The active space of sperm whale codas: inter-click information for intra-unit communication.

Sperm whales (Physeter macrocephalus) are social mega-predators who form stable matrilineal units that often associate within a larger vocal clan. Clan membership is defined by sharing a repertoire of coda types consisting of specific temporal spacings of multi-pulsed clicks. It has been hypothesized that codas communicate membership across socially segregated sympatric clans, but others propose that codas are primarily used for behavioral coordination and social cohesion within a closely spaced social unit. Here, we test these hypotheses by combining measures of ambient noise levels and coda click source levels with models of sound propagation to estimate the active space of coda communication. Coda clicks were localized off the island of Dominica with a four- or five-element 80 m vertical hydrophone array, allowing us to calculate the median RMS source levels of 1598 clicks from 444 codas to be 161 dB re. 1 µPa (IQR 153-167), placing codas among the most powerful communication sounds in toothed whales. However, together with measured ambient noise levels, these source levels lead to a median active space of coda communication of ∼4 km, reflecting the maximum footprint of a single foraging sperm whale unit. We conclude that while sperm whale codas may contain information about clan affiliation, their moderate active space shows that codas are not used for long range acoustic communication between units and clans, but likely serve to mediate social cohesion and behavioral transitions in intra-unit communication.

Thresholds for noise induced hearing loss in harbor porpoises and phocid seals.

Intense sound sources, such as pile driving, airguns, and military sonars, have the potential to inflict hearing loss in marine mammals and are, therefore, regulated in many countries. The most recent criteria for noise induced hearing loss are based on empirical data collected until 2015 and recommend frequency-weighted and species group-specific thresholds to predict the onset of temporary threshold shift (TTS). Here, evidence made available after 2015 in light of the current criteria for two functional hearing groups is reviewed. For impulsive sounds (from pile driving and air guns), there is strong support for the current threshold for very high frequency cetaceans, including harbor porpoises (Phocoena phocoena). Less strong support also exists for the threshold for phocid seals in water, including harbor seals (Phoca vitulina). For non-impulsive sounds, there is good correspondence between exposure functions and empirical thresholds below 10 kHz for porpoises (applicable to assessment and regulation of military sonars) and between 3 and 16 kHz for seals. Above 10 kHz for porpoises and outside of the range 3-16 kHz for seals, there are substantial differences (up to 35 dB) between the predicted thresholds for TTS and empirical results. These discrepancies call for further studies.

Echolocating Daubenton's bats call louder, but show no spectral jamming avoidance in response to bands of masking noise during a landing task.

Echolocating bats listen for weak echoes to navigate and hunt, which makes them prone to masking from background noise and jamming from other bats and prey. As for electrical fish that display clear spectral jamming avoidance responses (JAR), bats have been reported to mitigate the effects of jamming by shifting the spectral contents of their calls, thereby reducing acoustic interference to improve echo-to-noise ratio (ENR). Here, we tested the hypothesis that frequency-modulating bats (FM bats) employ a spectral JAR in response to six masking noise bands ranging from 15 to 90 kHz, by measuring the -3 dB endpoints and peak frequency of echolocation calls from five male Daubenton's bats (Myotis daubentonii) during a landing task. The bats were trained to land on a noise-generating spherical transducer surrounded by a star-shaped microphone array, allowing for acoustic localization and source parameter quantification of on-axis calls. We show that the bats did not employ spectral JAR as the peak frequency during jamming remained unaltered compared with that of silent controls (all P>0.05, 60.73±0.96 kHz, mean±s.e.m.), and -3 dB endpoints decreased in noise irrespective of treatment type. Instead, Daubenton's bats responded to acoustic jamming by increasing call amplitude via a Lombard response that was bandwidth dependent, ranging from a mean of 0.05 dB/dB (95% confidence interval 0.04-0.06 dB/dB) noise for the most narrowband noise (15-30 kHz) to 0.17 dB/dB (0.16-0.18 dB/dB) noise for the most broadband noise (30-90 kHz). We conclude that Daubenton's bats, despite having the vocal flexibility to do so, do not employ a spectral JAR, but defend ENRs via a bandwidth-dependent Lombard response.

