Diversity of temporal response patterns in midbrain auditory neurons of frogs Batrachyla and its relevance for male vocal responses.

We investigated response selectivities of single auditory neurons in the torus semicircularis of male frogs Batrachyla leptopus (72 neurons) and B. taeniata (57 neurons) to synthetic stimuli of different temporal structures. Series of stimuli in which note and pulse rate, note and pulse structure and call duration varied systematically were presented. Neuronal responses quantified in terms of proportions of units displaying diverse temporal transfer functions are related in different modes with patterns of evoked vocal responses studied previously in these frogs. Correspondences and mismatches occurred between the auditory and vocal domains. The analysis of this evidence together with corresponding information from previous neuronal and behavioral studies in the third species of this genus, B. antartandica, indicates that different modes of preferences for acoustic communication signals can coexist within this anuran group.

Geographic variation in the matching between call characteristics and tympanic sensitivity in the Weeping lizard.

Effective communication requires a match among signal characteristics, environmental conditions, and receptor tuning and decoding. The degree of matching, however, can vary, among others due to different selective pressures affecting the communication components. For evolutionary novelties, strong selective pressures are likely to act upon the signal and receptor to promote a tight match among them. We test this prediction by exploring the coupling between the acoustic signals and auditory sensitivity in Liolaemus chiliensis, the Weeping lizard, the only one of more than 285 Liolaemus species that vocalizes. Individuals emit distress calls that convey information of predation risk to conspecifics, which may respond with antipredator behaviors upon hearing calls. Specifically, we explored the match between spectral characteristics of the distress calls and the tympanic sensitivities of two populations separated by more than 700 km, for which previous data suggested variation in their distress calls. We found that populations differed in signal and receptor characteristics and that this signal variation was explained by population differences in body size. No precise match occurred between the communication components studied, and populations differed in the degree of such correspondence. We suggest that this difference in matching between populations relates to evolutionary processes affecting the Weeping lizard distress calls.

Lizard predation by spiders: A review from the Neotropical and Andean regions.

Vertebrate predation by invertebrates has been classically underexplored and thus underestimated, despite the fact that many arthropods consume vertebrates. To shed some light on the relevance that spider predation may have upon lizards in the Neotropical and Andean regions, we compiled the available information in the literature on this trophic interaction. We found 50 reports of spiders consuming lizards in these regions, and the 88% of these were from the Neotropical region. Spiders belong to eight families, but Ctenidae and Theraphosidae were the most frequently reported predators. Lizards belong to 12 families, and the most commonly consumed species corresponded to the families Dactyloidae (all Anolis lizards), Gymnophthalmidae, and Sphaerodactylidae. Data suggest trophic spider-lizard associations between Ctenidae and Dactyloidae, followed by Theraphosidae and Liolaemidae. The body sizes of the spiders and lizards showed a positive relationship, and spiders were smaller than their prey. We conclude that various spider taxa can be considered lizard predators and they may be ecologically important in the Neotropical and Andean regions. However, spiders of prime predation relevance seem to be those of the Ctenidae and Theraphosidae families.

Biotic and abiotic sounds affect calling activity but not plasma testosterone levels in male frogs (Batrachyla taeniata) in the field and in captivity.

In animals, the expression of diverse reproductive behaviors is hormonally regulated. In particular, vocalizing during courtship has been related to circulating androgen levels, and reciprocally, conspecific vocalizations are known to modulate androgen secretion in vertebrates. The effect of natural sounds of abiotic origin on hormonal status has virtually not received attention. Therefore, we evaluated the vocal responses of male Batrachyla taeniata frogs to conspecific chorus and rainfall sounds in natural and controlled laboratory settings, measuring the testosterone levels of exposed individuals. In field and laboratory conditions, testosterone levels of frogs exposed to 31.5 min of chorus and rain sounds and non-exposed individuals were similar. In the field, frogs increased their call rate in response to playbacks of chorus and rain sound, but the evoked calling activity was unrelated to plasma testosterone. In contrast to the field, frogs showed limited responsiveness to 31.5-min acoustic exposures in the laboratory. Similarly to the field, for vocally active males tested in the laboratory there was no association between call rate and testosterone levels. Additionally, in this group, testosterone levels were higher in vocally active males relative to non-calling individuals. Overall, these results indicate that in B. taeniata testosterone levels are not altered following a short-term exposure to conspecific biotic and to abiotic sounds. Our results are suggestive of a threshold influence of testosterone on the vocal activity of the species studied. Further explorations of the influence of abiotic sounds on endocrine activation are required to understand how animals respond to variable acoustic environmental conditions.

