Analysis of subcellular energy metabolism in five Lacertidae lizards across varied environmental conditions.

Aerobic respiration is the main energy source for most eukaryotes, and efficient mitochondrial energy transfer greatly influences organismal fitness. To survive environmental changes, cells have evolved to adjust their biochemistry. Thus, measuring energy metabolism at the subcellular level can enhance our understanding of individual performance, population dynamics, and species distribution ranges. We investigated three important metabolic traits at the subcellular level in five lacertid lizard species sampled from different elevations, from sea level up to 2000 m. We examined hemoglobin concentration, two markers of oxidative stress (catalase activity and carbonyl concentration) and maximum rate of metabolic respiration at the subcellular level (potential metabolic activity at the electron transport system). The traits were analysed in laboratory acclimated adult male lizards to investigate the adaptive metabolic responses to the variable environmental conditions at the local sampling sites. Potential metabolic activity at the cellular level was measured at four temperatures - 28 °C, 30 °C, 32 °C and 34 °C - covering the range of preferred body temperatures of the species studied. Hemoglobin content, carbonyl concentration and potential metabolic activity did not differ significantly among species. Interspecific differences were found in the catalase activity, Potential metabolic activity increased with temperature in parallel in all five species. The highest response of the metabolic rate with temperature (Q10) and Arrhenius activation energy (Ea) was recorded in the high-mountain species Iberolacerta monticola.

Melanistic coloration does not influence thermoregulation in the crepuscular gecko Eublepharis macularius.

Body coloration in ectotherms serves multiple biological functions, including avoiding predators, communicating with conspecific individuals, and involvement in thermoregulation. As ectotherms rely on environmental sources of heat to regulate their internal body temperature, stable melanistic body coloration or color change can be used to increase or decrease heat absorption and heat exchange with the environment. While melanistic coloration for thermoregulation functions to increase solar radiation absorption and consequently heating in many diurnal ectotherms, research on crepuscular and nocturnal ectotherms is lacking. Since crepuscular and nocturnal ectotherms generally absorb heat from the substrate, in these organisms melanistic coloration may have other primary functions beside thermoregulation. As such, in this work we hypothesized that the proportion of dorsal melanistic body coloration would not influence heating and cooling rates in the crepuscular gecko, Eublepharis macularius, and that changes in environmental temperature would not trigger color changes in this species. Temperature measurements of the geckos and of the environment were taken using infrared thermography and temperature loggers. Color data were obtained using objective photography and a newly developed custom software package. We found that body temperature reflected substrate temperatures, and that the proportion of melanistic coloration has no influence on heating or cooling rates or on color changes. These findings support that melanistic coloration in E. macularius may not be used for thermoregulation and strengthen the hypothesis that in animals active in low light conditions, melanistic coloration may be used instead for camouflage or other functions.

An integrative analysis of the short-term effects of tail autotomy on thermoregulation and dehydration rates in wall lizards.

Maintaining body temperature is essential for the optimal performance of physiological functions. Ectotherms depend on external heat sources to thermoregulate. However, thermoregulation may be constrained by body condition and hydration state. Autotomy (i.e., the voluntary shed of a body part) evolved in various animal lineages and allowed surviving certain events (such as predator attacks), but it may affect body condition and volume/surface ratios, increase dehydration and constrain thermoregulation. In the framework of a general analysis of the evolution of autotomy, here we assessed the effects of tail loss on the thermal preferences and evaporative water loss rates (EWL) in the lizard Podarcis bocagei, integrating the thermal and hydric factors. We did not observe shifts in the thermal preferences of experimentally autotomized lizards when compared to the controls, which contradicted the hypothesis that they would raise preferred temperature to increase metabolic rates and accelerate regeneration. Evaporative water loss rates were also similar for tailed and tailless individuals, suggesting negligible increase of water loss through the injury and no specific ecophysiological responses after autotomy. Therefore, the changes observed in autotomized lizards in the field are to be considered primarily behavioural, rather than physiological, and thermoregulation could be secondarily affected by behavioural compensations for an increased predation risk after autotomy. Functional studies are necessary to understand how lizards' interaction with the environment is altered after autotomy, and further studies including different dehydration levels would be useful to fully understand the effect of water shortage on lizards' performance after caudal autotomy.

