Revealing sound-induced motion patterns in fish hearing structures in 4D: a standing wave tube-like setup designed for high-resolution time-resolved tomography.

Modern bony fishes possess a high morphological diversity in their auditory structures and auditory capabilities. Yet, how auditory structures such as the otoliths in the inner ears and the swim bladder work together remains elusive. Gathering experimental evidence on the in situ motion of fish auditory structures while avoiding artifacts caused by surgical exposure of the structures has been challenging for decades. Synchrotron radiation-based tomography with high spatio-temporal resolution allows the study of morphofunctional issues non-invasively in an unprecedented way. We therefore aimed to develop an approach that characterizes the moving structures in 4D (=three spatial dimensions+time). We designed a miniature standing wave tube-like setup to meet both the requirements of tomography and those of tank acoustics. With this new setup, we successfully visualized the motion of isolated otoliths and the auditory structures in zebrafish (Danio rerio) and glass catfish (Kryptopterus vitreolus).

Temperature (but not acclimation) affects hearing in fishes adapted to different temperature regimes.

Temperature affects various metabolic and physiological processes in ectothermic animals, including auditory systems. The current study investigates the effect of temperature and thermal acclimation time on hearing sensitivities in a eurythermal and a stenothermal fish possessing accessory hearing structures. Using the auditory evoked potential (AEP) recording technique, we determined thresholds from 0.1 to 4 kHz and peak latencies of AEP-waveforms in response to a click stimulus. The goldfish Carassius auratus was chosen as a model for eurythermal and the Amazonian catfish Megalodoras uranoscopus as a model for stenothermal species. Both species were tested at two different temperatures (C. auratus: 15 °C and 25 °C, M. uranoscopus: 22 °C and 30 °C) and acclimation periods, within 22 h (unacclimated) or three to four weeks (acclimated) after reaching the target temperature. A frequency-dependent increase in auditory sensitivity and a decrease of peak latencies was recorded in both species at higher temperatures, independent of acclimation time. The change in hearing thresholds per degree Celsius was more pronounced in the stenothermal catfish. The data indicate that higher temperatures improved hearing (lower thresholds, shorter latencies), whereas acclimation had no effect on hearing in either species. The latter data contradict previous findings in the eurythermal channel catfish Ictalurus punctatus in which acclimation slightly improved hearing when raising the temperatures. A comparison of changes in hearing sensitivity per degree Celsius of all seven species tested so far revealed no differences between eurythermal and stenothermal species.

Both sexes produce sounds in vocal fish species: testing the hypothesis in the pygmy gourami (labyrinth fishes).

In vocal fish species, males possess larger sound-generating organs and signal acoustically with pronounced sex-specific differences. Sound production is known in two out of three species of croaking gouramis (Trichopsis vittata and T. schalleri). The present study investigates sex-specific differences in sonic organs, vocalizing behaviour and sounds emitted in the third species, the pygmy gourami, T. pumila, in order to test the hypothesis that females are able to vocalize despite their less-developed sonic organs, and despite contradictory reports. Croaking gouramis stretch and pluck two enhanced (sonic) pectoral fin tendons during alternate fin beating, resulting in a series of double-pulsed bursts, termed croaking sound. We measured the diameter of the first and second sonic tendon and showed that male tendons were twice as large as in similar-sized females. We also determined the duration of dyadic contests, visual displays, number of sounds and buttings. Sexes differ in all sound characteristics but in no behavioural variable. Male sounds consisted of twice as many bursts, a higher percentage of double-pulsed bursts and a higher burst period. Additionally, male sounds had a lower dominant frequency and a higher sound level. In summary, female pygmy gouramis possessed sonic organs and vocalized in most dyadic contests. The sexual dimorphism in sonic tendons is clearly reflected in sex-specific differences in sound characteristics, but not in agonistic behaviour, supporting the hypothesis that females are vocal.

Temperature affects sound production in fish with two sets of sonic organs: The Pictus cat.

