Anterior Swim Bladder Research Articles

Nitrate and Conductivity Effects in Fish Embryo-Larvae.

Nitrate concentrations have been rising in surface waters over the last century and now frequently exceed drinking water standards and environmental safety benchmarks globally. Health-wise, these trends are concerning because nitrate has been shown to disrupt endocrine function and developmental outcomes. This project investigated potential sublethal effects of nitrate on developing fathead minnows. Fish were exposed from fertilization through 21 days post fertilization (dpf) to environmentally relevant concentrations of nitrate (0, 2, 5, 10, 25, or 100 mg/L NO3 -N as NaNO3 ). Nitrate effects on hatch timing, heart rate and rhythm at 3 dpf, growth through 21 dpf, swim bladder inflation timing and size, scoliosis, pericardial edema, and mortality were assessed. Because adding NaNO3 increases water conductivity, two conductivity controls were included to match the ionic strength of the 10 and 100 mg/L NO3 -N treatments. Increasing nitrate delayed posterior swim bladder (PSB) inflation in a dose-dependent manner, with possible inhibition of anterior swim bladder (ASB) inflation at higher doses, although nitrate did not affect swim bladder size. Conversely, nitrate did not affect hatch timing or cardiac endpoints at 3 dpf or induce pericardial edema or scoliosis, although there was a noted brood effect on these latter defects. As was observed with increasing nitrate, higher ion concentrations in the conductivity controls caused dose-dependent increases in fish body size at 21 dpf. Increased ionic strength also hastened ASB inflation independent of nitrate. As in other published studies, the observed delay in PSB inflation suggests that nitrate disrupts the thyroid axis and warrants further investigation. Additionally, this work supports the need for conductivity controls in nitrate toxicity studies to distinguish nitrate-specific effects. This article is protected by copyright. All rights reserved. Environ Toxicol Chem 2023;00:0-0. © 2023 SETAC.

A multisensory perspective on near-field detection and localization of hydroacoustic sources

This paper gives a brief synopsis of the research career of S.C. in fish bioacoustics with an emphasis on dipole near fields. The hydroacoustic nature of the dipole near field and the effective stimuli to lateral line and auditory systems combine to produce a multisensory, range-fractionated region that is critically important to many fish behaviors. The mottled sculpin and goldfish lateral lines encode the spatial complexities of the near field as spatial excitation patterns along the body surface to provide instantaneous snapshots of various source features such as distance, orientation, and direction of movement. In contrast, the pressure-sensitive channel of the goldfish auditory system [the anterior swim bladder (SB)-saccule complex] encodes the spatial complexities in a temporal fashion whenever the position or orientation of the source changes with respect to the anterior SB. A full appreciation for how these somatotopic and egocentric representations guide fish behavior requires an understanding of how multisensory information, including vision, is combined in sensorimotor regions of the brain to effect behavior. A brief overview of vertebrate brain organization indicates that behaviors directed to or away from hydroacoustic sources likely involve a variety of mechanisms, behavioral strategies, and brain regions.

X-ray Microtomography Study of Cranium, Pectoral Fin and Girdle, and Weberian Apparatus Development in Juvenile Triplophysa stewarti (Nemacheilidae)

The stone loach genus Triplophysa is a major component of the Tibetan Plateau ichthyofauna, and its taxonomy and phylogenetic relationships are far from resolved. Skeletogeny patterns contain useful phylogenetic signals, but developmental data for Triplophysa are scarce. Here, we present an X-ray microtomography study of the development of the cranium, pectoral fin and girdle, and Weberian apparatus of juvenile Triplophysa stewarti (12–55 mm standard length). We reveal that the smallest juveniles possess a fontanelle and well-developed Weberian ossicles. Furthermore, the development of the cranium, pectoral fin and girdle, and bony capsule of the Weberian apparatus is illustrated, such as the suturation of preethmoid I to prevomer, the reduction of pectoral radials from four to three by fusion of the most internal two. A paired bony capsule fully enclosing the anterior swim bladder room characterizes the Weberian apparatus of nemacheilid and balitorid loaches. Our data demonstrate that the anterior wall of the bony capsule originates from a vertical plate expanded from the descending process 2, which is a homologue to the “anterior plate” in the superfamily Cobitoidea sensu lato, with the exception of Cobitidae. The lateral process 2 separates from this anterior plate. The horizontal process 4 expands anteriorly to meet the anterior wall, and when posteriorly expanded and curved downward, finally fuses with the bottom wall to close the bony capsule. Accelerated or retarded development of the aforementioned skeletal structures will result in differentiated features in Triplophysa and related nemacheilid genera.

