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Zebrafish is a well-established model organism in hearing research. Although the acoustic environment is known to shape the structure and sensitivity of auditory systems, there is no information on the natural soundscape of this species. Moreover, zebrafish are typically reared in large-scale housing systems (HS), although their acoustic properties and potential effects on hearing remain unknown. We characterized the soundscape of both zebrafish natural habitats and laboratory captive conditions, and discussed possible impact on auditory sensitivity. Sound recordings were conducted in five distinct zebrafish habitats (Southwest India), from quieter stagnant environments with diverse biological/abiotic sounds to louder watercourses characterized by current and moving substrate sounds. Sound pressure level (SPL) varied between 98 and 126 dB re 1 μPa. Sound spectra presented most energy below 3000 Hz and quieter noise windows were found in the noisiest habitats matching the species best hearing range. Contrastingly, recordings from three zebrafish HS revealed higher SPL (122-143 dB) and most energy below 1000 Hz with more spectral peaks, which might cause significant auditory masking. This study establishes an important ground for future research on the adaptation of zebrafish auditory system to the natural soundscapes, and highlights the importance of controlling noise conditions in captivity.

Noise pollution is increasingly present in aquatic ecosystems, causing detrimental effects on growth, physiology and behaviour of organisms. However, limited information exists on how this stressor affects animals in early ontogeny, a critical period for development and establishment of phenotypic traits. We tested the effects of chronic noise exposure to increasing levels (130 and 150 dB re 1 μPa, continuous white noise) and different temporal regimes on larval zebrafish (Danio rerio), an important vertebrate model in ecotoxicology. The acoustic treatments did not affect general development or hatching but higher noise levels led to increased mortality. The cardiac rate, yolk sac consumption and cortisol levels increased significantly with increasing noise level at both 3 and 5 dpf (days post fertilization). Variation in noise temporal patterns (different random noise periods to simulate shipping activity) suggested that the time regime is more important than the total duration of noise exposure to down-regulate physiological stress. Moreover, 5 dpf larvae exposed to 150 dB continuous noise displayed increased dark avoidance in anxiety-related dark/light preference test and impaired spontaneous alternation behaviour. We provide first evidence of noise-induced physiological stress and behavioural disturbance in larval zebrafish, showing that both noise amplitude and timing negatively impact key developmental endpoints in early ontogeny.

Exposure to continuous moderate noise levels is known to impair the auditory system leading to Noise-Induced Hearing Loss (NIHL) in animals including humans. The mechanism underlying noise-dependent auditory Temporary Threshold Shifts (TTS) is not fully understood. In fact, only limited information is available on vertebrates such as fishes, which share homologous inner ear structures to mammals and have the ability to regenerate hair cells. The zebrafish Danio rerio is a well-established model in hearing research providing an unmatched opportunity to investigate the molecular and physiological mechanisms of NIHL at the sensory receptor level. Here we investigated for the first time the effects of noise exposure on TTS and functional recovery in zebrafish, as well as the associated morphological damage and regeneration of the inner ear saccular hair cells. Adult specimens were exposed for 24h to white noise at various amplitudes (130, 140 and 150 dB re. 1 μPa) and their auditory sensitivity was subsequently measured with the Auditory Evoked Potential (AEP) recording technique. Sensory recovery was tested at different times post-treatment (after 3, 7 and 14 days) and compared to individuals kept under quiet lab conditions. Results revealed noise level-dependent TTS up to 33 dB and increase in response latency. Recovery of hearing function occurred within 7 days for fish exposed to 130 and 140 dB noise levels, while fish subject to 150 dB only returned to baseline thresholds after 14 days. Hearing impairment was accompanied by significant loss of hair cells only at the highest noise treatment. Full regeneration of the sensory tissue (number of hair cell receptors) occurred within 7 days, which was prior to functional recovery. We provide first baseline data of NIHL in zebrafish and validate this species as an effective vertebrate model to investigate the impact of noise exposure on the structure and function of the adult inner ear and its recovery process.

