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Biogeographic barriers such as rivers have been shown to shape spatial patterns of biodiversity in the Amazon basin, yet relatively little is known about the distribution of genetic variation across continuous rainforest. Here, we characterize the genetic structure of the brilliant-thighed poison frog (Allobates femoralis) across an 880-km-long transect along the Purus-Madeira interfluve south of the Amazon river, based on 64 individuals genotyped at 7609 single-nucleotide polymorphism (SNP) loci. A population tree and clustering analyses revealed 4 distinct genetic groups, one of which was strongly divergent. These genetic groups were concomitant with femoral spot coloration differences, which was intermediate within a zone of admixture between two of the groups. The location of these genetic groups did not consistently correspond to current ecological transitions between major forest types. A multimodel approach to quantify the relative influence of isolation-by-geographic distance (IBD) and isolation-by-environmental resistance (IBR) nevertheless revealed that, in addition to a strong signal of IBD, spatial genetic differentiation was explained by IBR primarily linked to dry season intensity (r2 = 8.4%) and canopy cover (r2 = 6.4%). We show significant phylogenetic divergence in the absence of obvious biogeographical barriers and that finer-scaled measures of genetic structure are associated with environmental variables also known to predict the density of A. femoralis.

Sex differences in vertebrate spatial abilities are typically interpreted under the adaptive specialization hypothesis, which posits that male reproductive success is linked to larger home ranges and better navigational skills. The androgen spillover hypothesis counters that enhanced male spatial performance may be a byproduct of higher androgen levels. Animal groups that include species where females are expected to outperform males based on life-history traits are key for disentangling these hypotheses. We investigated the association between sex differences in reproductive strategies, spatial behavior, and androgen levels in three species of poison frogs. We tracked individuals in natural environments to show that contrasting parental sex roles shape sex differences in space use, where the sex performing parental duties shows wider-ranging movements. We then translocated frogs from their home areas to test their navigational performance and found that the caring sex outperformed the non-caring sex only in one out of three species. In addition, males across species displayed more explorative behavior than females and androgen levels correlated with explorative behavior and homing accuracy. Overall, we reveal that poison frog reproductive strategies shape movement patterns but not necessarily navigational performance. Together this work suggests that prevailing adaptive hypotheses provide an incomplete explanation of sex differences in spatial abilities.

Predation risk is allegedly reduced in Batesian and Mullerian 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.

Longer-range movements of anuran amphibians such as mass migrations and habitat invasion have received a lot of attention, but fine-scale spatial behavior remains largely understudied. This gap is especially striking for species that show long-term site fidelity and display their whole behavioral repertoire in a small area. Studying fine- scale movement with conventional capture-mark-recapture techniques is difficult in inconspicuous amphibians: individuals are hard to find, repeated captures might affect their behavior and the number of data points is too low to allow a detailed interpretation of individual space use and time budgeting. In this study, we overcame these limitations by equipping females of the Brilliant-Thighed Poison Frog (Allobates femoralis) with a tag allowing frequent monitoring of their location and behavior. Neotropical poison frogs are well known for their complex behavior and diverse reproductive and parental care strategies. Although the ecology and behavior of the polygamous leaf-litter frog Allobates femoralis is well studied, little is known about the fine-scale space use of the non-territorial females who do not engage in acoustic and visual displays. We tracked 17 females for 6 to 17 days using a harmonic direction finder to provide the first precise analysis of female space use in this species. Females moved on average 1 m per hour and the fastest movement, over 20 m per hour, was related to a subsequent mating event. Traveled distances and activity patterns on days of courtship and mating differed considerably from days without reproduction. Frogs moved more on days with lower temperature and more precipitation, but mating seemed to be the main trigger for female movement. We observed 21 courtships of 12 tagged females. For seven females, we observed two consecutive mating events. Estimated home ranges after 14 days varied considerably between individuals and courtship and mating associated space use made up for ���30% of the home range. Allobates femoralis females spent large parts of their time in one to three small centers of use. Females did not adjust their time or space use to the density of males in their surroundings and did not show wide-ranging exploratory behavior. Our study demonstrates how tracking combined with detailed behavioral observations can reveal the patterns and drivers of fine-scale spatial behavior in sedentary species.

The distribution of biodiversity within the Amazon basin is often structured by sharp environmental boundaries, such as large rivers. The Amazon region is also characterized by subtle environmental clines, but how they might affect the distributions and abundance of organisms has so far received less attention. Here, we test whether soil and forest characteristics are associated with the occurrence and relative abundance of the forest-floor dwelling Aromobatid frog,Allobates femoralis. We applied a structured sampling regime along an 880 km long transect through forest of different density. High detection probabilities were estimated forA. femoralisin each of the sampling modules. Using generalized linear mixed-effects models and simple linear regressions that take detectability into account, we show thatA. femoralisis more abundant in open forests than in dense forests. The presence and relative abundance ofA. femoralisis also positively associated with clay-rich soils, which are poorly drained and therefore likely support the standing water bodies required for reproduction. Taken together, we demonstrate that relatively easy-to-measure environmental features can explain the distribution and abundance of a widespread species at different spatial scales. Such proxies are of clear value to ecologists and conservation managers working in large inaccessible areas such as the Amazon basin.

