Choosing a Mate in a Cocktail Party‐Like Situation: The Effect of Call Complexity and Call Timing between Two Rival Males on Female Mating Preferences in the Túngara Frog Physalaemus pustulosus

Signal detection, recognition, and localization are hampered when multiple signalers coincide in time and space, a problem known as ‘cocktail party effect’. In many taxa, senders utter complex calls consisting of two or more elements which often vary in the ease with which they can be assessed in different signaling environments. Receivers’ selective attention to different cues may increase the probability of correctly assigning a signal to its source (localization) in face of conspecific interference. Túngara frogs, Physalaemus pustulosus, produce complex calls consisting of an initial whine, followed by zero up to seven broad‐banded, amplitude‐modulated chucks. Under ideal conditions (without interference or noise), females prefer whines followed by chucks over whines alone, but the preference is not linear; females do not discriminate between whines with one or two chucks. When whines lack chucks, call overlap elicits random responses in females, with no preference for leading calls. In this study, I explored the combined effect of call timing and call complexity on female preferences in a two‐choice paradigm—a simplification of the cocktail party scenario. I tested the hypothesis that the effect of call overlap can be reduced when the calls of one of the two rivals have chucks, specifically more chucks than those of the rival. I gave females a choice between whines alone and with chucks (one or two) presented at three time relations (alternated, abutted, and partially overlapped) and two emission orders (whine with less chucks leading and whine with more chucks leading). I found that the preference for one chuck over no chuck was preserved in all the experimental treatments, but when a w + 2chk preceded a w + chk, either overlapped or abutted, a preference existed for the whine with more chucks. Therefore, an interaction between call order and the number of chucks was obtained. The results only partially supported the hypothesis, and call order emerges as an opportunistic component of signaling in P. pustulosus.     

ALLOZYME AND ADVERTISEMENT CALL VARIATION IN THE TÚNGARA FROG,PHYSALAEMUS PUSTULOSUS

We analyzed variation in advertisement calls and allozymes in 30 populations along a 5000-km transect throughout most of the range of the túngara frog, Physalaemus pustulosus. All 12 call variables measured show significant differences among populations despite the importance of the advertisement call in species recognition. Some call variables exhibited clinal variation, whereas most others differed between the two major allozyme groups that have invaded Panama at different times, perhaps 4–4.5 million yr apart. Call variables that primarily affect discrimination among conspecifics tended to exhibit greater variation than call variables that are crucial for species recognition. The proximate mechanism of production underlying a call variable, however, is a better predictor of its variation. Contrary to predictions of some sexual selection models, call variation exhibits predictable patterns of geographical variation, although a substantial portion of variation among populations is not explained by geographic position. Although allozymes, calls, and geography usually covary, closer populations can have more similar calls independent of allozyme similarity.     

Neural networks predict response biases of female túngara frogs.

Artificial neural networks have become useful tools for probing the origins of perceptual biases in the absence of explicit information on underlying neuronal substrates. Preceding studies have shown that neural networks selected to recognize or discriminate simple patterns may possess emergent biases toward pattern size of symmetry--preferences often exhibited by real females--and have investigated how these biases shape signal evolution. We asked whether simple neural networks could evolve to respond to an actual mate recognition signal, the call of the túngara frog, Physalaemus pustulosus. We found that not only were networks capable of recognizing the call of the túngara frog, but that they made remarkably accurate quantitative predictions about how well females generalized to many novel calls, and that these predictions were stable over several architectures. The data suggest that the degree to which P. pustulosus females respond to a call may often be an incidental by-product of a sensory system selected simply for species recognition.     

