The Role of Feeding Specialization on Post-Prandial Metabolic Rate in Snakes of the Genus Bothrops.

Feeding specialization is a recurrent issue in the evolution of snakes and is sometimes associated to morphological and/or behavioral adaptations that improve snake performance to exploit a particular food type. Despite its importance for animal fitness, the role of physiological traits has been much less studied than morphological and behavioral traits in the evolution of feeding specialization in snakes. In this context, the energetic cost of post-prandial period is an important physiological factor due to the remarkable effect on the snake energy budget. We collected data on post-prandial metabolic rate (SDA) in five species of pit vipers from the genus Bothrops with different degrees of mammal feeding specialization to test the hypothesis that feeding specialist species have lower energy costs during the digestion of their regular food item when compared to species with a more generalist diet. Our results support this hypothesis and suggest that ontogenetic changes in diet can be accompanied by changes in energy cost of the digestion process.

Assessing Antibacterial Potential of Components of Phyllomedusa distincta Skin and its Associated Dermal Microbiota.

The granular glands of anuran skin secrete an array of bioactive molecules that protect a frog against pathogens and predators. The skin also harbors a microbial community. Although there is evidence to suggest that the microbiota complement the innate immune defense systems against pathogen infection, the effect of the frog bioactive molecules on its resident microbiota has not yet been fully investigated. In the present study, the skin microbiota of Phyllomedusa distincta obtained from two different geographical areas was evaluated with molecular and culture-based approaches. The antagonistic effects exhibited by the host's microbiota and by a novel dermaseptin peptide isolated from P. distincta skin were investigated. Four isolated bacterial colonies displayed antimicrobial activity against known frog pathogens. Our results were consistent with the hypothesis that microbiota from P. distincta may interact with pathogenic microorganisms to protect a frog's health. On the other hand, the novel dermaseptin peptide exhibited an antimicrobial effect on pathogens as well as on some of the bacteria obtained from the skin microbiota. The richness of bacteria on P. distincta skin was further investigated by 16S rRNA gene clone libraries, which revealed that the family Enterobacteriaceae was prevalent, but a high variability at the species level was observed among individual frogs. Differences observed on the microbiota of frogs from contrasting habitats indicated an influence of the environment on the structure of the skin microbiota of P. distincta.

Upper thermal tolerance plasticity in tropical amphibian species from contrasting habitats: implications for warming impact prediction.

Tropical ectothermic species are currently depicted as more vulnerable to increasing temperatures because of the proximity between their upper thermal limits and environmental temperatures. Yet, the acclimatory capacity of thermal limits has rarely been measured in tropical species, even though they are generally predicted to be smaller than in temperate species. We compared critical thermal maximum (CTmax) and warming tolerance (WT: the difference between CTmax and maximum temperature, Tmax), as well as CTmax acclimatory capacity of toad species from the Atlantic forest (AF) and the Brazilian Caatinga (CAA), a semi-arid habitat with high temperatures. Acclimation temperatures represented the mean temperatures of AF and CAA habitats, making estimates of CTmax and WT more ecologically realistic. CAA species mean CTmax was higher compared to AF species in both acclimation treatments. Clutches within species, as well as between AF and CAA species, differed in CTmax plasticity and we discuss the potential biological meaning of these findings. We did not find a trade-off between absolute CTmax and CTmax plasticity, indicating that species can have both high CTmax and high CTmax plasticity. Although CTmax was highly correlated to Tmax, CTmax plasticity was not related to Tmax or Tmax coefficients of variation. CAA species mean WT was lower than for AF species, but still very high for all species, diverging from other studies with tropical species. This might be partially related to over-estimation of vulnerability due to under-appreciation of realistic acclimation treatments in CTmax estimation. Thus, some tropical species might not be as vulnerable to warming as previously predicted if CTmax is considered as a shifting population parameter.

A new species of Bachia Gray, 1845 (Squamata: Gymnophthalmidae) from the Eastern Brazilian Cerrado, and data on its ecology, physiology and behavior.

