Offshore wind energy development: Research priorities for sound and vibration effects on fishes and aquatic invertebrates.

There are substantial knowledge gaps regarding both the bioacoustics and the responses of animals to sounds associated with pre-construction, construction, and operations of offshore wind (OSW) energy development. A workgroup of the 2020 State of the Science Workshop on Wildlife and Offshore Wind Energy identified studies for the next five years to help stakeholders better understand potential cumulative biological impacts of sound and vibration to fishes and aquatic invertebrates as the OSW industry develops. The workgroup identified seven short-term priorities that include a mix of primary research and coordination efforts. Key research needs include the examination of animal displacement and other behavioral responses to sound, as well as hearing sensitivity studies related to particle motion, substrate vibration, and sound pressure. Other needs include: identification of priority taxa on which to focus research; standardization of methods; development of a long-term highly instrumented field site; and examination of sound mitigation options for fishes and aquatic invertebrates. Effective assessment of potential cumulative impacts of sound and vibration on fishes and aquatic invertebrates is currently precluded by these and other knowledge gaps. However, filling critical gaps in knowledge will improve our understanding of possible sound-related impacts of OSW energy development to populations and ecosystems.

Seasonal plasticity of auditory saccular sensitivity in "sneaker" type II male plainfin midshipman fish, Porichthys notatus.

Adult female and nesting (type I) male midshipman fish (Porichthys notatus) exhibit an adaptive form of auditory plasticity for the enhanced detection of social acoustic signals. Whether this adaptive plasticity also occurs in "sneaker" type II males is unknown. Here, we characterize auditory-evoked potentials recorded from hair cells in the saccule of reproductive and non-reproductive "sneaker" type II male midshipman to determine whether this sexual phenotype exhibits seasonal, reproductive state-dependent changes in auditory sensitivity and frequency response to behaviorally relevant auditory stimuli. Saccular potentials were recorded from the middle and caudal region of the saccule while sound was presented via an underwater speaker. Our results indicate saccular hair cells from reproductive type II males had thresholds based on measures of sound pressure and acceleration (re. 1 µPa and 1 ms-2, respectively) that were ~8-21 dB lower than non-reproductive type II males across a broad range of frequencies, which include the dominant higher frequencies in type I male vocalizations. This increase in type II auditory sensitivity may potentially facilitate eavesdropping by sneaker males and their assessment of vocal type I males for the selection of cuckoldry sites during the breeding season.

Effects of Exposure to the Sound from Seismic Airguns on Pallid Sturgeon and Paddlefish.

This study examined the effects of exposure to a single acoustic pulse from a seismic airgun array on caged endangered pallid sturgeon (Scaphirhynchus albus) and on paddlefish (Polyodon spathula) in Lake Sakakawea (North Dakota, USA). The experiment was designed to detect the onset of physiological responses including minor to mortal injuries. Experimental fish were held in cages as close as 1 to 3 m from the guns where peak negative sound pressure levels (Peak- SPL) reached 231 dB re 1 µPa (205 dB re 1 µPa2·s sound exposure level [SEL]). Additional cages were placed at greater distances in an attempt to develop a dose-response relationship. Treatment and control fish were then monitored for seven days, euthanized, and necropsied to determine injuries. Necropsy results indicated that the probability of delayed mortality associated with pulse pressure following the seven day monitoring period was the same for exposed and control fish of both species. Exposure to a single pulse from a small air gun array (10,160 cm3) was not lethal for pallid sturgeon and paddlefish. However, the risks from exposure to multiple sounds and to sound exposure levels that exceed those reported here remain to be examined.

Evaluation of Three Sensor Types for Particle Motion Measurement.

All fish sense acoustic particle motion; some species also sense pressure. Concern over the effects of anthropogenic sounds is increasing the need to monitor acoustic particle motion. Particle motion can be measured directly using vector sensors or calculated from pressure gradients. This article compares three devices that measure particle motion: a three-axis accelerometer, a three-axis velocity sensor, and two 4-element hydrophone arrays. A series of sounds (constant-wave tones, white noise, and Ricker wavelets) were played from a fixed-position projector. The particle motion of sounds from imploding light bulbs was also measured.

Pile Driving at the New Bridge at Tappan Zee: Potential Environmental Impacts.