The brilliant-thighed frog [Allobates femoralis (Boulenger, 1884)] is distributed across the Amazon basin and aggregates several allopatric evolutionary lineages, some of which present variation in their advertisement calls. In 2009, an unregistered call phenotype was discovered in the region of Altamira and Vitória do Xingu, State of Pará, Brazil, where males emit advertisement calls formed by six notes, differing from the typical four-note calls described for other A. femoralis populations. In this study, we describe in detail these untypical calls. Additionally, we test whether the aggressive responses of males of a 4-note reference population (Reserva Ducke - RFAD, in Manaus, States of Amazonas) is differential towards the 6-note calls of males recorded in Altamira (Pará State), and towards 4-note calls recorded in one location at the Tapajós-Xingu interfluve (Belterra, Pará State), and in RFAD. Playback experiments were conducted between 2011-2012, and used standardized stimuli produced from natural call recordings. A total of 30 independent experiments were conducted, 10 for each stimuli class. We measured the phonotaxis of focal males in relation to the loudspeaker, considering the time to orientation and the time to approach the loudspeaker. We found that not all A. femoralis males at RFAD promptly recognize calls from males recorded in Altamira. However, when considering only males who approached the loudspeaker, differences in aggressive reactions were not seen between stimuli classes. Our findings show that the ability to recognize calls from Altamira as belonging to co-specific males is not universal among males at RFAD. The new A. femoralis phenotype occurs in areas potentially impacted by the Belo Monte hydroelectric complex and complementary studies indicate that no gene flow exists between this group and A. femoralis from adjacent regions. Hence, developments in Altamira may put this incipient speciation process at risk. © 2017, Fundacao Zoobotanica do Rio Grande do Sul. All rights reserved.

Animals relying on uncertain, ephemeral and patchy resources have to regularly update their information about profitable sites. For many tropical amphibians, widespread, scattered breeding pools constitute such fluctuating resources. Among tropical amphibians, poison frogs (Dendrobatidae) exhibit some of the most complex spatial and parental behaviors—including territoriality and tadpole transport from terrestrial clutches to ephemeral aquatic deposition sites. Recent studies have revealed that poison frogs rely on spatial memory to successfully navigate through their environment. This raises the question of when and how these frogs gain information about the area and suitable reproductive resources. To investigate the spatial patterns of pool use and to reveal potential explorative behavior, we used telemetry to follow males of the territorial dendrobatid frogAllobates femoralisduring tadpole transport and subsequent homing. To elicit exploration, we reduced resource availability experimentally by simulating desiccated deposition sites. We found that tadpole transport is strongly directed towards known deposition sites and that frogs take similar direct paths when returning to their home territory. Frogs move faster during tadpole transport than when homing after the deposition, which probably reflects different risks and costs during these two movement phases. We found no evidence for exploration, neither during transport nor homing, and independent of the availability of deposition sites. We suggest that prospecting during tadpole transport is too risky for the transported offspring as well as for the transporting male. Relying on spatial memory of multiple previously discovered pools appears to be the predominant and successful strategy for the exploitation of reproductive resources inA. femoralis. Our study provides for the first time a detailed description of poison frog movement patterns during tadpole transport and corroborates recent findings on the significance of spatial memory in poison frogs. When these frogs explore and discover new reproductive resources remains unknown.

Life history trade-offs in reproductive strategy are often invoked as ecological agents of evolutionary change, despite a limited amount of experimental data from the field. The larval deposition strategy of Allobates femoralis was tested in Southeastern Peru using a blocked, fully crossed experimental design. Arrays of four pools were used to test the effects of pool size and the presence of a predatory insect (Belostomatid) on the deposition behavior of A. femoralis. Further experiments investigated the colonization of insect predators into potential larval habitats and interactions between predatory insects. Results suggest that pool size, the presence of predatory aquatic insects, and interactions between predators, influence larval deposition in A. femoralis. Similar ecological interactions may have driven toxic dendrobatids to the use of arboreal phytotelmata and associated derived reproductive strategies.

This Special Issue demonstrates the breadth of evolutionary ecology research occurring on Neotropical poison frogs (Dendrobatidae). It also draws attention to the incredible diversity of the poison frogs themselves (phenotypic, behavioral and taxonomic diversity) and this group's potential for research programs focusing on speciation, the evolution of toxicity, color pattern diversity, Mullerian and Batesian mimicry, sexual selection, mating systems and parental care. This Special Issue focused on two key research foci in the field of evolutionary ecology: (1) Understanding the interplay between ecology and the evolution of parental care and (2) Elucidating the ecological factors contributing to the evolution of toxicity, aposematism and color pattern diversity. In this paper, I summarize the major findings of studies presented in this Special Issue. Understanding the interplay between ecology and the evolution of parental care Parent poison frogs transport and subsequently deposit their tadpoles in pools of water. The evolutionary transition from terrestrial to smaller arboreal phytotelmata for tadpole deposition is thought to be driven by a reduction in competition and predation associated with smaller tadpole nurseries. This transition is considered a key evolutionary innovation, allowing access to relatively unexploited resources and facilitating the evolution of many novel breeding strategies. To date, there are only a few empirical examples in dendrobatids to support this hypothesis (Weygoldt 1980; Caldwell 1997; Brown et al. 2010). Field experimentation by McKeon and Summers (2013) present a more complete picture of the interactions between the natural history of Allobates femoralis and the tadpole predators present. Their results suggest that pool size, the presence of predatory insects and inter- actions between predators (and indirect effects on tadpoles) influence larval deposition in A. femoralis. Previous research on Ranitomeya species, a genus distantly related to Allobates, has demonstrated that adults also assess multiple aspects of reproductive resources to