Sound production and mechanism in the cryptic cusk-eel Parophidion vassali.

This study investigates the sounds and the anatomy of the sound-producing organ in the male and female sand-dwelling cusk-eel Parophidion vassali. Although both sexes have similar external phenotype, they can be distinguished by their sonic apparatus and sounds. As in many Ophioidei, Parophidion vassali presents a panel of highly derived characters. Fish possess three pairs of sonic muscles, and males have mineralized swimbladder caps on which inserts the ventral sonic muscle, a neural arch that pivots, a stretchable swimbladder fenestra, an osseous swimbladder plate and a rounded pressure-release membrane in the caudal swimbladder. Females, however, do not possess anterior swimbladder caps, a swimbladder fenestra and the caudal rounded membrane. Males possess the unusual ability to produce sounds starting with a set of low amplitude pulses followed by a second set with higher amplitudes clearly dividing each sound unit into two parts. Females do not vary their sound amplitude in this way: they produce shorter sounds and pulse periods but with a higher peak frequency. Morphology and sound features support the sound-producing mechanism is based on a rebound system (i.e. quick backward snap of the anterior swimbladder). Based on features of the sounds from tank recordings, we have putatively identified the sound of male Parophidion vassali at sea. As these species are ecologically cryptic, we hope this work will allow assessment and clarify the distribution of their populations.

Effect of Thyroperoxidase and Deiodinase Inhibition on Anterior Swim Bladder Inflation in the Zebrafish.

A set of adverse outcome pathways (AOPs) linking inhibition of thyroperoxidase and deiodinase to impaired swim bladder inflation in fish has recently been developed. These AOPs help to establish links between these thyroid hormone (TH) disrupting molecular events and adverse outcomes relevant to aquatic ecological risk assessment. Until now, very little data on the effects of TH disruption on inflation of the anterior chamber (AC) of the swim bladder were available. The present study used zebrafish exposure experiments with three model compounds with distinct thyroperoxidase and deiodinase inhibition potencies (methimazole, iopanoic acid, and propylthiouracil) to evaluate this linkage. Exposure to all three chemicals decreased whole body triiodothyronine (T3) concentrations, either through inhibition of thyroxine (T4) synthesis or through inhibition of Dio mediated conversion of T4 to T3. A quantitative relationship between reduced T3 and reduced AC inflation was established, a critical key event relationship linking impaired swim bladder inflation to TH disruption. Reduced inflation of the AC was directly linked to reductions in swimming distance compared to controls as well as to chemical-exposed fish whose ACs inflated. Together the data provide compelling support for AOPs linking TH disruption to impaired AC inflation in fish.

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.

Swim bladder morphology changes with female reproductive state in the mouth-brooding African cichlid Astatotilapia burtoni.

Mouth brooding is an extreme form of parental care in which the brooding parent carries the developing young in their buccal cavity for the duration of development. Brooding fish need to compensate for the brood weight on the anterior portion of their body. For fishes with a compartmentalized swim bladder, gas distribution between the chambers may aid in regulating buoyancy during brooding. To test this hypothesis, we took radiographs of Astatotilapia burtoni to compare the swim bladder morphology of gravid, mouth-brooding and recovering females. Following spawning, females carry developing fish in their buccal cavity for ∼2 weeks, resulting in a larger and rounder anterior swim bladder compartment. Comparatively, the swim bladder of gravid females is long and cylindrical. Using small beads to mimic brood weight and its effects on female buoyancy, swim bladder changes were induced that resembled those observed during brooding. Immediately after releasing their fry, brooding females swim at a positive angle of attack but correct their swimming posture to normal within 5min, suggesting a rapid change in swim bladder gas distribution. These data provide new insights into how swim bladder morphology and swimming behavior change during mouth brooding, and suggest a compartmentalized swim bladder may be a morphological adaptation for mouth brooding.

