Heavy Metal Concentration in Neotropical Aquatic Snakes (Helicops pastazae) and Its Potential as a Bioindicator of Water Pollution.

The purpose of this study was to test the potential role of the aquatic snake Helicops pastazae as an indicator of water pollution caused by heavy metals. In particular, we tested whether the total heavy metal concentration is related to (1) the position (upstream vs downstream) of the sampling point and its distance from the point where wastewater is discharged; (2) the taxonomic group studied: piscivorous snakes vs characid fish that occupy the same habitats; and (3) the organ or tissue examined: snake liver versus muscle. We used atomic absorption spectrophotometry with electrothermal atomization to quantify cadmium (Cd), chromium (Cr) and lead (Pb) and found significant differences between some of the sampling points, with particularly high metal concentrations detected upstream at point 1. However, we found no clear spatial pattern nor any significant differences in the concentration of any of the metals in fish and snake muscle, suggesting that both species accumulate similar amounts of the sampled elements. With regard to interactions, snake liver had the highest concentrations of Cd, while muscle had the highest concentrations of Pb and Cr, which may indicate tissue affinity differences for certain metals. Altogether, our results indicate that H. pastazae accumulates contaminants differentially, depending on the tissue and location, which highlights their potential as bioindicators of water contamination. Further research is necessary to understand their role as bioindicators based on extensive sampling and environmental contaminant data.

First characterization of toxic alkaloids and volatile organic compounds (VOCs) in the cryptic dendrobatid Silverstoneia punctiventris.

BACKGROUND: Poison frogs are known for the outstanding diversity of alkaloid-based chemical defences with promising therapeutic applications. However, current knowledge about chemical defences in Dendrobatoidea superfamily has two sources of bias. First, cryptic, brown-colored species have been neglected in comparison to those conspicuously colored, and second, there has been little interest in characterizing metabolites other than alkaloids mediating defensive functions. In an effort to contribute to fill the gap of knowledge about cryptic species and broadening the spectrum of compounds analyzed we have applied head-space solid phase microextraction coupled to gas chromatography and mass spectrometry (HS-SPME/GC-MS) for extracting amphibian alkaloids and volatile organic compounds (VOCs) from Silverstoneia punctiventris. RESULTS: Using the skin from 8 specimens in 4 biological replicates we have found 33 different compounds. Twenty of them were classified as VOCs into 15 chemical classes including alkanes, alcohols, carbonyl compounds, methylpyridines, benzothiazoles, N-alkylpyrrolidines, pyrazines, and sesquiterpenoids, some of which were previously reported as repellents, defence compounds or defence pheromones in other organisms, and as sex pheromones in a treefrog. Interestingly, six of the remaining compounds were identified as alkaloids previously reported in other toxic/unpalatable dendrobatid frogs. CONCLUSIONS: This is the first report of alkaloids and VOCs found in the Silverstoneia genus, which has been assumed for decades as non-chemically defended. This study establishes HS-SPME/GC-MS as a new application for a simultaneous approach to amphibian alkaloids and VOCs in poison frogs while opens up new research questions to assess the co-occurrence of both type of compounds and to investigate the evolutionary significance of a defence gradient that includes olfactory avoidance, unpalatability, and toxicity in dendrobatids. In addition, our results show that amphibian alkaloids could have a dual function (olfactory at distance, taste by contact) never explored before neither in Silverstonaeia nor in any other dendrobatid species.

Divergence, gene flow, and the origin of leapfrog geographic distributions: The history of colour pattern variation in Phyllobates poison-dart frogs.