Individual-based data sets tracking organisms over space and time are fundamental to answering broad questions in ecology and evolution. A “permanent” genetic tag circumvents a need to invasively mark or tag animals, especially if there are little phenotypic differences among individuals. However, genetic tracking of individuals does not come without its limits; correctly matching genotypes and error rates associated with lab work can make it difficult to parse out matched individuals. In addition, defining a sampling design that effectively matches individuals in the wild can be a challenge for researchers. Here, we combine the two objectives of defining sampling design and reducing genotyping error through an efficient Python-based computer-modeling program, WisePair. We describe the methods used to develop the computer program and assess its effectiveness through three empirical data sets; with and without reference genotypes. Our results show that WisePair outperformed similar genotype matching programs by using previously-published from reference genotype data of diurnal poison frogs (Allobates femoralis) and without-reference (fecal) genotype sample datasets of harbor seals (Phoca vitulina) and Eurasian otters (Lutra lutra). In addition, due to limited sampling effort in the harbor seal data, we present optimal sampling designs for future projects. WisePair allows for minimal sacrifice in the available methods as it incorporates sample re-run error data, allelic pairwise comparisons, and probabilistic simulations to determine matching thresholds. Our program is the lone tool available to researchers to define parameters a priori for genetic-tracking studies. This article is protected by copyright. All rights reserved.

Field observations and mark–recapture sampling carried out in the Reserva Ducke in Central Amazonia provided information on the occurrence of reproductive events of the Brilliant-Thighed Frog, Allobates femoralis, in the context of seasonal rainfall. In addition, we report on the daily pattern of calling activity and the use of space by marked individuals. Rainfall was a strong determinant of the calling and breeding activities at a seasonal scale. The phenology of A. femoralis was affected by interannual differences in both the length and severity of the dry season. This species showed a consistent diel pattern throughout the year, with calling activity peaking between 1500 and 1600 h. At the seasonal scale, calling activity was higher during the wet months and was correlated positively with monthly rainfall. Although recruitment is concentrated in the late wet season, our observations of juveniles in the smallest size classes throughout the year indicated that reproduction occurs sporadically e...

Natural hybridization between allopatric species following secondary contact has been poorly documented for Neotropical anurans inhabiting the Amazonian lowlands. We conducted a genetic survey across a contact zone between two species of litter frogs, Allobates hodli and Allobates femoralis (family Dendrobatidae), located on the left riverbank of the upper Madeira River, State of Rondonia, Brazil. We obtained tissue samples from 11 sampling sites on both riverbanks, covering approximately a 400 km long transect. We evaluated the genetic relationships between samples using haplotype networks and a distance-based phylogenetic tree obtained from a dataset of 16S rRNA mtDNA sequences. Estimates of genetic diversity, population structure, and the identification of sites where genetic admixture occurred were carried out by means of frequency-based methods and Bayesian inference on mtDNA and a set of four microsatellite loci, including samples collected throughout the study area. A reduced dataset including only microsatellite loci genotyped from samples on the left riverbank was applied in assignment tests for detecting levels of admixture at the contact zone and adjacent sampling sites, and for detecting and quantifying hybrid individuals. Our results suggest that genetic introgression between A. hodli and A. femoralis is restricted to the core area of the contact zone, where potential hybrids are less frequent than parental genotypes. Effects on the genetic variability of adjacent populations are only detected at sites located 1.5 km downstream and upstream of the core area, suggesting the existence of selection against hybrids, possibly mediated by postzygotic isolation mechanisms. The contact zone between A. femoralis and A. hodli is the first well delimited suture line between anuran species ever documented in the Brazilian Amazon. The settlement of two dams along the upper Madeira River poses an immediate threat to the gene flow and hybridization balance observed between the populations studied. Our results provided guidelines for a current monitoring program, aiming at the impacts of dams on this evolutionary system’s dynamics.

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.

Parental care is a common strategy among vertebrates to ensure successful reproduction. Anuran amphibians have evolved a remarkable diversity of reproductive methods including advanced levels of parental care. Among the most derived strategies are those of the Neotropical poison frogs (Dendrobatidae). These amphibians exhibit a wide array of behavioural traits such as egg guarding, larval transport by parental frogs and larval feeding with trophic (unfertilized) eggs. Ranitomeya variabilis from the upper Amazon basin in Peru deposits both eggs and tadpoles in phytotelmata. The exploitation of these small pools is advantageous as it lowers the risk of predation, but it is more costly because of limited resource availability. Additionally, poison frog larvae are often cannibalistic, so the identification and avoidance of conspecifics represents an adaptive behaviour for these amphibians. While studies have shown that poison frogs actively avoid depositing with conspecifics, the mechanism for assessing pool quality remains unknown. In field experiments, we found that parental R. variabilis frogs used chemical cues to recognize the presence of tadpoles in phytotelmata. Furthermore, they distinguished between cannibalistic and noncannibalistic tadpoles, a behaviour that supports the survival of their own offspring. 2011 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

The identification of divergence in reproductive traits may substantially improve integrative approaches to understand species limits within clades that are suspected to contain cryptic diversity. The frog Allobates femoralis has been regarded as a pan-Amazonian species, and widely used as a model for addressing evolutionary issues regarding patterns of intraspecific diversification, social organisation, and animal communication. Recent accumulation of genetic, morphological, and bioacoustic data gathered from different localities strongly supports the idea that it represents a species complex, but field behavioural observations related to courtship and mating are surprisingly scarce. Here, we provide a description of several aspects of the reproductive biology of A. femoralis from a Central Amazon site, and compare our results with the few published reports for the species. This study demonstrated that, besides the known divergence in the number of notes of the A. femoralis call, there are both quantitat...

