Physalaemus Pustulosus Research Articles

Urban sensory conditions alter rival interactions and mate choice in urban and forest túngara frogs.

Sexual communication often takes place in networks with multiple competing signalers being simultaneously assessed by mate choosers. Altered sensory conditions, such as noise and light pollution, can affect communication by altering signal production and perception. While evidence of sensory pollution affecting sexual signaling is widespread, few studies assess impacts on sexual signaling during rival interactions as well as mate choice, let alone whether urban and non-urban populations have diverged in their response. Here, we investigate the effects of urban sensory conditions on sexual communication in urban and forest túngara frogs (Engystomops pustulosus). We recorded dyadic vocal rival interactions and assessed mate choice with and without noise and light pollution in the lab. We show that urban sensory conditions can directly impact the intensity of rival interactions, differences between rivals, and mate choice, though changes were often in opposite directions for frogs of urban and forest origins. Moreover, we demonstrate that urban-induced changes in rival interactions can also indirectly affect how females choose between potential mates. Our study reveals origin-dependent direct and indirect effects of noise and light pollution and suggests local adaptation of sexual communication in urban populations.

Ten new records of anomalies for eight species of anurans – amphibians from Honduras and Colombia

ABSTRACT In recent decades, there has been a significant increase in reported anomalies among amphibians worldwide, especially malformations. These anomalies not only signify a decline in the quality of their habitats but also hold special significance among anuran species as vital indicators of ecosystem health. Our study presents ten previously undocumented cases of anomalies within eight anuran species found in Honduras and Colombia. These species include Ameerega bilinguis, A. hahneli and A. ingeri (Dendrobatidae), Craugastor sp. (Craugastoridae), Engystomops pustulosus (Leptodactylidae), Incilius valliceps (Bufonidae), Lithobates maculatus (Ranidae), and Smilisca baudinii (Hylidae). The observed anomalies encompass a range of abnormalities, including a protuberant tumor (1), adactyly (1), amely (2), anophthalmy (1), brachydactyly (1), ectrodactyly (1), ectromely (5), and phocomely (1), which affect the mobility and vision of these individuals. While the root causes may differ, we suspect that pollution stemming from agricultural and urban activities in the observed areas could be a contributing factor. However, we do not dismiss the possibility of other physicochemical or biological stressors.

Condition dependence in the sexual communication system of the túngara frog

Sexual selection can result in the evolution of extreme armaments and ornaments, and the development and maintenance of these traits can come at a considerable cost. These costs have been implicated in enforcing an upper limit on trait divergence and promoting condition-dependent traits, such that only individuals in sufficiently high condition can effectively wield these armaments and advertise these ornaments. Numerous studies demonstrate the condition-dependence of sexually selected traits, especially those used by males to advertise to females. In this study, we investigated condition-dependent mating calls in the túngara frog Physalaemus (= Engystomops) pustulosus. We manipulated male condition in the laboratory over a nine-day period by restricting food availability. We then documented: the relationship between male condition (the relative change in body mass from night 1 to night 9) and acoustic parameters of his mating call; how male condition influenced the male’s responses to call playbacks; and finally, how male condition influenced the attractiveness of the male’s calls to females. Males who were not fed during this period showed significant changes in call frequency, duration, and amplitude. In response to playbacks, unfed males called less, and made fewer complex calls. Finally, in phonotaxis experiments, females were more attracted to the calls of unfed males on night 1 to the calls of the same males on night 9. Fed males, on the other hand, showed no significant differences between nights 1 and 9 in call parameters, calling effort, and call attractiveness. This study shows the pervasive effects of condition on three aspects of sexual communication: signal parameters, behavioral response to vocal competition, and mating call attractiveness.

Supernumerary presence of the oribatid mite Archegozetes sp. Grandjean, 1931 in stomach contents of the túngara frog Engystomops pustulosus (Cope, 1864) encourages validation of interaction pathways: phoretic, parasitic, or prey?

ABSTRACT The trhypochthoniid mite Archegozetes sp. was identified at high abundances (N = 126 and 46) in the stomach contents of the túngara frog Engystomops pustulosus. This is the second report of the genus Archegozetes for Colombia. This species has been reported in Central and South America attached to the skin of some anurans, and these observations were assumed as either phoretic or parasitic interactions between both groups. However, based on our data and the fact that Archegozetes forms large aggregations, we hypothesize that 1) E. pustulosus exploits this ecological behaviour through a predator-prey interaction, and that 2) external presence previously reported in the frogs may have been accidental while the frog fed on these large groups of oribatid mites.

