Wild harbour porpoises startle and flee at low received levels from acoustic harassment device.

Acoustic Harassment Devices (AHD) are widely used to deter marine mammals from aquaculture depredation, and from pile driving operations that may otherwise cause hearing damage. However, little is known about the behavioural and physiological effects of these devices. Here, we investigate the physiological and behavioural responses of harbour porpoises (Phocoena phocoena) to a commercial AHD in Danish waters. Six porpoises were tagged with suction-cup-attached DTAGs recording sound, 3D-movement, and GPS (n = 3) or electrocardiogram (n = 2). They were then exposed to AHDs for 15 min, with initial received levels (RL) ranging from 98 to 132 dB re 1 µPa (rms-fast, 125 ms) and initial exposure ranges of 0.9-7 km. All animals reacted by displaying a mixture of acoustic startle responses, fleeing, altered echolocation behaviour, and by demonstrating unusual tachycardia while diving. Moreover, during the 15-min exposures, half of the animals received cumulative sound doses close to published thresholds for temporary auditory threshold shifts. We conclude that AHD exposure at many km can evoke both startle, flight and cardiac responses which may impact blood-gas management, breath-hold capability, energy balance, stress level and risk of by-catch. We posit that current AHDs are too powerful for mitigation use to prevent hearing damage of porpoises from offshore construction.

Effects of amplitude and duration of noise exposure on the hearing and anti-predator behaviour of common roach (Rutilus rutilus) and sand goby (Pomatoschistus minutus).

This study investigates whether an exposure to two different received sound pressure levels at equal cumulative energy affects anti-predator behaviour and auditory detection thresholds of common roach (Rutilus rutilus) and sand goby (Pomatoschistus minutus) differently. This was examined in regard to a vessel slowdown as a management strategy to decrease vessel noise impact on fishes. Using continuous broadband noise, we found significant temporary threshold shifts (TTS) in roach, with 11.9 and 13.4 dB at 250 and 1000 Hz respectively, for the louder exposure. In contrast, gobies exhibited a non-significant shift of 6.6 dB at 125 Hz. Group cohesion increased in roach exposed to an artificial predator in the control group, but not during noise exposures. Gobies showed an initial freezing reaction towards the predator stimulus remaining motionless regardless of treatment. Our results show that a reduction in vessel speed with a corresponding reduction in source level could mitigate the effects on the auditory senses of sensitive fish, but does not appear to have any mitigating effect on their noise-induced behavioural changes. Further studies should investigate the effects of multiple vessel passages, but also the ecological consequences of the described effects on hearing and behaviour at individual and population level.

Vessel noise exposures of harbour seals from the Wadden Sea.

The North Sea faces intense ship traffic owing to increasing human activities at sea. As harbour seals (Phoca vitulina) are abundant top predators in the North Sea, it is hypothesised that they experience repeated, high-amplitude vessel exposures. Here, we test this hypothesis by quantifying vessel noise exposures from deployments of long-term sound and movement tags (DTAGs) on nine harbour seals from the Wadden Sea. An automated tool was developed to detect intervals of elevated noise in the sound recordings. An assessment by multiple raters was performed to classify the source as either vessels or other sounds. A total of 133 vessel passes were identified with received levels > 97 dB re 1µPa RMS in the 2 kHz decidecade band and with ambient noise > 6 dB below this detection threshold. Tagged seals spent most of their time within Marine Protected Areas (89 ± 13%, mean ± SD) and were exposed to high-amplitude vessel passes 4.3 ± 1.6 times per day. Only 32% of vessel passes were plausibly associated with an AIS-registered vessel. We conclude that seals in industrialized waters are exposed repeatedly to vessel noise, even in areas designated as protected, and that exposures are poorly predicted by AIS data.

Effects of multiple exposures to pile driving noise on harbor porpoise hearing during simulated flights-An evaluation tool.

Exploitation of renewable energy from offshore wind farms is substantially increasing worldwide. The majority of wind turbines are bottom mounted, causing high levels of impulsive noise during construction. To prevent temporary threshold shifts (TTS) in harbor porpoise hearing, single strike sound exposure levels (SELSS) are restricted in Germany by law to a maximum of 160 dB re 1 µPa2s at a distance of 750 m from the sound source. Underwater recordings of pile driving strikes, recorded during the construction of an offshore wind farm in the German North Sea, were analyzed. Using a simulation approach, it was tested whether a TTS can still be induced under current protective regulations by multiple exposures. The evaluation tool presented here can be easily adjusted for different sound propagation, acoustic signals, or species and enables one to calculate a minimum deterrence distance. Based on this simulation approach, only the combination of SELSS regulation, previous deterrence, and soft start allow harbor porpoises to avoid a TTS from multiple exposures. However, deterrence efficiency has to be monitored.