Our knowledge about genetic mating systems and the underlying causes for and consequences of variation in reproductive success has substantially improved in recent years. When linked to longitudinal population studies, cross-generational pedigrees across wild populations can help answer a wide suite of questions in ecology and evolutionary biology. We used microsatellite markers and exhaustive sampling of two successive adult generations to obtain population-wide estimates of individual reproductive output of males and females in a natural population of the Neotropical frog Allobates femoralis (Aromobatidae), a pan-Amazonian species that features prolonged iteroparous breeding, male territoriality and male parental care. Parentage analysis revealed a polygynandrous mating system in which high proportions of males (35.5%) and females (56.0%) produced progeny that survived until adulthood. Despite contrasting reproductive strategies, successfully reproducing males and females had similar numbers of mating partners that sired the adult progeny (both sexes: median 2; range 1– 6); the numbers of their offspring that reached adulthood were also similar (both sexes: median 2; range 1–8). Measures of reproductive skew indicate selection on males only for their opportunity to breed. Reproductive success was significantly higher in territorial than in nonterritorial males, but unrelated to territory size in males or to body size in both sexes. We hypothesize that female polyandry in this species has evolved because of enhanced offspring survival when paternal care is allocated to multiple partners.

The call of the pan-Amazonian frog Allobates femoralis shows wide geographical variation, and males show a stereotyped and conspicuous phonotactic response to playback of conspecific calls. We evaluated the capacity of males of A. femoralis and a closely related species A. hodli to respond aggressively to natural conspecific and heterospecific calls varying in number of notes, by means of field playback experiments performed at two sites in the Brazilian Amazon. The first site, Cachoeira do Jirau (Porto Velho, Rondonia), is a parapatric contact zone between A. femoralis that use 4-note calls, and A. hodli with 2-note calls, where we performed cross-playbacks in both focal populations. The second site, the Reserva Florestal Adolpho Ducke (Manaus, Amazonas), contained only A. femoralis with 4-note calls. There, we broadcast natural stimuli of 2-note A. hodli, 3-note and 4-note A. femoralis, and 6-note A. myersi. We found that the phonotactic behaviour of A. femoralis and A. hodli males did not differ toward conspecific and heterospecific stimuli, even in parapatry. Our results indicated that the evolutionary rates of call design and call perception are different, because the geographical variation in calls was not accompanied by variation in the males' aggressive behaviour.

The social and reproductive behaviour of the dart-poison frog, Epipedobates femoralis, was studied in Amazonian Peru for 14 months. Males defended territories with advertisement calls and, ultimately, fighting. Territory size ranged from 0.25 to 26 m2 and was positively correlated with duration of residence and calling activity of the owner. Females were not territorial and were never attacked when approaching calling males. Males and females only mated once and females sampled calling males before mating. Male mating success was closely correlated with territory size and calling activity. No correlation was found between male body size and mating success. Territories provide residents with sufficient space for mate attraction and reproduction without interference from rivals. Since territory size is dependent on calling activity which involves high energetic costs, it is suggested that territory size reveals male quality.

Evolutionary divergence in behavioural traits related to mating may represent the initial stage of speciation. Direct selective forces are usually invoked to explain divergence in mate-recognition traits, often neglecting a role for neutral processes or concomitant differentiation in ecological traits. We adopted a multi-trait approach to obtain a deeper understanding of the mechanisms behind allopatric divergence in the Amazonian frog, Allobates femoralis. We tested the null hypothesis that geographic distance between populations correlates with genetic and phenotypic divergence, and compared divergence between mate-recognition (acoustic) and ecological (coloration, body-shape) traits. We quantified geographic variation in 39 phenotypic traits and a mitochondrial DNA marker among 125 individuals representing eight populations. Geographic variation in acoustic traits was pronounced and tracked the spatial genetic variation, which appeared to be neutral. Thus, the evolution of acoustic traits tracked the shared history of the populations, which is unexpected for pan-Amazonian taxa or for mate-recognition traits. Divergence in coloration appeared uncorrelated with genetic distance, and might be partly attributed to local selective pressures, and perhaps to Batesian mimicry. Divergence in body-shape traits was low. The results obtained depict a complex evolutionary scenario and emphasize the importance of considering multiple traits when disentangling the forces behind allopatric divergence. ©2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98, 826-838.

We studied movement and site fidelity of males and females of the territorial frog Allobates femoralis (Aromobatidae) in a population in the Nature Reserve “Les Nouragues” in French Guiana, South America. Observations during 3 months in 2006 ascertained intra-seasonal site fidelity for males and females. Males actively defend large multi-purpose territories whereas females retreat to small resting sites from where they commute to neighbouring males for courtship and mating. Female short-term movement corroborates the previous assumption of a polygynous or promiscuous resource-defence mating system. Year-to-year recaptures from 2005 until 2008 revealed distinct patterns of inter-annual movement for males and regional site fidelity for females. Males abandon their territories and have to re-negotiate them when reproduction starts again at the end of the dry season. Females are not subject to intra- or inter-sexual territorial competition and as a result move significantly less between reproductive seasons than males. Male long-term movement reflects spatial structure and prevailing social interactions and is a reliable indicator for tadpole deposition sites. The combined effects of intra- and inter-seasonal movement promote the diversity of mates for both sexes.