Conserved chromatin and repetitive patterns reveal slow genome evolution in frogs

Frogs are an ecologically diverse and phylogenetically ancient group of anuran amphibians that include important vertebrate cell and developmental model systems, notably the genus Xenopus. Here we report a high-quality reference genome sequence for the western clawed frog, Xenopus tropicalis, along with draft chromosome-scale sequences of three distantly related emerging model frog species, Eleutherodactylus coqui, Engystomops pustulosus, and Hymenochirus boettgeri. Frog chromosomes have remained remarkably stable since the Mesozoic Era, with limited Robertsonian (i.e., arm-preserving) translocations and end-to-end fusions found among the smaller chromosomes. Conservation of synteny includes conservation of centromere locations, marked by centromeric tandem repeats associated with Cenp-a binding surrounded by pericentromeric LINE/L1 elements. This work explores the structure of chromosomes across frogs, using a dense meiotic linkage map for X. tropicalis and chromatin conformation capture (Hi-C) data for all species. Abundant satellite repeats occupy the unusually long (~20 megabase) terminal regions of each chromosome that coincide with high rates of recombination. Both embryonic and differentiated cells show reproducible associations of centromeric chromatin and of telomeres, reflecting a Rabl-like configuration. Our comparative analyses reveal 13 conserved ancestral anuran chromosomes from which contemporary frog genomes were constructed.

Genomic footprints of (pre) colonialism: Population declines in urban and forest túngara frogs coincident with historical human activity.

Urbanisation is rapidly altering ecosystems, leading to profound biodiversity loss. To mitigate these effects, we need a better understanding of how urbanisation impacts dispersal and reproduction. Two contrasting population demographic models have been proposed that predict that urbanisation either promotes (facilitation model) or constrains (fragmentation model) gene flow and genetic diversity. Which of these models prevails likely depends on the strength of selection on specific phenotypic traits that influence dispersal, survival, or reproduction. Here, we a priori examined the genomic impact of urbanisation on the Neotropical túngara frog (Engystomops pustulosus), a species known to adapt its reproductive traits to urban selective pressures. Using whole-genome resequencing for multiple urban and forest populations we examined genomic diversity, population connectivity and demographic history. Contrary to both the fragmentation and facilitation models, urban populations did not exhibit substantial changes in genomic diversity or differentiation compared with forest populations, and genomic variation was best explained by geographic distance rather than environmental factors. Adopting an a posteriori approach, we additionally found both urban and forest populations to have undergone population declines. The timing of these declines appears to coincide with extensive human activity around the Panama Canal during the last few centuries rather than recent urbanisation. Our study highlights the long-lasting legacy of past anthropogenic disturbances in the genome and the importance of considering the historical context in urban evolution studies as anthropogenic effects may be extensive and impact nonurban areas on both recent and older timescales.

Ripple effects in a communication network: anti-eavesdropper defence elicits elaborated sexual signals in rival males.

Emitting conspicuous signals into the environment to attract mates comes with the increased risk of interception by eavesdropping enemies. As a defence, a commonly described strategy is for signallers to group together in leks, diluting each individual's risk. Lekking systems are often highly social settings in which competing males dynamically alter their signalling behaviour to attract mates. Thus, signalling at the lek requires navigating fluctuations in risk, competition and reproductive opportunities. Here, we investigate how behavioural defence strategies directed at an eavesdropping enemy have cascading effects across the communication network. We investigated these behaviours in the túngara frog (Engystomops pustulosus), examining how a calling male's swatting defence directed at frog-biting midges indirectly affects the calling behaviour of his rival. We found that the rival responds to swat-induced water ripples by increasing his call rate and complexity. Then, performing phonotaxis experiments, we found that eavesdropping fringe-lipped bats (Trachops cirrhosus) do not exhibit a preference for a swatting male compared to his rival, but females strongly prefer the rival male. Defences to minimize attacks from eavesdroppers thus shift the mate competition landscape in favour of rival males. By modulating the attractiveness of signalling prey to female receivers, we posit that eavesdropping micropredators likely have an unappreciated impact on the ecology and evolution of sexual communication systems.