Effects of traffic noise exposure on corticosterone, glutathione and tonic immobility in chicks of a precocial bird.

Repeated exposure to traffic noise may be perceived as a succession of stressors, and therefore, noisy urban environments could lead to a state of chronic stress. In developing animals, glucocorticoids can have organizational effects on the hypothalamic-pituitary-adrenal axis in addition to the classic activation effects, so evaluating the effect of traffic noise during development is urgently needed. To our knowledge, to date six studies have investigated the effects of traffic noise on baseline corticosterone (CORT) and/or the stress response in birds during development; however, these studies were performed in nestling (altricial species), where confounding factors (e.g. communication between nestlings and parents) could mask the real impact of traffic noise on stress. In this study, we evaluated the effect of traffic noise (traffic noise group vs. rural noise group) on baseline levels of CORT and stress responses in chicks of a precocial bird species, the Japanese quail (Coturnix japonica). Because CORT can also decrease glutathione (GSH) levels (antioxidant and neurotransmitter/modulator), secondly by means of path analysis we investigated whether the strength of the association between CORT levels, GSH levels and tonic immobility (TI) varied in relation to treatment. We observed (i) similar baseline levels of CORT in both groups, (ii) a trend toward higher stress response in the traffic noise group (P = 0.08), (iii) similar TI duration in both groups, (iv) higher GSH levels in the traffic noise group and (v) differences in the strength and sign of the associations in relation to the treatment (traffic vs. rural). We conclude that the acoustic environment perceived during development has implications for physiology and behaviour; as more research is done on this topic, the need for sustainable urban planning will become clearer.

The acoustic adaptation hypothesis in a widely distributed South American frog: Southernmost signals propagate better.

Animal communication occurs in environments that affect the properties of signals as they propagate from senders to receivers. We studied the geographic variation of the advertisement calls of male Pleurodema thaul individuals from eight localities in Chile. Furthermore, by means of signal propagation experiments, we tested the hypothesis that local calls are better transmitted and less degraded than foreign calls (i.e. acoustic adaptation hypothesis). Overall, the advertisement calls varied greatly along the distribution of P. thaul in Chile, and it was possible to discriminate localities grouped into northern, central and southern stocks. Propagation distance affected signal amplitude and spectral degradation in all localities, but temporal degradation was only affected by propagation distance in one out of seven localities. Call origin affected signal amplitude in five out of seven localities and affected spectral and temporal degradation in six out of seven localities. In addition, in northern localities, local calls degraded more than foreign calls, and in southern localities the opposite was observed. The lack of a strict optimal relationship between signal characteristics and environment indicates partial concordance with the acoustic adaptation hypothesis. Inter-population differences in selectivity for call patterns may compensate for such environmental constraints on acoustic communication.

Comparing the antipredator behaviour of two sympatric, but not syntopic, Liolaemus lizards.

The microhabitat preferences of prey animals can modulate how they perceive predation risk, and therefore, their antipredator behaviour. We tested under standardized conditions how microhabitat preferences of two Liolaemus lizards affected their responses when confronted with two types of ambush predators (raptor vs. snake), under two levels of predation risk (low vs. high). These lizard species are sympatric, but not syntopic; L. chiliensis basks on bushes, a complex microhabitat that may provide protection against visual predators, while L. nitidus prefers open microhabitats, basking on the top of large bare rocks, highly exposed to visual predators. If microhabitat complexity modulates the antipredator response, L. chiliensis may perceive lower predation risk, exhibiting lower intensity of antipredator responses than L. nitidus. Both species reduced their activity after being exposed to both predators, but lizards differed in the assessment of predation risk; L. nitidus reduced its activity independently of the predation risk experienced, while L. chiliensis only reduced its activity in the high-risk condition. The microhabitat preferences shaped during the evolution of these species seem to modulate their perception of predation risk, which may cause interspecific differences in the associated costs of their antipredator responses.