Dendrobatid frogs perform a unique mode of parental care, ranging from egg attendance and tadpole transport to tadpole feeding. It is hypothesized that a behaviour in which the egg attending parent remains with the cluch is the most primitive condition. In more advanced forms or parental care, the male is able to attend several clutches of eggs. Tadpole attendance and feeding, finally, started as deceit; the male induced the female to lay eggs into a bromeliad leaf axil already occupied by a larva. Costs and benefits of the different modes of parental care and possible alternative reproductive tactics are discussed.

We studied patterns of call acoustics and external morphological differentiation in populations of the dart-poison frog Allobates femoralis occurring in forested areas along a 250-km stretch of the upper Madeira River, Brazil. Multivariate analyses of variance using principal components representing shared acoustic and morphological parameters distinguished three groups in relation to call structure and external morphology: (1) populations belonging to a two-note call morphotype; (2) populations with four-note calls inhabiting the left riverbank; and (3) populations with four-note calls inhabiting the right riverbank. Our results report a case of Amazonian anuran diversity hidden by current taxonomy and provide evidence for the upper Madeira River being a boundary between distinct populations of A. femoralis, and suggest a new taxonomic interpretation for these groups. Samples that did not fit into the general differentiation pattern and the existence of a well-defined contact zone between two morphotypes on the left riverbank indicate that mechanisms complementary to river-barrier hypotheses are necessary to explain the phenotypic differentiation between populations. Our study shows that at least one anuran species shows congruence between population differentiation and separation by a large Amazonian river, as documented for birds and mammals. Conservation efforts should not consider the taxon now known as A. femoralis as a homogeneous entity. There is much within-taxon variability, which can be probably explained partly by the existence of cryptic species, partly by geological barriers and part of which currently has no obvious explanation.

One or both sexes exhibit territorial behavior in all dendrobatids that have been studied. Two types of territoriality are identified. In some Colosthethus species only females or both sexes defend small nonreproductive territories that offer access to water, food, and shelter. In other species, only males defend large reproductive territories against conspecific calling males, suggesting that females are the limited resource. However, in no case has the defended resource been unambiguously identified. I hypothesize that, in at least some species, males compete for calling sites in areas with high female density and with favorable sound transmission features. I found little support for the hypothesis that dendrobatids exhibit resource defense polygyny. More research is necessary to understand the function of territoriality and to identify limiting resources in poison frogs.

The phenology of calling activity and reproduction of a neotropical anuran community in French Guiana was studied during one rainy season. We investigated the correlation between calling activity, rainfall, temperature and water level in two ponds and recorded the occurrence of tadpoles of pond-breeding species. The study site contained 31 calling frog species, which were divided into groups according to reproductive mode. Increased rainfall was associated with increased reproductive activity in all groups, but temporal patterns in calling activity varied significantly between groups. Species with aquatic oviposition exhibited sporadic acoustic activity, aggregating into explosive breeding events following heavy rainfall. Species laying eggs in foam nests had the peak of calling activity at the start of the rainy season. Taxa with embryonic development on vegetation called mainly from middle to late wet season, being the only group which showed a significant correlation of calling with increasing water level. Dendrobatids with terrestrial oviposition and subsequent parental tadpole transportation were continuously active. Species with direct development or with non-feeding larvae were mainly active at the beginning of the rainy season. It is concluded that phenologies of calling activity in South American tropical anuran species are strongly influenced by abiotic factors like rainfall and availability of breeding sites. The temporal limitation of the rainy season forces species to adjust calling and reproductive activity according to their reproductive modes.

The distribution of many vertebrate species in the Amazon basin is delimited by large rivers, which are frequently regarded as geographic barriers related with speciation and are therefore of high conservation value. Rapid development in the region includes plans to dam one of its thirteen major rivers, the Xingu. Environmental impact assessment is required for large scale development within the Brazilian portion of the Amazon Basin. However, impacts on biodiversity are only considered at the species level, and taxonomic work is far from described for many groups. In particular, anuran diversity is underestimated, in part owing to the scale of the region and, for some taxa, by morphological conservatism. Here we describe genetic and phenotypic variation in the brilliant-thighed frog, Allobates femoralis. We show that a unique genetic lineage, with a vocal repertoire distinct from that described for the species throughout its remaining geographic range, is located within the region to be directly and indirectly impacted by damming the Xingu River. Further, genetic variation within the A. femoralis group is largely structured in accordance to river systems, despite morphological conservatism. Our data add support for conservation policy to be amended to include intraspecific measures of diversity in order to more effectively conserve current biodiversity and evolutionary processes. © Pedro Ivo Simões, Adam Stow, Walter Hödl, Adolfo Amézquita, Izeni P. Farias and Albertina P. Lima.