Noise and light pollution elicit endocrine responses in urban but not forest frogs

Urban areas are characterised by the presence of sensory pollutants, such as anthropogenic noise and artificial light at night (ALAN). Animals can quickly adapt to novel environmental conditions by adjusting their behaviour, which is proximately regulated by endocrine systems. While endocrine responses to sensory pollution have been widely reported, this has not often been linked to changes in behaviour, hampering the understanding of adaptiveness of endocrine responses. Our aim was, therefore, to investigate the effects of urbanisation, specifically urban noise and light pollution, on hormone levels in male urban and forest túngara frogs (Engystomops pustulosus), a species with reported population divergence in behaviour in response to urbanisation. We quantified testosterone and corticosterone release rates in the field and in the lab before and after exposure to urban noise and/or light. We show that urban and forest frogs differ in their endocrine phenotypes under field as well as lab conditions. Moreover, in urban frogs exposure to urban noise and light led, respectively, to an increase in testosterone and decrease in corticosterone, whereas in forest frogs sensory pollutants did not elicit any endocrine response. Our results show that urbanisation, specifically noise and light pollution, can modulate hormone levels in urban and forest populations differentially. The observed endocrine responses are consistent with the observed behavioural changes in urban frogs, providing a proximate explanation for the presumably adaptive behavioural changes in response to urbanisation.

Purification, conformational analysis and cytotoxic activities of host-defense peptides from the Tungara frog Engystomops pustulosus (Leptodactylidae; Leiuperinae)

The amphibian family Leptodactylidae is divided into three sub-families: Leiuperinae, Leptodactylinae, and Paratelmatobiinae. Host-defense peptides (HDPs) present in the skins of frogs belonging to the Leptodactylinae have been studied extensively, but information is limited regarding peptides from Leiuperinae species. Peptidomic analysis of norepinephrine-stimulated skin secretions from the Tungara frog Engystomops pustulosus (Leiuperinae) collected in Trinidad led to the isolation and structural characterization of previously undescribed pustulosin-1 (FWKADVKEIG KKLAAKLAEELAKKLGEQ), [Q28E] pustulosin-1 (pustulosin-2), and pustulosin-3 (DWKETAKELLKKIGAKVAQVISDKLNPAPQ). The primary structures of these peptides do not resemble those of previously described frog skin HDPs. In addition, the secretions contained tigerinin-1EP (GCKTYLIEPPVCT) with structural similarity to the tigerinins previously identified in skin secretions from frogs from the family Dicroglossidae. Pustulosin-1 and -3 adopted extended α-helical conformations in 25% trifluoroethanol–water and in the presence of cell membrane models (sodium dodecylsulfate and dodecylphosphocholine micelles). Pustulosin-1 and -3 displayed cytotoxic activity against a range of human tumor-derived cell lines (A549, MDA-MB-231, and HT29), but their therapeutic potential for development into anti-cancer agents is limited by their comparable cytotoxic activity against non-neoplastic human umbilical vein endothelial cells. The peptides also displayed weak antimicrobial activity against Escherichia coli (MIC = 125 µM) but were inactive against Staphylococcus aureus. Tigerinin-1EP was inactive against both the tumor-derived cells and bacteria.

The time course of acclimation of critical thermal maxima is modulated by the magnitude of temperature change and thermal daily fluctuations

Plasticity in the critical thermal maximum (CTmax) helps ectotherms survive in variable thermal conditions. Yet, little is known about the environmental mechanisms modulating its time course. We used the larvae of three neotropical anurans (Boana platanera, Engystomops pustulosus and Rhinella horribilis) to test whether the magnitude of temperature changes and the existence of fluctuations in the thermal environment affected both the amount of change in CTmax and its acclimation rate (i.e., its time course). For that, we transferred tadpoles from a pre-treatment temperature (23 °C, constant) to two different water temperatures: mean (28 °C) and hot (33 °C), crossed with constant and daily fluctuating thermal regimes, and recorded CTmax values, daily during six days. We modeled changes in CTmax as an asymptotic function of time, temperature, and the daily thermal fluctuation. The fitted function provided the asymptotic CTmax value (CTmax∞) and CTmax acclimation rate (k). Tadpoles achieved their CTmax∞ between one and three days. Transferring tadpoles to the hot treatment generated higher CTmax∞ at earlier times, inducing faster acclimation rates in tadpoles. In contrast, thermal fluctuations equally led to higher CTmax∞ values but tadpoles required longer times to achieve CTmax∞ (i.e., slower acclimation rates). These thermal treatments interacted differently with the studied species. In general, the thermal generalist Rhinella horribilis showed the most plastic acclimation rates whereas the ephemeral-pond breeder Engystomops pustulosus, more exposed to heat peaks during larval development, showed less plastic (i.e., canalized) acclimation rates. Further comparative studies of the time course of CTmax acclimation should help to disentangle the complex interplay between the thermal environment and species ecology, to understand how tadpoles acclimate to heat stress.