Fish represent the largest group of vertebrates and display the greatest diversity of auditory structures. However, studies addressing how the form and function of the auditory system change during development to enhance perception of the acoustic environment are rather sparse in this taxon compared to other vertebrate groups. An ontogenetic perspective of the auditory system in fishes provides a readily testable framework for understanding structure–function relationships. Additionally, studying ancestral models such as fish can convey valuable comparable information across vertebrates, as early developmental events are often evolutionary conserved. This chapter reviews the literature on the morphological development of the fish auditory system, with particular focus on the inner ear structures that evolve from an otic placode during early embryonic development and then continue to undergo differentiation and maturation in the postembryonic phase. Moreover, the chapter provides a systematic overview of how auditory sensitivity develops during ontogeny. Although most studies indicate a developmental improvement in auditory sensitivity, there is considerably species-specific variation. Lastly, the paucity of information and literature concerning the development of auditory capabilities for social communication in fishes is also discussed. Further investigation on the development of structure and function of the fish auditory system is recommended in order to obtain a deeper understanding of how ontogenetic morphological changes in the auditory pathway relate to modifications in acoustic reception, auditory processing, and the capacity to communicate acoustically.

Anthropogenic noise of variable temporal patterns is increasing in aquatic environments, causing physiological stress and sensory impairment. However, scarce information exists on exposure effects to continuous versus intermittent disturbances, which is critical for noise sustainable management. We tested the effects of different noise regimes on the auditory system and behaviour in the zebrafish (Danio rerio). Adult zebrafish were exposed for 24 h to either white noise (150 ± 10 dB re 1 μPa) or silent control. Acoustic playbacks varied in temporal patterns—continuous, fast and slow regular intermittent, and irregular intermittent. Auditory sensitivity was assessed with Auditory Evoked Potential recordings, revealing hearing loss and increased response latency in all noise-treated groups. The highest mean threshold shifts (c. 13 dB) were registered in continuous and fast intermittent treatments, and no differences were found between regular and irregular regimes. Inner ear saccule did not reveal significant hair cell loss but showed a decrease in presynaptic Ribeye b protein especially after continuous exposure. Behavioural assessment using the standardized Novel Tank Diving assay showed that all noise-treated fish spent > 98% time in the bottom within the first minute compared to 82% in control, indicating noise-induced anxiety/stress. We provide first data on how different noise time regimes impact a reference fish model, suggesting that overall acoustic energy is more important than regularity when predicting noise effects.

Fish acoustic signals associated with mating behaviour are typically low-frequency sounds produced by males when in close proximity to females. However, some species make sounds that serve the function and follow the design of advertisement calls, well known in insects, anurans, and birds. Close-range courtship acoustic signals may be used by females in mate assessment as they contain information of male quality such as size and condition. For example, sound-dominant frequency, amplitude, and fatigue resistance may signal body size whereas pulse period (i.e. muscle contraction rate) and calling activity are related with body condition in some species. Some signal features, such as sound pulse number, may carry multiple messages including size and condition. Playback experiments on mate choice of a restricted number of species suggest that females prefer vocal to silent males and may use sound frequency, amplitude, and mainly calling rateCalling ratewhen assessing males. The assessment of males by females becomes more challenging when males engage in choruses or when sounds are otherwise masked by anthropogenic noise but almost nothing is known about how these aspects affect mating decisions and fish reproductive success.

Studies addressing structure-function relationships of the fish auditory system during development are sparse compared to other taxa. The Batrachoididae has become an important group to investigate mechanisms of auditory plasticity and evolution of auditory-vocal systems. A recent study reported ontogenetic improvements in the inner ear saccule sensitivity of the Lusitanian toadfish, Halobatrachus didactylus, but whether this results from changes in the sensory morphology remains unknown. We investigated how the macula and organization of auditory receptors in the saccule and utricle change during growth in this species. Inner ear sensory epithelia were removed from the end organs of previously PFA-fixed specimens, from non-vocal posthatch fry (<1.4 cm, standard length) to adults (>23 cm). Epithelia were phalloidin-stained and analysed for area, shape, number and orientation patterns of hair cells (HC), and number and size of saccular supporting cells (SC). Saccular macula area expanded 41x in total, and significantly more (relative to body length) among vocal juveniles (2.3–2.9 cm). Saccular HC number increased 25x but HC density decreased, suggesting that HC addition is slower relative to epithelial growth. While SC density decreased, SC apical area increased, contributing to the epithelial expansion. The utricule revealed increased HC density (striolar region) and less epithelial expansion (5x) with growth, contrasting with the saccule that may have a different developmental pattern due to its larger size and main auditory functions. Both macula shape and HC orientation patterns were already established in the posthatch fry and retained throughout growth in both end organs. We suggest that previously reported ontogenetic improvements in saccular sensitivity might be associated with changes in HC number (not density), size and/or molecular mechanisms controlling HC sensitivity. This is one of the first studies investigating the ontogenetic development of the saccule and utricle in a vocal fish and how it potentially relates to auditory enhancement for acoustic communication.