Phylogeny of the túngara frog genus Engystomops (= Physalaemus pustulosus species group; Anura: Leptodactylidae)

We present a phylogeny of the Neotropical genus Engystomops (= Physalaemus pustulosus species group) based on sequences of approximately 2.4 kb of mtDNA, (12S rRNA, valine-tRNA, and 16S rRNA) and propose a phylogenetic nomenclature. The phylogeny includes all described taxa and two unnamed species. All analyses indicate that Engystomops is monophyletic and contains two basal allopatric clades. Clade I (Edentulus) includes E. pustulosus and the Amazonian E. petersi + E. cf. freibergi. Clade II (Duovox) includes all species distributed in W Ecuador and NW Peru. Brevivox, a clade of small-sized species is strongly supported within Duovox. Populations of Engystomops pustulosus fall into two well-supported clades, each of which occupies two disjunct portions of the species range. Overall, our phylogeny is congruent with most previous hypotheses. This study is among the few published species-level phylogenies of Neotropical amphibians derived from molecular datasets. A review of the proportion of new species detected by similar studies suggests that the increasing use of molecular techniques will lead to the discovery of a vast number of species of Neotropical amphibians.     

Generalization in response to mate recognition signals

Females usually exhibit strong and unequivocal recognition of conspecific mating signals and reject those of other sympatric heterospecifics. However, most species are allopatric with one another, and the degree to which females recognize mating signals of allopatric species is more varied. Such mating signals are often rejected but are sometimes falsely recognized as conspecific. We studied the dynamics of mate recognition in female túngara frogs (Physalaemus pustulosus) in response to a series of calls that were intermediate between the conspecific and each of five allopatric-heterospecific calls: two that elicited recognition from females in previous studies and three that did not. This study shows that females perceive variation in allopatric mating signals in acontinuous manner with no evidence of perceptual category formation. The strength of recognition is predicted by how different the target stimulus is from the conspecific call within a series of calls. But the differences in recognition responses among call series are not predicted by the similarity of the call series to the conspecific call. The latter result suggests that the strength of recognition of allopatric signals might be influenced by processes of stimulus generalization and past evolutionary history.     

Do frog-eating bats perceptually bind the complex components of frog calls?

The mating calls of male túngara frogs, Physalaemus pustulosus, attract intended (conspecific females) and unintended (eavesdropping predators and parasites) receivers. The calls are complex, having two components: a frequency-modulated “whine” followed by 0–7 harmonic bursts or “chucks”. The whine is necessary and sufficient to elicit phonotaxis from females and the chuck enhances call attractiveness when it follows a whine. Although chucks are never made alone, females perceptually bind the whine and chuck when they are spatially separated. We tested whether an unintended receiver with independent evolution of phonotaxis, the frog-eating bat, Trachops cirrhosus, has converged with frogs in its auditory grouping of the call components. In contrast to frogs, bats approached chucks broadcast alone; when the chuck was spatially separated from the whine the bats preferentially approached the whine, and bats were sensitive to whine–chuck temporal sequence. This contrast suggests that although disparate taxa may be selected to respond to the same signals, different evolutionary histories, selective regimes, and neural and cognitive architectures may result in different weighting and grouping of signal components between generalist predators and conspecific mates.     

Acoustic preferences and localization performance of blood-sucking flies (Corethrella Coquillett) to tungara frog calls

Mating signals that increase attractiveness of males to femalescan also increase conspicuousness of the signaler to predatorsand parasites. We investigated the acoustic preference of speciesof blood-sucking flies of the genus Corethrella (Diptera: Corethrellidae),which eavesdrop on the sexual advertisement signals of túngarafrogs (Physalaemus pustulosus). Male frogs of this species facultativelyproduce 2 types of mating calls: simple (whines alone) and complex(whines and chucks). We tested the acoustic preference of theflies and their ability to locate their host when the frogsproduce simple or complex calls. The flies exhibited phonotaxisto both types of calls but were preferentially attracted tocomplex calls. We show that acoustic information alone is sufficientfor the flies' accurate localization of calling frogs. Complexcalls, however, were not approached at closer distance or withdecreased landing error (i.e., proportion of landings outsidethe target) than simple calls, suggesting that call structuredoes not influence localization performance. Female túngarafrogs and frog-eating bats (Trachops cirrhosus) also prefercomplex to simple túngara frog calls. Thus, intendedand unintended receivers with different ear morphologies exhibitthe same preference for a specific túngara frog calltype. This result is discussed in the context of the evolutionof call attractiveness in a communication network.     