The effect of heterospecific and conspecific competition on inter-individual differences in tungara frog tadpole (Engystomops pustulosus) behavior.

Repeated social interactions with conspecifics and/or heterospecifics during early development may drive the differentiation of behavior among individuals. Competition is a major form of social interaction and its impacts can depend on whether interactions occur between conspecifics or heterospecifics and the directionality of a response could be specific to the ecological context that they are measured in. To test this, we reared tungara frog tadpoles (Engystomops pustulosus) either in isolation, with a conspecific tadpole or with an aggressive heterospecific tadpole, the whistling frog tadpole (Leptodactylus fuscus). In each treatment, we measured the body size and distance focal E. pustulosus tadpoles swam in familiar, novel and predator risk contexts six times during development. We used univariate and multivariate hierarchical mixed effect models to investigate the effect of treatment on mean behavior, variance among and within individuals, behavioral repeatability and covariance among individuals in their behavior between contexts. There was a strong effect of competition on behavior, with different population and individual level responses across social treatments. Within a familiar context, the variance in the distance swam within individuals decreased under conspecific competition but heterospecific competition caused more variance in the average distance swam among individuals. Behavioral responses were also context specific as conspecific competition caused an increase in the distance swam within individuals in novel and predator risk contexts. The results highlight that the impact of competition on among and within individual variance in behavior is dependent on both competitor species identity and context.

Anthropogenic noise and light alter temporal but not spatial breeding behavior in a wild frog.

Increasing urbanization has led to large-scale land-use changes, exposing persistent populations to drastically altered environments. Sensory pollutants, including low-frequency anthropogenic noise and artificial light at night (ALAN), are typically associated with urban environments and known to impact animal populations in a variety of ways. Both ALAN and anthropogenic noise can alter behavioral and physiological processes important for survival and reproduction, including communication and circadian rhythms. Although noise and light pollution typically co-occur in urbanized areas, few studies have addressed their combined impact on species' behavior. Here, we assessed how anthropogenic noise and ALAN can influence spatial and temporal variation in breeding activity of a wild frog population. By exposing artificial breeding sites inside a tropical rainforest to multiple sensory environments, we found that both anthropogenic noise and ALAN impact breeding behavior of túngara frogs (Engystomops pustulosus), albeit in different ways. Males arrived later in the night at their breeding sites in response to anthropogenic noise. ALAN, on the other hand, led to an increase in calling effort. We found no evidence that noise or light pollution either attracted frogs to or repelled frogs from breeding sites. Thus, anthropogenic noise may negatively affect calling males by shifting the timing of sexual signaling. Conversely, ALAN may increase the attractiveness of calling males. These changes in breeding behavior highlight the complex ways that urban multisensory pollution can influence behavior and suggest that such changes may have important ecological implications for the wildlife that are becoming increasingly exposed to urban multisensory pollution.

Interactive and independent effects of light and noise pollution on sexual signaling in frogs

Urbanization drastically changes environmental conditions, including the introduction of sensory pollutants, such as artificial light at night (ALAN) and anthropogenic noise. To settle in urban habitats, animals need to cope with this new sensory environment. On a short timescale, animals might cope with sensory pollutants via behavioral adjustments, such as changes in sexual signaling, which can have important fitness consequences. While ALAN and anthropogenic noise generally co-occur in urban habitats and are known to be able to interact to modify behavioral responses, few studies have addressed their combined impact. Our aim was, therefore, to assess the effects of ALAN, anthropogenic noise, and their interaction on sexual signaling in túngara frogs (Engystomops pustulosus). We observed the calling behavior of frogs in urban and forest areas, and subsequently recorded these frogs in a laboratory set-up while independently manipulating light and noise levels. Frogs in urban areas called with a higher call rate and complexity, which was correlated with local sensory conditions. Furthermore, our lab experiment revealed that ALAN can directly alter sexual signaling independently as well as in combination with anthropogenic noise. Exposure to ALAN alone increased call amplitude, whereas a combination of ALAN and anthropogenic noise interacted to lead to a higher call complexity and amplitude. Overall, the response patterns consistently showed that exposure to ALAN and anthropogenic noise led to more conspicuous sexual signals than expected based on the additive effects of single pollutants. Our results support the notion that urban and forest population differences in sexual signaling can be partially explained by exposure to ALAN and anthropogenic noise. Furthermore, by demonstrating interactive effects between light and noise pollution, our study highlights the importance of examining the effects of multisensory pollution, instead of single pollutants, when trying to understand phenotypic divergence in urbanized vs. natural areas.