Thyroid disrupting effects of halogenated and next generation chemicals on the swim bladder development of zebrafish

Endocrine disrupting chemicals (EDCs) can alter thyroid function and adversely affect growth and development. Halogenated compounds, such as perfluorinated chemicals commonly used in food packaging, and brominated flame retardants used in a broad range of products from clothing to electronics, can act as thyroid disruptors. Due to the adverse effects of these compounds, there is a need for the development of safer next generation chemicals. The objective of this study was to test the thyroid disruption potential of old use and next generation halogenated chemicals. Zebrafish embryos were exposed to three old use compounds, perfluorooctanoic acid (PFOA), tetrabromobisphenol A (TBBPA) and tris (1,3-dichloro-2-propyl) phosphate (TDCPP) and two next generation chemicals, 9,10-dihydro-9-oxa-10-phosphaphenanthrene-10-oxdie (DOPO) and perfluorobutyric acid (PFBA). Sub-chronic (0–6days post fertilization (dpf)) and chronic (0–28dpf) exposures were conducted at 1% of the concentration known to kill 50% (LC50) of the population. Changes in the surface area of the swim bladder as well as in expression levels of genes involved in the thyroid control of swim bladder inflation were measured. At 6dpf, zebrafish exposed to all halogenated chemicals, both old use and next generation, had smaller posterior swim bladder and increased expression in the gene encoding thyroid peroxidase, tpo and the genes encoding two swim bladder surfactant proteins, sp-a and sp-c. These results mirrored the effects of thyroid hormone-exposed positive controls. Fish exposed to a TPO inhibitor (methimazole, MMI) had a decrease in tpo expression levels at 28dpf. Effects on the anterior swim bladder at 28dpf, after exposure to MMI as well as both old and new halogenated chemicals, were the same, i.e., absence of SB in ∼50% of fish, which were also of smaller body size. Overall, our results suggest thyroid disruption by the halogenated compounds tested via the swim bladder surfactant system. However, with the exception of TBBPA and TDCPP, the concentrations tested (∼5–137ppm) are not likely to be found in the environment.

Impaired swim bladder inflation in early life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid.

Inflation of the posterior and/or anterior swim bladder is a process previously demonstrated to be regulated by thyroid hormones. We investigated whether inhibition of deiodinases, which convert thyroxine (T4) to the more biologically active form, 3,5,3'-triiodothyronine (T3), would impact swim bladder inflation. Two experiments were conducted using a model deiodinase inhibitor, iopanoic acid (IOP). First, fathead minnow embryos were exposed to 0.6, 1.9, or 6.0 mg/L or control water until 6 d postfertilization (dpf), at which time posterior swim bladder inflation was assessed. To examine anterior swim bladder inflation, a second study was conducted with 6-dpf larvae exposed to the same IOP concentrations until 21 dpf. Fish from both studies were sampled for T4/T3 measurements and gene transcription analyses. Incidence and length of inflated posterior swim bladders were significantly reduced in the 6.0 mg/L treatment at 6 dpf. Incidence of inflation and length of anterior swim bladder were significantly reduced in all IOP treatments at 14 dpf, but inflation recovered by 18 dpf. Throughout the larval study, whole-body T4 concentrations increased and T3 concentrations decreased in all IOP treatments. Consistent with hypothesized compensatory responses, deiodinase-2 messenger ribonucleic acid (mRNA) was up-regulated in the larval study, and thyroperoxidase mRNA was down-regulated in all IOP treatments in both studies. These results support the hypothesized adverse outcome pathways linking inhibition of deiodinase activity to impaired swim bladder inflation. Environ Toxicol Chem 2017;36:2942-2952. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Sound production by a recoiling system in the pempheridae and terapontidae.