一种泡蟾的配偶识别:脑、行为和进化研究

综述了对泡蟾(Physalaemuspus pustulosus)的性通讯所进行的各种研究。本文作者研究了通讯系统的功能重要性、通讯的神经机制以及通讯进化的格局,特别研究了雌蛙如何使用简单和复杂的叫声来识别可以交配的物种,以及如何识别同种的不同雄性个体。本文确定了复杂叫声的反选择力、一种食蛙蝠——缨唇蝠(Trachops cirrhosus)以及叫声的能量消耗,并阐述了引起雌性反应的重要特征。作者的比较研究表明,对复杂叫声的偏好在复杂叫声进化前就有了,这支持感官开发假说(Hypothesis of sensory exploitation,雄性特征进化的结果是开发雌性预先存在的对雄性的偏好)。结合比较研究和人工神经网络的模拟,作者认为雌蛙用来识别雄性简单叫声的解码策略受其祖先解码策略的影响[动物学报49(6)：713～726,2003]。     

THE SENSORY BASIS OF SEXUAL SELECTION FOR COMPLEX CALLS IN THE TÚNGARA FROG,PHYSALAEMUS PUSTULOSUS (SEXUAL SELECTION FOR SENSORY EXPLOITATION)

Male túngara frogs (Physalaemus pustulosus) vocalize to attract females, and enhance the attractiveness of their simple, whine-only call by adding chucks to produce complex calls. Complex calls contain more total energy and are of longer duration. By virtue of the greater frequency range of the chuck, complex calls also simultaneously stimulate both the amphibian papilla and the basilar papilla of the frog's inner ear. Female phonotaxis experiments using synthetic stimuli demonstrate that an increase in the call's acoustic energy is not sufficient to account for the enhanced attractiveness of the complex call. However, the stimulation of either or both of the female's sound-sensitive inner-ear organs is sufficient to elicit her preference. We suggest that the female's sensory system generates selection that equally favors at least three evolutionary alternatives for enhancing call attractiveness and that historical constraints imposed by the male's morphology determined which of the alternatives was more likely to evolve. These data are consistent with our hypothesis of sensory exploitation, which states that selection favors those traits that elicit greater stimulation from the female's sensory system and which emphasizes the nonadaptive nature of female preference.     

Acoustic Competition in Physalaemus pustulosus, a Differential Response to Calls of Relative Frequency

The response of the male túngara frog ( Physalaemus pustulosus ) to conspecific whines of different frequencies was examined. In the first series of playback experiments (fixed frequency), three types of synthetic stimuli were used, corresponding to calls of high frequency (HFF; x + 2 SD), mid-frequency (MFF; x ), and low frequency (LFF; x − 2 SD) for the study population. In the second series of interactive playback experiments (relative frequency), whines of frequency relative to that of the male subject were used: male frequency + 2 SD (HRF), male frequency (MRF), and male frequency −2 SD (LRF). In the fixed frequency experiments, male vocal response did not vary among treatments. However, in the relative frequency experiments, males responded with more whines, and above all, with more chucks, to stimuli of similar or higher frequency than to stimuli of lower frequency than their own. In other words, male vocal competition escalates when competitors have whines with similar or relatively higher frequencies to their own, but does not increase when competitors have call frequencies that are at the mean or higher for the population. This differential response might result from competition between males of different sizes, since the frequency of the whine and male size is significantly correlated.     

