Exploring the effects of methodological choices on the estimation and biological interpretation of life history parameters for harbour porpoises in Norway and beyond.

This study investigates effects of subtle methodological choices on the estimation and biological interpretation of age, growth and reproductive parameters for harbour porpoises. The core analyses are based on a focal Norwegian data set built on samples from 134 harbour porpoises caught incidentally in gillnet fisheries along the Norwegian coast during autumn 2016 and spring 2017. Two contrasting practices for interpretation of seasonal and ontogenetic characteristics of tooth growth layer formation resulted in significant age differences among spring samples of young porpoises and for older animals across seasons. In turn, these differences affected estimates of age at maturity and asymptotic lengths, respectively. We also found significant differences in male age at maturity between two well-documented maturity criteria and between mathematical estimators of age at maturity for both sexes. Two different criteria for corpus albicans classification furthermore resulted in different patterns of ovarian corpora accumulation, which may affect some estimates of fecundity rates and contaminant loads. Both corpora accumulation patterns were also found in reanalysed data from German and Greenlandic porpoises. Based on tabulated overviews of methodological choices made in previous harbour porpoise studies, we argue that several of the issues mentioned above have wider relevance and may affect the validity of meta-analyses as a tool for estimating harbour porpoise sensitivity to extrinsic pressures. Differences in cause of death (COD) composition between data sets can have a similar effect. We demonstrate this in a meta-analysis of published harbour porpoise pregnancy rates, showing significantly higher values for trauma-killed samples compared to samples comprising mixed COD categories. COD also affected the estimated impacts of three previously analysed extrinsic predictors as well as an added predictor for vessel noise levels. We discuss the potential contributions of methodological, biological and anthropogenic factors in shaping observed regional differences in estimates of harbour porpoise life history parameters.

Low hunting costs in an expensive marine mammal predator.

Many large terrestrial mammalian predators use energy-intensive, high-risk, high-gain strategies to pursue large, high-quality prey. However, similar-sized marine mammal predators with even higher field metabolic rates (FMRs) consistently target prey three to six orders of magnitude smaller than themselves. Here, we address the question of how these active and expensive marine mammal predators can gain sufficient energy from consistently targeting small prey during breath-hold dives. Using harbor porpoises as model organisms, we show that hunting small aquatic prey is energetically cheap (<20% increase in FMR) for these marine predators, but it requires them to spend a large proportion (>60%) of time foraging. We conclude that this grazing foraging strategy on small prey is viable for marine mammal predators despite their high FMR because they can hunt near continuously at low marginal expense. Consequently, cessation of foraging due to human disturbance comes at a high cost, as porpoises must maintain their high thermoregulation costs with a reduced energy intake.

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.

Directional hydrophone clusters reveal evasive responses of small cetaceans to disturbance during construction at offshore windfarms.

Mitigation measures to disperse marine mammals prior to pile-driving include acoustic deterrent devices and piling soft starts, but their efficacy remains uncertain. We developed a self-contained portable hydrophone cluster to detect small cetacean movements from the distributions of bearings to detections. Using an array of clusters within 10 km of foundation pile installations, we tested the hypothesis that harbour porpoises (Phocoena phocoena) respond to mitigation measures at offshore windfarm sites by moving away. During baseline periods, porpoise movements were evenly distributed in all directions. By contrast, animals showed significant directional movement away from sound sources during acoustic deterrent device use and piling soft starts. We demonstrate that porpoises respond to measures aimed to mitigate the most severe impacts of construction at offshore windfarms by swimming directly away from these sound sources. Portable directional hydrophone clusters now provide opportunities to characterize responses to disturbance sources across a broad suite of habitats and contexts.

Emergent interactions in the management of multiple threats to the conservation of harbour porpoises.

