidative state. Here, we examine a study animal ideally suited to test for the oxidative costs of reproduction: the Allen Cays Rock Iguana. Female rock iguanas reproduce at varying frequencies, often skipping years, allowing for a comparison between reproductive and non-reproductive females during the same narrow, annual breeding season. This feature of iguana life history enabled us to address not just sex and seasonal differences in physiology, but also potential oxidative costs of reproduction in females. Male and female iguanas were sampled during the early (vitellogenic), late (gravid) and post-reproductive seasons. Ultrasound examinations were performed on females to quantify reproductive investment, and blood samples were collected for physiology assays, which included reactive oxygen metabolites (d-ROMs), antioxidants, triglycerides, free glycerol and glucose. The early reproductive season was characterized by significant increases in reproductive female's triglycerides, free glycerol and oxidative stress compared to late and post-reproductive periods and non-reproductive females and males during all sampling periods. Antioxidants were significantly elevated during the early reproductive season for reproductive females, non-reproductive females and males when compared to late and post-season. Follicle number in early reproductive females was positively related to d-ROMs, triglycerides and free glycerol, negatively related to antioxidants and showed no relationship with glucose. Measures of oxidative stress, d-ROMs and oxidative index were positively correlated with circulating levels of the lipid metabolite free glycerol during the early reproductive period, but this relationship weakened in the late season and disappeared in the post-season. Broadly, this study supports the hypothesis that the relationship between reproduction and oxidative stress is driven by energy investment, being greatest during early reproduction when vitellogenesis is occurring.

Cane toads (Rhinella marina) rely on water access, not drought tolerance, to invade xeric Australian environments.

The invasion of habitats with novel environmental challenges may require physiological tolerances not seen in conspecifics from the native range. We used a combination of field and laboratory-based experiments to assess physiological tolerance to limited water access at four sites distributed across the historical invasion path of cane toads (Rhinella marina) in Australia that, from east to west, alternated between mesic and seasonally xeric habitats. Toads from all locations were well hydrated at the time of capture. However, experimental dehydration caused greater mass loss, higher plasma osmolality, and inhibition of lytic ability in toads from xeric compared to mesic locations. These results suggest somewhat surprisingly that toads from xeric environments are physiologically more vulnerable to water loss. In contrast, bactericidal ability was not sensitive to hydric state and was greater in toads from eastern (long-colonized) areas. Similar patterns in lytic ability in hydrated toads and agglutination ability in wild toads suggest that toads along the invasion front face a tradeoff between enhanced dispersal ability and physiological responses to dehydration. The ability of this invasive species to spread into drier environments may be underpinned by a combination of phenotypic plasticity and evolved (heritable) traits.

Dehydration during egg production alters egg composition and yolk immune function.

Parent-offspring conflicts occur when resources are limited for allocation, and, historically, energy has been the primary currency of focus when examining these trade-offs. Water is a fundamental resource that has received far less consideration for parent-offspring conflicts. Previous research suggests that, when water is limited, reproductive females are compromised in favor of developing embryos. However, these studies limited their assessments to standard metrics such as clutch size and mass. We tested the hypothesis that the mother-offspring conflict over limited water resources leads to finer scale morphological and physiological impacts on the eggs in Children's pythons (Antaresia childreni). We predicted that water deprivation during gravidity alters female investment into her eggs, impacting egg water content and shell development. Additionally, we predicted that the yolk in these dehydrated eggs would have enhanced immune performance metrics, as has been documented in dehydrated adults. We found that eggs from water-deprived females were dehydrated as indicated by reduced percent water and greater yolk osmolality compared to eggs from females that received ad libitum water. We also found that eggs from dehydrated mothers had thinner shells and higher water loss rates. The impacts were not entirely negative as dehydrated eggs had higher antimicrobial capabilities. Also, thinner and more permeability eggshells might allow for elevated rates of rehydration from nest substrate. Overall, by examining an array of egg traits, we demonstrated that dehydration of gravid females impacts the eggs, not just the females as previously reported. As a result, the mother-offspring conflicts are indeed two-sided.

Muscles provide an internal water reserve for reproduction.

The use of fat to support the energy needs of reproduction (i.e. capital breeding) has been studied in a diversity of taxa. However, despite reproductive output (i.e. young or eggs) being approximately 70% water, little is known about the availability of internal resources to accommodate the hydric demands of reproduction. Recent research suggests that dehydration increases the catabolism of muscle as a means of maintaining water balance. Accordingly, we investigated the interactive effects of reproductive investment and water deprivation on catabolism and reproductive output in female Children's pythons (Antaresia childreni). Both reproductive and non-reproductive females were either provided water ad libitum or were water-deprived for three weeks at the time when reproductive females were gravid. We found that water-deprived reproductive females had, in general, greater body mass loss, epaxial muscle loss, plasma osmolality and plasma uric acid concentrations relative to the other groups. Furthermore, water-deprived females had similar clutch sizes compared with females with access to water, but produced lighter eggs and lower total clutch masses. Our results provide the first evidence that selective protein catabolism can be used to support water demands during reproduction, and, as a result, these findings extend the capital breeding concept to non-energetic resources.