Sound communication is affected by ambient temperature in ectothermic animals including fishes. The present study examines the effects of temperature on acoustic signaling in a fish species possessing two different sound-generating mechanisms. The Amazonian Pictus catfish Pimelodus pictus produces low-frequency harmonic sounds (swimbladder drumming muscles) and high-frequency stridulation sounds (rubbing pectoral fin spines in the pectoral girdle). Sounds of 15 juveniles were recorded when hand-held after three weeks of acclimation at 30 °C, 22 °C and again 30 °C. The following sound characteristics were investigated: calling activity, sound duration, fundamental frequency of drumming sounds and dominant frequency of stridulation sounds. The number of both sound types produced within the first minute of experiments did not change with temperature. In contrast, sound duration was significantly shorter at 30 °C than at 22 °C (drumming: 78-560 ms; stridulation: 23-96 ms). The fundamental frequency of drumming sounds and thus the drumming muscle contraction rate varied from 127 Hz to 242 Hz and increased with temperature. The dominant frequency of broadband stridulation sounds ranged from 1.67 kHz to 3.39 kHz and was unaffected by temperature changes. Our data demonstrate that temperature affects acoustic signaling in P. pictus, although the changes differed between sound characteristics and sound type. The effects vary from no change in calling activity and dominant frequency, to an increase in fundamental frequency and shortened duration of both sound types. Together with the known effects of temperature on hearing in the Pictus cat, the present results indicate that global warming may affect acoustic communication in fishes.

Sound production in female Trichopsis schalleri (Labyrinth fishes): comparison to males and evolutionary considerations.

Croaking gouramis (genus Trichopsis, Anabantoidei) generate series of two-pulsed bursts (croaks) during agonistic interactions. Sex-specific differences are minor in T. vittata which raises the question whether sexes differ in the other two species. The current study analyses sounds recorded in female T. schalleri, compares the sound characteristics to those of males investigated earlier and correlates these characteristics to female body size. Sex-specific differences were found in three out of six sound characteristics. In females, sounds were lower in burst number, burst period and SPL. Pulse period, dominant frequency and peak-to-peak amplitude ratios of pulses did not differ between sexes. Burst period and SPL increased significantly with female body weight, whereas dominant frequency decreased. The present acoustic data indicate the sex-specific differences are more pronounced in T. schalleri than T. vittata. The results also demonstrate that both sexes are vocal, which remains to be shown for females of the third species, T. pumila, which have poorly developed sonic organs. The evolution of the pectoral sound-producing mechanism in Trichopsis is most likely based on an exaptation process during which acoustic signals are generated by fin tendons initially related to other functions as is evident in closely related genera lacking this organ.

Acoustic communication in terrestrial and aquatic vertebrates.

Sound propagates much faster and over larger distances in water than in air, mainly because of differences in the density of these media. This raises the question of whether terrestrial (land mammals, birds) and (semi-)aquatic animals (frogs, fishes, cetaceans) differ fundamentally in the way they communicate acoustically. Terrestrial vertebrates primarily produce sounds by vibrating vocal tissue (folds) directly in an airflow. This mechanism has been modified in frogs and cetaceans, whereas fishes generate sounds in quite different ways mainly by utilizing the swimbladder or pectoral fins. On land, vertebrates pick up sounds with light tympana, whereas other mechanisms have had to evolve underwater. Furthermore, fishes differ from all other vertebrates by not having an inner ear end organ devoted exclusively to hearing. Comparing acoustic communication within and between aquatic and terrestrial vertebrates reveals that there is no 'aquatic way' of sound communication, as compared with a more uniform terrestrial one. Birds and mammals display rich acoustic communication behaviour, which reflects their highly developed cognitive and social capabilities. In contrast, acoustic signaling seems to be the exception in fishes, and is obviously limited to short distances and to substrate-breeding species, whereas all cetaceans communicate acoustically and, because of their predominantly pelagic lifestyle, exploit the benefits of sound propagation in a dense, obstacle-free medium that provides fast and almost lossless signal transmission.

Peripheral Hearing Structures in Fishes: Diversity and Sensitivity of Catfishes and Cichlids.