A new species of Bachia of the bresslaui group, Bachia geralista sp. nov., is described from Planalto dos Gerais, an old and partially dissected plateau extending along the Cerrados of Bahia, Minas Gerais and Tocantins states, Brazil. The new species is morphologically similar to B. bresslaui, with which it has been confused; however head scalation resembles other species from sandy spots within the Cerrado (B. psamophila and B. oxyrhina). Like in B. psamophila and B. oxyrhina, the shovel-shaped snout of the new species is highly prominent, a typical trait of psammophilous habits in other gymnophthalmids. The examination of specimens of B. bresslaui from several populations within the Cerrado revealed great variation among localities, leading to the reidentification of a specimen from Utiariti, Mato Grosso, previously referred to in the literature as the second record of B. bresslaui, as the recently described B. didactyla, suggesting that cryptic diversity might remain still undiscovered within this genus in the Cerrado. Despite occurring in a relatively open Cerrado, thermal physiology of Bachia geralista sp. nov. restricts its occurrence to shaded microhabitats within this habitat.

Vocal and territorial behavior in the Smith frog (Hypsiboas faber): relationships with plasma levels of corticosterone and testosterone.

The possible trade-off between the roles of glucocorticoids as facilitators of energy substrate mobilization and neural inhibitors of sexual behavior during breeding season is under debate. We studied the relationship between calling and territorial behavior with plasma levels of corticosterone (CORT) and plasma levels of testosterone (T) across the breeding season of Hypsiboas faber, a large and territorial Neotropical treefrog. We investigated these relationships through focal observations of males calling naturally, followed by blood sampling for hormonal radioimmunoassay. We additionally used an experimental approach, which consisted of broadcasting recorded advertisement calls for 10 min to simulate an invasion in the territory of the focal subjects, followed by behavioral observation and blood sampling for hormonal radioimmunoassay. Results showed a pattern of co-variation between CORT and T across the breeding season. Furthermore, individual variation in CORT and T was related to different aspects of behavior: individuals with higher CORT showed higher calling rates, and individuals with higher steroid levels, mainly T, showed higher responsivity to social stimulation by other males in the chorus. Experimental simulation of territorial intrusion by using playback of advertisement calls of this species did not elicit consistent changes in agonistic behavior and CORT, but decreased T in focal males.

Water balance and locomotor performance in three species of neotropical toads that differ in geographical distribution.

Water availability in the environment is a fundamental factor in determining the limits of geographical distribution and the evolution of the physiological characters associated to water balance in anurans. In this paper, we compare some aspects of water balance and the sensitivity of locomotor performance to dehydration at different temperatures for three species of toads from the genus Rhinella, with different levels of dependence on forested environments. Results show patterns associated to interspecific differences in both geographical distribution and time of seasonal reproduction. Sensitivity of locomotor performance to dehydration was lower at low temperatures for R. icterica, the species that are reproductively active during winter and lower at intermediate temperatures for R. schneideri, the species that reproduces mostly during spring, suggesting a pattern of thermal adaptation of locomotor performance for these species. Otherwise, R. ornata, a species with broader reproductive season, shows high sensitivity of locomotor performance to dehydration at all temperatures tested, suggesting a stronger relation of breeding activity with patterns of rainfall than temperature variation. Furthermore, the low rates of water uptake of R. ornata may pose restrictions on the occupation of open areas by this species.

Phylogenetic analysis of standard metabolic rate of snakes: a new proposal for the understanding of interspecific variation in feeding behavior.

