ABSTRACT In piranhas, sounds are produced through the vibration of the swim bladder wall caused by the contraction of bilateral sonic muscles. Because they are solely innervated by spinal nerves, these muscles likely evolved from the locomotor hypaxial musculature. The transition from a neuromuscular system initially shaped for slow movements (locomotion) to a system that requires a high contraction rate (sound production) was accompanied with major peripheral structural modifications, yet the associated neural adjustments remain to this date unclear. To close this gap, we investigated the activity of both the locomotor and the sonic musculature using electromyography. The comparison between the activation patterns of both systems highlighted modifications of the neural motor pathway: (1) a transition from a bilateral alternating pattern to a synchronous activation pattern, (2) a switch from a slow- to a high-frequency regime, and (3) an increase in the synchrony of motor neuron activation. Furthermore, our results demonstrate that sound features correspond to the activity of the sonic muscles, as both the variation patterns of periods and amplitudes of sounds highly correspond to those seen in the sonic muscle electromyograms (EMG sonic ). Assuming that the premotor network for sound production in piranhas is of spinal origin, our results show that the neural circuit associated with spinal motor neurons transitioned from the slow alternating pattern originally used for locomotion to a much faster simultaneous activation pattern to generate vocal signals.

ABSTRACT The /Kwa/ vocalization dominates the soundscape of Posidonia oceanica meadows but the identity of the species emitting this peculiar fish sound remains a mystery. Information from sounds recorded in the wild indicates that the emitting candidates should be abundant, nocturnal and benthic. Scorpaena spp. combine all these characteristics. This study used an interdisciplinary approach to investigate the vocal abilities of Scorpaena spp.; morphological, histological and electrophysiological examinations were interpreted together with visual and acoustic recordings conducted in semi-natural conditions. All observed Scorpaena spp. ( S. porcus , S. scrofa and S. notata ) share the same sonic apparatus at the level of the abdominal region. This apparatus, present in both males and females, consists of 3 bilaterally symmetrical muscular bundles, having 3–5 long tendons, which insert on ventral bony apophyses of the vertebral bodies. In all chordophones (stringed instruments), the frequency of the vibration is dependent on the string properties and not on the rate at which the strings are plucked. Similarly, we suggest that each of the 3–5 tendons found in the sonic mechanism of Scorpaena spp. acts as a frequency multiplier of the muscular bundle contractions, where the resonant properties of the tendons determine the peak frequency of the /Kwa/, its frequency spectra and pseudo-harmonic profile. The variability in the length and number of tendons found between and within species could explain the high variability of /Kwa/ acoustic features recorded in the wild. Finally, acoustic and behavioural experiments confirmed that Scorpaena spp. can emit the /Kwa/ sound.

ABSTRACT The ability to produce sounds for acoustic communication is known in different Balistidae species but the eventual synapomorphic aspect of the mechanism remains to be shown. In Rhinecanthus aculeatus , sounds result from alternate sweeping movements of the right and left pectoral fins, which push a system of three scutes against the swim bladder wall. In this study, we made a comparison between the sounds produced by this species and two additional ones ( Balistapus undulatus and Rhinecanthus rectangulus ) using hand-held specimens to provide a description of the sound mechanism. The results highlighted that the sound production mechanism is similar in the three species. According to recent phylogenetic data and shared morphological features, this mechanism could be common to the majority of Balistidae family members and all species could be capable of sound production using pectoral fins.

ABSTRACT Fish acoustic signals play a major role during agonistic and reproductive interactions. Among the sound-generating fish, Gobiidae, a large fish family with 1866 valid species, is one of the most studied groups of acoustic fishes, with sound production being documented in a number of species. Paradoxically, the sound-producing mechanism remains poorly studied in this group. The painted goby, Pomatoschistus pictus , produces two distinct sounds called drums and thumps . A combination of morphological and experimental analyses involving high-speed videos synchronized with sound recordings supports that drums are produced during lateral head movements involving at least the alternate contractions of the levator pectoralis muscles originating on the skull and inserting on the pectoral girdle. These movements are reported in many Gobiidae species, suggesting the pectoral-girdle-based mechanism is common in the family and could have evolved from locomotory movements.