The vocal sac increases call rate in the Tungara frog Physalaemus pustulosus

In most anurans, the production of advertisement calls is accompanied by the inflation of a vocal sac. Current functions of the vocal sac, however, are not fully understood, although several hypotheses have been proposed. One hypothesis suggests that the vocal sac decreases the intercall interval (i.e., increases call rate) by reinflating the lungs more rapidly than is possible with the buccal pump. We investigate this hypothesis by analyzing audio and video recordings of calling tungara frogs. We compare the first two call bouts emitted by an originally uninflated male. The first call bout requires lung inflation via buccal pumping, but in the second, the male is already inflated because of capture of air and reinflation of the lungs by the vocal sac. Lung inflation to typical field levels requires 26-51 buccal pumps, which takes at least 4.4 s. This estimate is more than 2.5 times the typical intercall interval with lung reinflation via a vocal sac (ca. 1.7 s). Evidence from phonotaxis tests demonstrates that these differences in intercall intervals are salient to females and that female Physalaemus pustulosus prefer the shorter intercall interval/higher call rate. Acoustic analyses demonstrate that the first call of bout 1, which requires buccal pumping, is usually shorter, of lower amplitude, and spans a smaller frequency range than the first call of bout 2, which does not require buccal pumping. Because females prefer longer, more intense calls, these results suggest that the vocal sac not only increases call rate but also allows males to produce more calls of increased attractiveness to females.     

Sexual selection in female perceptual space: how female túngara frogs perceive and respond to complex population variation in acoustic mating signals

Female preferences for male mating signals are often evaluated on single parameters in isolation or small suites of characters. Most signals, however, are composites of many individual parameters. In this study we quantified multivariate traits in the advertisement call of the túngara frog, Physalaemus pustulosus. We represented the calls in multidimensional scaling space and chose nine test calls to represent the range of population variation. We then tested females for phonotactic preference between calls in each pair of the nine test calls. We used statistics developed for paired comparisons in such "round robin" competitions to evaluate the null hypothesis of equal attractiveness, and to examine the degree to which females responded to calls as being different from or similar to one another in attractiveness. We then examined the attractiveness of each test call relative to all other test calls as a function of their location in multivariate acoustic space (the acoustic landscape) to visualize sexual selection on calls. Finally, we used methods from cognitive psychology to illustrate the females' perception of call attractiveness in multivariate space, and compared this perceptual landscape to the acoustic landscape of quantitative call variation. We show that correlations between individual call characters are not strong and thus there are few biomechanical constraints on their independent evolution. Most call variables differed among males, and there was high repeatability of call characters within males. Females often discriminated between pairs of calls from the population, and there were significant differences among calls in their attractiveness. Female preferences for calls were not stabilizing. The region of the acoustic landscape that was most attractive to females included the mean call but was not centered around it. The females' perceptual or preference landscape did not correlate with the call's acoustic landscape, and female perception of calls decreased rather than enhanced call differences.     

The influence of geographic heterogeneity in predation pressure on sexual signal divergence in an Amazonian frog species complex

Sexual selection plays an important role in mating signal divergence, but geographic variation in ecological factors can also contribute to divergent signal evolution. We tested the hypothesis that geographic heterogeneity in predation causes divergent selection on advertisement call complexity within the Engystomops petersi (previously Physalaemus petersi) frog species complex. We conducted predator phonotaxis experiments at two sites where female choice is consistent with call trait divergence. Engystomops at one site produces complex calls, whereas the closely related species at the other site produces simple calls. Bats approached complex calls more than simple calls at both sites, suggesting selection against complex calls. Moreover, bat predation pressure was greater at the site with simple calls, suggesting stronger selection against complex calls and potentially precluding evolution of complex calls at this site. Our results show that geographic variation in predation may play an important role in the evolution and maintenance of mating signal divergence.     