Evidence for atypical nest overwintering by hatchling lizards, Heloderma suspectum.

The timing of reproductive events (e.g. oviposition and hatching) to coincide with favourable seasonal conditions is critical for successful reproduction. However, developmental time may not match the duration between the optimal time for oviposition and the optimal time for hatchling survival. Thus, strategies that alter the time between oviposition and hatchling emergence can be highly advantageous. Arrested development and the resulting extension of the duration between oviposition and hatching has been widely documented across oviparous amniotes, but nest overwintering by hatchlings has only been documented in aquatic chelonians that live where winters are quite cold. Herein, we present a compilation of evidence regarding reproductive phenology by hatchlings of the Gila monster (Heloderma suspectum), a lizard inhabiting the Sonoran Desert of North America. Our data demonstrate that (i) Gila monster hatchlings from eggs oviposited in July do not emerge from their nests until late spring or summer of the following year, yet (ii) Gila monster eggs artificially incubated at field-relevant temperatures hatch in 4-5 months. Furthermore, we describe a fortuitous excavation of a hatching Gila monster nest in late October, which coincides with the artificial incubation results. Together, these results provide strong support for the existence of overwintering in the nest by a lizard, and suggest that this reproductive strategy should be explored in a broader array of taxa.

When less means more: dehydration improves innate immunity in rattlesnakes.

Immune function can vary based on availability of resources, and most studies of such influences have focused on the co-investment of energy into immune and other physiological functions. When energy resources are limited, trade-offs exist, which can compromise immunity for other functions. As with energy, water limitation can also alter various physiological processes, yet water has received little consideration for its possible role in modulating immune functions. We examined the relationship between immunocompetence and hydration state using the western diamond-backed rattlesnake (Crotalus atrox). This species is known to undergo substantial seasonal fluctuations in water availability with extreme limitations during the hot-dry season. We collected blood samples from free-ranging C. atrox to compare osmolality and innate immune function (lysis, agglutination and bacterial growth inhibition) during the milder and relatively moister early spring season, the hot-dry season and the hot-wet season. To isolate effects of dehydration from other possible seasonal influences, we complemented this field study with a laboratory study in which we withheld food and water from individually housed adult C. atrox for up to 16 weeks. We collected blood samples from each snake as it dehydrated and collected a final sample after the snake was given water ad libitum at the end of the experiment. Our results demonstrate that C. atrox experience significant dehydration during the hot-dry season, and that, in general, innate immune function is highly correlated with osmolality, whether natural or artificially manipulated.

Dehydration enhances multiple physiological defense mechanisms in a desert lizard, Heloderma suspectum.

The physiological challenges associated with dehydration can induce an increase in plasma glucocorticoid concentrations, a response thought to provide the mechanism for dehydration suppressing immune function. However, a comprehensive examination of the inter-relationship of dehydration, stress and immune function has not been conducted within a single species. We previously demonstrated that Gila monsters (Heloderma suspectum), which inhabit a xeric environment with a predictable seasonal drought, have enhanced measures of innate immunity when dehydrated. These results suggest that, in this species, dehydration may not induce a glucocorticoid response, but, instead, enhances physiological defense mechanisms. To explore this possibility, we examined multiple measures of innate immunity as well as initial and reactive plasma concentrations of glucocorticoids in captive and free-ranging Gila monsters at various hydration states. Our results show that, in this species, dehydration alone does not cause a substantial increase in plasma glucocorticoids, and we provide broader evidence that dehydration enhances defensive mechanisms including stress reactivity and various measures of innate immune function. These findings suggest that physiological responses to dehydration may depend heavily on an organism's ecology. More research on the effects of dehydration on the glucocorticoid response and immunity will help clarify the interactive roles they play in response to hydration challenges and whether adaptations to water-limited environments influence these interactions.

Modeling the costs and benefits associated with the evolution of endothermy using a robotic python.