ABSTRACT The ability to produce sound has been known for decades in Balistidae. Sounds of many species have been recorded and a variety of sound-producing mechanisms have been proposed, including teeth stridulation, collision of the buccal teeth and movements of the fins. The best-supported hypothesis involves movements of the pectoral fin against the lateral part of the swimbladder, called a drumming membrane. In this study, we describe for the first time the sounds made by the blackbar triggerfish Rhinecanthus aculeatus , which are like short drum rolls with an average duration of 85 ms, 193 Hz dominant frequency and 136 dB SPL level at 3 cm distance. The sounds are a series of pulses that result from alternate sweeping movements of the right and left pectoral fins, which push a system of three scutes that are forced against the swimbladder wall. Pulses from each fin occur in consecutive pairs. High-speed videos indicate that each pulse consists of two cycles. The first part of each cycle corresponds to the inward buckling of the scutes, whereas the second part of the cycle corresponds to an apparent passive recoil of the scutes and swimbladder wall. This novel sound production mechanism is probably found in many members of Balistidae because these peculiar scutes occur in other species in the family. Comparison of sound characteristics from fishes of different sizes shows that dominant frequency decreases with size in juveniles but not in adults.

ABSTRACT A variety of teleost fishes produce sounds for communication by vibrating the swim bladder with fast contracting muscles. Doradid catfishes have an elastic spring apparatus (ESA) for sound production. Contractions of the ESA protractor muscle pull the anterior transverse process of the 4th vertebra or Müllerian ramus (MR) to expand the swim bladder and elasticity of the MR returns the swim bladder to the resting state. In this study, we examined the sound characteristics and associated fine structure of the protractor drumming muscles of three doradid species: Acanthodoras cataphractus , Platydoras hancockii and Agamyxis pectinifrons . Despite large variations in size, sounds from all three species had similar mean dominant rates ranging from 91 to 131 Hz and showed frequencies related to muscle contraction speed rather than fish size. Sounds differed among species in terms of waveform shape and their rate of amplitude modulation. In addition, multiple distinguishable sound types were observed from each species: three sound types from A. cataphractus and P. hancockii , and two sound types from A. pectinifrons . Although sounds differed among species, no differences in muscle fiber fine structure were observed at the species level. Drumming muscles from each species bear features associated with fast contractions, including sarcoplasmic cores, thin radial myofibrils, abundant mitochondria and an elaborated sarcoplasmic reticulum. These results indicate that sound differences between doradids are not due to swimbladder size, muscle anatomy, muscle length or Müllerian ramus shape, but instead result from differences in neural activation of sonic muscles.

Onuxodon species are well known for living inside pearl oysters. As in other carapids, their anatomy highlights their ability to make sounds but sound production has never been documented in Onuxodon . This paper describes sound production in Onuxodon fowleri as well as the anatomy of the sound production apparatus. Single-pulsed sounds and multiple-pulsed sounds that sometimes last more than 3 s were recorded in the field and in captivity (Makemo Island, French Polynesia). These pulses are characterized by a broadband frequency spectrum from 100 to 1000 Hz. Onuxodon fowleri is mainly characterized by its ability to modulate the pulse period, meaning that this species can produce pulsed sounds and tonal-like sounds using the same mechanism. In addition, the sound can be remarkably amplified by the shell cavity (peak gain can exceed 10 dB for some frequencies). The sonic apparatus of O. fowleri is characterized by a rocker bone in front of the swimbladder, modified vertebrae and epineurals, and two pairs of sonic muscles, one of which (primary sonic muscle) inserts on the rocker bone. The latter structure, which is absent in other carapid genera, appears to be sexually dimorphic suggesting differences in sound production in males and females. Sound production in O. fowleri could be an example of adaptation where an animal exploits features of its environment to enhance communication.

Grunts are fish that are well known to vocalize, but how they produce their grunting sounds has not been clearly identified. In addition to characterizing acoustic signals and hearing in the French grunt Haemulon flavolineatum , the present study investigates the sound-production mechanism of this species by means of high-speed X-ray videos and scanning electron microscopy of the pharyngeal jaw apparatus. Vocalizations consist of a series of stridulatory sounds: grunts lasting ~ 47 ms with a mean period of 155 ms and a dominant frequency of ~ 700 Hz. Auditory capacity was determined to range from 100 to 600 Hz, with greatest sensitivity at 300 Hz (105.0±11.8 dB re. 1 μPa). This suggests that hearing is not tuned exclusively to detect the sounds of conspecifics. High-speed X-ray videos revealed how pharyngeal jaws move during sound production. Traces of erosion on teeth in the fourth ceratobranchial arch suggest that they are also involved in sound production. The similarity of motor patterns of the upper and lower pharyngeal jaws between food processing and sound production indicates that calling is an exaptation of the food-processing mechanism.