The geographic distribution of phenotypic variation among closely related populations is a valuable source of information about the evolutionary processes that generate and maintain biodiversity. Leapfrog distributions, in which phenotypically similar populations are disjunctly distributed and separated by one or more phenotypically distinct populations, represent geographic replicates for the existence of a phenotype, and are therefore especially informative. These geographic patterns have mostly been studied from phylogenetic perspectives to understand how common ancestry and divergent evolution drive their formation. Other processes, such as gene flow between populations, have not received as much attention. Here, we investigate the roles of divergence and gene flow between populations in the origin and maintenance of a leapfrog distribution in Phyllobates poison frogs. We found evidence for high levels of gene flow between neighbouring populations but not over long distances, indicating that gene flow between populations exhibiting the central phenotype may have a homogenizing effect that maintains their similarity, and that introgression between 'leapfroging' taxa has not played a prominent role as a driver of phenotypic diversity in Phyllobates. Although phylogenetic analyses suggest that the leapfrog distribution was formed through independent evolution of the peripheral (i.e. leapfrogging) populations, the elevated levels of gene flow between geographically close populations poise alternative scenarios, such as the history of phenotypic change becoming decoupled from genome-averaged patterns of divergence, which we cannot rule out. These results highlight the importance of incorporating gene flow between populations into the study of geographic variation in phenotypes, both as a driver of phenotypic diversity and as a confounding factor of phylogeographic inferences.

The evolution of polymorphism in the warning coloration of the Amazonian poison frog Adelphobates galactonotus.

While intraspecific variation in aposematic signals can be selected for by different predatory responses, their evolution is also contingent on other processes shaping genetic variation. We evaluate the relative contributions of selection, geographic isolation, and random genetic drift to the evolution of aposematic color polymorphism in the poison frog Adelphobates galactonotus, distributed throughout eastern Brazilian Amazonia. Dorsal coloration was measured for 111 individuals and genetic data were obtained from 220 individuals at two mitochondrial genes (mtDNA) and 7963 Single Nucleotide Polymorphisms (SNPs). Four color categories were described (brown, blue, yellow, orange) and our models of frog and bird visual systems indicated that each color was distinguishable for these taxa. Using outlier and correlative analyses we found no compelling genetic evidence for color being under divergent selection. A time-calibrated mtDNA tree suggests that the present distribution of dorsal coloration resulted from processes occurring during the Pleistocene. Separate phylogenies based on SNPs and mtDNA resolved the same well supported clades, each containing different colored populations. Ancestral character state analysis provided some evidence for evolutionary transitions in color type. Genetic structure was more strongly associated with geographic features, than color category, suggesting that the distribution of color is explained by localized processes. Evidence for geographic isolation together with estimates of low effective population size implicates drift as playing a key role in color diversification. Our results highlight the relevance of considering the neutral processes involved with the evolution of traits with important fitness consequences.

Genotyping of Fusarium Isolates from Onychomycoses in Colombia: Detection of Two New Species Within the Fusarium solani Species Complex and In Vitro Antifungal Susceptibility Testing.

Fusariosis have been increasing in Colombia in recent years, but its epidemiology is poorly known. We have morphologically and molecularly characterized 89 isolates of Fusarium obtained between 2010 and 2012 in the cities of Bogotá and Medellín. Using a multi-locus sequence analysis of rDNA internal transcribed spacer, a fragment of the translation elongation factor 1-alpha (Tef-1α) and of the RNA-dependent polymerase subunit II (Rpb2) genes, we identified the phylogenetic species and circulating haplotypes. Since most of the isolates studied were from onychomycoses (nearly 90 %), we carried out an epidemiological study to determine the risk factors associated with such infections. Five phylogenetic species of the Fusarium solani species complex (FSSC), i.e., F. falciforme, F. keratoplasticum, F. lichenicola, F. petroliphilum, and FSSC 6 as well as two of the Fusarium oxysporum species complex (FOSC), i.e., FOSC 3 and FOSC 4, were identified. The most prevalent species were FOSC 3 (38.2%) followed by F. keratoplasticum (33.7%). In addition, our isolates were distributed into 23 haplotypes (14 into FOSC and nine into FSSC). Two of the FSSC phylogenetic species and two haplotypes of FSSC were not described before. Our results demonstrate that recipients of pedicure treatments have a lower probability of acquiring onychomycosis than those not receiving such treatments. The antifungal susceptibility of all the isolates to five clinically available agents showed that amphotericin B was the most active drug, while the azoles exhibited lower in vitro activity.

Thermal ecology of montane Atelopus (Anura: Bufonidae): A study of intrageneric diversity.