Echolocating Daubenton's bats are resilient to broadband, ultrasonic masking noise during active target approaches.

Echolocating bats hunt prey on the wing under conditions of poor lighting by emission of loud calls and subsequent auditory processing of weak returning echoes. To do so, they need adequate echo-to-noise ratios (ENRs) to detect and distinguish target echoes from masking noise. Early obstacle avoidance experiments report high resilience to masking in free-flying bats, but whether this is due to spectral or spatiotemporal release from masking, advanced auditory signal detection or an increase in call amplitude (Lombard effect) remains unresolved. We hypothesized that bats with no spectral, spatial or temporal release from masking noise defend a certain ENR via a Lombard effect. We trained four bats (Myotis daubentonii) to approach and land on a target that broadcasted broadband noise at four different levels. An array of seven microphones enabled acoustic localization of the bats and source level estimation of their approach calls. Call duration and peak frequency did not change, but average call source levels (SLRMS, at 0.1 m as dB re. 20 µPa) increased, from 112 dB in the no-noise treatment, to 118 dB (maximum 129 dB) at the maximum noise level of 94 dB re. 20 µPa root mean square. The magnitude of the Lombard effect was small (0.13 dB SLRMS dB-1 of noise), resulting in mean broadband and narrowband ENRs of -11 and 8 dB, respectively, at the highest noise level. Despite these poor ENRs, the bats still performed echo-guided landings, making us conclude that they are very resilient to masking even when they cannot avoid it spectrally, spatially or temporally.

Hunting bats adjust their echolocation to receive weak prey echoes for clutter reduction.

How animals extract information from their surroundings to guide motor patterns is central to their survival. Here, we use echo-recording tags to show how wild hunting bats adjust their sensory strategies to their prey and natural environment. When searching, bats maximize the chances of detecting small prey by using large sensory volumes. During prey pursuit, they trade spatial for temporal information by reducing sensory volumes while increasing update rate and redundancy of their sensory scenes. These adjustments lead to very weak prey echoes that bats protect from interference by segregating prey sensory streams from the background using a combination of fast-acting sensory and motor strategies. Counterintuitively, these weak sensory scenes allow bats to be efficient hunters close to background clutter broadening the niches available to hunt for insects.

Do echolocating toothed whales direct their acoustic gaze on- or off-target in a static detection task?

Echolocating mammals produce directional sound beams with high source levels to improve echo-to-noise ratios and reduce clutter. Recent studies have suggested that the differential spectral gradients of such narrow beams are exploited to facilitate target localization by pointing the beam slightly off targets to maximize the precision of angular position estimates [maximizing bearing Fisher information (FI)]. Here, we test the hypothesis that echolocating toothed whales focus their acoustic gaze askew during target detection to maximize spectral cues by investigating the acoustic gaze direction of two trained delphinids (Tursiops truncatus and Pseudorca crassidens) echolocating to detect an aluminum cylinder behind a hydrophone array in a go/no-go paradigm. The animals rarely placed their beam axis directly on the target, nor within the narrow range around the off-axis angle that maximizes FI. However, the target was, for each trial, ensonified within the swath of the half-power beam width, and hence we conclude that the animals solved the detection task using a strategy that seeks to render high echo-to-noise ratios rather than maximizing bearing FI. We posit that biosonar beam adjustment and acoustic gaze strategies are likely task-dependent and that maximizing bearing FI by pointing off-axis does not improve target detection performance.

Sodium bicarbonate cotransporter NBCn1/Slc4a7 affects locomotor activity and hearing in mice.