Sound-producing mechanisms in fishes are extraordinarily diversified. We report here original mechanisms of three species from two families: the pempherid Pempheris oualensis, and the terapontids Terapon jarbua and Pelates quadrilineatus. All sonic mechanisms are built on the same structures. The rostral part of the swimbladder is connected to a pair of large sonic muscles from the head whereas the posterior part is fused with bony widenings of vertebral bodies. Two bladder regions are separated by a stretchable fenestra that allows forward extension of the anterior bladder during muscle contraction. A recoiling apparatus runs between the inner face of the anterior swimbladder and a vertebral body expansion. The elastic nature of the recoiling apparatus supports its role in helping the swimbladder to recover its initial position during sonic muscle relaxation. This system should aid fast contraction (between 100 and 250Hz) of sonic muscles. There are many differences between species in terms of the swimbladder and its attachments to the vertebral column, muscle origins, and morphology of the recoiling apparatus. The recoiling apparatus found in the phylogenetically-related families (Glaucosomatidae, Pempheridae, Terapontidae) could indicate a new character within the Percomorpharia. J. Morphol. 277:717-724, 2016. © 2016 Wiley Periodicals, Inc.

Impaired anterior swim bladder inflation following exposure to the thyroid peroxidase inhibitor 2-mercaptobenzothiazole part II: Zebrafish

Disruption of the thyroid hormone (TH) system, an important mode of action, can lead to ecologically relevant adverse outcomes, especially during embryonic development. The present study characterizes the effects of disruption of TH synthesis on swim bladder inflation during zebrafish early-life stages using 2-mercaptobenzothiazole (MBT), a thyroid peroxidase (TPO) inhibitor. Zebrafish were exposed to different MBT concentrations until 120/168h post fertilization (hpf) and 32days post fertilization (dpf), in two sets of experiments, to investigate the effects of TPO inhibition on posterior and anterior swim bladder inflation respectively, as well as whole body thyroid hormone concentrations (triiodothyronine (T3) and its prohormone, thyroxine (T4)). At 120hpf, MBT did not directly impair posterior chamber inflation or size, while anterior chamber inflation and size was impaired at 32dpf. As previously shown in amphibians and mammals, we confirmed that MBT inhibits TPO in fish. Whole-body T4 decreased after MBT exposure at both time points, while T3 levels were unaltered. There was a significant relationship between T4 levels and the anterior chamber surface at 32dpf. The absence of effects on posterior chamber inflation can possibly be explained by maternal transfer of T4 into the eggs. These maternally derived THs are depleted at 32dpf and cannot offset TPO inhibition, resulting in impaired anterior chamber inflation. Therefore, we hypothesize that TPO inhibition only inhibits swim bladder inflation during late development, after depletion of maternally derived T4. In a previous study, we showed that iodothyronine deiodinase (ID) knockdown impaired posterior chamber inflation during early development. Our findings, in parallel with similar effects observed in fathead minnow (see part I, this issue) suggest that thyroid disruption impacts swim bladder inflation, and imply an important distinction among specific subtypes of TH disrupting chemicals. However, the existence of another – yet unknown – mode of action of MBT impacting swim bladder inflation cannot be excluded. These results can be helpful for delineating adverse outcome pathways (AOPs) linking TPO inhibition, ID inhibition and other TH related molecular initiating events, to impaired swim bladder inflation in fish during early life stages. Such AOPs can support the use of in vitro enzyme inhibition assays for predicting reduced survival due to impaired posterior and anterior chamber inflation.

Impaired anterior swim bladder inflation following exposure to the thyroid peroxidase inhibitor 2-mercaptobenzothiazole part I: Fathead minnow

In the present study, a hypothesized adverse outcome pathway linking inhibition of thyroid peroxidase (TPO) activity to impaired swim bladder inflation was investigated in two experiments in which fathead minnows (Pimephales promelas) were exposed to 2-mercaptobenzothiazole (MBT). Continuous exposure to 1mg MBT/L for up to 22 days had no effect on inflation of the posterior chamber of the swim bladder, which typically inflates around 6 days post fertilization (dpf), a period during which maternally-derived thyroid hormone is presumed to be present. In contrast, inflation of the anterior swim bladder, which occurs around 14dpf, was impacted. Specifically, at 14dpf, approximately 50% of fish exposed to 1mg MBT/L did not have an inflated anterior swim bladder. In fish exposed to MBT through 21 or 22dpf, the anterior swim bladder was able to inflate, but the ratio of the anterior/posterior chamber length was significantly reduced compared to controls. Both abundance of thyroid peroxidase mRNA and thyroid follicle histology suggest that fathead minnows mounted a compensatory response to the presumed inhibition of TPO activity by MBT. Time-course characterization showed that fish exposed to MBT for at least 4 days prior to normal anterior swim bladder inflation had significant reductions in anterior swim bladder size, relative to the posterior chamber, compared to controls. These results, along with similar results observed in zebrafish (see part II, this issue) are consistent with the hypothesis that thyroid hormone signaling plays a significant role in mediating anterior swim bladder inflation and development in cyprinids, and that role can be disrupted by exposure to thyroid hormone synthesis inhibitors. Nonetheless, possible thyroid-independent actions of MBT on anterior swim bladder inflation cannot be ruled out based on the present results. Overall, although anterior swim bladder inflation has not been directly linked to survival as posterior swim bladder inflation has, potential links to adverse ecological outcomes are plausible given involvement of the anterior chamber in sound production and detection.