Harlequin frogs (Bufonidae: Atelopus) are among the most threatened frog genus in the world and reach very high elevations in the tropical Andes and the Sierra Nevada de Santa Marta (SNSM). Learning about their thermal ecology is essential to infer sensitivity to environmental changes, particularly climate warming. We report on the activity temperature and thermoregulatory behavior of three high-elevation species of harlequin frogs, Atelopus nahumae, Atelopus laetissimus and Atelopus carrikeri. The first two mentioned live in streams in Andean rain forests, whereas A. carrikeri inhabits paramo streams in the SNSM. We studied the thermal ecology of these species in tree localities differing in altitude, and focused on activity body, operative, substrate and air temperature. A main trend was lower body temperature as elevation increased, so that differences among species were largely explained by differences in substrate temperature. However, this temperature variation was much lower in forest species than paramo species. The Atelopus species included in this work proved to be thermoconformers, a trend that not extended to all congenerics at high elevation. This diversity in thermal ecology poses important questions when discussing the impact of climate warming for high-elevation harlequin frogs. For example, forest species show narrow thermal ranges and, if highly specialized, may be more susceptible to temperature change. Paramo species such as A. carrikeri, in contrast, may be more resilient to temperature change.

Acoustic interference and recognition space within a complex assemblage of dendrobatid frogs.

In species-rich assemblages of acoustically communicating animals, heterospecific sounds may constrain not only the evolution of signal traits but also the much less-studied signal-processing mechanisms that define the recognition space of a signal. To test the hypothesis that the recognition space is optimally designed, i.e., that it is narrower toward the species that represent the higher potential for acoustic interference, we studied an acoustic assemblage of 10 diurnally active frog species. We characterized their calls, estimated pairwise correlations in calling activity, and, to model the recognition spaces of five species, conducted playback experiments with 577 synthetic signals on 531 males. Acoustic co-occurrence was not related to multivariate distance in call parameters, suggesting a minor role for spectral or temporal segregation among species uttering similar calls. In most cases, the recognition space overlapped but was greater than the signal space, indicating that signal-processing traits do not act as strictly matched filters against sounds other than homospecific calls. Indeed, the range of the recognition space was strongly predicted by the acoustic distance to neighboring species in the signal space. Thus, our data provide compelling evidence of a role of heterospecific calls in evolutionarily shaping the frogs' recognition space within a complex acoustic assemblage without obvious concomitant effects on the signal.

Molecular phylogenetics uncovers two new species in the genus Phyllobates (Anura, Dendrobatidae): the terrible frog gets two new sisters.

True poison-dart frogs (Phyllobates, Dendrobatidae) evolved the ability to secrete batrachotoxins, the most powerful alkaloids known to date. The genus comprises five species whose systematics, at first glance, appeared clear. The most derived clade would include two Colombian species (P.terribilis and P.bicolor) with the highest toxicity, the largest body size, and predominantly yellow body colouration. The other three species (P.aurotaenia, P.vittatus, and P.lugubris) are less toxic on average, have smaller size, and are predominantly black with bright dorsolateral stripes. Recent research has revealed the existence of two major lineages among the three Colombian species. The northern lineage appears to result from a complex evolutionary history, including perhaps introgression among yellow and black taxa. The southern lineage instead revealed the existence of new clades closely related to P.terribilis, black and yellow, that arguably deserve their recognition as new species. Here, available evidence is combined to support the erection of southern populations of P.aurotaenia as a new highly toxic species, sister to P.terribilis, and much closer to it than to any other yellow or black-bodied species, Phyllobatessamperi sp. nov. Their common ancestor is sister to an additional yellow species, which we also describe here as Phyllobatesbezosi sp. nov. Both new species can be externally diagnosed using colouration. Our previous and current analyses also suggest the existence of additional taxa and corroborate multiple transitions in colouration across these hypertoxic taxa.