Despite a widespread expression pattern in the central nervous system, the role of the sodium bicarbonate cotransporter NBCn1/Slc4a7 has not been investigated for locomotor activity, emotion and cognition. Here, we addressed the behavioral consequences of NBCn1 knockout and evaluated hearing and vision that are reportedly impaired in an earlier line of NBCn1 knockout mice and may contribute to behavioral changes. In a circular open field, the knockout mice traveled a shorter distance, especially in the periphery of the chamber, than wildtype littermates. The knockout mice also traveled a shorter total distance in a home cage-like open field. Rearing and grooming behaviors were reduced. The knockout and control mice displayed similar time spent and number of open and closed arms in the elevated plus maze test, indicating negligible change in anxiety. In the Morris water maze test, both groups of mice learned the location of an escape platform within comparable time on the training trials and showed similar platform identification on the probe trial. The knockout mice maintained normal visual responses in the optokinetic drum and produced evoked potentials in response to light stimuli. However, these mice failed to produce auditory evoked potentials. qPCR revealed a robust expression of an alternatively transcribed NBCn1 variant in the knockout mouse retina. These results indicate that NBCn1 deletion leads to reduced locomotor activity in mice by affecting their exploratory behaviors or emotionality. The deletion also causes hearing loss, but its effect on vision varies between different lines of knockout mice.

Lateralized sound production in the beluga whale (Delphinapterus leucas).

Like other toothed whales, belugas produce sound through pneumatic actuation of two phonic lip pairs, but it is unclear whether both pairs are actuated concurrently to generate a single sound (the dual actuation hypothesis) or laterally in the production of their rich vocal repertoires. Here, using suction cup hydrophones on the head of a trained beluga whale, we measured seven different communication signal types and echolocation clicks in order to test the hypothesis that belugas produce distinct sounds unilaterally. We show that, like other delphinoids, belugas produce echolocation clicks with the right phonic lips and tonal sounds from the left. We also demonstrate for the first time that the left phonic lips are responsible for generating communication signals other than tonal sounds. Thus, our findings provide empirical support for functionalized laterality in delphinoid sound production, in keeping with the functional laterality hypothesis of vocal-motor control in toothed whales.

Energy compensation and received echo level dynamics in constant-frequency bats during active target approaches.

Bats have been reported to adjust the energy of their outgoing vocalizations to target range (R) in a logarithmic fashion close to 20log10R which has been interpreted as providing one-way compensation for increasing echo levels during target approaches. However, it remains unknown how species using high-frequency calls, which are strongly affected by absorption, adjust their vocal outputs during approaches to point targets. We hypothesized that such species should compensate less than the 20log10R model predicts at longer distances and more at shorter distances as a consequence of the significant influence of absorption at longer ranges. Using a microphone array and an acoustic recording tag, we show that the output adjustments of two Hipposideros pratti and one Hipposiderosarmiger do not decrease logarithmically during approaches to different-sized targets. Consequently, received echo levels increase dramatically early in the approach phase with near-constant output levels, but level off late in the approach phase as a result of substantial output reductions. To improve echo-to-noise ratio, we suggest that bats using higher frequency vocalizations compensate less at longer ranges, where they are strongly affected by absorption. Close to the target, they decrease their output levels dramatically to mitigate reception of very high echo levels. This strategy maintains received echo levels between 6 and 40 dB re. 20 µPa2 s across different target sizes. The bats partially compensated for target size, but not in a one-to-one dB fashion, showing that these bats do not seek to stabilize perceived echo levels, but may instead use them to gauge target size.

Recreational vessels without Automatic Identification System (AIS) dominate anthropogenic noise contributions to a shallow water soundscape.

Recreational boating is an increasing activity in coastal areas and its spatiotemporal overlap with key habitats of marine species pose a risk for negative noise impacts. Yet, recreational vessels are currently unaccounted for in vessel noise models using Automatic Identification System (AIS) data. Here we conduct a case study investigating noise contributions from vessels with and without AIS (non-AIS) in a shallow coastal area within the Inner Danish waters. By tracking vessels with theodolite and AIS, while recording ambient noise levels, we find that non-AIS vessels have a higher occurrence (83%) than AIS vessels, and that motorised recreational vessels can elevate third-octave band noise centred at 0.125, 2 and 16 kHz by 47-51 dB. Accordingly, these vessels dominated the soundscape in the study site due to their high numbers, high speeds and proximity to the coast. Furthermore, recreational vessels caused 49-85% of noise events potentially eliciting behavioural responses in harbour porpoises (AIS vessels caused 5-24%). We therefore conclude that AIS data would poorly predict vessel noise pollution and its impacts in this and other similar marine environments. We suggest to improve vessel noise models and impact assessments by requiring that faster and more powerful recreational vessels carry AIS-transmitters.