Can you hear/see me? Multisensory integration of signals does not always facilitate mate choice

Abstract Females of many species choose mates using multiple sensory modalities. Multimodal noise may arise, however, in dense aggregations of animals communicating via multiple sensory modalities. Some evidence suggests multimodal signals may not always improve receiver decision-making performance. When sensory systems process input from multimodal signal sources, multimodal noise may arise and potentially complicate decision-making due to the demands on cognitive integration tasks. We tested female túngara frog, Physalaemus (=Engystomops) pustulosus, responses to male mating signals in noise from multiple sensory modalities (acoustic and visual). Noise treatments were partitioned into three categories: acoustic, visual, and multimodal. We used natural calls from conspecifics and heterospecifics for acoustic noise. Robotic frogs were employed as either visual signal components (synchronous vocal sac inflation with call) or visual noise (asynchronous vocal sac inflation with call). Females expressed a preference for the typically more attractive call in the presence of unimodal noise. However, during multimodal signal and noise treatments (robofrogs employed with background noise), females failed to express a preference for the typically attractive call in the presence of conspecific chorus noise. We found that social context and temporal synchrony of multimodal signaling components are important for multimodal communication. Our results demonstrate that multimodal signals have the potential to increase the complexity of the sensory scene and reduce the efficacy of female decision making.

Tuned in to communication sounds: Neuronal sensitivity in the túngara frog midbrain to frequency modulated signals

For complex communication signals, it is often difficult to identify the information-bearing elements and their parameters necessary to elicit functional behavior. Consequently, it may be difficult to design stimuli that test how neurons contribute to communicative processing. For túngara frogs (Physalaemus pustulosus), however, previous behavioral testing with numerous stimuli showed that a particular frequency modulated (FM) transition in the male call is required to elicit phonotaxis and vocal responses. Modeled on such behavioral experiments, we used awake in vivo recordings of single units in the midbrain to determine if their excitation was biased to behaviorally important FM parameters. Comparisons of stimulus driven action potentials revealed greatest excitation to the behaviorally important FM transition: a downward FM sweep or step that crosses ~600 Hz. Previous studies using long-duration acoustic exposure found immediate early gene expression in many midbrain neurons to be most sensitive to similar FM. However, those data could not determine if FM coding was accomplished by the population and/or individual neurons. Our data suggest both coding schemes could operate, as 1) individual neurons are more sensitive to the behaviorally significant FM transition and 2) when single unit recordings are analytically combined across cells, the combined code can produce high stimulus discrimination (FM vs. noise driven excitation), approaching that found in behavioral discrimination of call vs. noise.

Non-destructive methods to assess health of wild tropical frogs (túngara frogs: Engystomops pustulosus) in Trinidad reveal negative impacts of agricultural land

Amphibians are threatened globally with at least 43% of species declining and the most important stressor being habitat loss or degradation. Amphibians inhabiting highly biodiverse tropical regions are disproportionately threatened; however, the effects of landscape alterations on amphibian health are virtually unknown. In this study, we utilised non-destructive techniques to compare size (weight, snout-vent length [SVL]), body condition, male secondary sexual features (forelimb width, nuptial pad length) and breeding success (egg number, fertility [percentage fertilised eggs], hatching success) in túngara frogs (Engystomops pustulosus) collected from reference (n = 5), suburban (n = 6) and agricultural (n = 4) sites in Trinidad; characterised by presence/absence of crops/houses. All measured endpoints were negatively impacted in frogs collected from agricultural sites. The largest effect was observed for hatching success (2.77-fold lower) and egg number (2.5-fold lower). Less pronounced effects were observed on male frogs (weight: 1.77-fold lower; SVL: 1.18-fold lower; forelimb width: 1.33-fold lower; nuptial pad length: 1.15-fold lower). Our findings demonstrate negative impacts of agricultural sites on túngara frog health, with the number of viable offspring reduced by almost one third. The methods outlined here are technically simple and low-cost and thereby have potential for application to other species in order to investigate the potential impacts of habitat degradation on amphibian health. Furthermore, as these methods are non-destructive, they could be used to investigate the potential contribution of frog size and/or reproductive capability as a causative factor contributing to population declines in threatened species, which is particularly pressing in tropical regions.