Arginine Vasotocin, the Social Neuropeptide of Amphibians and Reptiles.

Arginine vasotocin (AVT) is the non-mammalian homolog of arginine vasopressin (AVP) and, like vasopressin, serves as an important modulator of social behavior in addition to its peripheral functions related to osmoregulation, reproductive physiology, and stress hormone release. In amphibians and reptiles, the neuroanatomical organization of brain AVT cells and fibers broadly resembles that seen in mammals and other taxa. Both parvocellular and magnocellular AVT-containing neurons are present in multiple populations located mainly in the basal forebrain from the accumbens-amygdala area to the preoptic area and hypothalamus, from which originate widespread fiber connections spanning the brain with a particularly heavy innervation of areas associated with social behavior and decision-making. As for mammalian AVP, AVT is present in greater amounts in males in many brain areas, and its presence varies seasonally, with hormonal state, and in males with differing social status. AVT's social influence is also conserved across herpetological taxa, with significant effects on social signaling and aggression, and, based on the very small number of studies investigating more complex social behaviors in amphibians and reptiles, AVT may also modulate parental care and social bonding when it is present in these vertebrates. Within this conserved pattern, however, both AVT anatomy and social behavior effects vary significantly across species. Accounting for this diversity represents a challenge to understanding the mechanisms by which AVT exerts its behavioral effects, as well are a potential tool for discerning the structure-function relationships underlying AVT's many effects on behavior.

Vocal responses of austral forest frogs to amplitude and degradation patterns of advertisement calls.

Degradation phenomena affecting animal acoustic signals may provide cues to assess the distance of emitters. Recognition of degraded signals has been extensively demonstrated in birds, and recently studies have also reported detection of degraded patterns in anurans that call at or above ground level. In the current study we explore the vocal responses of the syntopic burrowing male frogs Eupsophus emiliopugini and E. calcaratus from the South American temperate forest to synthetic conspecific calls differing in amplitude and emulating degraded and non-degraded signal patterns. The results show a strong dependence of vocal responses on signal amplitude, and a general lack of differential responses to signals with different pulse amplitude modulation depths in E. emiliopugini and no effect of relative amplitude of harmonics in E. calcaratus. Such limited discrimination of signal degradation patterns from non-degraded signals is likely related to the burrowing habits of these species. Shelters amplify outgoing and incoming conspecific vocalizations, but do not counteract signal degradation to an extent comparable to calling strategies used by other frogs. The limited detection abilities and resultant response permissiveness to degraded calls in these syntopic burrowing species would be advantageous for animals communicating in circumstances in which signal alteration prevails.

Synthetic rainfall vibrations evoke toad emergence.

Toads occupy underground refugia during periods of daily or seasonal inactivity, emerging only during rainfall [1]. We test the hypothesis that rainfall-induced vibrations in soil are the cues that trigger the emergence of toads from underground. Using playback experiments in the absence of natural rainfall in native habitats, we observed that two Iberian toad species (Pelobates cultripes and Bufo calamita) emerged significantly earlier than controls when exposed to low-frequency soil vibrations that closely mimic those of rainfall. Our results suggest that detection of abiotic seismic events are biologically relevant and widespread in arid-zone anurans. These findings provide insights into the evolutionary role played by the two low-frequency-tuned inner-ear organs in anuran amphibians - the amphibian papilla and sacculus, both detectors of weak environmental vibrational cues.

Extended amplification of acoustic signals by amphibian burrows.