The current proposal about the variation of standard metabolic rates (SMR) in snakes predicts that SMR is influenced by the feeding frequency (frequent or infrequent feeders). However, feeding frequency in snakes is poorly studied and hard to quantify under natural conditions. Alternatively, foraging strategy was studied for a large number of species and is usually related to the feeding frequency. In this work, we performed a meta-analysis on the SMR of compiled data from 74 species of snakes obtained from the literature and five more different species of lanceheads (genus Bothrops), after categorization according to the foraging mode (ambush or active foraging) and regarding their phylogenetic history. We tested the hypothesis that foraging mode (FM) is a determinant factor on the interspecific variation of SMR despite the phylogenetic relationship among species. We demonstrated that FM predicted SMR, but there is also a partial phylogenetic structuration of SMR in snakes. We also detected that evolution rates of SMR in active foragers seem to be higher than ambush-hunting snakes. We suggested that foraging mode has a major effect over the evolution of SMR in snakes, which could represent an ecophysiological co-adaptation, since ambush hunters (with low feeding rates) present a lower maintenance energetic cost (SMR) when compared to active foragers. The higher SMR evolution rates for active foraging snakes could be related to a higher heterogeny in the degree of activity during hunting by active foragers when compared to ambush-hunting snakes.

Skin microbiota in frogs from the Brazilian Atlantic Forest: Species, forest type, and potential against pathogens.

The cutaneous microbiota of amphibians can be defined as a biological component of protection, since it can be composed of bacteria that produce antimicrobial compounds. Several factors influence skin microbial structure and it is possible that environmental variations are among one of these factors, perhaps through physical-chemical variations in the skin. This community, therefore, is likely modified in habitats in which some ecophysiological parameters are altered, as in fragmented forests. Our research goal was to compare the skin bacterial community of four anuran species of the Atlantic Forest of Brazil in landscapes from two different environments: continuous forest and fragmented forest. The guiding hypotheses were: 1) microbial communities of anuran skin vary among sympatric frog species of the Atlantic forest; 2) the degree to which forested areas are intact affects the cutaneous bacterial community of amphibians. If the external environment influences the skin microbiota, and if such influences affect microorganisms capable of inhibiting the colonization of pathogens, we expect consequences for the protection of host individuals. We compared bacterial communities based on richness and density of colony forming units; investigated the antimicrobial potential of isolated strains; and did the taxonomic identification of isolated morphotypes. We collected 188 individual frogs belonging to the species Proceratophrys boiei, Dendropsophus minutus, Aplastodiscus leucopygius and Phyllomedusa distincta, and isolated 221 bacterial morphotypes. Our results demonstrate variation in the skin microbiota of sympatric amphibians, but only one frog species exhibited differences in the bacterial communities between populations from fragmented and continuous forest. Therefore, the variation we observed is probably derived from both intrinsic aspects of the host amphibian species and extrinsic aspects of the environment occupied by the host. Finally, we detected antimicrobial activity in 27 morphotypes of bacteria isolated from all four amphibian species.

Caloric Restriction Promotes Structural and Metabolic Changes in the Skin.

Caloric restriction (CR) is the most effective intervention known to enhance lifespan, but its effect on the skin is poorly understood. Here, we show that CR mice display fur coat remodeling associated with an expansion of the hair follicle stem cell (HFSC) pool. We also find that the dermal adipocyte depot (dWAT) is underdeveloped in CR animals. The dermal/vennule annulus vasculature is enlarged, and a vascular endothelial growth factor (VEGF) switch and metabolic reprogramming in both the dermis and the epidermis are observed. When the fur coat is removed, CR mice display increased energy expenditure associated with lean weight loss and locomotion impairment. Our findings indicate that CR promotes extensive skin and fur remodeling. These changes are necessary for thermal homeostasis and metabolic fitness under conditions of limited energy intake, suggesting a potential adaptive mechanism.

Temperature effects on a whole metabolic reaction cannot be inferred from its components.

Changes in temperature affect the kinetic energy of the constituents of a system at the molecular level and have pervasive effects on the physiology of the whole organism. A mechanistic link between these levels of organization has been assumed and made explicit through the use of values of organismal Q10 to infer control of metabolic rate. To be valid this postulate requires linearity and independence of the isolated reaction steps, assumptions not accepted by all. We address this controversy by applying dynamic systems theory and metabolic control analysis to a metabolic pathway model. It is shown that temperature effects on isolated steps cannot rigorously be extrapolated to higher levels of organization.

Temperature effects on energy metabolism: a dynamic system analysis.