In teleosts, superfast muscles are generally associated with the swimbladder wall, whose vibrations result in sound production. In Ophidion rochei , three pairs of muscles were named ‘sonic’ because their contractions affect swimbladder position: the dorsal sonic muscle (DSM), the intermediate sonic muscle (ISM), and the ventral sonic muscle (VSM). These muscles were investigated thanks to electron microscopy and electromyography in order to determine their function in sound production. Fibers of the VSM and DSM were much thinner than the fibers of the ISM and epaxial musculature. However, only VSM fibers had the typical ultrastructure of superfast muscles: low proportion of myofibrils, and high proportions of sarcoplasmic reticulum and mitochondria. In females, each sound onset was preceded by the onset of electrical activity in the VSM and the DSM (ISM was not tested). The electromyograms of the VSM were very similar to the waveforms of the sounds: means for the pulse period were 3.6±0.5 and 3.6±0.7 ms, respectively. This shows that the fast VSM (ca. 280 Hz) is responsible for the pulse period and fundamental frequency of female sounds. DSM electromyograms were generally characterized by one or two main peaks followed by periods of lower electrical activity, which suggests a sustained contraction over the course of the sound. The fiber morphology of the ISM and its antagonistic position relative to the DSM are not indicative of a muscle capable of superfast contractions. Overall, this study experimentally shows the complexity of the sound production mechanism in the nocturnal fish O. rochei.

Numerous studies have highlighted the diversity of fish inner ear morphology. However, the function of the shape, size and orientation of the different structures remains poorly understood. The saccule (otolithic endorgan) is considered to be the principal hearing organ in fishes and it has been hypothesized that sagitta (saccular otolith) shape and size affect hearing capacities: large sagittae are thought to increase sensitivity. The sagittae of many ophidiids and carapids occupy a large volume inside the neurocranium. Hence they are a good structure with which to test the size hypothesis. The main aim of this study was to investigate hearing capacities and inner ear morphology in two ophidiiform species: Ophidion rochei and Carapus acus. We used a multidisciplinary approach that combines dissections, μCT-scan examinations and auditory evoked potential techniques. Carapus acus and O. rochei sagittae have similar maximal diameters; both species have larger otoliths than many non-ophidiiform species, especially compared with the intra-neurocranium volume. Both species are sensitive to sounds up to 2100 Hz. Relative to the skull, O. rochei has smaller sagittae than the carapid, but better hearing capacities from 300 to 900 Hz and similar sensitivities at 150 Hz and from 1200 to 2100 Hz. Results show that hearing capacities of a fish species cannot be predicted only based on sagitta size. Larger otoliths (in size relative to the skull) may have evolved mainly for performing vestibular functions in fishes, especially those species that need to execute precise and complex movements.

SUMMARY Gobiidae, the largest fish family (>1500 species), has species from at least 10 genera that produce sounds for communication. Studies focused on goby sound production mechanisms have suggested that sounds are produced by the forcible ejection of water through small apertures in the opercles (hydrodynamic mechanism). The present study was a multidisciplinary investigation (morphology, muscle histology, high-speed video, sound analysis and electromyography) of the sound emission mechanism in Gobius paganellus , which produces both pulsed and tonal calls. Two populations were used, from Brittany and Venice. In the French population, sounds were accompanied by a suite of coordinated movements of the buccal, branchial and opercular regions. This was not the case in the Venetian population, and thus the direct role of head movements in sound production was rejected. The hydrodynamic mechanism hypothesis was also rejected in G. paganellus on the basis of sound oscillogram shape and because sounds are still produced after the opercles and hyohyoid muscles are cut. The use of both electromyography and electron microscopy showed that the levator pectoralis muscle, which originates on the skull and inserts on the dorsal tip of the cleithrum, is involved in sound production. We propose that the contraction of this muscle and associated vibration of the large radials is used to make sounds. In addition, we propose that different sound types (pulsed sounds and tonal calls) could occur because of differences in fish size.