ResumenLas verdaderas ranas de dardo venenoso (género Phyllobates) evolucionaron la capacidad de secretar batracotoxinas, los alcaloides más poderosos que se conocen hasta hoy. El género comprende cinco especies, cuya taxonomía parecía clara a primera vista. El clado más derivado incluye dos especies colombianas (P.terribilis and P.bicolor) con la mayor toxicidad, mayor tamaño y coloración predominantemente amarilla. Las otras tres especies (P.aurotaenia, P.vittatus and P.lugubris) son en general menos tóxicas, menores en tamaño y predominantemente negras con conspicuas manchas dorsolaterales.Una reciente investigación confirmó la existencia dos linajes mayores entre las tres especies colombianas. El del norte parece ser el producto de una historia evolutiva compleja, incluyendo quizás introgresión entre especies amarillas y negras. El del sur reveló en cambio la existencia de nuevos clados filogenéticamente cercanos a P.terribilis, uno negro y dos amarillos, que requieren reconocimiento formal como nuevas especies. Esta investigación combina la evidencia disponible para apoyar el reconocimiento de las poblaciones del sur de P.aurotaenia como una nueva especie negra, hermana de P.terribilis, y mucho más cercana a ella que a las otras especies negras o amarillas, Phyllobatessamperi sp. nov. Su ancestro común es a su vez el grupo hermano de una especie adicional, amarilla, que también describimos como Phyllobatesbezosi sp. nov. Las dos nuevas especies pueden ser externamente reconocidas por la coloración. Nuestros análisis previos y actuales también sugieren la existencia de otras especies no descritas y corroboran la existencia de múltiples transiciones de coloración en la evolución de las especies hipertóxicas.

A new species of Andean poison frog, Andinobates (Anura: Dendrobatidae), from the northwestern Andes of Colombia.

The poison frogs of the Colombian Andes, Pacific lowlands and Panama have been recently recognized as a new, monophyletic and well-supported genus: Andinobates. The species richness and distribution within Andinobates remain poorly understood due to the paucity of geographic, genetic and phenotypic data. Here we use a combination of molecular, bioacoustic and morphometric evidence to describe a new species of Andean poison frog: Andinobates cassidyhornae sp. nov. from the high elevation cloud forests of the Colombian Cordillera Occidental, in the northwestern Andes. The new species is associated to the bombetes group and characterized by a unique combination of ventral and dorsal color patterns. Data on 1119 bp from two mitochondrial markers allowed us to reject the null hypotheses that A. cassidyhornae sp. nov. is part of the phenotypically similar and geographically less distant species: A. opisthomelas, A. virolinensis or A. bombetes. The best available phylogenetic trees and the genetic distance to other Andinobates species further support this decision. Altogether, the advertisement call parameters unambiguously separated A. cassidyhornae sp. nov. calls from the calls of the three closest species. The new species adds to a poorly known and highly endangered genus of poison frogs that requires further studies and urgent conservation measures.

Two new species of Salamanders, Genus Bolitoglossa (Amphibia: Plethodontidae), from the Eastern Colombian Andes.

The salamander fauna of Colombia is very poorly known, probably because most research efforts have been devoted to anurans during the last two decades. Here, we describe two new species of the genus Bolitoglossa (Eladinea) from the eastern flank of the Eastern Colombian Andes (Cordillera Oriental), near the border with Venezuela. Bolitoglossa tamaense sp. nov. is distributed between 2000 to 2700 m.a.s.l. and Bolitoglossa leandrae sp. nov. is distributed in the low-lands at about 600 m. The new species are diagnosed by a combination of molecular (16S rRNA sequences), coloration, body size, and morphometric (number of maxillary and vomerine teeth and differences in foot webbing) characters. Both species face threats such as chytridiomycosis infections and habitat fragmentation that have already affected other sala-manders in the country. Thus, intensive field and museum work is needed to better document and perhaps protect the local salamander diversity.

Species richness under a vertebral stripe: integrative taxonomy uncovers three additional species of Pholidobolus lizards (Sauria, Squamata, Gymnophthalmidae) from the north-western Colombian Andes.