Introduction Individuals should aim to adjust their parental behaviours in order to maximize the success of their offspring but minimize associated costs. Plasticity in parental care is well documented from various bird, mammal and fish species, whereas amphibians were traditionally assumed as being highly instinct-bound. Therefore, little is known about ‘higher’ cognitive abilities of amphibians, such as strategic planning and behavioural flexibility. Dendrobatid frogs have evolved a remarkable diversity of parental behaviours. The most noticeable of these behaviours is tadpole transport, which is obligatory in almost all species. Nonetheless, there is limited knowledge about spatial and temporal patterns of tadpole transport and the possible existence of behavioural plasticity on the individual level. In this study, we investigated correlates of tadpole transport behaviour in a natural population of the dendrobatid frog Allobates femoralis during five years. Results Tadpole transport was predominantly observed during morning hours. Although tadpoles were carried almost exclusively by males (N = 119), we also observed ten females performing this task. The parentage analysis revealed that in all cases females transported their own offspring. In contrast, four tadpole-carrying males were not the genetic fathers of the larvae they were transporting. The average clutch size of 20 eggs and our observation of an average of 8 tadpoles on the back of transporting individuals indicate that frogs do not carry entire clutches at once, and/or that they distribute their larvae across several water bodies. Contrary to the predictions from a hypothetical random search for deposition sites, the number of transported tadpoles was higher in males that travelled over longer distances. Conclusions Our results suggest a strong selective pressure on males to shift the time invested in tadpole transport to periods of low intra-specific competition. The number of tadpoles on the back of the males significantly correlated with displacement distance from the respective home territories, indicating a strategic non-random tadpole transport rather than random search for suitable tadpole deposition sites during tadpole transport. The observation of females who occasionally transported larvae supports the prevalence of adaptive plasticity in parental behaviours even in a species with a rather low level of parental care.

The adaptive significance of sequential polyandry is a challenging question in evolutionary and behavioral biology. Costs\ud and benefits of different mating patterns are shaped by the spatial distribution of individuals and by genetic parameters\ud such as the pairwise relatedness between potential mating partners. Thus, females should become less choosy as costs of\ud mating and searching for mates increase. We used parentage assignments to investigate spatial and genetic patterns of\ud mating across a natural population of the Neotropical frog Allobates femoralis, a species characterized by male territoriality\ud and care and female iteroparity. There was no correlation between genetic and spatial distances between adult individuals\ud across the population. In 72% of cases, females mated with males available within a radius of 20 m. Mean pairwise\ud relatedness coefficients of successful reproducers did not differ from random mating but had a lower variance than\ud expected by chance, suggesting maximal reproductive output at intermediate genetic divergence. We also found evidence\ud for selection in favor of more heterozygous individuals between the embryo and adult stage. The level of sequential\ud polyandry significantly increased with the number of spatially available males. Females that had more candidate males also\ud produced more adult progeny. We hypothesize that the benefits associated with female multiple mating outweigh the\ud costs of in- and outbreeding depression, and consequently precluded the evolution of ‘choosy’ mate selection in this\ud species.

Territoriality is a widespread behaviour in animals and its analysis is crucial in several areas of behavioural, ecological and evolutionary research. Commonly, territory size is assessed through territory mapping and the application of simple area estimators such as minimum convex polygons. In the present study we demonstrate that territory size can be determined adequately with an active approach through intrusion experiments, a technique that is commonly used in behavioural research in other contexts. Tests with simulated data indicate that a minimum of twelve trials needs to be performed to establish reliable orders of relative territory size. To estimate absolute territory size, detailed hull techniques are most appropriate when analyzing point patterns of intrusion experiments, while the local convex hull estimator enables the construction of internal utilization distributions based on such point patterns. Additionally we suggest a 'stretch the centre' approach to emphasize the actual process of intrusion experiments in the construction of internal utilization distributions. To demonstrate the utility of the method, we apply all findings from the simulations to data from fieldwork with the model species Allobates femoralis, a territorial aromobatid frog from the lowland rainforest of French Guiana.

Linking specific ecological factors to the evolution of parental care pattern and mating system is a difficult task of key importance. We provide evidence from comparative analyses that an ecological factor (breeding pool size) is associated with the evolution of parental care across all frogs. We further show that the most intensive form of parental care (trophic egg feeding) evolved in con- cert with the use of small pools for tadpole deposition and that egg feeding was associated with the evolution of biparental care. Previous research on two Peruvian poison frogs (Ranitomeya imitator and Ranitomeya variabilis) revealed similar life histories, with the excep- tion of breeding pool size. This key ecological difference led to di- vergence in parental care patterns and mating systems. We present ecological field experiments that demonstrate that biparental care is essential to tadpole survival in small (but not large) pools. Field observations demonstrate social monogamy in R. imitator, the species that uses small pools. Molecular analyses demonstrate genetic mo- nogamy in R. imitator, the first example of genetic monogamy in an amphibian. In total, this evidence constitutes the most complete documentation to date that a single ecological factor drove the evo- lution of biparental care and genetic and social monogamy in an animal.

Animal acoustic signals play seminal roles in mate attraction and regulation of male spacing, maintenance of pairbonds, localization of hosts by parasites, and feeding behavior. Among vertebrate signals, it is becoming clear that no single stereotyped signal feature reliably elicits species-specific behavior, but rather, that a suite of characters is involved. Within the largely nocturnal clade of anuran amphibians, the dart-poison frog, Epipedobates femoralis , is a diurnal species that physically and vigorously defends its calling territory against conspecific intruders. Here we report that physical attacks by a territorial male are provoked only in response to dynamic bimodal stimuli in which the acoustic playback of vocalizations is coupled with vocal sac pulsations, but not by either unimodal cues presented in isolation or static bimodal stimuli. These results suggest that integration of dynamic bimodal cues is necessary to elicit aggression in this species.

The presence of the dendrobatid frog Allobates femoralis in Venezuela is herein confirmed though recorded and analized advertisement calls taken at two Venezuelan localities.

