Novel rebreathing adaptation extends dive time in a semi-aquatic lizard.

Bubble use evolved in many small invertebrates to enable underwater respiration, but, until recently, there has been no evidence that vertebrate animals use bubbles in a similar manner. Only one group of vertebrates, semi-aquatic Anolis lizards, may be an exception: these lizards dive underwater when threatened and, while underwater, rebreathe a bubble of air over their nostrils. Although it seems that rebreathing should be adaptive, possibly functioning to extend the time that lizards remain in underwater refugia, this has not been empirically tested. Here, I demonstrate that rebreathing serves to extend dive time in a semi-aquatic anole, Anolis aquaticus. I prevented the formation of normal rebreathing bubbles by applying a commercial emollient on the skin surface where bubbles form to assess the impact of bubbles on rebreathing cycles, gular pumps, and dive times. Lizards that were allowed to rebreathe normally remained underwater an average of 32% longer than those with impaired rebreathing, suggesting a functional role of rebreathing in underwater respiration. Unlike rebreathing, gular pumping was unaffected by treatment and may warrant further research regarding its role in supplementing underwater respiration. This study provides evidence that vertebrates can use bubbles to respire underwater and raises questions about adaptive mechanisms and potential bio-inspired applications.

Ambient light spectrum affects larval Mexican jumping bean moth (Cydia saltitans) behavior despite light obstruction from host seed.

Spectral differences in ambient light can affect animal behavior and convey crucial information about an individual's environment. The ability to perceive and respond to differences in ambient light varies widely by taxa and is shaped by a species' ecology. Mexican jumping bean moths, Cydia saltitans, spend their entire larval period encased in fallen host seeds and contend with potentially lethal environmental temperatures when host seeds are in direct sunlight. We investigate if and how C. saltitans larvae in host seeds respond to lighting conditions associated with these thermal risks. In a temperature-controlled experiment, we identified that larvae demonstrated distinct behavioral ("jumping") responses corresponding to four lighting treatments (white, red, green, and purple), despite extremely minimal light penetration through host seed walls. Red light induced the greatest larval activity (measured by probability of movement and by displacement from origin), suggesting that larvae have mechanisms to perceive low levels of red light and/or to detect subtle increases in heat produced by red/near infrared-biased light spectra, possibly providing them with an early-warning mechanism against thermal stress. Our findings highlight the interplay of environmental lighting, behavior, and potential thermosensory adaptations in a species with a visually constrained environment.

Keeping it cool to take the heat: tropical lizards have greater thermal tolerance in less disturbed habitats.

Global climate change has profound effects on species, especially those in habitats already altered by humans. Tropical ectotherms are predicted to be at high risk from global temperature increases, particularly those adapted to cooler temperatures at higher altitudes. We investigated how one such species, the water anole (Anolis aquaticus), is affected by temperature stress similar to that of a warming climate across a gradient of human-altered habitats at high elevation sites. We conducted a field survey on thermal traits and measured lizard critical thermal maxima across the sites. From the field survey, we found that (1) lizards from the least disturbed site and (2) operative temperature models of lizards placed in the least disturbed site had lower temperatures than those from sites with histories of human disturbance. Individuals from the least disturbed site also demonstrated greater tolerance to high temperatures than those from the more disturbed sites, in both their critical thermal maxima and the time spent at high temperatures prior to reaching critical thermal maxima. Our results demonstrate within-species variability in responses to high temperatures, depending on habitat type, and provide insight into how tropical reptiles may fare in a warming world.

Rapid Body Color Change Provides Lizards with Facultative Crypsis in the Eyes of Their Avian Predators.

AbstractColor change serves many antipredator functions and may allow animals to better match environments or disrupt outlines to prevent detection. Rapid color change could potentially provide camouflage to animals that frequently move among microhabitats. Determining the adaptiveness of whole-animal rapid color changes in natural habitats with respect to predator visual systems would greatly broaden our fundamental understanding of the evolution of rapid color change. We tested whether whole-body color change provides water anoles (Anolis aquaticus) with camouflage against avian predators and whether these rapid changes allow them to shift between environment matching and edge disruption. We manipulated A. aquaticus placement in natural microhabitats and used digital image analysis to quantify color matching, pattern matching, and edge disruption produced by microhabitat-induced color change. Color change reduced lizard detectability to predators in microhabitat-specific ways. Environment matching was favored when lizards were in solid-colored microhabitats, regardless of exposure to predators. Edge disruption was instead induced by high exposure and varied by body region. We provide the first evidence that rapid color change permits a tetrapod to flexibly employ the most optimal camouflaging strategy by form (e.g., color matching vs. edge disruption) to minimize detection in the eyes of its predators.