Q(10) factors are widely used as indicators of the magnitude of temperature-induced changes in physico-chemical and physiological rates. However, there is a long-standing debate concerning the extent to which Q(10) values can be used to derive conclusions about energy metabolism regulatory control. The main point of this disagreement is whether or not it is fair to use concepts derived from molecular theory in the integrative physiological responses of living organisms. We address this debate using a dynamic systems theory, and analyse the behaviour of a model at the organismal level. It is shown that typical Q(10) values cannot be used unambiguously to deduce metabolic rate regulatory control. Analytical constraints emerge due to the more formal and precise equation used to compute Q(10), derived from a reference system composed from the metabolic rate and the Q(10). Such an equation has more than one unknown variable and thus is unsolvable. This problem disappears only if the Q(10) is assumed to be a known parameter. Therefore, it is concluded that typical Q(10) calculations are inappropriate for addressing questions about the regulatory control of a metabolism unless the Q(10) values are considered to be true parameters whose values are known beforehand. We offer mathematical tools to analyse the regulatory control of a metabolism for those who are willing to accept such an assumption.

Inter-individual variation and temperature-dependent antipredator behavior in the snake Tomodon dorsatus (Dipsadidae).

Although many studies assessed the influence of temperature on the behavior of ectotermic vertebrates, little attention has been given to interindividual variation in the defensive responses of reptiles. In the present study we investigated the defensive behavior of the snake Tomodon dorsatus, in order to test the hypotheses that (1) individuals differ in their antipredator behavior consistently with the concept of behavioral syndromes, (2) temperature influences the defensive behavior, and (3) these two factors interact with each other. There was significant interindividual variation in defensive behavior, as well as consistently aggressive, passive or evasive behaviors. Temperature influenced aggressiveness, which was slightly higher when body temperature was lower, but this trend was only evident in animals with aggressive disposition. Our results corroborate the hypothesis of interaction between individuality of behavior and temperature-dependent defensive behavior in T. dorsatus. These results, together with results from previous studies, suggest that the evolution of temperature-dependent defensive behavior differs among lineages of ectothermic tetrapods. This article is part of a Special Issue entitled: insert SI title.

Considerations for assessing maximum critical temperatures in small ectothermic animals: insights from leaf-cutting ants.

The thermal limits of individual animals were originally proposed as a link between animal physiology and thermal ecology. Although this link is valid in theory, the evaluation of physiological tolerances involves some problems that are the focus of this study. One rationale was that heating rates shall influence upper critical limits, so that ecological thermal limits need to consider experimental heating rates. In addition, if thermal limits are not surpassed in experiments, subsequent tests of the same individual should yield similar results or produce evidence of hardening. Finally, several non-controlled variables such as time under experimental conditions and procedures may affect results. To analyze these issues we conducted an integrative study of upper critical temperatures in a single species, the ant Atta sexdens rubropiosa, an animal model providing large numbers of individuals of diverse sizes but similar genetic makeup. Our specific aims were to test the 1) influence of heating rates in the experimental evaluation of upper critical temperature, 2) assumptions of absence of physical damage and reproducibility, and 3) sources of variance often overlooked in the thermal-limits literature; and 4) to introduce some experimental approaches that may help researchers to separate physiological and methodological issues. The upper thermal limits were influenced by both heating rates and body mass. In the latter case, the effect was physiological rather than methodological. The critical temperature decreased during subsequent tests performed on the same individual ants, even one week after the initial test. Accordingly, upper thermal limits may have been overestimated by our (and typical) protocols. Heating rates, body mass, procedures independent of temperature and other variables may affect the estimation of upper critical temperatures. Therefore, based on our data, we offer suggestions to enhance the quality of measurements, and offer recommendations to authors aiming to compile and analyze databases from the literature.

Energy substrate utilization during nightly vocal activity in three species of Scinax (Anura/Hylidae).