SUMMARY It was recently demonstrated that clownfishes produce aggressive sounds by snapping their jaw teeth. To date, only the onset of the sound has been studied, which raises the question, what structure is involved in sound radiation? Here, a combination of different approaches has been used to determine the anatomical structure(s) responsible for the size-related variations observed in sound duration and frequency. Filling the swimbladder with physiological liquid specifically modified size-related acoustic features by inducing a significant decrease in pulse duration of approximately 3 ms and a significant increase in dominant frequency of approximately 105 Hz. However, testing the acoustics of the swimbladder by striking it with a piezoelectric impact hammer showed that this structure is a highly damped sound source prevented from prolonged vibrations. In contrast, the resonant properties of the rib cage seems to account for the size-related variations observed in acoustic features. For an equivalent strike on the rib cage, the duration and dominant frequency of induced sounds changed with fish size: sound duration and dominant frequency were positively and negatively correlated with fish size, respectively. Such relationships between sonic features and fish size are consistent with those observed in natural sounds emitted by fish. Therefore, the swimbladder itself does not act as a resonator; its wall just seems to be driven by the oscillations of the rib cage. This set of observations suggests the need for reassessment of the acoustic role of swimbladders in various fish species.

SUMMARY Piranhas are known to be sound-producing animals. Nevertheless, the biological significance of piranha calls remains unclear because sounds have been recorded only when specimens were held by hand or trapped in a gill net. These sounds are generated by rapid contractions of sonic muscles that insert on a broad tendon surrounding ventrally the cranial sac of the swimbladder. The piranha swimbladder is thought to play an important role in sound production as an impedance-matching device and as a resonator. However, the vibratory capacities of the cranial and caudal sacs and the exact role of both sacs in sound production remain poorly understood. In this study, three sounds were each associated to a specific behaviour. The first sound (type 1) was produced during frontal display; it had numerous pulses and lasted 140!±17 ms, with a fundamental frequency of 120±4 Hz. It corresponded to the sound made by hand-held fishes. The second sound (type 2) was produced during circling and fighting behaviour; it was a single pulse lasting 36±8 ms, with a fundamental frequency of 43±10 Hz. The third sound (type 3) corresponded to chasing behaviour and comprised three to four pulses, each lasting 3±1 ms, with a fundamental frequency of 1739±18 Hz. Using a laser vibrometer to study the swimbladder displacement when stimulated at different frequencies, it was demonstrated that the first two sounds corresponded to the swimbladder mechanism. By contrast, the third sound was associated with the jaw mechanism. The vibrometer indicated that the swimbladder is a highly damping structure, simply copying the sonic muscle contraction rate. This study provides two interesting insights. First, it shows the relationships between three kinds of piranha sound and three specific behaviours. Second, using muscle stimulation at different rates, it shows which simultaneous conditions are required for production of sound in this species. Swimbladder calls were produced by a muscle contraction rate of approximately 100 Hz because this periodicity allowed the swimbladder to vibrate. At this frequency range, the contraction–relaxation cycles of the swimbladder muscles engendered wall displacements that had short amplitudes and with only a small variability between them.

SUMMARY The diversity in calls and sonic mechanisms appears to be important in Chaetodontidae. Calls in Chaetodon multicinctus seem to include tail slap, jump, pelvic fin flick and dorsal–anal fin erection behaviors. Pulsatile sounds are associated with dorsal elevation of the head, anterior extension of the ventral pectoral girdle and dorsal elevation of the caudal skeleton in Forcipiger flavissiumus . In Hemitaurichthys polylepis , extrinsic swimbladder muscles could be involved in sounds originating from the swimbladder and correspond to the inward buckling of tissues situated dorsally in front of the swimbladder. These examples suggest that this mode of communication could be present in other members of the family. Sounds made by the pennant bannerfish ( Heniochus chrysostomus ) were recorded for the first time on coral reefs and when fish were hand held. In hand-held fishes, three types of calls were recorded: isolated pulses (51%), trains of four to 11 pulses (19%) and trains preceded by an isolated pulse (29%). Call frequencies were harmonic and had a fundamental frequency between 130 and 180 Hz. The fundamental frequency, sound amplitude and sound duration were not related to fish size. Data from morphology, sound analysis and electromyography recordings highlight that the calls are made by extrinsic sonic drumming muscles in association with the articulated bones of the ribcage. The pennant bannerfish system differs from other Chaetodontidae in terms of sound characteristics, associated body movements and, consequently, mechanism.