Animals relying on acoustic signals for communication must cope with the constraints imposed by the environment for sound propagation. A resource to improve signal broadcast is the use of structures that favor the emission or the reception of sounds. We conducted playback experiments to assess the effect of the burrows occupied by the frogs Eupsophus emiliopugini and E. calcaratus on the amplitude of outgoing vocalizations. In addition, we evaluated the influence of these cavities on the reception of externally generated sounds potentially interfering with conspecific communication, namely, the vocalizations emitted by four syntopic species of anurans (E. emiliopugini, E. calcaratus, Batrachyla antartandica, and Pleurodema thaul) and the nocturnal owls Strix rufipes and Glaucidium nanum. Eupsophus advertisement calls emitted from within the burrows experienced average amplitude gains of 3-6 dB at 100 cm from the burrow openings. Likewise, the incoming vocalizations of amphibians and birds were amplified on average above 6 dB inside the cavities. The amplification of internally broadcast Eupsophus vocalizations favors signal detection by nearby conspecifics. Reciprocally, the amplification of incoming conspecific and heterospecific signals facilitates the detection of neighboring males and the monitoring of the levels of potentially interfering biotic noise by resident frogs, respectively.

Individual and sex distinctiveness in bark calls of domestic chinchillas elicited in a distress context.

Animals obtain information about their social environment by means of communication signals, which provide relevant subtle cues for individual recognition. An important requisite for this process is the existence of larger between- than within-emitter signal variation. Acoustic signals are complex traits susceptible of variation in their spectral and temporal components, implying that signal distinctiveness can result from differences in single or various acoustic components. In this study, domestic chinchillas were induced to vocalize in a distress context to describe the acoustic characteristics of the bark calls, and to determine features that denote the potential value of this vocalization for individual and/or sexual recognition. The results demonstrate that the variation in spectral and temporal components of the bark calls of chinchillas elicited under a distress context is larger between than within individuals, suggesting the potential of these signals for distinctiveness between individual signalers, although the potential of this call type for sex distinctiveness is quite limited. These results combined with previous studies on auditory capabilities of chinchillas contribute to position this rodent as a valuable model species for studying auditory-vocal interactions.

Contrasting Propagation of Natural Calls of Two Anuran Species from the South American Temperate Forest.

The acoustic adaptation hypothesis predicts that sound communication signals have an optimal relationship with animals' native environments. However, species sharing a habitat produce signals stratified in the spectral domain and exhibit different temporal patterns resulting in acoustic niche partitioning. The diversity generated is likely to affect differently the characteristics of propagating signals. We recorded at various distances from the sound source calls of the frogs Eupsophus calcaratus and E. emiliopugini in the austral temperate forest where they communicate and breed syntopically. The calls of E. calcaratus have higher frequency components and lower amplitude relative to calls of E. emiliopugini, and the acoustic active space for the signals of E. calcaratus is restricted relative to E. emiliopugini. The signals of both species experience similar attenuation patterns, but calls of E. calcaratus are affected by spectral degradation to a larger extent, with linear decreases in spectral cross-correlation and in the amplitude ratio between the first two harmonics. The calls of E. emiliopugini are affected by temporal degradation as a linear decrease in amplitude modulation depth of their pulsed structure. Further studies are needed to assess the relative importance of selective and phylogenetic factors on the divergent propagation patterns reported.

Lack of phonotactic preferences of female frogs and its consequences for signal evolution.

Sexual selection is one of the main evolutionary forces that drive signal evolution. In previous studies, we have found out that males of Pleurodema thaul, a frog with an extensive latitudinal distribution in Chile, emits advertisement calls that show remarkable variation among populations. In addition, this variation is related to intense inter-male acoustic competition (intra-sexual selection) occurring within each population. However, the extent to which female preferences contribute to the signal divergence observed is unclear. To study the responsiveness of females in each population, we stimulated females with synthetic calls designed with the acoustic structure of their own population and subsequently responsive females were subjected to a two-choice experiment, where they were stimulated with synthetic calls of their own population versus a call of a foreign population. Females do not show phonotactic preferences for calls of their own or foreign populations as measured with both linear and circular variables. The lack of phonotactic preferences suggests an absence of participation of inter-sexual selection processes in the divergence of the acoustic signals of P. thaul, highlighting the importance of intra-sexual selection for the evolution of these signals. These results concur with studies in other vertebrates emphasizing the relevance of interactions among males for the evolution of acoustic communication systems.