The ability to associate environmental cues with valuable resources strongly increases the chances of finding them again, and thus memory often guides animal movement. For example, many temperate region amphibians show strong breeding site fidelity and will return to the same areas even after the ponds have been destroyed. In contrast, many tropical amphibians depend on exploitation of small, scattered and fluctuating resources such as ephemeral pools for reproduction. It remains unknown whether tropical amphibians rely on spatial memory for effective exploitation of their reproductive resources. Poison frogs (Dendrobatidae) routinely shuttle their tadpoles from terrestrial clutches to dispersed aquatic deposition sites. We investigated the role of spatial memory for relocating previously discovered deposition sites in an experimental population of the brilliant-thighed poison frog, Allobates femoralis, a species with predominantly male tadpole transport. We temporarily removed an array of artificial pools that served as the principal tadpole deposition resource for the population. In parallel, we set up an array of sham sites and sites containing conspecific tadpole odour cues. We then quantified the movement patterns and site preferences of tadpole-transporting males by intensive sampling of the area and tracking individual frogs. We found that tadpole-carrier movements were concentrated around the exact locations of removed pools and most individuals visited several removed pool sites. In addition, we found that tadpole-transporting frogs were attracted to novel sites that contained high concentrations of conspecific olfactory tadpole cues. Our results suggest that A. femoralis males rely heavily on spatial memory for efficient exploitation of multiple, widely dispersed deposition sites once they are discovered. Additionally, olfactory cues may facilitate the initial discovery of the new sites.

Spreading reproduction across time or space can optimize fitness by minimizing the risks for offspring survival in varying and unpredictable environments. Poison frogs (Dendrobatidae) are characterized by complex spatial and reproductive behaviour, such as territoriality, prolonged courtship and parental care. The partitioning of larvae from terrestrial clutches across several water bodies is mainly known from species with carnivorous tadpoles that allocate their tadpoles in very small pools, where limited food availability is accompanied by an increased risk of cannibalism. However, little is known about the deposition behaviour of non-carnivorous species that use medium-sized to large pools. In the present study, we investigated whether the Neotropical poison frog Allobates femoralis exhibits brood-partitioning behaviour when males transport tadpoles 3 weeks after oviposition. We sampled 30 artificial water bodies for tadpoles, which we genotyped at seven highly polymorphic microsatellite loci. Based on the reconstructed pedigree, we show that A. femoralis males distribute larvae of single and of successive clutches across several water bodies. The number of pools used was significantly associated with the number of clutches per male. Ninety-three percent of the males that were assigned to more than one clutch spread their tadpoles across several water bodies. Given the highly variable and unpredictable biotic and abiotic conditions in tropical rainforest, at the spatial scale of the study species’ behaviour, we interpret this behaviour as bet-hedging to improve offspring survival.

During acoustic communication, an audible message is transmitted from a sender to a receiver, often producing changes in behavior. In a system where evolutionary changes of the sender do not result in a concomitant adjustment in the receiver, communication and species recognition could fail. However, the possibility of an evolutionary decoupling between sender and receiver has rarely been studied. Frog populations in the Allobates femoralis cryptic species complex are known for their extensive morphological, genetic and acoustic variation. We hypothesized that geographic variation in acoustic signals of A. femoralis was correlated with geographic changes in communication through changes in male-male recognition. To test this hypothesis, we quantified male call recognition using phonotactic responses to playback experiments of advertisement calls with two, three and four notes in eight localities of the Amazonian basin. Then, we reconstructed the ancestral states of call note number in a phylogenetic framework and evaluated whether the character state of the most recent common ancestor predicted current relative responses to two, three and four notes. The probability of a phonotactic response to advertisement calls of A. femoralis males was strongly influenced by the call mid-frequency and the number of notes in most populations. Positive phonotaxis was complete for calls from each individual's population, and in some populations, it was also partial for allotopic calls; however, in two populations, individuals equally recognized calls with two, three or four notes. This evidence, in conjunction with our results from phylogenetic comparative methods, supports the hypothesis of decoupled evolution between sender and receiver in the male-male communication system of the A. femoralis complex. Thus, signal recognition appears to evolve more slowly than the calls.

“Ecosystem engineering” describes habitat alteration by an organism that affects another organism; such nontrophic interactions between organisms are a current focus in ecological research. Our study quantifies the actual impact an ecosystem engineer can have on another species by using a previously identified model system—peccaries and rainforest frogs. In a 4-year experiment, we simulated the impact of peccaries on a population of Allobates femoralis (Dendrobatidae) by installing an array of artificial pools to mimic a forest patch modified by peccaries. The data were analyzed using a gradual before-after control-impact (gBACI) model. Following the supplementation, population size almost doubled as a result of increased autochthonous recruitment driven by a higher per-capita reproduction of males and a higher proportion of reproducing females. The effect was evenly distributed across the population. The differential response of males and females reflects the reproductive behavior of A. femoralis, as only the males use the aquatic sites for tadpole deposition. Our study shows that management and conservation must consider nontrophic relationships and that human “ecosystem engineering” can play a vital role in efforts against the “global amphibian decline.”

Among vertebrates, comparable spatial learning abilities have been found in birds, mammals, turtles and fishes, but virtually nothing is known about such abilities in amphibians. Overall, amphibians are the most sedentary vertebrates, but poison frogs (Dendrobatidae) routinely shuttle tadpoles from terrestrial territories to dispersed aquatic deposition sites. We hypothesize that dendrobatid frogs rely on learning for flexible navigation. We tested the role of experience with the local cues for poison frog way-finding by (i) experimentally displacing territorial males of Allobates femoralis over several hundred metres, (ii) using a harmonic direction finder with miniature transponders to track these small frogs, and (iii) using a natural river barrier to separate the translocated frogs from any familiar landmarks. We found that homeward orientation was disrupted by the translocation to the unfamiliar area but frogs translocated over similar distances in their local area showed significant homeward orientation and returned to their territories via a direct path. We suggest that poison frogs rely on spatial learning for way-finding in their local area.