The Psychoactive Drug Escitalopram Affects Foraging Behavior in Zebrafish (Danio rerio).

Selective serotonin reuptake inhibitors (SSRIs) are psychoactive pharmaceuticals that have been detected intact in natural waters globally. Laboratory experiments have reported that several SSRIs inhibit fish foraging behavior, but data for the SSRI escitalopram are lacking. The objectives of the present study were to determine whether escitalopram affects feeding behavior in zebrafish and whether possible sex differences exist. We exposed female and male zebrafish (Danio rerio) to 0.00, 0.10, and 1.50 µg/L of escitalopram in flow-through tanks for a 3-wk exposure period. We used a video tracking system with high temporal and spatial resolution to collect data on zebrafish swimming patterns in test tanks containing a food source. The results show a more pronounced effect of escitalopram in males compared with females. At the assumed most environmentally relevant concentration (0.10 µg/L), male average feeding time/visit and maximum feeding duration were significantly reduced by 27 and 42%, respectively. In addition, male total feeding duration was also significantly reduced (by 73%) at the highest concentration (1.50 µg/L). In females, only the maximum feeding duration was significantly reduced (by 41%) in the 0.10 µg/L treatment group. Hence, we reject our initial hypothesis that female feeding behavior is more vulnerable to escitalopram. There was no effect of escitalopram on length or weight among the experimental groups. The present study demonstrates that escitalopram, like other SSRIs, can inhibit fish foraging behavior and therefore potentially disturb natural food chains. Finally, our study suggests that SSRIs can both be sex and behavior specific. Environ Toxicol Chem 2019;38:1902-1910. © 2019 SETAC.

Evolution and plasticity of thermal performance: an analysis of variation in thermal tolerance and fitness in 22 Drosophila species.

The thermal biology of ectotherms is often used to infer species' responses to changes in temperature. It is often proposed that temperate species are more cold-tolerant, less heat-tolerant, more plastic, have broader thermal performance curves (TPCs) and lower optimal temperatures when compared to tropical species. However, relatively little empirical work has provided support for this using large interspecific studies. In the present study, we measure thermal tolerance limits and thermal performance in 22 species of Drosophila that developed under common conditions. Specifically, we measure thermal tolerance (CTmin and CTmax) as well as the fitness components viability, developmental speed and fecundity at seven temperatures to construct TPCs for each of these species. For 10 of the species, we also measure thermal tolerance and thermal performance following developmental acclimation to three additional temperatures. Using these data, we test several fundamental hypotheses about the evolution and plasticity of heat and cold resistance and thermal performance. We find that cold tolerance (CTmin) varied between the species according to the environmental temperature in the habitat from which they originated. These data support the idea that the evolution of cold tolerance has allowed species to persist in colder environments. However, contrary to expectation, we find that optimal temperature ( Topt) and the breadth of thermal performance ( Tbreadth) are similar in temperate, widespread and tropical species and we also find that the plasticity of TPCs was constrained. We suggest that the temperature range for optimal thermal performance is either fixed or under selection by the more similar temperatures that prevail during growing seasons. As a consequence, we find that Topt and Tbreadth are of limited value for predicting past, present and future distributions of species. This article is part of the theme issue 'Physiological diversity, biodiversity patterns and global climate change: testing key hypotheses involving temperature and oxygen'.

Variability of the inter-pulse interval in sperm whale clicks with implications for size estimation and individual identification.