Phylogenetic influence on mating call preferences in female túngara frogs, Physalaemus pustulosus

We evaluated how various phylogenetic models for estimating ancestral characters can influence studies of behavioural evolution. Previously we used a single model of evolution to estimate the values of call characters at ancestral nodes for the Physalaemus pustulosus species group and some close relatives (Ryan & Rand 1995, Science, 269, 390-392). We then synthesized these ancestral calls and measured the females' responses to such calls in phonotaxis experiments. We repeated the above procedure to determine the sensitivity of these results and conclusions to various models used to estimate the ancestral call characters. We asked whether: (1) different models gave different call estimates for the same nodes; (2) different call estimates at the same node were perceived as different by females; and (3) differences in female responses influenced previous conclusions. We used seven different models that varied in at least one of the following parameters: tree topology (bifurcating versus pectinate in-group trees), algorithms (local squared-change versus squared-change parsimony), tempo (gradual or punctuated evolution), and outgroups (two or three outgroup taxa used). Although different models often gave different call estimates for the same node, these different estimates often were not perceived as different by the females. These data reinforce our previous conclusions that: (1) the range of female preferences exceeds the known variation of the conspecific call; (2) females do not discriminate between the conspecific call and the call of their most recent ancestor; and (3) female responses may be context dependent, given that females differ in their responses to the same signal variation in discrimination and recognition experiments. Copyright 1999 The Association for the Study of Animal Behaviour.     

Frogs Have Rules: Selective Attention Algorithms Regulate Chorusing in Physalaemus pustulosus (Leptodactylidae)

In various acoustic insects and frogs, females preferentially orient towards the leading of two or more males' advertisement signals that occur closely in time. Such preferences in receivers have apparently selected for timing mechanisms whereby male signallers actively refrain from calling immediately following the onset of a neighbour's call and thereby increase their production of leading calls. However, indiscriminate application of this inhibitory mechanism to all neighbours might severely reduce a male's calling rate, particularly in high density. Consequently, mechanisms of selective attention to only a subset of signalling neighbours are expected.
Female Túngara frogs ( Physalaemus pustulosus ) exhibit strong preferences for leading male calls, and males refrain from calling immediately following a neighbour. Four-loudspeaker playback experiments demonstrated that males do selectively apply this inhibitory mechanism to only a subset of close signalling neighbours. Selective attention is regulated by a combination of sliding threshold and fixed number rules: (i) Attend to the loudest (nearest) conspecific neighbour and those additional ones whose calls are within 6–8 dB of the loudest one; (ii) attend to only two neighbours in total when the calls are weak or the second one is much farther than the first; (iii) attend to three neighbours when the calls are loud or all neighbours are approximately equidistant. The means by which such plasticity may be achieved and its potential adaptiveness are discussed.     

Phylogeny of frogs of the Physalaemus pustulosus species group,with an examination of data incongruence

Characters derived from advertisement calls, morphology, allozymes, and the sequences of the small subunit of the mitochondrial ribosomal gene (12S) and the cytochrome oxidase I (COI) mitochondrial gene were used to estimate the phylogeny of frogs of the Physalaemus pustulosus group (Leptodactylidae). The combinability of these data partitions was assessed in several ways: measures of phylogenetic signal, character support for trees, congruence of tree topologies, compatibility of data partitions with suboptimal trees, and homogeneity of data partitions. Combined parsimony analysis of all data equally weighted yielded the same tree as the 12S partition analyzed under parsimony and maximum likelihood. The COI, allozyme, and morphology partitions were generally congruent and compatible with the tree derived from combined data. The call data were significantly different from all other partitions, whether considered in terms of tree topology alone, partition homogeneity, or compatibility of data with trees derived from other partitions. The lack of effect of the call data on the topology of the combined tree is probably due to the small number of call characters. The general incongruence of the call data with other data partitions is consistent with the idea that the advertisement calls of this group of frogs are under strong sexual selection.     