Fishes have evolved an astonishing diversity of peripheral (accessory/ancillary) auditory structures to improve hearing based on their ability to transmit oscillations of gas bladder walls to the inner ears. So far it is unclear to what degree the size of the bladder and the linkage to the ear affect hearing in fishes. An interfamilial study in catfishes revealed that families which possess large, single swim bladders and one to four Weberian ossicles were more sensitive at higher frequencies (≥1 kHz) than families which have small, paired, and encapsulated bladders and one to two ossicles. An intrafamilial investigation in thorny catfishes (family Doradidae) revealed that small differences in bladder morphology did not affect hearing similarly. Members of the cichlid family possess an even larger variation in peripheral auditory structures than catfishes. The linkage between the swim bladder and ear can either be present via anterior extensions of the bladder or be completely absent (in contrast to catfishes). Representatives having large bladders with extensions had the best sensitivities. Cichlids lacking extensions had lower sensitivities above 0.3 kHz. Species with a vestigial swim bladder exhibited a smaller hearing bandwidth than those with larger swim bladder (maximum frequency: 0.7 kHz vs. 3 kHz). Catfishes and cichlids reveal that larger gas bladders and more pronounced connections between the swim bladder and the inner ear result in improved hearing at higher frequencies. The lack of a connection between a large bladder and the inner ear does not necessarily result in a smaller detectable frequency range.

Diversity of Inner Ears in Fishes: Possible Contribution Towards Hearing Improvements and Evolutionary Considerations.

Fishes have evolved the largest diversity of inner ears among vertebrates. While G. Retzius introduced us to the diversity of the gross morphology of fish ears in the late nineteenth century, it was A. N. Popper who unraveled the large variety of the fine structure during the last four decades. Modifications of the basic inner ear structure-consisting of three semicircular canals and their sensory epithelia, the cristae and three otolithic end organs (utricle, saccule, lagena) including the maculae-mainly relate to the saccule and lagena and the respective sensory epithelia, the macula sacculi and macula lagenae. Despite the profound morphological knowledge of inner ears and the morphological variability, the functional significance of this diversity is still largely unknown. The aims of this review are therefore twofold. First it provides an update of the state of the art of inner ear diversity in bony fishes. Second it summarizes and discusses hypotheses on the evolution of this diversity as well as formulates open questions and promising approaches to tackle these issues.

Effect of temperature on acoustic communication: sound production in the croaking gourami (labyrinth fishes).

Sound communication comprising the production and detection of acoustic signals is affected by ambient temperature in ectothermic animals. In the present study we investigated the effects of temperature on sound production and characteristics in the croaking gourami Trichopsis vittata, a freshwater fish from Southeast Asia possessing a highly specialized sound-generating mechanism found only in a single genus. The croaking gourami produces pulsed sounds by stretching and plucking two enhanced pectoral fin tendons during rapid pectoral fin beating. Croaking sounds typically consist of a series of double-pulsed bursts with main energies between 1 and 1.5 kHz. Sounds were recorded during dyadic contests between two males at three different temperatures (25°, 30° and 35°C). The mean dominant frequency increased with rising temperature from 1.18 to 1.33 kHz, whereas temporal characteristics decreased. The sound interval dropped from 492 to 259 ms, the burst period from 51 to 35 ms and the pulse period from 5.8 to 5.1 ms. In contrast, the number of sounds and number of bursts within a sound were not affected by temperature. The current study shows that spectral and temporal characteristics of sounds are affected in different ways by temperature in the croaking gourami, whereas the numbers of sounds and bursts remain unaffected. We conclude that acoustic communication in gouramis is affected by changes in ambient temperature.

Are accessory hearing structures linked to inner ear morphology? Insights from 3D orientation patterns of ciliary bundles in three cichlid species.