Acoustic Communication in Butterflyfishes: Anatomical Novelties, Physiology, Evolution, and Behavioral Ecology.

Coral reef fishes live in noisy environments that may challenge their capacity for acoustic communication. Butterflyfishes (Family Chaetodontidae) are prominent and ecologically diverse members of coral reef communities worldwide. The discovery of a novel association of anterior swim bladder horns with the lateral line canal system in the genus Chaetodon (the laterophysic connection) revealed a putative adaptation for enhancement of sound reception by the lateral line system and/or the ear. Behavioral studies show that acoustic communication is an important component of butterflyfish social behavior. All bannerfish (Forcipiger, Heniochus, and Hemitaurichthys) and Chaetodon species studied thus far produce several sound types at frequencies of <1 to >1000 Hz. Ancestral character state analyses predict the existence of both shared (head bob) and divergent (tail slap) acoustic behaviors in these two clades. Experimental auditory physiology shows that butterflyfishes are primarily sensitive to stimuli associated with hydrodynamic particle accelerations of ≤500 Hz. In addition, the gas-filled swim bladder horns in Chaetodon are stimulated by sound pressure, which enhances and extends their auditory sensitivity to 1700-2000 Hz. The broadband spectrum of ambient noise present on coral reefs overlaps with the frequency characteristics of their sounds, thus both the close social affiliations common among butterflyfishes and the evolution of the swim bladder horns in Chaetodon facilitate their short-range acoustic communication. Butterflyfishes provide a unique and unexpected opportunity to carry out studies of fish bioacoustics in the lab and the field that integrate the study of sensory anatomy, physiology, evolution, and behavioral ecology.

Sound pressure enhances the hearing sensitivity of Chaetodon butterflyfishes on noisy coral reefs

Butterflyfishes are conspicuous members of coral reefs that communicate with acoustic signals during social interactions with mates and other conspecifics. Members of the genus Chaetodon have a laterophysic connection (LC) - a unique association of anterior swim bladder horns and the cranial lateral line - but the action of the LC system on auditory sensitivity is unexplored. Here, we show in baseline auditory evoked potential threshold experiments that Forcipiger flavissimus (which lacks swim bladder horns and LC) is sensitive to sound tones from 100 Hz up to 1000 Hz, and that thresholds for three species of Chaetodon are 10-15 dB lower, with extended hearing ranges up to 1700-2000 Hz. The relatively high thresholds to sound pressure and low pass response near 500 Hz for all four species are consistent with a primary sensitivity to hydrodynamic particle acceleration rather than sound pressure. Deflation of the swim bladder in F. flavissimus had no measurable effect on auditory sensitivity. In contrast, displacement of gas from the swim bladder horns in Chaetodon multicinctus and Chaetodon auriga increased thresholds (decreased sensitivity) by 5-20 dB, with the greatest effect at 600 Hz. The evolution of swim bladder horns associated with the LC system in Chaetodon species has increased hearing sensitivity through sound pressure transduction in the frequency bands used for social acoustic communication. The close affiliative behaviors that are common in Chaetodon species and other butterflyfish facilitate sound perception and acoustic communication at close distances relative to the high background noise levels found in their natural reef environment.

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

BackgroundCichlid 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 resultsWe 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.ConclusionsOur 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.