Time and tide: Seasonal, diel and tidal rhythms in Wadden Sea Harbour porpoises (Phocoena phocoena).

Odontocetes have evolved a rich diversity of prey- and habitat-specific foraging strategies, which allows them to feed opportunistically on locally and temporally abundant prey. While habitat-specific foraging strategies have been documented for some odontocete species, this is less known for the harbour porpoise (Phocoena phocoena). We collected multiple years of acoustic data using echolocation click loggers to analyse porpoise occurrence and buzzing behaviour, indicating feeding, in the German Wadden Sea (North Sea). Seasonal, diel and tidal effects were studied using Generalised Estimating Equations (GEE-GAMs). Locally season, time of day and tidal time significantly influenced the probability of porpoise detections and detection of foraging sequences (buzzes). Hunting strategies, and therefore frequency of buzzes, were likely affected by prey distribution and large differences between POD locations indicated that porpoises used highly specific behaviour adapted to tide and time of day to efficiently feed on the available prey. Strong seasonal and spatial variation in diel and tidal effects underline the importance of long-term observations. Studies on porpoise behaviour are often based on short-term observations and might rather reflect a seasonal than a general pattern. The results of this study show clearly that significant changes in porpoise behaviour can be found in short and long-term observations. Here some features are based on short term determinants and others are stable over years and care should be taken about drawing general conclusions based on local patterns. Highly variable spatio-temporal patterns indicate a high flexibility of porpoises in a highly variable environment and address a challenge for complex conservation management plans.

The use of seal scarers as a protective mitigation measure can induce hearing impairment in harbour porpoises.

Acoustic deterrent devices (ADDs) are used to deter seals from aquacultures but exposure of harbour porpoises (Phocoena phocoena) occurs as a side-effect. At construction sites, by contrast, ADDs are used to deter harbour porpoises from the zone in which pile driving noise can induce temporary threshold shifts (TTSs). ADDs emit such high pressure levels that there is concern that ADDs themselves may induce a TTS. A harbour porpoise in human care was exposed to an artificial ADD signal with a peak frequency of 14 kHz. A significant TTS was found, measured by auditory evoked potentials, with an onset of 142 dB re 1 µPa2s at 20 kHz and 147 dB re 1 µPa2s at 28 kHz. The authors therefore strongly recommend to gradually increase and down regulate source levels of ADDs to the desired deterrence range. However, further research is needed to develop a reliable relationship between received levels and deterrence.

Supporting evidence for PCB pollution threatening global killer whale population.

A recent Science report predicted the global killer whale population to collapse due to PCB pollution. Here we present empirical evidence, which supports and extends the reports' statement. In 2016, a neonate male killer whale stranded on the German island of Sylt. Neonatal attributes indicated an age of at least 3 days. The stomach contained ∼20 mL milk residue and no pathologies explaining the cause of death could be detected. Blubber samples presenting low lipid concentrations were analysed for persistent organic pollutants. Skin samples were collected for genotyping of the mitochondrial control region. The blubber PCB concentrations were very high [SPCBs, 225 mg/kg lipid weight (lw)], largely exceeding the PCB toxicity thresholds reported for the onset of immunosuppression [9 mg/kg lw ∑PCB] and for severe reproductive impairment [41 mg/kg lw ∑PCB] reported for marine mammals. Additionally, this individual showed equally high concentrations in p,p'-DDE [226 mg/kg lw], PBDEs [5 mg/kg lw] and liver mercury levels [1.1 µg/g dry weight dw]. These results suggest a high placental transfer of pollutants from mother to foetus. Consequently, blubber and plasma PCB concentrations and calf mortality rates are both high in primiparous females. With such high pollutant levels, this neonate had poor prerequisites for survival. The neonate belonged to Ecotype I (generalist feeder) and carried the mitochondrial haplotype 35 present in about 16% of the North Atlantic killer whale from or close to the North Sea. The relevance of this data becomes apparent in the UK West Coast Community, the UK's only residentorca population, which is currently composed of only eight individuals (each four males and females) and no calves have been reported over the last 19 years.Despite worldwide regulations, PCBs persist in the environment and remain a severe concern for killer whale populations, placing calves at high risk due to the mother-offspring PCB-transfer resulting in a high toxicological burden of the neonates.

Size and shape variations of the bony components of sperm whale cochleae.

Several mass strandings of sperm whales occurred in the North Sea during January and February 2016. Twelve animals were necropsied and sampled around 48 h after their discovery on German coasts of Schleswig Holstein. The present study aims to explore the morphological variation of the primary sensory organ of sperm whales, the left and right auditory system, using high-resolution computerised tomography imaging. We performed a quantitative analysis of size and shape of cochleae using landmark-based geometric morphometrics to reveal inter-individual anatomical variations. A hierarchical cluster analysis based on thirty-one external morphometric characters classified these 12 individuals in two stranding clusters. A relative amount of shape variation could be attributable to geographical differences among stranding locations and clusters. Our geometric data allowed the discrimination of distinct bachelor schools among sperm whales that stranded on German coasts. We argue that the cochleae are individually shaped, varying greatly in dimensions and that the intra-specific variation observed in the morphology of the cochleae may partially reflect their affiliation to their bachelor school. There are increasing concerns about the impact of noise on cetaceans and describing the auditory periphery of odontocetes is a key conservation issue to further assess the effect of noise pollution.