BACKGROUND: Cichlid fishes show considerable diversity in swim bladder morphology. In members of the subfamily Etroplinae, the connection between anterior swim bladder extensions and the inner ears enhances sound transmission and translates into an improved hearing ability. We tested the hypothesis that those swim bladder modifications coincide with differences in inner ear morphology and thus compared Steatocranus tinanti (vestigial swim bladder), Hemichromis guttatus (large swim bladder without extensions), and Etroplus maculatus (intimate connection between swim bladder and inner ears). METHODOLOGY AND RESULTS: We applied immunostaining together with confocal imaging and scanning electron microscopy for the investigation of sensory epithelia, and high-resolution, contrast-enhanced microCT imaging for characterizing inner ears in 3D, and evaluated otolith dimensions. Compared to S. tinanti and H. guttatus, inner ears of E. maculatus showed an enlargement of all three maculae, and a particularly large lacinia of the macula utriculi. While our analysis of orientation patterns of ciliary bundles on the three macula types using artificially flattened maculae uncovered rather similar orientation patterns of ciliary bundles, interspecific differences became apparent when illustrating the orientation patterns on the 3D models of the maculae: differences in the shape and curvature of the lacinia of the macula utriculi, and the anterior arm of the macula lagenae resulted in an altered arrangement of ciliary bundles. CONCLUSIONS: Our results imply that improved audition in E. maculatus is associated not only with swim bladder modifications but also with altered inner ear morphology. However, not all modifications in E. maculatus could be connected to enhanced auditory abilities, and so a potential improvement of the vestibular sense, among others, also needs to be considered. Our study highlights the value of analyzing orientation patterns of ciliary bundles in their intact 3D context in studies of inner ear morphology and physiology.

Distress sounds of thorny catfishes emitted underwater and in air: characteristics and potential significance.

Thorny catfishes produce stridulation (SR) sounds using their pectoral fins and drumming (DR) sounds via a swimbladder mechanism in distress situations when hand held in water and in air. It has been argued that SR and DR sounds are aimed at different receivers (predators) in different media. The aim of this study was to analyse and compare sounds emitted in both air and water in order to test different hypotheses on the functional significance of distress sounds. Five representatives of the family Doradidae were investigated. Fish were hand held and sounds emitted in air and underwater were recorded (number of sounds, sound duration, dominant and fundamental frequency, sound pressure level and peak-to-peak amplitudes). All species produced SR sounds in both media, but DR sounds could not be recorded in air for two species. Differences in sound characteristics between media were small and mainly limited to spectral differences in SR. The number of sounds emitted decreased over time, whereas the duration of SR sounds increased. The dominant frequency of SR and the fundamental frequency of DR decreased and sound pressure level of SR increased with body size across species. The hypothesis that catfish produce more SR sounds in air and more DR sounds in water as a result of different predation pressure (birds versus fish) could not be confirmed. It is assumed that SR sounds serve as distress sounds in both media, whereas DR sounds might primarily be used as intraspecific communication signals in water in species possessing both mechanisms.

A unique swim bladder-inner ear connection in a teleost fish revealed by a combined high-resolution microtomographic and three-dimensional histological study.

BACKGROUND: In most modern bony fishes (teleosts) hearing improvement is often correlated with a close morphological relationship between the swim bladder or other gas-filled cavities and the saccule or more rarely with the utricle. A connection of an accessory hearing structure to the third end organ, the lagena, has not yet been reported. A recent study in the Asian cichlid Etroplus maculatus provided the first evidence that a swim bladder may come close to the lagena. Our study was designed to uncover the swim bladder-inner ear relationship in this species. We used a new approach by applying a combination of two high-resolution techniques, namely microtomographic (microCT) imaging and histological serial semithin sectioning, providing the basis for subsequent three-dimensional reconstructions. Prior to the morphological study, we additionally measured auditory evoked potentials at four frequencies (0.5, 1, 2, 3 kHz) to test the hearing abilities of the fish. RESULTS: E. maculatus revealed a complex swim bladder-inner ear connection in which a bipartite swim bladder extension contacts the upper as well as the lower parts of each inner ear, a condition not observed in any other teleost species studied so far. The gas-filled part of the extension is connected to the lagena via a thin bony lamella and is firmly attached to this bony lamella with connective material. The second part of the extension, a pad-like structure, approaches the posterior and horizontal semicircular canals and a recessus located posterior to the utricle. CONCLUSIONS: Our study is the first detailed report of a link between the swim bladder and the lagena in a teleost species. We suggest that the lagena has an auditory function in this species because the most intimate contact exists between the swim bladder and this end organ. The specialized attachment of the saccule to the cranial bone and the close proximity of the swim bladder extension to the recessus located posterior to the utricle indicate that the saccule and the utricle also receive parallel inputs from the swim bladder extension. We further showed that a combination of non-destructive microCT imaging with histological analyses on the same specimen provides a powerful tool to decipher and interpret fine structures and to compensate for methodological artifacts.