Sperm whales generate multi-pulsed clicks for echolocation and communication with an inter-pulse interval (IPI) determined by the size of their hypertrophied sound producing nose. The IPI has therefore been used to estimate body size and distinguish between individuals, and it has been hypothesized that conspecifics may use IPIs to recognize each other. However, the degree to which IPIs vary within individuals has not explicitly been tested, and therefore the inherent precision of this measure and its applicability for size estimation for researchers and sperm whales alike remain unknown. Here, the variability in IPI from both animal-borne Dtags and far-field recordings from echolocating and communicating sperm whales is quantified. Three different automatic methods (envelope, cepstrum, and cross-correlation) are tested and it is found that the envelope approach results in the least dispersion. Furthermore, it is shown that neither growth, depth, nor recording aspect fully explains the observed variability among clicks recorded from the same individual. It is proposed that dynamics in the soft structures of the nose are affecting IPIs, resulting in a variation of approximately 0.2 ms. Therefore, it is recommended that this variation be considered in IPI studies and that IPIs may have limited functionality as an identity cue among large groups of conspecifics.

Allosteric mechanisms underlying the adaptive increase in hemoglobin-oxygen affinity of the bar-headed goose.

The high blood-O2 affinity of the bar-headed goose (Anser indicus) is an integral component of the biochemical and physiological adaptations that allow this hypoxia-tolerant species to undertake migratory flights over the Himalayas. The high blood-O2 affinity of this species was originally attributed to a single amino acid substitution of the major hemoglobin (Hb) isoform, HbA, which was thought to destabilize the low-affinity T state, thereby shifting the T-R allosteric equilibrium towards the high-affinity R state. Surprisingly, this mechanistic hypothesis has never been addressed using native proteins purified from blood. Here, we report a detailed analysis of O2 equilibria and kinetics of native major HbA and minor HbD isoforms from bar-headed goose and greylag goose (Anser anser), a strictly lowland species, to identify and characterize the mechanistic basis for the adaptive change in Hb function. We find that HbA and HbD of bar-headed goose have consistently higher O2 affinities than those of the greylag goose. The corresponding Hb isoforms of the two species are equally responsive to physiological allosteric cofactors and have similar Bohr effects. Thermodynamic analyses of O2 equilibrium curves according to the two-state Monod-Wyman-Changeaux model revealed higher R-state O2 affinities in the bar-headed goose Hbs, associated with lower O2 dissociation rates, compared with the greylag goose. Conversely, the T state was not destabilized and the T-R allosteric equilibrium was unaltered in bar-headed goose Hbs. The physiological implication of these results is that increased R-state affinity allows for enhanced O2 saturation in the lungs during hypoxia, but without impairing O2 delivery to tissues.

Echolocation click source parameters of Australian snubfin dolphins (Orcaella heinsohni).

The Australian snubfin dolphin (Orcaella heinsohni) is endemic to Australian waters, yet little is known about its abundance and habitat use. To investigate the feasibility of Passive Acoustic Monitoring for snubfin dolphins, biosonar clicks were recorded in Cygnet Bay, Australia, using a four-element hydrophone array. Clicks had a mean source level of 200 ± 5 dB re 1 µPa pp, transmission directivity index of 24 dB, mean centroid frequency of 98 ± 9 kHz, and a root-mean-square bandwidth of 31 ± 3 kHz. Such properties lend themselves to passive acoustic monitoring, but are comparable to similarly-sized delphinids, thus requiring additional cues to discriminate between snubfins and sympatric species.

Simulated seal scarer sounds scare porpoises, but not seals: species-specific responses to 12 kHz deterrence sounds.

Acoustic harassment devices (AHD) or 'seal scarers' are used extensively, not only to deter seals from fisheries, but also as mitigation tools to deter marine mammals from potentially harmful sound sources, such as offshore pile driving. To test the effectiveness of AHDs, we conducted two studies with similar experimental set-ups on two key species: harbour porpoises and harbour seals. We exposed animals to 500 ms tone bursts at 12 kHz simulating that of an AHD (Lofitech), but with reduced output levels (source peak-to-peak level of 165 dB re 1 µPa). Animals were localized with a theodolite before, during and after sound exposures. In total, 12 sound exposures were conducted to porpoises and 13 exposures to seals. Porpoises were found to exhibit avoidance reactions out to ranges of 525 m from the sound source. Contrary to this, seal observations increased during sound exposure within 100 m of the loudspeaker. We thereby demonstrate that porpoises and seals respond very differently to AHD sounds. This has important implications for application of AHDs in multi-species habitats, as sound levels required to deter less sensitive species (seals) can lead to excessive and unwanted large deterrence ranges on more sensitive species (porpoises).