Assessing auditory evoked potentials of wild harbor porpoises (Phocoena phocoena).

Testing the hearing abilities of marine mammals under water is a challenging task. Sample sizes are usually low, thus limiting the ability to generalize findings of susceptibility towards noise influences. A method to measure harbor porpoise hearing thresholds in situ in outdoor conditions using auditory steady state responses of the brainstem was developed and tested. The method was used on 15 live-stranded animals from the North Sea during rehabilitation, shortly before release into the wild, and on 12 wild animals incidentally caught in pound nets in Denmark (inner Danish waters). Results indicated that although the variability between individuals is wide, the shape of the hearing curve is generally similar to previously published results from behavioral trials. Using 10-kHz frequency intervals between 10 and 160 kHz, best hearing was found between 120 and 130 kHz. Additional testing using one-third octave frequency intervals (from 16 to 160 kHz) allowed for a much faster hearing assessment, but eliminated the fine scale threshold characteristics. For further investigations, the method will be used to better understand the factors influencing sensitivity differences across individuals and to establish population-level parameters describing hearing abilities of harbor porpoises.

Impacts of Underwater Noise on Marine Vertebrates: Project Introduction and First Results.

The project conducts application-oriented research on impacts of underwater noise on marine vertebrates in the North and Baltic Seas. In distinct subprojects, the hearing sensitivity of harbor porpoises and gray seals as well as the acoustic tolerance limit of harbor porpoises to impulsive noise from pile driving and stress reactions caused by anthropogenic noise is investigated. Animals are equipped with DTAGs capable of recording the actual surrounding noise field of free-swimming harbor porpoises and seals. Acoustic noise mapping including porpoise detectors in the Natura 2000 sites of the North and Baltic Seas will help to fully understand current noise impacts.

In-air evoked potential audiometry of grey seals (Halichoerus grypus) from the North and Baltic Seas.

In-air anthropogenic sound has the potential to affect grey seal (Halichoerus grypus) behaviour and interfere with acoustic communication. In this study, a new method was used to deliver acoustic signals to grey seals as part of an in-air hearing assessment. Using in-ear headphones with adapted ear inserts allowed for the measurement of auditory brainstem responses (ABR) on sedated grey seals exposed to 5-cycle (2-1-2) tone pips. Thresholds were measured at 10 frequencies between 1-20 kHz. Measurements were made using subcutaneous electrodes on wild seals from the Baltic and North Seas. Thresholds were determined by both visual and statistical approaches (single point F-test) and good agreement was obtained between the results using both methods. The mean auditory thresholds were ≤40 dB re 20 µPa peak equivalent sound pressure level (peSPL) between 4-20 kHz and showed similar patterns to in-air behavioural hearing tests of other phocid seals between 3 and 20 kHz. Below 3 kHz, a steep reduction in hearing sensitivity was observed, which differed from the rate of decline in sensitivity obtained in behavioural studies on other phocids. Differences in the rate of decline may reflect influence of the ear inserts on the ability to reliably transmit lower frequencies or interference from the structure of the distal end of the ear canal.

Determination of DPPH Radical Oxidation Caused by Methanolic Extracts of Some Microalgal Species by Linear Regression Analysis of Spectrophotometric Measurements.

The demonstrated modified spectrophotometric method makes use of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and its specific absorbance properties. Theabsorbance decreases when the radical is reduced by antioxidants. In contrast to otherinvestigations, the absorbance was measured at a wavelength of 550 nm. This wavelengthenabled the measurements of the stable free DPPH radical without interference frommicroalgal pigments. This approach was applied to methanolic microalgae extracts for twodifferent DPPH concentrations. The changes in absorbance measured vs. the concentrationof the methanolic extract resulted in curves with a linear decrease ending in a saturationregion. Linear regression analysis of the linear part of DPPH reduction versus extractconcentration enabled the determination of the microalgae's methanolic extractsantioxidative potentials which was independent to the employed DPPH concentrations. Theresulting slopes showed significant differences (6 - 34 µmol DPPH g-1 extractconcentration) between the single different species of microalgae (Anabaena sp.,Isochrysis galbana, Phaeodactylum tricornutum, Porphyridium purpureum, Synechocystissp. PCC6803) in their ability to reduce the DPPH radical. The independency of the signal on the DPPH concentration is a valuable advantage over the determination of the EC50 value.