Hearing in catfishes: 200 years of research.

Ernst Weber stated in 1819, based on dissections, that the swimbladder in the European wels (Silurus glanis, Siluridae) and related cyprinids serves as an eardrum and that the ossicles connecting it to the inner ear function as hearing ossicles similar to mammals. In the early 20th century, K. von Frisch showed experimentally that catfishes and cyprinids (otophysines) indeed hear excellently compared to fish taxa lacking auxiliary hearing structures (ossicles, eardrums). Knowledge on hearing in catfishes progressed in particular in the 21st century. Currently, hearing abilities (audiograms) are known in 28 species out of 13 families. Recent ontogenetic and comparative studies revealed that the ability to detect sounds of low-level and high frequencies (4-6 kHz) depends on the development of Weberian ossicles. Species with a higher number of ossicles and larger bladders hear better at higher frequencies (>1 kHz). Hearing sensitivities are furthermore affected by ecological factors. Rising temperatures increase, whereas various noise regimes decrease hearing. Exposure to high-noise levels (>150 dB) for hours result in temporary thresholds shifts (TTS) and recovery of hearing after several days. Low-noise levels reduce hearing abilities due to masking without a TTS. Furthermore, auditory evoked potential (AEP) experiments reveal that the temporal patterns of fish-produced pulsed stridulation and drumming sounds are represented in their auditory pathways, indicating that catfishes are able to extract important information for acoustic communication. Further research should concentrate on inner ears to determine whether the diversity in swimbladders and ossicles is paralleled in the inner ear fine structure.

Sex-specific difference in agonistic sounds depends on size of sonic organs in fishes: Testing the hypothesis in the croaking gourami (Labyrinth fishes).

In most vocal fish species, females possess smaller sound-generating organs and vocalize less than males. In certain cases females lack sonic organs, in others differences between sexes are unknown. This study analyzes in detail the relationship between sexual dimorphism of sonic organs and the characteristics of agonistic behavior and of sounds recorded under the same behavioral conditions in a vocal fish species, the croaking gourami Trichopsis vittata. During agonistic contests both sexes stretch and pluck two enhanced (sonic) tendons when beating pectoral fins alternately, resulting in a series of double-pulsed bursts, termed croaking sound. The following anatomical, behavioral, and acoustic variables were analyzed: diameter of enhanced tendons in each specimen, duration of same-sex dyadic contests, number and duration of lateral display bouts and of sounds, number of single- and double-pulsed bursts, burst period, peak-to-peak amplitudes of pulses, dominant frequency and sound pressure level (SPLrms). Female sonic tendons were approximately one-fifth smaller than male's of the same size. Six out of seven behavioral variables did not differ between sexes. Sound characteristics were similar in both sexes except for SPLs, which were on average 5 dB lower in females. The degree of sexual dimorphisms in sonic organs may explain differences in sound characteristics. Sounds differ only in one sound characteristic (SPLrms) in T. vittata, in contrast with the congeneric Trichopsis pumila which possesses a more pronounced sexual dimorphism in sonic organs and in which agonistic sounds differ in all sound properties between sexes.

Different sound characteristics produced by the left and right pectoral fins constitute a new form of lateralization in a vocal fish.