The systematic study of biodiversity underlies appropriate inference in most other fields of biological research, yet it remains hampered by disagreements on both theoretical and empirical issues such as the species concept and the operational diagnosis of a species. Both become particularly challenging in those lineages where morphological traits are evolutionarily constrained by their adaptive value. For instance, cryptic organisms often conserve or converge in their external appearance, which hinders the recognition of species boundaries. An integrative approach has been adopted to study microgeographic variation in the leaf-litter lizard Pholidobolusvertebralis and test three predictions derived from the evolutionary species concept. Molecular data provided unambiguous evidence of divergence among the three recovered new clades and a common evolutionary history for each of them. The broadly sympatric clades were indeed diagnosable from externally visible traits, such as head scales, adult size, and sexually dimorphic ventral colouration. Also, they barely overlapped on the phenotypic space that summarised 39 morphometric and meristic traits. These clades are described as three species and an available name is suggested for a recovered fourth clade. The geographic distribution of the new and proximate species suggests a role for elevation on evolutionary divergence; it also raises interesting questions on the speciation pattern of an otherwise underestimated cryptic lineage.

Auto-deconvolution and molecular networking of gas chromatography-mass spectrometry data.

We engineered a machine learning approach, MSHub, to enable auto-deconvolution of gas chromatography-mass spectrometry (GC-MS) data. We then designed workflows to enable the community to store, process, share, annotate, compare and perform molecular networking of GC-MS data within the Global Natural Product Social (GNPS) Molecular Networking analysis platform. MSHub/GNPS performs auto-deconvolution of compound fragmentation patterns via unsupervised non-negative matrix factorization and quantifies the reproducibility of fragmentation patterns across samples.

Adjusted phonotactic reactions to sound amplitude and pulse number mediate territoriality in the harlequin poison frog.

The information content of signals remains one of the central questions in animal communication. Auditory signals might contain information that allows receivers to estimate the distance as well as the size or the motivational state of senders. Proper differential reactions by receivers could be especially important for territorial species and lead to behavioural adjustments towards intruders according to the threat level: the perceived risk of losing territory. Therefore, territorial individuals should decode signal parameters that indicate the sender's size, distance, and motivation/treat level, reacting accordingly. To test this hypothesis, we studied the phonotactic reaction in territorial males of the poison frog Oophaga histrionica, after analyzing the variability of spectral and temporal parameters of natural calls. Next, we constructed synthetic calls by manipulating sound amplitude (sound pressure level: SPL), gross (inter-call intervals), and fine-temporal (number of pulses) structure of synthetic signals. In this way, we simulated near and far intruders with potentially variable sizes, motivational states, or threat levels. Then, we conducted playback experiments using these synthetic calls to determine 1) how perceived proximity of vocal competitors (determined by SPL) affect the behavior of receivers, and 2) how variation in the temporal structure of calls (inter-call interval and call pulse number) impact the behavior of receivers. We also asked whether signals convey body-size related information by analyzing the relationship between call parameters and body size of the males receiving the experimental stimuli. Consistent with our hypothesis, males recognized the variations in the SPL of calls. Males attacking the simulated opponent increased the number of pulses per call, while those retreating, kept pulse number unchanged (a graded aggressive signal). On the other hand, despite call traits resulted as poor predictors of body size, both SPL and fine temporal call traits might aid to predict whether a contestant will attack an opponent. Our data demonstrate that males discriminate and use SPL to estimate the sender's distance and use the call's fine temporal traits and to adjust the competitive/aggressive reaction using bimodal signals. They further suggest that body size assessment is not always an important factor in vocally- mediated agonistic interactions.

Analysis of Malassezia Lipidome Disclosed Differences Among the Species and Reveals Presence of Unusual Yeast Lipids.