SUMMARY The air-filled swimbladder acts as an acoustic amplifier for some fish by converting sound pressure into particle motion, which is transmitted to the inner ear. Here, we describe in detail the specialized connection between the swimbladder and ear in the mojarra, as well as a modified cone on the anal fin in which the posterior end of the swimbladder sits. Hearing tests show the mojarra has better hearing sensitivity than other species of fish without a connection. However, mojarras do not seem to use this adaptation for communication. Furthermore, the inclined position of the swimbladder may help the fish to catch their prey more easily, as the swimbladder will be horizontal when they are picking up benthic prey.

SUMMARY The anatomical structures of the sound-producing organ in Ophidion rochei males present an important panel of highly derived characters: three pairs of putatively slow sonic muscles; a neural arch that pivots; a rocker bone at the front pole of the swimbladder; a stretchable swimbladder fenestra; a swimbladder plate; and an internal cone that terminates in a pair of membranes in the caudal swimbladder. Male courtship calls are produced nocturnally and consist of trains of 10 to 40 pulses that increase in amplitude and decrease in rate before exhibiting alternating periods of ca. 84 and 111 ms. Each pulse includes an unusual waveform with two parts. Pulse part 1 is a single cycle followed by a longer duration pulse part that exhibits gradual damping. Sounds and morphology suggest two hypotheses on the sound-producing mechanism. The ‘pulley’ hypothesis would require an alternate contraction of the ventral and dorsal muscles to form the two parts of each pulse. The ‘bow’ hypothesis involves a release mechanism with the sustained contraction of the dorsal muscle during all of the call, and the rapid contraction/relaxation of the ventral muscle to form each pulse.

SUMMARY Although acoustic communication is an integral part of cichlid behaviour,its mechanism has never been identified before. In the present study, a combination of approaches was used to investigate the sound-producing mechanism of Oreochromis niloticus . Synchronisation of high-speed video data (500 frames s –1 ) and cineradiographies (250 frames s –1 ) with the sound recordings made it possible to locate the different body parts involved in sound production in territorial males. Sounds are made during a backward movement of the pelvic and pectoral girdles and a forward movement of the second pterygiophore of the anal fin. Various electrostimulation experiments, dissections and observation of histological cross-sections revealed a set of bundles (that we call the vesica longitudinalis) situated in the hypaxial musculature, ventro-laterally to the swimbladder. Contraction of these bundles should result in compression of the rib cage and also of the swimbladder, because of its close association with the serosa and ribs. Deflation of the swimbladder resulted in a reduced sound intensity.

SUMMARY Clownfish live in social groups in which there is a size-based dominance hierarchy. In such a context, sonic cues could play a role in social organisation because dominant frequency and pulse length of sounds are strongly correlated with fish size. Data on the hearing ability of these fish are, however, needed to show that they have the sensory ability to detect the frequencies in their sounds. The present study determines the hearing sensitivity in three different anemonefish species ( Amphiprion frenatus,Amphiprion ocellaris and Amphiprion clarkii ), and compares it with the frequencies in their calls. The frequency range over which the three species can detect sounds was between 75 and 1800 Hz, and they were most sensitive to frequencies below 200 Hz. During sound production, dominant frequency is clearly related ( R =0.95) to the fish size, whatever the species. Dominant frequency extends from 370 to 900 Hz for specimens having a size between 55 and 130 mm. The best hearing sensitivity of small specimens were found to be lower than the dominant frequency of their own calls. However, they were found to be close to the dominant frequency of larger fish calls. The interest of juveniles lies in localising the adults and thus their location on the reef.

SUMMARY Fish sonic swimbladder muscles are the fastest muscles in vertebrates and have fibers with numerous biochemical and structural adaptations for speed. Carapid fishes produce sounds with a complex swimbladder mechanism, including skeletal components and extrinsic sonic muscle fibers with an exceptional helical myofibrillar structure. To study this system we stimulated the sonic muscles, described their insertion and action and generated sounds by slowly pulling the sonic muscles. We find the sonic muscles contract slowly, pulling the anterior bladder and thereby stretching a thin fenestra. Sound is generated when the tension trips a release system that causes the fenestra to snap back to its resting position. The sound frequency does not correspond to the calculated resonant frequency of the bladder, and we hypothesize that it is determined by the snapping fenestra interacting with an overlying bony swimbladder plate. To our knowledge this tension release mechanism is unique in animal sound generation.