Human activities at sea are intensifying and diversifying. This is leading to more complex interactions of anthropogenic impacts requiring adaptable management interventions to mitigate their cumulative effects on biodiversity conservation and restoration objectives. Bycatch remains the dominant conservation threat for coastal cetaceans. Additionally, the indirect impact of repeated exposure to disturbances, particularly acoustic disturbances, can affect cetacean population growth and therefore conservation objectives. Pingers are used to ensonify nets to provide an effective mitigation of bycatch risk. As those become more prevalent across fisheries at risk to catch for example harbour porpoises, pingers become contributors to the anthropogenic noise landscape which may affect the vital rates of this species as well. Currently, we do not know how to best balance pinger prevalence to minimise both bycatch rate and the population consequences of acoustic disturbance (PCoD). Here we use an agent-based model to determine how pinger prevalence in nets can be adjusted to minimise bycatch rate and noise disturbance propagating to affect population growth for harbour porpoises. We show that counter-intuitively bycatch rate can increase at lower pinger prevalence. When ecological conditions are such that PCOD can emerge, higher prevalence of pingers can lead to indirect effects on population growth. This would result from condition-mediated decreased reproductive potential. Displacing fishing effort, via time-area closure, can be an effective mitigation strategy in these circumstances. These findings have important implications for current management plans which, for practical consideration, may lead to lower overall pinger prevalence at sea. This study also shows that estimating the reproductive potential of the species should be incorporated in bycatch monitoring programmes. We now need to better understand how physiological condition affect reproductive decisions and behavioural responses to noise in cetaceans to better appraise and estimate the cumulative impacts of bycatch and its mitigations.

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.

High heart rates in hunting harbour porpoises.

The impressive breath-hold capabilities of marine mammals are facilitated by both enhanced O2 stores and reductions in the rate of O2 consumption via peripheral vasoconstriction and bradycardia, called the dive response. Many studies have focused on the extreme role of the dive response in maximizing dive duration in marine mammals, but few have addressed how these adjustments may compromise the capability to hunt, digest and thermoregulate during routine dives. Here, we use DTAGs, which record heart rate together with foraging and movement behaviour, to investigate how O2 management is balanced between the need to dive and forage in five wild harbour porpoises that hunt thousands of small prey daily during continuous shallow diving. Dive heart rates were moderate (median minimum 47-69 bpm) and relatively stable across dive types, dive duration (0.5-3.3 min) and activity. A moderate dive response, allowing for some perfusion of peripheral tissues, may be essential for fuelling the high field metabolic rates required to maintain body temperature and support digestion during diving in these small, continuously feeding cetaceans. Thus, despite having the capacity to prolong dives via a strong dive response, for these shallow-diving cetaceans, it appears to be more efficient to maintain circulation while diving: extreme heart rate gymnastics are for deep dives and emergencies, not everyday use.

Heart rate and startle responses in diving, captive harbour porpoises (Phocoena phocoena) exposed to transient noise and sonar.

Anthropogenic noise can alter marine mammal behaviour and physiology, but little is known about cetacean cardiovascular responses to exposures, despite evidence that acoustic stressors, such as naval sonars, may lead to decompression sickness. Here, we measured heart rate and movements of two trained harbour porpoises during controlled exposure to 6-9 kHz sonar-like sweeps and 40 kHz peak-frequency noise pulses, designed to evoke acoustic startle responses. The porpoises initially responded to the sonar sweep with intensified bradycardia despite unaltered behaviour/movement, but habituated rapidly to the stimuli. In contrast, 40 kHz noise pulses consistently evoked rapid muscle flinches (indicative of startles), but no behavioural or heart rate changes. We conclude that the autonomous startle response appears decoupled from, or overridden by, cardiac regulation in diving porpoises, whereas certain novel stimuli may motivate oxygen-conserving cardiovascular measures. Such responses to sound exposure may contribute to gas mismanagement for deeper-diving cetaceans.

Toothed whale auditory brainstem responses measured with a non-invasive, on-animal tag.

Empirical measurements of odontocete hearing are limited to captive individuals, constituting a fraction of species across the suborder. Data from more species could be available if such measurements were collected from unrestrained animals in the wild. This study investigated whether electrophysiological hearing data could be recorded from a trained harbor porpoise (Phocoena phocoena) using a non-invasive, animal-attached tag. The results demonstrate that auditory brainstem responses to external and self-generated stimuli can be measured from a stationary odontocete using an animal-attached recorder. With additional development, tag-based electrophysiological platforms may facilitate the collection of hearing data from freely swimming odontocetes in the wild.

No need to shout? Harbor porpoises (Phocoena phocoena) echolocate quietly in confined murky waters of the Wadden Sea.

Porpoise echolocation parameters may vary depending on their acoustic habitat and predominant behavior. Research was conducted in the Wadden Sea, an acoustically complex, tidally driven habitat with high particle resuspension. Source levels and echolocation parameters of wild harbor porpoises were estimated from time-of-arrival-differences of a six-element hydrophone array. The back-calculated peak-to-peak apparent source level of 169 ± 5 dB re 1 µPa was significantly lower than reported from Inner Danish Waters (-20 dB) and British Columbia (-9 dB) with narrower bandwidth. Porpoises therefore reduce their source level in the Wadden Sea under acoustically complex conditions suggesting an avoidance of cluttering.