Changes of loggerhead turtle (Caretta caretta) dive behavior associated with tropical storm passage during the inter-nesting period.

To improve conservation strategies for threatened sea turtles, more knowledge on their ecology, behavior, and how they cope with severe and changing weather conditions is needed. Satellite and animal motion datalogging tags were used to study the inter-nesting behavior of two female loggerhead turtles in the Gulf of Mexico, which regularly has hurricanes and tropical storms during nesting season. We contrast the behavioral patterns and swimming energetics of these two turtles, the first tracked in calm weather and the second tracked before, during and after a tropical storm. Turtle 1 was highly active and swam at the surface or submerged 95% of the time during the entire inter-nesting period, with a high estimated specific oxygen consumption rate (0.95 ml min-1 kg-0.83). Turtle 2 was inactive for most of the first 9 days of the inter-nesting period, during which she rested at the bottom (80% of the time) with low estimated oxygen consumption (0.62 ml min-1 kg-0.83). Midway through the inter-nesting period, turtle 2 encountered a tropical storm and became highly active (swimming 88% of the time during and 95% after the storm). Her oxygen consumption increased significantly to 0.97 ml min-1 kg-0.83 during and 0.98 ml min-1 kg-0.83 after the storm. However, despite the tropical storm, turtle 2 returned to the nesting beach, where she successfully re-nested 75 m from her previous nest. Thus, the tropical storm had a minor effect on this female's individual nesting success, even though the storm caused 90% loss nests at Casey Key.

Amazon river dolphins (Inia geoffrensis) modify biosonar output level and directivity during prey interception in the wild.

Toothed whales have evolved to live in extremely different habitats and yet they all rely strongly on echolocation for finding and catching prey. Such biosonar-based foraging involves distinct phases of searching for, approaching and capturing prey, where echolocating animals gradually adjust sonar output to actively shape the flow of sensory information. Measuring those outputs in absolute levels requires hydrophone arrays centred on the biosonar beam axis, but this has never been done for wild toothed whales approaching and capturing prey. Rather, field studies make the assumption that toothed whales will adjust their biosonar in the same manner to arrays as they will when approaching prey. To test this assumption, we recorded wild botos (Inia geoffrensis) as they approached and captured dead fish tethered to a hydrophone in front of a star-shaped seven-hydrophone array. We demonstrate that botos gradually decrease interclick intervals and output levels during prey approaches, using stronger adjustment magnitudes than predicted from previous boto array data. Prey interceptions are characterised by high click rates, but although botos buzz during prey capture, they do so at lower click rates than marine toothed whales, resulting in a much more gradual transition from approach phase to buzzing. We also demonstrate for the first time that wild toothed whales broaden biosonar beamwidth when closing in on prey, as is also seen in captive toothed whales and bats, thus resulting in a larger ensonified volume around the prey, probably aiding prey tracking by decreasing the risk of prey evading ensonification.

Time-varying auditory gain control in response to double-pulse stimuli in harbour porpoises is not mediated by a stapedial reflex.

Echolocating animals reduce their output level and hearing sensitivity with decreasing echo delays, presumably to stabilize the perceived echo intensity during target approaches. In bats, this variation in hearing sensitivity is formed by a call-induced stapedial reflex that tapers off over time after the call. Here, we test the hypothesis that a similar mechanism exists in toothed whales by subjecting a trained harbour porpoise to a series of double sound pulses varying in delay and frequency, while measuring the magnitudes of the evoked auditory brainstem responses (ABRs). We find that the recovery of the ABR to the second pulse is frequency dependent, and that a stapedial reflex therefore cannot account for the reduced hearing sensitivity at short pulse delays. We propose that toothed whale auditory time-varying gain control during echolocation is not enabled by the middle ear as in bats, but rather by frequency-dependent mechanisms such as forward masking and perhaps higher-order control of efferent feedback to the outer hair cells.