Songbirds and toothed whales are able to produce different sounds with the left and right part of their sonic organs, a phenomenon termed lateralized sound production. In fishes this phenomenon is poorly known, with lateralization having been observed solely in the channel catfish (Ictalurus punctatus). They produce more sounds with their right pectoral fins. Croaking gouramis Trichopsis vittata beat their pectoral fins alternately, resulting in a series of two-pulsed sound bursts termed croaking sounds. This study investigates lateralized sound production by comparing temporal and amplitude characteristics of sound bursts generated by pectoral fins in T. vittata. Croaking sounds, produced during dyadic contests, were analyzed in 19 females. We investigated the following characteristics of sound bursts: burst period, pulse period within bursts, the relative peak-to-peak amplitudes of bursts, and the ratio of peak-to-peak amplitudes of the first and second pulse within bursts. Sound bursts produced by the right and left sonic organ differed in 17 out of 19 females in at least one to four measured sound characteristics. The number of females whose temporal characteristics differed between pectoral fins was significantly higher than the number of females lacking such differences (16 out of 19). This was not the case for amplitude characteristics. Our data demonstrated that the sound characteristics produced by the left and right sonic organ in T. vittata differed significantly in most specimens. These differences in sound properties may constitute a new form of lateralized sound production in vocal fishes.

Noise-induced masking of hearing in a labyrinth fish: effects on sound detection in croaking gouramis.

An increasing level of anthropogenic underwater noise (shipping, drilling, sonar use, etc.) impairs acoustic orientation and communication in fish by hindering signal transmission or detection. Different noise regimes can reduce the ability to detect sounds of conspecifics due to an upward shift of the hearing threshold, a phenomenon termed masking. We therefore investigated the masking effect of white noise on the auditory thresholds in female croaking gouramis (Trichopsis vittata, Osphronemidae). We hypothesized that noise would influence the detection of conspecific vocalizations and thus acoustic communication. The auditory evoked potentials (AEP) thresholds were measured at six different frequencies between 0.1 and 4 kHz using the AEP recording technique. Sound pressure level audiograms were determined under quiet laboratory conditions (no noise) and continuous white noise of 110 dB RMS. Thresholds increased in the presence of white noise at all tested frequencies by 12-18 dB, in particular at 1.5 kHz. Moreover, hearing curves were compared to spectra of conspecific sounds to assess sound detection in the presence of noise in various contexts. We showed that masking hinders the detection of conspecific sounds, which have main energies between 1.0 and 1.5 kHz. We predict that this will particularly affect hearing of female's low-intensity purring sounds during mating. Accordingly, noise will negatively affect acoustic communication and most likely reproductive success.

Acoustic and visual adaptations to predation risk: a predator affects communication in vocal female fish.

Predation is an important ecological constraint that influences communication in animals. Fish respond to predators by adjusting their visual signaling behavior, but the responses in calling behavior in the presence of a visually detected predator are largely unknown. We hypothesize that fish will reduce visual and acoustic signaling including sound levels and avoid escalating fights in the presence of a predator. To test this we investigated dyadic contests in female croaking gouramis (Trichopsis vittata, Osphronemidae) in the presence and absence of a predator (Astronotus ocellatus, Cichlidae) in an adjoining tank. Agonistic behavior in T. vittata consists of lateral (visual) displays, antiparallel circling, and production of croaking sounds and may escalate to frontal displays. We analyzed the number and duration of lateral display bouts, the number, duration, sound pressure level, and dominant frequency of croaking sounds as well as contest outcomes. The number and duration of lateral displays decreased significantly in predator when compared with no-predator trials. Total number of sounds per contest dropped in parallel but no significant changes were observed in sound characteristics. In the presence of a predator, dyadic contests were decided or terminated during lateral displays and never escalated to frontal displays. The gouramis showed approaching behavior toward the predator between lateral displays. This is the first study supporting the hypothesis that predators reduce visual and acoustic signaling in a vocal fish. Sound properties, in contrast, did not change. Decreased signaling and the lack of escalating contests reduce the fish's conspicuousness and thus predation threat.

Representation of complex vocalizations in the Lusitanian toadfish auditory system: evidence of fine temporal, frequency and amplitude discrimination.