Anuran amphibians exhibit different patterns of energy substrate utilization that correlate with the intensity of vocal and locomotor activities. Given the remarkable differences among species in breeding and feeding strategies, and the different ways energy is used in the whole animal, the suggested correlations between calling and locomotor behavior and the level of energy substrates in the muscles responsible for such activities are more complex than previously reported. We explored the relationships between calling and locomotor behavior and energy supply to trunk and hindlimb muscles, respectively, within the ecologically diverse tree-frog genus Scinax. Specifically, we measured the relative amount of carbohydrates and lipids in these two groups of muscles, and in the liver of three species of Scinax that differ in vocal and locomotor performance, and compared our results with those of two other species for which comparable data are available. We also compared the contents of lipids and carbohydrates of conspecific males collected at the beginning and after 4 h of calling activity. The stomach content to potential feeding opportunities across species was also assessed in both groups of males. Scinax hiemalis and S. rizibilis exhibit comparatively low and episodic calling during long periods of activity whereas S. crospedospilus calls at higher rates over shorter periods. Male S. hiemalis had highest levels of trunk muscle glycogen followed by those of S. rizilbilis and S. crospedospilus, respectively. There was no correlation between total lipid content in trunk muscle and calling rate among different species, suggesting that other metabolic aspects may be responsible for the energetic support for vocal activity. The levels of lipids and carbohydrates in trunk and hindlimb muscles and liver of males collected at the beginning and 4 h into the calling period were similar across species, so the extent of energetic reserves does not appear to constrain vocal or locomotor activity. Finally, we found exceptionally high levels of carbohydrates and lipids in the liver of S. rizibilis, a trait perhaps related to a long and demanding breeding period.

Comparative biochemistry and physiology in Brazil: a critical appraisal.

Brazil stood out as the country with the highest number of submissions to the editorial project dedicated to Latin America by the journal Comparative Biochemistry and Physiology. Therefore, we felt that it was important to critically discuss the state of comparative biochemistry and physiology in this country. Our study is based on data collected from the ISI Web-of-Science. We analyzed publication trends through time, availability of novel approaches and techniques, patterns of collaboration among different geographical regions, patterns of collaboration with researchers abroad, and relative efforts dedicated to the study of biochemical and physiological adaptation of native fauna representing different terrestrial Brazilian biomes. Overall, our data shows that comparative biochemistry and physiology is a lively and productive discipline, but that some biases limit the scope of the field in Brazil. Some important limitations are the very heterogeneous distribution of research nuclei throughout the country and the absence of some important approaches, such as remote sensing and the use of molecular biology techniques in a comparative or evolutionary context. We also noticed that international collaboration far surpasses interregional collaboration, and discuss the possible causes and consequences of this situation. Finally, we found that Brazilian comparative biochemistry and physiology is biome-biased, as the Amazonian fauna has received far more attention than the whole pool of fauna representing other terrestrial biomes. We discuss the possible causes of these biases, and propose some directions that may contribute to invigorate the field in the country.

Control of metabolic rate is a hidden variable in the allometric scaling of homeotherms.

The allometric scaling exponent of the relationship between standard metabolic rate (SMR) and body mass for homeotherms has a long history and has been subject to much debate. Provided the external and internal conditions required to measure SMR are met, it is tacitly assumed that the metabolic rate (B) converges to SMR. If SMR does indeed represent a local minimum, then short-term regulatory control mechanisms should not operate to sustain it. This is a hidden assumption in many published articles aiming to explain the scaling exponent in terms of physical and morphological constraints. This paper discusses the findings of a minimalist body temperature (Tb) control model in which short-term controlling operations, related to the difference between Tb and the set-point temperatures by specific gains and time delays in the control loops, are described by a system of differential equations of Tb, B and thermal conductance. We found that because the gains in the control loops tend to increase as body size decreases (i.e. changes in B and thermal conductance are speeded-up in small homeotherms), the equilibrium point of the system potentially changes from asymptotically stable to a centre, transforming B and Tb in oscillating variables. Under these specific circumstances the very concept of SMR no longer makes sense. A series of empirical reports of metabolic rate in very small homeotherms supports this theoretical prediction, because in these animals B seems not to converge to a SMR value. We conclude that the unrestricted use of allometric equations to relate metabolic rate to body size might be misleading because metabolic control itself experiences size effects that are overlooked in ordinary allometric analysis.