A new bridge will be constructed to replace the aging Tappan Zee Bridge over the Hudson River in New York. Construction will potentially result in hydroacoustic impacts to the local fish fauna. As a consequence, a substantial environmental impact analysis had to be conducted to obtain construction permits. This paper describes the process of environmental analysis and some of the results of the studies that led up to the final permitting. The process included modeling of pile-driving acoustics, analysis of river ambient noise, analysis of test piling, and observations on fish behavior during these tests.

Effects of Seismic Air Guns on Pallid Sturgeon and Paddlefish.

Pallid sturgeon and paddlefish were placed at different distances from a seismic air gun array to determine the potential effects on mortality and nonauditory body tissues from the sound from a single shot. Fish were held 7 days postexposure and then necropsied. No fish died immediately after sound exposure or over the postexposure period. Statistical analysis of injuries showed no differences between the experimental and control animals in either type or severity of injuries. There was also no difference in injuries between fish exposed closest to the source compared with those exposed furthest from the source.

Whistle source levels of free-ranging bottlenose dolphins and Atlantic spotted dolphins in the Gulf of Mexico.

Whistles of bottlenose dolphins (Tursiops truncatus) and Atlantic spotted dolphins (Stenella frontalis) in the eastern Gulf of Mexico were recorded and measured with a calibrated towed hydrophone array. Surveys encountered groups of both bottlenose (N = 10) and spotted dolphins (N = 5). Analysis of those data produced 1695 bottlenose dolphin whistles and 1273 spotted dolphin whistles with a high signal-to-noise ratio. Whistle frequency metrics were lower in bottlenose than spotted dolphins, while whistle duration was longer in spotted dolphins, data that may help inform automatic classification algorithms. Source levels were estimated by determining the range and bearing of an individual dolphin from the array and then adding the predicted transmission loss to the calculated received level. The median bottlenose dolphin source level was 138 dB re 1µPa at 1 m with a range of 114-163 dB re 1µPa at 1 m. The median spotted dolphin source level was 138 dB re 1µPa at 1 m with a range of 115-163 dB re 1µPa at 1 m. These source level measurements, in conjunction with estimates of vocalization rates and transmission loss models, can be used to improve passive acoustically determined dolphin abundance estimates in the Gulf of Mexico.

Use of the swim bladder and lateral line in near-field sound source localization by fish.

We investigated the roles of the swim bladder and the lateral line system in sound localization behavior by the plainfin midshipman fish (Porichthys notatus). Reproductive female midshipman underwent either surgical deflation of the swim bladder or cryoablation of the lateral line and were then tested in a monopolar sound source localization task. Fish with nominally 'deflated' swim bladders performed similar to sham-deflated controls; however, post-experiment evaluation of swim bladder deflation revealed that a majority of 'deflated' fish (88%, seven of the eight fish) that exhibited positive phonotaxis had partially inflated swim bladders. In total, 95% (21/22) of fish that localized the source had at least partially inflated swim bladders, indicating that pressure reception is likely required for sound source localization. In lateral line experiments, no difference was observed in the proportion of females exhibiting positive phonotaxis with ablated (37%) versus sham-ablated (47%) lateral line systems. These data suggest that the lateral line system is likely not required for sound source localization, although this system may be important for fine-tuning the approach to the sound source. We found that midshipman can solve the 180 deg ambiguity of source direction in the shallow water of our test tank, which is similar to their nesting environment. We also found that the potential directional cues (phase relationship between pressure and particle motion) in shallow water differs from a theoretical free-field. Therefore, the general question of how fish use acoustic pressure cues to solve the 180 deg ambiguity of source direction from the particle motion vector remains unresolved.

Auditory sensitivity of larval zebrafish (Danio rerio) measured using a behavioral prepulse inhibition assay.

Zebrafish (Danio rerio) have become a valuable model for investigating the molecular genetics and development of the inner ear in vertebrates. In this study, we employed a prepulse inhibition (PPI) paradigm to assess hearing in larval wild-type (AB) zebrafish during early development at 5-6 days post-fertilization (d.p.f.). We measured the PPI of the acoustic startle response in zebrafish using a 1-dimensional shaker that simulated the particle motion component of sound along the fish's dorsoventral axis. The thresholds to startle-inducing stimuli were determined in 5-6 d.p.f. zebrafish, and their hearing sensitivity was then characterized using the thresholds of prepulse tone stimuli (90-1200 Hz) that inhibited the acoustic startle response to a reliable startle stimulus (820 Hz at 20 dB re. 1 m s(-2)). Hearing thresholds were defined as the minimum prepulse tone level required to significantly reduce the startle response probability compared with the baseline (no-prepulse) condition. Larval zebrafish showed greatest auditory sensitivity from 90 to 310 Hz with corresponding mean thresholds of -19 to -10 dB re. 1 m s(-2), respectively. Hearing thresholds of prepulse tones were considerably lower than previously predicted by startle response assays. The PPI assay was also used to investigate the relative contribution of the lateral line to the detection of acoustic stimuli. After aminoglycoside-induced neuromast hair-cell ablation, we found no difference in PPI thresholds between treated and control fish. We propose that this PPI assay can be used to screen for novel zebrafish hearing mutants and to investigate the ontogeny of hearing in zebrafish and other fishes.