Amphibian breeding phenology influences offspring size and response to a common wetland contaminant.

BACKGROUND: Increases in temperature variability associated with climate change have critical implications for the phenology of wildlife across the globe. For example, warmer winter temperatures can induce forward shifts in breeding phenology across taxa ("false springs"), which can put organisms at risk of freezing conditions during reproduction or vulnerable early life stages. As human activities continue to encroach on natural ecosystems, it is also important to consider how breeding phenology interacts with other anthropogenic stressors (e.g., pollutants). Using 14 populations of a widespread amphibian (wood frog; Rana sylvatica), we compared 1) growth; 2) tolerance to a common wetland contaminant (NaCl); and 3) the ability of tadpoles to acclimate to lethal NaCl exposure following sublethal exposure earlier in life. We evaluated these metrics across two breeding seasons (2018 and 2019) and across populations of tadpoles whose parents differed in breeding phenology (earlier- versus later-breeding cohorts). In both years, the earlier-breeding cohorts completed breeding activity prior to a winter storm and later-breeding cohorts completed breeding activities after a winter storm. The freezing conditions that later-breeding cohorts were exposed to in 2018 were more severe in both magnitude and duration than those in 2019. RESULTS: In 2018, offspring of the later-breeding cohort were larger but less tolerant of NaCl compared to offspring of the earlier-breeding cohort. The offspring of the earlier-breeding cohort additionally were able to acclimate to a lethal concentration of NaCl following sublethal exposure earlier in life, while the later-breeding cohort became less tolerant of NaCl following acclimation. Interestingly, in 2019, the warmer of the two breeding seasons, we did not detect the negative effects of later breeding phenology on responses to NaCl. CONCLUSIONS: These results suggest that phenological shifts that expose breeding amphibians to freezing conditions can have cascading consequences on offspring mass and ability to tolerate future stressors but likely depends on the severity of the freeze event.

Repeated evolution of underwater rebreathing in diving Anolis lizards.

Air-based respiration limits the use of aquatic environments by ancestrally terrestrial animals. To overcome this challenge, diving arthropods have evolved to respire without resurfacing using air held between their cuticle and surrounding water.1-4 Inspired by natural history observations in Haiti (unpublished data) and Costa Rica,5,6 we conducted experiments documenting routine air-based underwater respiration in several distantly related semi-aquatic Anolis lizard species. Semi-aquatic anoles live along neotropical streams and frequently dive for refuge or food,7-12 remaining underwater for up to 18 min. While submerged, these lizards iteratively expire and re-inspire narial air bubbles-underwater "rebreathing." Rebreathed air is used in respiration, as the partial pressure of oxygen in the bubbles decreases with experimental submersion time in living anoles, but not in mechanical controls. Non-aquatic anoles occasionally rebreathe when submerged but exhibit more rudimentary rebreathing behaviors. Anole rebreathing is facilitated by a thin air layer (i.e., a "plastron," sensu Brocher13) supported by the animal's rugose skin upon submergence. We suggest that hydrophobic skin, which we observed in all sampled anoles,14,15 may have been exaptative, facilitating the repeated evolution of specialized rebreathing in species that regularly dive. Phylogenetic analyses strongly suggest that specialized rebreathing is adaptive for semi-aquatic habitat specialists. Air-based rebreathing may enhance dive performance by incorporating dead space air from the buccal cavity or plastron into the lungs, facilitating clearance of carbon dioxide, or allowing uptake of oxygen from surrounding water (i.e., a "physical gill" mechanism4,16).

Dewlap size in male water anoles associates with consistent inter-individual variation in boldness.

Male sexually selected signals can indicate competitive ability by honestly signaling fitness-relevant traits such as condition or performance. However, behavior can also influence contest outcomes; in particular, boldness often predicts dominance rank and mating success. Here, we sought to determine whether male ornament size is associated with consistent individual differences in boldness in water anoles Anolis aquaticus. We measured the relative size of the dewlap, a flap of skin under the chin that is a sexually selected ornament in Anolis lizards, and tested for associations with responses to a novel and potentially risky environment: time to emerge from a refuge into an arena and number of head scans post-emergence. We found that individuals consistently differed in both time to emerge and head scanning (i.e., individual responses were repeatable), and that dewlap size was negatively related to number of head scans. This suggests that ornament size could indicate male boldness if scanning represents antipredator vigilance. We found that males that had larger relative dewlaps were also in better body condition, but boldness (i.e., head scanning) was not related to condition. Lastly, we found consistent differences in behavior between trials, showing that anoles were becoming habituated or sensitized to the testing arena. Overall, our study shows that in addition to indicating condition and performance, dewlap size could also honestly indicate male boldness in Anolis lizards.