Hearing in Cichlid Fishes under Noise Conditions

BackgroundHearing thresholds of fishes are typically acquired under laboratory conditions. This does not reflect the situation in natural habitats, where ambient noise may mask their hearing sensitivities. In the current study we investigate hearing in terms of sound pressure (SPL) and particle acceleration levels (PAL) of two cichlid species within the naturally occurring range of noise levels. This enabled us to determine whether species with and without hearing specializations are differently affected by noise.Methodology/Principal FindingsWe investigated auditory sensitivities in the orange chromide Etroplus maculatus, which possesses anterior swim bladder extensions, and the slender lionhead cichlid Steatocranus tinanti, in which the swim bladder is much smaller and lacks extensions. E. maculatus was tested between 0.2 and 3kHz and S. tinanti between 0.1 and 0.5 kHz using the auditory evoked potential (AEP) recording technique. In both species, SPL and PAL audiograms were determined in the presence of quiet laboratory conditions (baseline) and continuous white noise of 110 and 130 dB RMS. Baseline thresholds showed greatest hearing sensitivity around 0.5 kHz (SPL) and 0.2 kHz (PAL) in E. maculatus and 0.2 kHz in S. tinanti. White noise of 110 dB elevated the thresholds by 0–11 dB (SPL) and 7–11 dB (PAL) in E. maculatus and by 1–2 dB (SPL) and by 1–4 dB (PAL) in S. tinanti. White noise of 130 dB elevated hearing thresholds by 13–29 dB (SPL) and 26–32 dB (PAL) in E. maculatus and 6–16 dB (SPL) and 6–19 dB (PAL) in S. tinanti.ConclusionsOur data showed for the first time for SPL and PAL thresholds that the specialized species was masked by different noise regimes at almost all frequencies, whereas the non-specialized species was much less affected. This indicates that noise can limit sound detection and acoustic orientation differently within a single fish family.

Fast drum strokes: Novel and convergent features of sonic muscle ultrastructure, innervation, and motor neuron organization in the pyramid butterflyfish (hemitaurichthys polylepis)

Sound production that is mediated by intrinsic or extrinsic swim bladder musculature has evolved multiple times in teleost fishes. Sonic muscles must contract rapidly and synchronously to compress the gas-filled bladder with sufficient velocity to produce sound. Muscle modifications that may promote rapid contraction include small fiber diameter, elaborate sarcoplasmic reticulum (SR), triads at the A-I boundary, and cores of sarcoplasm. The diversity of innervation patterns indicate that sonic muscles have independently evolved from different trunk muscle precursors. The analysis of sonic motor pathways in distantly related fishes is required to determine the relationships between sonic muscle evolution and function in acoustic signaling. We examined the ultrastructure of sonic and adjacent hypaxial muscle fibers and the distribution of sonic motor neurons in the coral reef Pyramid Butterflyfish (Chaetodontidae: Hemitaurichthys polylepis) that produces sound by contraction of extrinsic sonic muscles near the anterior swim bladder. Relative to adjacent hypaxial fibers, sonic muscle fibers were sparsely arranged among the endomysium, smaller in cross-section, had longer sarcomeres, a more elaborate SR, wider t-tubules, and more radially arranged myofibrils. Both sonic and non-sonic muscle fibers possessed triads at the Z-line, lacked sarcoplasmic cores, and had mitochondria among the myofibrils and concentrated within the peripheral sarcoplasm. Sonic muscles of this derived eutelost possess features convergent with other distant vocal taxa (other euteleosts and non-euteleosts): small fiber diameter, a well-developed SR, and radial myofibrils. In contrast with some sonic fishes, however, Pyramid Butterflyfish sonic muscles lack sarcoplasmic cores and A-I triads. Retrograde nerve label experiments show that sonic muscle is innervated by central and ventrolateral motor neurons associated with spinal nerves 1-3. This restricted distribution of sonic motor neurons in the spinal cord differs from many euteleosts and likely reflects the embryological origin of sonic muscles from hypaxial trunk precursors rather than occipital somites.