Effects of low-frequency naval sonar exposure on three species of fish.

To address growing concern over the impact of anthropogenic sound on fishes, a series of experiments was conducted that exposed several fish species to high-intensity low-frequency naval sonar. This study extends auditory findings by adding largemouth bass, yellow perch, and channel catfish. No effects on hearing were found in largemouth bass and yellow perch and only small effects in channel catfish (a fish with morphological adaptations for enhanced pressure reception). Together with prior findings, these results suggest limited impact on hearing from high-intensity sonar. Susceptibility may be due to genetic stock, developmental conditions, seasonal variation, and/or buoyancy during exposure.

Effects of mid-frequency active sonar on hearing in fish.

Caged fish were exposed to sound from mid-frequency active (MFA) transducers in a 5 × 5 planar array which simulated MFA sounds at received sound pressure levels of 210 dB SPL(re 1 µPa). The exposure sound consisted of a 2 s frequency sweep from 2.8 to 3.8 kHz followed by a 1 s tone at 3.3 kHz. The sound sequence was repeated every 25 s for five repetitions resulting in a cumulative sound exposure level (SEL(cum)) of 220 dB re 1 µPa(2) s. The cumulative exposure level did not affect the hearing sensitivity of rainbow trout, a species whose hearing range is lower than the frequencies in the presented MFA sound. In contrast, one cohort of channel catfish showed a statistically significant temporary threshold shift of 4-6 dB at 2300 Hz, but not at lower tested frequencies, whereas a second cohort showed no change. It is likely that this threshold shift resulted from the frequency spectrum of the MFA sound overlapping with the upper end of the hearing frequency range of the channel catfish. The observed threshold shifts in channel catfish recovered within 24 h. There was no mortality associated with the MFA sound exposure used in this test.

Local acoustic particle motion guides sound-source localization behavior in the plainfin midshipman fish, Porichthys notatus.

Sound-source localization behavior was studied in the plainfin midshipman fish (Porichthys notatus) by making use of the naturally occurring phonotaxis response of gravid females to playback of the male's advertisement call. The observations took place outdoors in a circular concrete tank. A dipole sound projector was placed at the center of the tank and an 80-90 Hz tone (the approximate fundamental frequency to the male's advertisement call) was broadcast to gravid females that were released from alternative sites approximately 100 cm from the source. The phonotaxic responses of females to the source were recorded, analyzed and compared with the sound field. One release site was approximately along the vibratory axis of the dipole source, and the other was approximately orthogonal to the vibratory axis. The sound field in the tank was fully characterized through measurements of the sound pressure field using hydrophones and acoustic particle motion using an accelerometer. These measurements confirmed that the sound field was a nearly ideal dipole. When released along the dipole vibratory axis, the responding female fish took essentially straight paths to the source. However, when released approximately 90 deg to the source's vibratory axis, the responding females took highly curved paths to the source that were approximately in line with the local particle motion axes. These results indicate that the acoustic cues used by fish during sound-source localization include the axes of particle motion of the local sound field.

Sound source localization by the plainfin midshipman fish, Porichthys notatus.

The aim of this study was to use plainfin midshipman fish (Porichthys notatus) as a general model to explore how fishes localize an underwater sound source in the relatively simple geometry of a monopole sound field. The robust phonotaxic responses displayed by gravid females toward a monopole sound projector (J-9) broadcasting a low-frequency (90 Hz) tone similar to the fundamental frequency of the male's advertisement call were examined. The projector's sound field was mapped at 5 cm resolution azimuth using an eight-hydrophone array. Acoustic pressure was measured with the array and acoustic particle motion was calculated from pressure gradients between hydrophones. The response pathways of the fish were analyzed from video recordings and compared to the sound field. Gravid females at initial release were directed toward the sound source, and the majority (73%) swam to the playback projector with straight to slightly curved tracks in the direction of the source and in line with local particle motion vectors. In contrast, the initial direction of the control (sound-off) group did not differ from random. This paper reports on a comparison of fish localization behavior with directional cues available in the form of local particle motion vectors.