Divergence of acoustic signals in a widely distributed frog: relevance of inter-male interactions.

Divergence of acoustic signals in a geographic scale results from diverse evolutionary forces acting in parallel and affecting directly inter-male vocal interactions among disjunct populations. Pleurodema thaul is a frog having an extensive latitudinal distribution in Chile along which males' advertisement calls exhibit an important variation. Using the playback paradigm we studied the evoked vocal responses of males of three populations of P. thaul in Chile, from northern, central and southern distribution. In each population, males were stimulated with standard synthetic calls having the acoustic structure of local and foreign populations. Males of both northern and central populations displayed strong vocal responses when were confronted with the synthetic call of their own populations, giving weaker responses to the call of the southern population. The southern population gave stronger responses to calls of the northern population than to the local call. Furthermore, males in all populations were stimulated with synthetic calls for which the dominant frequency, pulse rate and modulation depth were varied parametrically. Individuals from the northern and central populations gave lower responses to a synthetic call devoid of amplitude modulation relative to stimuli containing modulation depths between 30-100%, whereas the southern population responded similarly to all stimuli in this series. Geographic variation in the evoked vocal responses of males of P. thaul underlines the importance of inter-male interactions in driving the divergence of the acoustic traits and contributes evidence for a role of intra-sexual selection in the evolution of the sound communication system of this anuran.

Ample active acoustic space of a frog from the South American temperate forest.

The efficiency of acoustic communication depends on the power generated by the sound source, the attributes of the environment across which signals propagate, the environmental noise and the sensitivity of the intended receivers. Eupsophus emiliopugini, an anuran from the temperate austral forest communicates by means of an advertisement call of moderate intensity within the range for anurans. To estimate the range over which these frogs communicate effectively, we conducted measurements of call sound levels and of auditory thresholds to pure tones and to synthetic conspecific calls. The results show that E. emiliopugini produces advertisement calls of about 84 dB SPL at 0.25 m from the caller. The signals are affected by attenuation as they propagate, reaching average values of about 47 dB SPL at 8 m from the sound source. Midbrain multi-unit recordings show quite sensitive audiograms within the anuran range, with thresholds of about 44 dB SPL for synthetic imitations of conspecific calls, which would allow communication at distances beyond 8 m. This is an extended range as compared to E. calcaratus, a related syntopic species for which a previous study has shown to be restricted to active acoustic spaces shorter than 2 m. The comparison reveals divergent strategies for related taxa communicating amid the same environment.

Call transmission efficiency in native and invasive anurans: competing hypotheses of divergence in acoustic signals.

Invasive species are a leading cause of the current biodiversity decline, and hence examining the major traits favouring invasion is a key and long-standing goal of invasion biology. Despite the prominent role of the advertisement calls in sexual selection and reproduction, very little attention has been paid to the features of acoustic communication of invasive species in nonindigenous habitats and their potential impacts on native species. Here we compare for the first time the transmission efficiency of the advertisement calls of native and invasive species, searching for competitive advantages for acoustic communication and reproduction of introduced taxa, and providing insights into competing hypotheses in evolutionary divergence of acoustic signals: acoustic adaptation vs. morphological constraints. Using sound propagation experiments, we measured the attenuation rates of pure tones (0.2-5 kHz) and playback calls (Lithobates catesbeianus and Pelophylax perezi) across four distances (1, 2, 4, and 8 m) and over two substrates (water and soil) in seven Iberian localities. All factors considered (signal type, distance, substrate, and locality) affected transmission efficiency of acoustic signals, which was maximized with lower frequency sounds, shorter distances, and over water surface. Despite being broadcast in nonindigenous habitats, the advertisement calls of invasive L. catesbeianus were propagated more efficiently than those of the native species, in both aquatic and terrestrial substrates, and in most of the study sites. This implies absence of optimal relationship between native environments and propagation of acoustic signals in anurans, in contrast to what predicted by the acoustic adaptation hypothesis, and it might render these vertebrates particularly vulnerable to intrusion of invasive species producing low frequency signals, such as L. catesbeianus. Our findings suggest that mechanisms optimizing sound transmission in native habitat can play a less significant role than other selective forces or biological constraints in evolutionary design of anuran acoustic signals.