No pre-existing biases for heterospecific call traits in the frog Physalaemus enesefae

Previous phonotaxis studies with two species of the Physalaemuspustulosus species group indicated that female preferences for several heterospecific call traits resulted from sensory biases inherited from a common ancestor. In phonotaxis experiments, we determined whether Physalaemus enesefae, a distant relative of the P. pustulosus group, showed similar preferences for call traits not present in conspecific males. We presented females with a choice between the typical conspecific advertisement call and the same call to which we digitally appended a chuck from P. pustulosus, a squawk from P. freibergi, and an amplitude-modulated prefix from P. pustulatus. In addition we presented the advertisement call in doublets, a trait peculiar to P. coloradorum. We also analysed male vocal behaviour evoked in response to the same suite of stimuli. Physalaemus enesefae females did not prefer the calls of their own males with appended heterospecific traits over unmodified calls, nor conspecific calls in doublets over single calls. The lack of preference among females was not the result of a behavioural polymorphism. Female responses to repeated presentations of the same stimulus pair were not consistent. Males also did not show an enhanced vocal response to altered calls relative to the typical conspecific call. Consequently, there are no pre-existing biases for these heterospecific call traits in P. enesefae; pre-existing preferences in the P. pustulosus group could have been inherited from an ancestor not shared with P. enesefae, but data from other closely related species are needed to confirm this conclusion. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Tests of biogeographic hypotheses for diversification in the Amazonian forest frog, Physalaemus petersi

Several hypotheses have been proposed to explain the biogeographic processes that generate the high species richness of the Amazon basin. We tested two of them in a terra firme (upland) forest frog species, Physalaemus petersi: (1) the riverine barrier hypothesis; and (2) the elevational gradient hypothesis. Mitochondrial DNA sequence data (2.4 kb) from the 12S, 16S, and intervening valine tRNA genes were obtained from 65 P. petersi individuals and 4 outgroup taxa and analyzed with a combination of phylogenetic and population genetic approaches. Moderate support for the riverine barrier hypothesis was found for one of the three rivers examined, but little evidence was found for the elevational gradient hypothesis. Phylogenetic analyses revealed that high levels of sequence divergence (an average of 4.57-4.79%) separate three well-supported clades from the northwestern, southwestern, and eastern Amazon. Strong evidence for recent population expansion in P. petersi in the southwestern region of the Amazon basin was also uncovered.     

Sexual selection drives speciation in an Amazonian frog

One proposed mechanism of speciation is divergent sexual selection, whereby divergence in female preferences and male signals results in behavioural isolation. Despite the appeal of this hypothesis, evidence for it remains inconclusive. Here, we present several lines of evidence that sexual selection is driving behavioural isolation and speciation among populations of an Amazonian frog (Physalaemus petersi). First, sexual selection has promoted divergence in male mating calls and female preferences for calls between neighbouring populations, resulting in strong behavioural isolation. Second, phylogenetic analysis indicates that populations have become fixed for alternative call types several times throughout the species' range, and coalescent analysis rejects genetic drift as a cause for this pattern, suggesting that this divergence is due to selection. Finally, gene flow estimated with microsatellite loci is an average of 30 times lower between populations with different call types than between populations separated by a similar geographical distance with the same call type, demonstrating genetic divergence and incipient speciation. Taken together, these data provide strong evidence that sexual selection is driving behavioural isolation and speciation, supporting sexual selection as a cause for speciation in the wild.     

SPECIES RECOGNITION AND SEXUAL SELECTION AS A UNITARY PROBLEM IN ANIMAL COMMUNICATION

We investigated patterns of mating call preference and mating call recognition by examining phonotaxis of female túngara frogs, Physalaemus pustulosus, in response to conspecific and heterospecific calls. There are four results: females always prefer conspecific calls; most heterospecific calls do not elicit phonotaxis; some heterospecific calls do elicit phonotaxis and thus are effective mate recognition signals; and females prefer conspecific calls to which a component of a heterospecific call has been added to a normal conspecific call. We use these data to illustrate how concepts of species recognition and sexual selection can be understood in a unitary framework by comparing the distribution of signal traits to female preference functions.