Many fishes rely on their auditory skills to interpret crucial information about predators and prey, and to communicate intraspecifically. Few studies, however, have examined how complex natural sounds are perceived in fishes. We investigated the representation of conspecific mating and agonistic calls in the auditory system of the Lusitanian toadfish Halobatrachus didactylus, and analysed auditory responses to heterospecific signals from ecologically relevant species: a sympatric vocal fish (meagre Argyrosomus regius) and a potential predator (dolphin Tursiops truncatus). Using auditory evoked potential (AEP) recordings, we showed that both sexes can resolve fine features of conspecific calls. The toadfish auditory system was most sensitive to frequencies well represented in the conspecific vocalizations (namely the mating boatwhistle), and revealed a fine representation of duration and pulsed structure of agonistic and mating calls. Stimuli and corresponding AEP amplitudes were highly correlated, indicating an accurate encoding of amplitude modulation. Moreover, Lusitanian toadfish were able to detect T. truncatus foraging sounds and A. regius calls, although at higher amplitudes. We provide strong evidence that the auditory system of a vocal fish, lacking accessory hearing structures, is capable of resolving fine features of complex vocalizations that are probably important for intraspecific communication and other relevant stimuli from the auditory scene.

Ontogenetic development of auditory sensitivity and sound production in the squeaker catfish Synodontis schoutedeni.

BACKGROUND: Surveys of ontogenetic development of hearing and sound production in fish are scarce, and the ontogenetic development of acoustic communication has been investigated in only two fish species so far. Studies on the labyrinth fish Trichopsis vittata and the toadfish Halobatrachus didactylus show that the ability to detect conspecific sounds develops during growth. In otophysine fish, which are characterized by Weberian ossicles and improved hearing sensitivities, the ontogenetic development of sound communication has never been investigated. We analysed the ontogeny of the auditory sensitivity and vocalizations in the mochokid catfish Synodontis schoutedeni. Mochokid catfishes of the genus Synodontis are commonly called squeakers because they produce broadband stridulation sounds during abduction and adduction of pectoral fin spines. Fish from six different size groups - from 22 mm standard length to 126 mm - were studied. Hearing thresholds were measured between 50 Hz and 6 kHz using the auditory evoked potentials recording technique; stridulation sounds were recorded and their sound pressure levels determined. Finally, absolute sound power spectra were compared to auditory sensitivity curves within each size group. RESULTS: The smallest juveniles showed the poorest hearing abilities of all size groups between 50 and 1,000 Hz and highest hearing sensitivity at 5 and 6 kHz. The duration of abduction and adduction sounds and the pulse period increased and sound pressure level (in animals smaller than 58 mm) increased, while the dominant frequency of sounds decreased with size in animals larger than 37 mm. Comparisons between audiograms and sound spectra revealed that the most sensitive frequencies correlate with the dominant frequencies of stridulation sounds in all S. schoutedeni size groups and that all specimens are able to detect sounds of all size groups. CONCLUSIONS: This study on the squeaker catfish S. schoutedeni is the first to demonstrate that absolute hearing sensitivity changes during ontogeny in an otophysine fish. This contrasts with prior studies on two cypriniform fish species in which no such change could be observed. Furthermore, S. schoutedeni can detect conspecific sounds at all stages of development, again contrasting with prior findings in fishes.

Auditory chain reaction: Effects of sound pressure and particle motion on auditory structures in fishes.

Despite the diversity in fish auditory structures, it remains elusive how otolith morphology and swim bladder-inner ear (= otophysic) connections affect otolith motion and inner ear stimulation. A recent study visualized sound-induced otolith motion; but tank acoustics revealed a complex mixture of sound pressure and particle motion. To separate sound pressure and sound-induced particle motion, we constructed a transparent standing wave tube-like tank equipped with an inertial shaker at each end while using X-ray phase contrast imaging. Driving the shakers in phase resulted in maximised sound pressure at the tank centre, whereas particle motion was maximised when shakers were driven out of phase (180°). We studied the effects of two types of otophysic connections-i.e. the Weberian apparatus (Carassius auratus) and anterior swim bladder extensions contacting the inner ears (Etroplus canarensis)-on otolith motion when fish were subjected to a 200 Hz stimulus. Saccular otolith motion was more pronounced when the swim bladder walls oscillated under the maximised sound pressure condition. The otolith motion patterns mainly matched the orientation patterns of ciliary bundles on the sensory epithelia. Our setup enabled the characterization of the interplay between the auditory structures and provided first experimental evidence of how different types of otophysic connections affect otolith motion.