Detour behaviour, an individual's ability to reach its goal by taking an indirect route, has been used to test spatial cognitive abilities across a variety of taxa. Although many amphibians show a strong homing ability, there is currently little evidence of amphibian spatial cognitive flexibility. We tested whether a territorial frog, Allobates femoralis, can flexibly adjust its homing path when faced with an obstacle. We displaced male frogs from their calling sites into the centre of circular arenas and recorded their escape routes. In the first experiment we provided an arena with equally high walls. In the second experiment we doubled the height of the homeward facing wall. Finally, we provided a tube as a shortcut through the high wall. In the equal-height arena, most frogs chose to escape via the quadrant facing their former calling site. However, when challenged with different heights, nearly all frogs chose the low wall, directing their movements away from the calling site. In the "escape tunnel" experiment most frogs still chose the low wall. Our results show that displaced A. femoralis males can flexibly adjust their homing path and avoid (presumably energetically costly) obstacles, providing experimental evidence of spatial cognitive flexibility in an amphibian.

Parental care systems are shaped by costs and benefits to each sex of investing into current versus future progeny. Flexible compensatory parental care is mainly known in biparental species, particularly where parental desertion or reduction of care by 1 parent is common. The other parent can then compensate this loss by either switching parental roles and/or by increasing its own parental effort. In uniparental species, desertion of the caregiver usually leads to total brood loss. In the poison frog, Allobates femoralis, obligatory tadpole transport (TT) is generally performed by males, whereas females abandon their clutches after oviposition. Nevertheless, in a natural population we previously observed 7.8% of TT performed by females, which we could link to the absence of the respective fathers. In the following experiment, under laboratory conditions, all tested A. femoralis females flexibly took over parental duties, but only when their mates were removed. Our findings provide clear evidence for compensatory flexibility in a species with unisexual parental care. Contrary to the view of amphibian parental care as being stereotypical and fixed, these results demonstrate behavioral flexibility as an adaptive response to environmental and social uncertainty. Behavioral flexibility might actually represent a crucial step in the evolutionary transition from uniparental to biparental care in poison frogs. We suspect that across animal species flexible parental roles are much more common than previously thought and suggest the idea of a 3-dimensional continuum regarding flexibility, parental involvement, and timing, when thinking about the evolution of parental care.

Dendrobatidae (dart-poison frogs) exhibit some of the most complex spatial behaviors among amphibians, such as territoriality and tadpole transport from terrestrial clutches to widely distributed deposition sites. In species that exhibit long-term territoriality, high homing performance after tadpole transport can be assumed, but experimental evidence is lacking, and the underlying orientation mechanisms are unknown. We conducted a field translocation experiment to test whether male Allobates femoralis, a dendrobatid frog with paternal extra-territorial tadpole transport, are capable of homing after experimental removal, as well as to quantify homing success and speed. Translocated individuals showed a very high homing success for distances up to 200 m and successfully returned from up to 400 m. We discuss the potential orientation mechanisms involved and selective forces that could have shaped this strong homing ability.

The ability to differentiate between one's own and foreign offspring ensures the exclusive allocation of costly parental care to only related progeny. The selective pressure to evolve offspring discrimination strategies is largely shaped by the likelihood and costs of offspring confusion. We hypothesize that males and females with different reproductive and spatial behaviours face different risks of confusing their own with others' offspring, and this should favour differential offspring discrimination strategies in the two sexes. In the brilliant-thighed poison frog, Allobates femoralis, males and females are highly polygamous, terrestrial clutches are laid in male territories and females abandon the clutch after oviposition. We investigated whether males and females differentiate between their own offspring and unrelated young, whether they use direct or indirect cues and whether the concurrent presence of their own clutch is essential to elicit parental behaviours. Males transported tadpoles regardless of location or parentage, but to a lesser extent in the absence of their own clutch. Females discriminated between clutches based on exact location and transported tadpoles only in the presence of their own clutch. This sex-specific selectivity of males and females during parental care reflects the differences in their respective costs of offspring confusion, resulting from differences in their spatial and reproductive behaviours.

Understanding the external stimuli and natural contexts that elicit complex behaviors, such as parental care, is key in linking behavioral mechanisms to their real-life function. Poison frogs provide obligate parental care by shuttling their tadpoles from terrestrial clutches to aquatic nurseries, but little is known about the proximate mechanisms that control these behaviors. In this study, we used Allobates femoralis, a poison frog with predominantly male parental care, to investigate whether tadpole transport can be induced in both sexes by transferring unrelated tadpoles to the backs of adults in the field. Specifically, we asked if the presence of tadpoles on an adult's back can override the decision-making rules preceding tadpole pick-up and induce the recall of spatial memory necessary for finding tadpole deposition sites. We used telemetry to facilitate accurate tracking of individual frogs and spatial analyses to compare movement trajectories. All tested individuals transported their foster-tadpoles to water pools outside their home area. Contrary to our expectation, we found no sex difference in the likelihood to transport nor in the spatial accuracy of finding tadpole deposition sites. We reveal that a stereotypical cascade of parental behaviors that naturally involves sex-specific offspring recognition strategies and the use of spatial memory can be manipulated by experimental placement of unrelated tadpoles on adult frogs. As individuals remained inside their home area when only the jelly from tadpole-containing clutches was brushed on the back, we speculate that tactile rather than chemical stimuli are triggering these parental behaviors.