Evidence from Tarentola mauritanica (Gekkota: Phyllodactylidae) helps validate thermography as a tool to infer internal body temperatures of lizards.

Infrared (IR) thermal imaging has become an increasingly popular tool to measure body temperature of animals. The high-resolution data it provides with short lag and minimum disturbance makes it an appealing tool when studying reptile thermal ecology. However, due to the common phenomenon of regional heterothermy and surface-to-core temperature gradients, it is essential to select the appropriate body part to measure and provide calibrations to accurately infer internal body temperatures. This work follows from a previous study on lacertid lizards to assess the reliability of thermography-measured body temperatures, from several body locations, as a proxy for internal body temperature in lizards. This study focuses on the Moorish gecko, Tarentola mauritanica, due to its distant phylogenetic relationship and its different ecology and morphology from the previously tested species. A total of 60 adult geckos of both sexes and of a range of sizes were tested in thermal gradients and subjected to a sequence of randomly assorted treatments of heating and cooling. The temperatures of the animals were periodically measured with a thermal camera at six different body parts and, immediately after, the cloacal temperature was then measured with a thermocouple probe. Body parts' temperatures, obtained thermographically, were regressed against cloacal temperature using OLS regression and the pairwise correlations were tested using Spearman coefficients. Relationships among all body parts and between all body parts and the cloaca were strong in all cases (R2 > 0.87, Spearman Correlation > 0.95). The observed pattern was very similar to those previously obtained from lacertid lizards. Ultimately, the eye proved to provide the best overall proxy for internal temperature, when accounting for both the slope and intercept of the regression. Hence, this study provides further support for the establishment of the eye as the standard location to infer internal body temperatures of lizards through thermography.

Physiological differences in preferred temperatures and evaporative water loss rates in two sympatric lacertid species.

Sister species living in sympatry offer the opportunity to study the degree of divergence in their ecological, physiological and life-history traits. It has been hypothesized that closely related species with overlapping distribution should differ in their niche to reduce competition for resources. Furthermore, the investigation of sympatric species may shed light on how they may coexist without outcompeting each other. In the present study, we assess the degree of physiological divergence in two sympatric lacertid lizards, Podarcis bocagei and Podarcis guadarramae lusitanicus. These species share a Pliocenic ancestry and overlap at a both geographical and ecological scale. We assessed their thermal preferences and water loss rates, two physiological traits considered stable across congeneric species. We found that the two species differ in both traits, with P. bocagei selecting higher temperature than P. g. lusitanicus and losing more water than the latter at and above its preferred temperature. The results also showed that for both species body size has a relevant impact on thermal and hydric traits, with bigger individuals losing proportionally less water and selecting higher temperatures. These results, combined with previous evidence, suggest that physiological mechanisms, ecological preferences and morphology probably allow these two species to overlap in their distribution while selecting different microhabitats and thus decreasing possible competition between them.

Assessing the reliability of thermography to infer internal body temperatures of lizards.

For many years lizard thermal ecology studies have relied on the use of contact thermometry to obtain internal body temperature (Tb) of the animals. However, with progressing technology, an interest grew in using new, less invasive methods, such as InfraRed (IR) pyrometry and thermography, to infer Tb of reptiles. Nonetheless few studies have tested the reliability of these new tools. The present study tested the use of IR cameras as a non-invasive tool to infer Tb of lizards, using three differently body-sized lacertid species (Podarcis virescens, Lacerta schreiberi and Timon lepidus). Given the occurrence of regional heterothermy, we pairwise compared thermography readings of six body parts (snout, eye, head, dorsal, hind limb, tail base) to cloacal temperature (measured by a thermometer-associated thermocouple probe) commonly employed to measure Tb in field and lab studies. The results showed moderate to strong correlations (R2=0.84-0.99) between all body parts and cloacal temperature. However, despite the readings on the tail base showed the strongest correlation in all three species, it was the eye where the absolute values and pattern of temperature change most consistently followed the cloacal measurements. Hence, we concluded that the eye would be the body location whose IR camera readings more closely approximate that of the animal's internal environment. Alternatively, other body parts can be used, provided that a careful calibration is carried out. We provide guidelines for future research using thermography to infer Tb of lizards.