Endothermy provides considerable benefits to an organism but requires large energy investment. To understand potential driving forces that would lead to the evolution of endothermy, it is important to understand the energy costs and potential benefits of intermediate steps between ectothermy and homeothermic endothermy as well as the influences of environmental conditions on energetic costs. However, efforts to examine intermediate conditions are greatly limited by the predominant natural dichotomy between ectothermy and endothermy. Facultative endothermy by brooding pythons provides a fortunate study system where endothermy is beneficial but not essential. As one cannot control the extent of energy investment in heat production by a female python, we created an artificial snake with controllable heating capability. This enabled us to determine the energetic costs of maintaining a clutch at a preferred temperature, and to determine the relative thermal benefit of limited energy-producing capability (i.e. 50% of the required energy to maintain the preferred developmental temperature). We manipulated the pseudoserpent's clutch size (5, 10, 15 eggs), diel ambient temperature cycle (2, 4, 6°C) and insulation (with and without) at each of these power levels: unlimited power, half required power and no power. We found no significant effect of clutch size on either power requirements or developmental temperature. Energy requirements increased with the amplitude of the diel cycle and decreased with the addition of insulation, while the quality of the thermal environment decreased with the amplitude of the diel cycle. Interestingly, the quality of the thermal environment also decreased with the addition of insulation. We discuss these results within the context of the reproductive model of the evolution of endothermy.

Water availability and environmental temperature correlate with geographic variation in water balance in common lizards.

Water conservation strategies are well documented in species living in water-limited environments, but physiological adaptations to water availability in temperate climate environments are still relatively overlooked. Yet, temperate species are facing more frequent and intense droughts as a result of climate change. Here, we examined variation in field hydration state (plasma osmolality) and standardized evaporative water loss rate (SEWL) of adult male and pregnant female common lizards (Zootoca vivipara) from 13 natural populations with contrasting air temperature, air humidity, and access to water. We found different patterns of geographic variation between sexes. Overall, males were more dehydrated (i.e. higher osmolality) than pregnant females, which likely comes from differences in field behaviour and water intake since the rate of SEWL was similar between sexes. Plasma osmolality and SEWL rate were positively correlated with environmental temperature in males, while plasma osmolality in pregnant females did not correlate with environmental conditions, reproductive stage or reproductive effort. The SEWL rate was significantly lower in populations without access to free standing water, suggesting that lizards can adapt or adjust physiology to cope with habitat dryness. Environmental humidity did not explain variation in water balance. We suggest that geographic variation in water balance physiology and behaviour should be taken account to better understand species range limits and sensitivity to climate change.

Water deprivation increases maternal corticosterone levels and enhances offspring growth in the snake Vipera aspis.

Circulating glucocorticoid (GC) levels may increase as a result of reproductive effort or in response to unpredictable events. However, GC secretion can vary with the availability of vital trophic resources such as energy. While water represents another critical resource, the impact of water deprivation on GC secretion during reproduction has not yet been thoroughly investigated. Here, we examined the effects of water deprivation on plasma corticosterone (CORT) concentrations of female aspic vipers (Vipera aspis), and determined the impacts of water deprivation on offspring traits. We exposed both pregnant and non-reproductive females to a 20-day water deprivation and compared their pre- and post-deprivation CORT levels with those of control females. At the end of the treatment, only water-deprived pregnant females showed a significant increase in CORT levels. In pregnant females, changes in baseline CORT level were correlated with changes in female hydration state. Changes in baseline CORT levels were also negatively influenced by maternal reproductive effort in pregnant control females, while such a relationship was not apparent in pregnant water-deprived females. Finally, we found that offspring from water-deprived females had higher growth rates than offspring from control females. Offspring growth was also positively correlated with changes in both maternal osmolality and baseline CORT levels. Together, our results suggest that dehydration increases maternal CORT levels, which may subsequently influence offspring development. Further long-term field studies are therefore required to assess whether there is an adaptive significance of this response.

Facultative thermogenesis during brooding is not the norm among pythons.

Facultative thermogenesis is often attributed to pythons in general despite limited comparative data available for the family. While all species within Pythonidae brood their eggs, only two species are known to produce heat to enhance embryonic thermal regulation. By contrast, a few python species have been reported to have insignificant thermogenic capabilities. To provide insight into potential phylogenetic, morphological, and ecological factors influencing thermogenic capability among pythons, we measured metabolic rates and clutch-environment temperature differentials at two environmental temperatures-python preferred brooding temperature (31.5 °C) and a sub-optimal temperature (25.5 °C)-in six species of pythons, including members of two major phylogenetic branches currently devoid of data on the subject. We found no evidence of facultative thermogenesis in five species: Aspidites melanocephalus, A. ramsayi, Morelia viridis, M. spilota cheynei, and Python regius. However, we found that Bothrochilus boa had a thermal metabolic sensitivity indicative of facultative thermogenesis (i.e., a higher metabolic rate at the lower temperature). However, its metabolic rate was quite low and technical challenges prevented us from measuring temperature differential to make conclusions about facultative endothermy in this species. Regardless, our data combined with existing literature demonstrate that facultative thermogenesis is not as widespread among pythons as previously thought.