Eavesdropping Micropredators as Dynamic Limiters of Sexual Signal Elaboration and Intrasexual Competition.

AbstractTo thoroughly understand the drivers of dynamic signal elaboration requires assessing the direct and indirect effects of naturally interacting factors. Here, we use structural equation modeling to test multivariate data from in situ observations of sexual signal production against a model of causal processes hypothesized to drive signal elaboration. We assess direct and indirect effects, and relative impacts, of male-male competition and attacks by eavesdropping frog-biting midges (Diptera: Corethrellidae) on call elaboration of male túngara frogs (Engystomops pustulosus). We find that the intensity of attacks by these micropredator flies drives the extent to which frogs elaborate their calls, likely due to a temporal trade-off between signaling and antimicropredator defense. Micropredator attacks appear to dynamically limit a male's call rate and complexity and consequently dampen the effects of intrasexual competition. In accounting for naturally interacting drivers of signal elaboration, this study presents a counterpoint to the mechanisms traditionally thought to drive sexual selection in this system. Moreover, the results shed light on the relatively unexamined and potentially influential role of eavesdropping micropredators in the evolution of sexual communication systems.

Serous Glands in the Skin of the Tungara Frog, <i>Engystomops pustulosus</i> (Cope, 1864) (Anura, Leptodactylidae): Degenerated Secretory Units are Selectively Removed by Macrophages

The cutaneous apparatus of Engystomops pustulosus (Cope, 1864) (the Tungara frog) includes serous glands that show impressive patterns of degeneration in their syncytial secretory units, and thus represent suitable organ models to investigate the role of macrophages in renewal processes of multicellular structures. The present case report exploits this chance and highlights that: (a) degenerating glands pertain to the Ia line of the polymorphic serous gland assortment in Tungara skin; (b) resident macrophages migrate from spongy dermis and remove syncytium debris; (c) secretory syncytium collapse results from impairment of the equilibrium between serous product manufacturing/storage and merocrine release into the dermal environment; (d) Intercalated tract (or gland neck) and myoepithelium (included its ortho-sympathetic nerve supply), are neither involved in degeneration nor affected by macrophage response. According to present evidence and current literature, it is concluded that the scavenger activity of macrophages prepares secretory unit renewal, performed by stem cells from the neck. In addition, gland functional rehabilitation may rely on effectiveness of the preexisting neuromuscular apparatus to achieve secretory bulk release onto the cutaneous surface.

Diurnal Calling in a Nocturnal Frog: Exceptional Calling Activity of Túngara Frogs (Engystomops pustulosus) on the Panamanian Island of Taboga

Túngara Frogs, Engystomops pustulosus, are known to reduce the complexity of their calls in the presence of predators. Although complex calls are more attractive to females, they also attract predators, particularly frog-eating bats, and are rarely emitted by solitary males. Therefore, if Túngara Frogs were to be released from predation pressure, as on the island of Taboga in the Gulf of Panama where frog-eating bats are absent, such constraints on calling should be alleviated. We compared the calling behavior of Túngara Frogs on Taboga with the calling behavior of those on Barro Colorado Island, located in central Panama where frog-eating bats are present, using timed video recordings. Compared with Túngara Frogs on Barro Colorado Island, male Túngara Frogs on Taboga called more, emitted consistently more calls in choruses, and called both day and night. On Taboga, even solitary males routinely embellished their calls with chuck elements. These results are consistent with a hypothesis of predator release positively affecting calling behavior.

Frog nest foams exhibit pharmaceutical foam-like properties.

Foams have frequently been used as systems for the delivery of cosmetic and therapeutic molecules; however, there is high variability in the foamability and long-term stability of synthetic foams. The development of pharmaceutical foams that exhibit desirable foaming properties, delivering appropriate amounts of the active pharmaceutical ingredient (API) and that have excellent biocompatibility is of great interest. The production of stable foams is rare in the natural world; however, certain species of frogs have adopted foam production as a means of providing a protective environment for their eggs and larvae from predators and parasites, to prevent desiccation, to control gaseous exchange, to buffer temperature extremes, and to reduce UV damage. These foams show great stability (up to 10 days in tropical environments) and are highly biocompatible due to the sensitive nature of amphibian skin. This work demonstrates for the first time that nests of the túngara frog (Engystomops pustulosus) are stable ex situ with useful physiochemical and biocompatible properties and are capable of encapsulating a range of compounds, including antibiotics. These protein foam mixtures share some properties with pharmaceutical foams and may find utility in a range of pharmaceutical applications such as topical drug delivery systems.