Relationship between Swim Bladder Morphology and Hearing Abilities–A Case Study on Asian and African Cichlids

BackgroundSeveral teleost species have evolved anterior extensions of the swim bladder which come close to or directly contact the inner ears. A few comparative studies have shown that these morphological specializations may enhance hearing abilities. This study investigates the diversity of swim bladder morphology in four Asian and African cichlid species and analyzes how this diversity affects their hearing sensitivity.Methodology/Principal FindingsWe studied swim bladder morphology by dissections and by making 3D reconstructions from high-resolution microCT scans. The auditory sensitivity was determined in terms of sound pressure levels (SPL) and particle acceleration levels (PAL) using the auditory evoked potential (AEP) recording technique. The swim bladders in Hemichromis guttatus and Steatocranus tinanti lacked anterior extensions and the swim bladder was considerably small in the latter species. In contrast, Paratilapia polleni and especially Etroplus maculatus possessed anterior extensions bringing the swim bladder close to the inner ears. All species were able to detect frequencies up to 3 kHz (SPL) except S. tinanti which only responded to frequencies up to 0.7 kHz. P. polleni and E. maculatus showed significantly higher auditory sensitivities at 0.5 and 1 kHz than the two species lacking anterior swim bladder extensions. The highest auditory sensitivities were found in E. maculatus, which possessed the most intimate swim bladder-inner ear relationship (maximum sensitivity 66 dB re 1 µPa at 0.5 kHz).ConclusionsOur results indicate that anterior swim bladder extensions seem to improve mean absolute auditory sensitivities by 21–42 dB (SPLs) and 21–36 dB (PALs) between 0.5 and 1 kHz. Besides anterior extensions, the size of the swim bladder appears to be an important factor for extending the detectable frequency range (up to 3 kHz).

Convergent evolution and intermediate status of the sonic mechanism in a perciform fish Glaucosoma buergeri (Glaucosomatidae).

Little is known about evolution of swimbladder sound production in fishes. Typical fish sound production utilizes fast muscles that drive the swimbladder to produce sound as a forced but rapidly‐damping response; a muscle contraction rate of 200 times/s will generate a sound with a fundamental frequency of 200 Hz. Recently, slow muscles have been demonstrated in a carapid fish. These muscles slowly extend the anterior swimbladder by stretching a swimbladder fenestra until the bladder snaps back exciting sound production. Here we describe sounds produced by a similar but phylogenetically‐unrelated mechanism in a perciform fish. A pair of extrinsic sonic muscles originates on the pterotic bones and inserts on the anterodorsal swimbladder just forward of a stretchable swimbladder fenestra. A fan‐shaped tendon that ends in a smooth muscle attaches to the bladder just forward of the fenestra. The tendon‐smooth muscle pair will be stretched by contraction of the rostral sonic muscles. Strain energy stored in th...

Pulse sound generation, anterior swim bladder buckling and associated muscle activity in the pyramid butterflyfish, Hemitaurichthys polylepis

Acoustic behaviors are widespread among diverse fish taxa but mechanisms of sound production are known from relatively few species, vary widely and convergent mechanisms are poorly known. We examined the sound production mechanism in the pyramid butterflyfish, Hemitaurichthys polylepis, a member of the socially and ecologically diverse reef fish family Chaetodontidae. In the field, fish produce pulse trains at dusk during social interactions that are probably related to mate attraction and courtship. In laboratory experiments, sound production was synchronized to high-speed video to determine body movement associated with sound generation. In addition, electromyography (EMG) recordings tested the activity of six candidate muscles. Fish produced individual pulses with a mean peak frequency of 97 Hz in rapid succession. EMG experiments show that anterior hypaxial muscles contract at high bilaterally synchronous rates (up to 120 Hz) in near perfect association with rapid inward buckling visible outside the body over the anterior swim bladder. Muscle activity often showed EMG doublets that occurred within the time of a single sound pulse but was not sustained. Buckling and sound pulse rates correlated strongly (R(2)≈1.00) and sound pulse rate measured over two successive pulses (maximum of 38 pulses s(-1)) was lower than muscle firing rate. These results show that the extrinsic swim bladder muscles of pyramid butterflyfish involve single contractions that produce pulses in a manner similar to distantly related teleosts, but involve a novel doublet motor-neuron firing pattern. Thus, the sound production mechanism in pyramid butterflyfish is likely convergent with several percomorph taxa and divergent from the related chaetodontid genus Forcipiger.