Depth dependent variation of the echolocation pulse rate of bottlenose dolphins (Tursiops truncatus).

Trained odontocetes appear to have good control over the timing (pulse rate) of their echolocation clicks; however, there is comparatively little information about how free-ranging odontocetes modify their echolocation in relation to their environment. This study investigates echolocation pulse rate in 14 groups of free-ranging bottlenose dolphins (Tursiops truncatus) at a variety of depths (2.4-30.1 m) in the Gulf of Mexico. Linear regression models indicated a significant decrease in mean pulse rate with mean water depth. Pulse rates for most groups were multi-modal. Distance to target estimates were as high as 91.8 m, assuming that echolocation was produced at a maximal rate for the target distance. A 5.29-ms processing lag time was necessary to explain the pulse rate modes observed. Although echolocation is likely reverberation limited, these results support the hypotheses that free-ranging bottlenose dolphins in this area are adapting their echolocation signals for a variety of target detection and ranging purposes, and that the target distance is a function of water depth.

Zebrafish pax5 regulates development of the utricular macula and vestibular function.

The zebrafish otic vesicle initially forms with only two sensory epithelia, the utricular and saccular maculae, which primarily mediate vestibular and auditory function, respectively. Here, we test the role of pax5, which is preferentially expressed in the utricular macula. Morpholino knockdown of pax5 disrupts vestibular function but not hearing. Neurons of the statoacoustic ganglion (SAG) develop normally. Utricular hair cells appear to form normally but a variable number subsequently undergo apoptosis and are extruded from the otic vesicle. Dendrites of the SAG persist in the utricle but become disorganized after hair cell loss. Hair cells in the saccule develop and survive normally. Otic expression of pax5 requires pax2a and fgf3, mutations in which cause vestibular defects, albeit by distinct mechanisms. Thus, pax5 works in conjunction with fgf3 and pax2a to establish and/or maintain the utricular macula and is essential for vestibular function.

Development of the acoustically evoked behavioral response in zebrafish to pure tones.

Zebrafish (Danio rerio) were placed in small wells that could be driven vertically with a series of calibrated sinusoids. Video images of the fish were obtained and analyzed to determine the levels and frequencies at which the fish responded to the stimulus tones. It was found that fish 4 days post fertilization (dpf) did not respond to the stimulus tones, whereas fish 5 dpf to adult did respond. It was further found that the stimulus thresholds and frequency bandwidth to which the fish responded did not change from 5 dpf to adult; indicating that the otolithic organ adaptations for high-frequency hearing are already present in larval fish. Deflating the swimbladders in adult fish eliminated their response, which is consistent with sensing sound pressure. Deflating the swimbladder in larval fish did not affect their thresholds, which is consistent with sensing the particle motion of the fluid directly. Because adult fish with Weberian ossicles have a greater input to the inner ear for a given sound pressure level (SPL), the finding that the adult and larval fish respond at the same SPL with intact swimbladders suggests that the acoustic startle response threshold is adjusted as the fish develop in order to maintain appropriate reactions to relevant stimuli.

A biophysical model of an inner hair cell.

Whole-cell patch-clamp recordings on isolated inner hair cells (IHCs) of guinea pig cochleae have revealed the presence of voltage-gated potassium channels. A biophysical model of an IHC is presented that indicates activation of slow voltage-gated potassium channels may lead to receptor potentials whose dc component decreases during the stimulus, and membrane potential hyperpolarizes when the stimulus is turned off. Both the decreasing dc and the hyperpolarization are, respectively, consistent with rapid adaptation and suppression of spontaneous rate in the auditory nerve. Receptor potentials recorded in vivo do not show these features, and when a nonspecific leak is included in the model to simulate microelectrode impalement, the model's receptor potentials become similar to those in vivo. The nonspecific leak creates an electrical shunt that masks slow channel activity and allows the cell to depolarize. Both the decreasing dc and the hyperpolarization are sensitive to the resting potential. Because the reported resting potentials in vivo and in vitro differ greatly, the model is used to investigate homeostatic mechanisms responsible for the resting potential. It is found that the voltage-gated potassium channels have the greatest influence on the resting potential, but that the standing transducer current may be sufficient to eliminate the decreasing dc and after-stimulus hyperpolarization.