Malassezia yeasts are lipid dependent and part of the human and animal skin microbiome. However, they are also associated with a variety of dermatological conditions and even cause systemic infections. How these yeasts can live as commensals on the skin and switch to a pathogenic stage has long been a matter of debate. Lipids are important cellular molecules, and understanding the lipid metabolism and composition of Malassezia species is crucial to comprehending their biology and host-microbe interaction. Here, we investigated the lipid composition of Malassezia strains grown to the stationary phase in a complex Dixon medium broth. In this study, we perform a lipidomic analysis of a subset of species; in addition, we conducted a gene prediction analysis for the detection of lipid metabolic proteins. We identified 18 lipid classes and 428 lipidic compounds. The most commonly found lipids were triglycerides (TAG), sterol (CH), diglycerides (DG), fatty acids (FAs), phosphatidylcholine (PC), phosphatidylethanolamine (PE), ceramides, cholesteryl ester (CE), sphingomyelin (SM), acylcarnitine, and lysophospholipids. Particularly, we found a low content of CEs in Malassezia furfur, atypical M. furfur, and Malassezia pachydermatis and undetectable traces of these components in Malassezia globosa, Malassezia restricta, and Malassezia sympodialis. Remarkably, uncommon lipids in yeast, like diacylglyceryltrimethylhomoserine and FA esters of hydroxyl FAs, were found in a variable concentration in these Malassezia species. The latter are bioactive lipids recently reported to have antidiabetic and anti-inflammatory properties. The results obtained can be used to discriminate different Malassezia species and offer a new overview of the lipid composition of these yeasts. We could confirm the presence and the absence of certain lipid-biosynthesis genes in specific species. Further analyses are necessary to continue disclosing the complex lipidome of Malassezia species and the impact of the lipid metabolism in connection with the host interaction.

The relative roles of vicariance versus elevational gradients in the genetic differentiation of the high Andean tree frog, Dendropsophus labialis.

There are two main competing hypotheses (vicariance and vertical ecotones) that attempt to explain the tremendous diversity of the tropical Andes. We test these hypotheses at the intraspecific level by analyzing mitochondrial and nuclear DNA sequences from 24 populations of the high Andean frog, Dendropsophus labialis (Anura: Hylidae). This species displays geographic variation in a number of phenotypic traits. Most of these traits covary with elevation, while few vary along the horizontal (latitudinal) axis. We found that, both, vicariance and elevation had important effects on the genetic differentiation in this species. We detected two highly divergent clades along the south-north axis using independent information from mitochondrial and nuclear genes, suggesting that this differentiation was the result of long-term barriers to gene flow rather than stochastic processes. We hypothesize mechanisms for D. labialis strong differentiation in light of geological and paleoenvironmental models of evolution in the northern Andean highlands.

Does batrachotoxin autoresistance coevolve with toxicity in Phyllobates poison-dart frogs?

Toxicity is widespread among living organisms, and evolves as a multimodal phenotype. Part of this phenotype is the ability to avoid self-intoxication (autoresistance). Evolving toxin resistance can involve fitness tradeoffs, so autoresistance is often expected to evolve gradually and in tandem with toxicity, resulting in a correlation between the degrees of toxicity and autoresistance among toxic populations. We investigate this correlation in Phyllobates poison frogs, notorious for secreting batrachotoxin (BTX), a potent neurotoxin that targets sodium channels, using ancestral sequence reconstructions of BTX-sensing areas of the muscular voltage-gated sodium channel. Reconstructions suggest that BTX resistance arose at the root of Phyllobates, coinciding with the evolution of BTX secretion. After this event, little or no further evolution of autoresistance seems to have occurred, despite large increases in toxicity throughout the history of these frogs. Our results, therefore, provide no evidence in favor of an evolutionary correlation between toxicity and autoresistance, which conflicts with previous work. Future research on the functional costs and benefits of mutations putatively involved in BTX resistance, as well as their prevalence in natural populations, should shed light on the evolutionary mechanisms driving the relationship between toxicity and autoresistance in Phyllobates frogs.

A new species of Pristimantis (Anura: Craugastoridae) from the cloud forests of Colombian western Andes.

The frogs of the genus Pristimantis are recognized for their characteristic reproductive mode and for their incredible diversity of species, becoming the genus with the highest number of species within tetrapod vertebrates. We describe here a new species of Pristimantis from the northwestern Andes of Colombia. The species was found between 2000-2500 m.a.s.l., mostly within moss hanging of tree branches in a tropical cloud forest. It can be easily distinguished from other Pristimantis species of the western Andes by the unique black and white patterning in the ventral surface and the flanks, the hourglass-shaped dorsal folds, and the prominent conical tubercles on eyelids, heels, and outer edge of tarsus. A phylogenetic analysis further supports its status of a lineage reciprocally monophyletic to P. satagius and separated by a genetic distance of 0.03; the latter species bear whitish rather than predominantly black ventral coloration. To the best of our knowledge, this species is only known from the 2,500 ha nature reserve Mesenia-Paramillo, despite other research on this area of the country. Therefore, the species is declared vulnerable while new evidence on its distribution range is accumulated.