Frogs adapt to physiologically costly anthropogenic noise.

Human activities impose novel pressures on amphibians, which are experiencing unprecedented global declines, yet population-level responses are poorly understood. A growing body of literature has revealed that noise is an anthropogenic stressor that impacts ecological processes spanning subcellular to ecosystem levels. These consequences can impose novel selective pressures on populations, yet whether populations can adapt to noise is unknown. We tested for adaptation to traffic noise, a widespread sensory 'pollutant'. We collected eggs of wood frogs (Rana sylvatica) from populations from different traffic noise regimes, reared hatchlings under the same conditions, and tested frogs for differences in sublethal fitness-relevant effects of noise. We show that prolonged noise impaired production of antimicrobial peptides associated with defence against disease. Additionally, noise and origin site interacted to impact immune and stress responses. Noise exposure altered leucocyte production and increased baseline levels of the stress-relevant glucocorticoid, corticosterone, in frogs from quiet sites, but noise-legacy populations were unaffected. These results suggest noise-legacy populations have adapted to avoid fitness-relevant physiological costs of traffic noise. These findings advance our understanding of the consequences of novel soundscapes and reveal a pathway by which anthropogenic disturbance can enable adaptation to novel environments.

Does repeated human handling of study animals during the mating season affect their offspring?

Minimizing disturbance of study animals is a major consideration in ethological and ecological research design. One nearly universal type of disturbance is the handling of study animals as a component of trial setup. Even low to moderate levels of handling can be a substantial stressor to study animals, which may negatively affect their offspring via maternal effects. Understanding how routine human handling and manipulation may affect the outcome of research studies is therefore critical for interpreting study outcomes. We tested whether repeatedly handling and manipulating (i.e., manually disengaging) amplexed pairs of wood frogs (Rana sylvatica [Lithobates sylvaticus]), which have an explosive breeding season, would affect their reproductive output and offspring fitness. Handling and manipulation did not alter any parameter that we measured: reproductive timing, hatching success, and offspring larval duration, survival, and size at metamorphosis. These results suggest that handling and manipulation by researchers may have a negligible effect on wood frog reproduction and offspring fitness. It is possible that many species that are commonly used in reproductive studies because they suppress behavioral and physiological responses during the mating season are likewise unaffected by human handling. Nevertheless, researchers should examine possible consequences of methodological interventions on their study species in order to determine any potential influence on their results. Having a broad understanding of these effects on species that have robust or dampened stress responsiveness during the breeding season would be useful for making generalizations about potential effects.

Trait allometries generate super-honesty in Anolis dewlaps and may underlie sexual dimorphism.

Whether or not sexually selected traits consistently exhibit positive allometry (i.e. are disproportionately large in larger individuals) is an ongoing debate. Multiple models and exceptions to this rule suggest that the underlying drivers of sexual trait allometry are nuanced. Here, we compare allometries of sexual and non-sexual traits of a species (Anolis aquaticus) within a well-studied lizard genus to test the competing hypotheses that sexual traits are, or are not, defined by positive allometry. We further consider the relationships of trait functions, which are relatively well understood in the genus Anolis, and allometry to identify potential drivers of allometric patterns. In particular, we explore how trait allometries interact to influence total organism function and generate sexual dimorphism. We quantified size (of targeted traits) and color of a sexual signal (the dewlap) in Anolis aquaticus in the field. The dewlap conveyed information relevant to intra-sexual combat and exhibited positive allometry. Overall, our results suggest that using single-trait allometries as indicators of past selection provides only an incomplete understanding of trait evolution. Although the function of positive allometry in some individual sexual signals (e.g. those conveying "super-honest" information) may be straightforward, we illustrate how scaling relationships interact synergistically to influence the function of phenotypes and propose avenues for future research.

Bearded ladies: females suffer fitness consequences when bearing male traits.

A central assumption in evolutionary biology is that females of sexually dimorphic species suffer costs when bearing male secondary sexual traits, such as ornamentation. Nevertheless, it is common in nature to observe females bearing rudimentary versions of male ornaments (e.g. 'bearded ladies'), as ornaments can be under similar genetic control in both sexes. Here, we provide evidence that masculinized females incur both social and reproductive costs in nature. Male fence lizards (Sceloporus undulatus) discriminated against ornamented females during mate choice. Ornamented females had lower reproductive output, and produced eggs that were laid and hatched later than those of non-ornamented females. These findings support established theories of the evolution of sexual dimorphism and intralocus sexual conflict, and raise questions regarding the persistence of masculinizing ornamentation in females.