Skip to Next Section Acoustic communication is an important part of social behaviour of fish species that live or breed in shallow noisy waters. Previous studies have shown that some fish species exploit a quiet window in the background noise for communication. However, it remains to be examined whether hearing abilities and sound production of fish are adapted to marine habitats presenting high hydrodynamism. Here, we investigated whether the communication system of the painted (Pomatoschistus pictus) and the marbled (Pomatoschistus marmoratus) gobies is adapted to enhance sound transmission and reception in Atlantic shallow water environments. We recorded and measured the sound pressure levels of social vocalisations of both species, as well as snapshots of ambient noise of habitats characterised by different hydrodynamics. Hearing thresholds (in terms of both sound pressure and particle acceleration) and responses to conspecific signals were determined using the auditory evoked potential recording technique. We found that the peak frequency range (100–300 Hz) of acoustic signals matched the best hearing sensitivity in both species and appeared well adapted for short-range communication in Atlantic habitats. Sandy/rocky exposed beaches presented a quiet window, observable even during the breaking of moderate waves, coincident with the main sound frequencies and best hearing sensitivities of both species. Our data demonstrate that the hearing abilities of these gobies are well suited to detect conspecific sounds within typical interacting distances (a few body lengths) in Atlantic shallow waters. These findings lend support to the acoustic adaptive hypothesis, under the sensory drive framework, proposing that signals and perception systems coevolve to be effective within local environment constraints.

Vocal differentiation is widely documented in birds and mammals but has been poorly investigated in other vertebrates, including fish, which represent the oldest extant vertebrate group. Neural circuitry controlling vocal behaviour is thought to have evolved from conserved brain areas that originated in fish, making this taxon key to understanding the evolution and development of the vertebrate vocal-auditory systems. This study examines ontogenetic changes in the vocal repertoire and whether vocal differentiation parallels auditory development in the Lusitanian toadfish Halobatrachus didactylus (Batrachoididae). This species exhibits a complex acoustic repertoire and is vocally active during early development. Vocalisations were recorded during social interactions for four size groups (fry: &lt;2 cm; small juveniles: 2–4 cm; large juveniles: 5–7 cm; adults &gt;25 cm, standard length). Auditory sensitivity of juveniles and adults was determined based on evoked potentials recorded from the inner ear saccule in response to pure tones of 75–945 Hz. We show an ontogenetic increment in the vocal repertoire from simple broadband-pulsed ‘grunts’ that later differentiate into four distinct vocalisations, including low-frequency amplitude-modulated ‘boatwhistles’. Whereas fry emitted mostly single grunts, large juveniles exhibited vocalisations similar to the adult vocal repertoire. Saccular sensitivity revealed a three-fold enhancement at most frequencies tested from small to large juveniles; however, large juveniles were similar in sensitivity to adults. We provide the first clear evidence of ontogenetic vocal differentiation in fish, as previously described for higher vertebrates. Our results suggest a parallel development between the vocal motor pathway and the peripheral auditory system for acoustic social communication in fish.

Anthropogenic noise can be hazardous for the auditory system and wellbeing of animals, including humans. However, very limited information is known on how this global environmental pollutant affects auditory function and inner ear sensory receptors in early ontogeny. The zebrafish (Danio rerio) is a valuable model in hearing research, including investigations of developmental processes of the vertebrate inner ear. We tested the effects of chronic exposure to white noise in larval zebrafish on inner ear saccular sensitivity and morphology at 3 and 5 days post-fertilization (dpf), as well as on auditory-evoked swimming responses using the prepulse inhibition (PPI) paradigm at 5 dpf. Noise-exposed larvae showed a significant increase in microphonic potential thresholds at low frequencies, 100 and 200 Hz, while the PPI revealed a hypersensitization effect and a similar threshold shift at 200 Hz. Auditory sensitivity changes were accompanied by a decrease in saccular hair cell number and epithelium area. In aggregate, the results reveal noise-induced effects on inner ear structure–function in a larval fish paralleled by a decrease in auditory-evoked sensorimotor responses. More broadly, this study highlights the importance of investigating the impact of environmental noise on early development of sensory and behavioural responsiveness to acoustic stimuli.