Microbiota and skin defense peptides may facilitate coexistence of two sympatric Andean frog species with a lethal pathogen.

Management of hyper-virulent generalist pathogens is an emergent global challenge, yet for most disease systems we lack a basic understanding as to why some host species suffer mass mortalities, while others resist epizootics. We studied two sympatric species of frogs from the Colombian Andes, which coexist with the amphibian pathogen Batrachochytrium dendrobatidis (Bd), to understand why some species did not succumb to the infection. We found high Bd prevalence in juveniles for both species, yet infection intensities remained low. We also found that bacterial community composition and host defense peptides are specific to amphibian life stages. We detected abundant Bd-inhibitory skin bacteria across life stages and Bd-inhibitory defense peptides post-metamorphosis in both species. Bd-inhibitory bacteria were proportionally more abundant in adults of both species than in earlier developmental stages. We tested for activity of peptides against the skin microbiota and found that in general peptides did not negatively affect bacterial growth and in some instances facilitated growth. Our results suggest that symbiotic bacteria and antimicrobial peptides may be co-selected for, and that together they contribute to the ability of Andean amphibian species to coexist with the global pandemic lineage of Bd.

The effect of captivity on the skin microbial symbionts in three Atelopus species from the lowlands of Colombia and Ecuador.

Many amphibian species are at risk of extinction in their natural habitats due to the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). For the most highly endangered species, captive assurance colonies have been established as an emergency measure to avoid extinction. Experimental research has suggested that symbiotic microorganisms in the skin of amphibians play a key role against Bd. While previous studies have addressed the effects of captivity on the cutaneous bacterial community, it remains poorly studied whether and how captive conditions affect the proportion of beneficial bacteria or their anti-Bd performance on amphibian hosts. In this study we sampled three amphibian species of the highly threatened genus, Atelopus, that remain in the wild but are also part of ex situ breeding programs in Colombia and Ecuador. Our goals were to (1) estimate the diversity of culturable bacterial assemblages in these three species of Atelopus, (2) describe the effect of captivity on the composition of skin microbiota, and (3) examine how captivity affects the bacterial ability to inhibit Bd growth. Using challenge assays we tested each bacterial isolate against Bd, and through sequencing of the 16S rRNA gene, we identified species from thirteen genera of bacteria that inhibited Bd growth. Surprisingly, we did not detect a reduction in skin bacteria diversity in captive frogs. Moreover, we found that frogs in captivity still harbor bacteria with anti-Bd activity. Although the scope of our study is limited to a few species and to the culturable portion of the bacterial community, our results indicate that captive programs do not necessarily change bacterial communities of the toad skins in a way that impedes the control of Bd in case of an eventual reintroduction.

Conspicuousness, color resemblance, and toxicity in geographically diverging mimicry: The pan-Amazonian frog Allobates femoralis.

Predation risk is allegedly reduced in Batesian and Müllerian mimics, because their coloration resembles the conspicuous coloration of unpalatable prey. The efficacy of mimicry is thought to be affected by variation in the unpalatability of prey, the conspicuousness of the signals, and the visual system of predators that see them. Many frog species exhibit small colorful patches contrasting against an otherwise dark body. By measuring toxicity and color reflectance in a geographically variable frog species and the syntopic toxic species, we tested whether unpalatability was correlated with between-species color resemblance and whether resemblance was highest for the most conspicuous components of coloration pattern. Heterospecific resemblance in colorful patches was highest between species at the same locality, but unrelated to concomitant variation in toxicity. Surprisingly, resemblance was lower for the conspicuous femoral patches compared to the inconspicuous dorsum. By building visual models, we further tested whether resemblance was affected by the visual system of model predators. As predicted, mimic-model resemblance was higher under the visual system of simulated predators compared to no visual system at all. Our results indicate that femoral patches are aposematic signals and support a role of mimicry in driving phenotypic divergence or mimetic radiation between localities.