Severe constraints for sound communication in a frog from the South American temperate forest.

The efficiency of acoustic communication depends on the power generated by the sound source, the quality of the environment across which signals propagate, the environmental noise and the sensitivity of the intended receivers. Eupsophus calcaratus, an anuran from the temperate austral forest, communicates by means of an advertisement call of weak intensity in a sound-attenuating environment. To estimate the range over which these frogs communicate effectively, we conducted measurements of sound level and degradation patterns of propagating advertisement calls in the field, and measurements of auditory thresholds to pure tones and to natural calls in laboratory conditions. The results show that E. calcaratus produces weak advertisement calls of about 72 dB sound pressure level (SPL) at 0.25 m from the caller. The signals are affected by attenuation and degradation patterns as they propagate in their native environment, reaching average values of 61 and 51 dB SPL at 1 and 2 m from the sound source, respectively. Midbrain multi-unit recordings show a relatively low auditory sensitivity, with thresholds of about 58 dB SPL for conspecific calls, which are likely to restrict communication to distances shorter than 2 m, a remarkably short range as compared to other anurans.

Propagation of natural toad calls in a Mediterranean terrestrial environment.

Propagation patterns of animal acoustic signals provide insights into the evolution of signal design to convey signaler's information to potential recipients. However, propagation properties of vertebrate calls have been rarely studied using natural calls from individuals; instead playback calls broadcast through loudspeakers have been used extensively, a procedure that may involve acoustical and physical features differing from natural sounds. Measurements of the transmission characteristics of natural advertisement calls, which are simple tonal sounds, of the Iberian midwife toad, Alytes cisternasii, were carried out, and the results were compared with previously published results broadcasting recorded calls of the same species. Measurements of sound pressure level (SPL) of calls from individual male A. cisternasii revealed that the call amplitude decreases at distances of 1-8 m from the source at rates averaging 1-5 dB above spherical transmission loss in an omni-directional pattern. A comparison between SPLs of natural calls in the current study and of playback calls from a previous study showed that patterns of propagation did not differ in average values, but variance was significantly higher for natural calls. Results suggest that using broadcast signals for transmission experiments may result in a simplification of the conditions in which actual animals communicate in nature.

When signal meets noise: immunity of the frog ear to interference.

Sound stimulates the tympanic membrane (TM) of anuran amphibians through multiple, poorly understood pathways. It is conceivable that interactions between the internal and external inputs to the TM contribute to the nonlinear effects that noise is known to produce at higher levels of the auditory pathway. To explore this issue, we conducted measurements of TM vibration in response to tones in the presence of noise in the frog Eupsophus calcaratus. Laser vibrometry revealed that the power spectra (n = 16) of the TM velocity in response to pure tones at a constant level of 80 dB sound-pressure level (SPL) had a maximum centered at an average frequency of 2,344 Hz (range 1,700-2,990 Hz) and a maximum velocity of 61.1 dB re 1 microm/s (range 42.9-66.6 dB re 1 microm/s). These TM-vibration velocity response profiles in the presence of increasing levels of 4-kHz band-pass noise were unaltered up to noise levels of 90 dB SPL. For the relatively low spectral densities of the noise used, the TM remains in its linear range. Such vibration patterns facilitate the detection of tonal signals in noise at the tympanic membrane and may underlie the remarkable vocal responsiveness maintained by males of E. calcaratus under noise interference.