The diel pattern in harbour porpoise clicking behaviour is not a response to prey activity.

Wild harbour porpoises (Phocoena phocoena) mainly forage during the night and, because they rely on echolocation to detect their prey, this is also when they are most acoustically active. It has been hypothesised that this activity pattern is a response to the diel behaviour of their major prey species. To test this hypothesis, we monitored the acoustic activity of two captive harbour porpoises held in a net pen continuously during a full year and fed by their human keepers during daylight hours, thus removing the influence of prey activity. The porpoises were exposed to similar temperature and ambient light conditions as free-ranging animals living in the same region. Throughout the year, there was a pronounced diel pattern in acoustic activity of the porpoises, with significantly greater activity at night, and a clear peak around sunrise and sunset throughout the year. Clicking activity was not dependent on lunar illumination or water level. Because the porpoises in the pen are fed and trained during daylight hours, the results indicate that factors other than fish behaviour are strongly influencing the diel clicking behaviour pattern of the species.

Asymmetric and Spiraled Genitalia Coevolve with Unique Lateralized Mating Behavior.

Asymmetric genitalia and lateralized mating behaviors occur in several taxa, yet whether asymmetric morphology in one sex correlates or coevolves with lateralized mating behavior in the other sex remains largely unexplored. While lateralized mating behaviors are taxonomically widespread, among mammals they are only known in the harbor porpoise (Phocoena phocoena). Males attempt copulation by approaching a female exclusively on her left side. To understand if this unusual lateralized behavior may have coevolved with genital morphology, we quantified the shape of female and male harbor porpoise reproductive tracts using 2D geometric morphometrics and 3D models of the vaginal lumen and inflated distal penis. We found that the vaginas varied individually in shape and that the vaginas demonstrated both significant directional and fluctuating asymmetry. This asymmetry resulted from complex 3D spirals and vaginal folds with deep recesses, which may curtail the depth or direction of penile penetration and/or semen movement. The asymmetric shapes of the vaginal lumen and penis tip were both left-canted with similar angular bends that mirrored one another and correspond with the left lateral mating approach. We suggest that the reproductive anatomy of both sexes and their lateral mating behavior coevolved.

Hyperbaric tracheobronchial compression in cetaceans and pinnipeds.

Assessment of the compressibility of marine mammal airways at depth is crucial to understanding vital physiological processes such as gas exchange during diving. Very few studies have directly assessed changes in cetacean and pinniped tracheobronchial shape, and none have quantified changes in volume with increasing pressure. A harbor seal, gray seal, harp seal, harbor porpoise and common dolphin were imaged promptly post mortem via computed tomography in a radiolucent hyperbaric chamber. Volume reconstructions were performed of segments of the trachea and bronchi of the pinnipeds and bronchi of the cetaceans for each pressure treatment. All specimens examined demonstrated significant decreases in airway volume with increasing pressure, with those of the harbor seal and common dolphin nearing complete collapse at the highest pressures. The common dolphin bronchi demonstrated distinctly different compression dynamics between 50% and 100% lung inflation treatments, indicating the importance of air in maintaining patent airways, and collapse occurred caudally to cranially in the 50% treatment. Dynamics of the harbor seal and gray seal airways indicated that the trachea was less compliant than the bronchi. These findings indicate potential species-specific variability in airway compliance, and cessation of gas exchange may occur at greater depths than those predicted in models assuming rigid airways. This may potentially increase the likelihood of decompression sickness in these animals during diving.

Acoustic reflectivity of a harbor porpoise (Phocoena phocoena).

Acoustic backscatter measurements were conducted on a stationary harbor porpoise (Phocoena phocoena) under controlled conditions. The measurements were made with the porpoise in the broadside aspect using three different types of signals: (1) a 475 µs linear frequency-modulated (FM) pulse with a frequency range from 23 to 160 kHz; (2) a simulated bottlenose dolphin (Tursiops "truncates") click with a peak frequency of 120 kHz; and (3) a simulated killer whale (Orcinus orca) click with a peak frequency of 60 kHz. The measurement with the FM pulse indicated that the mean target strength at the broadside aspect decreased from -26 to -50 dB as the frequency increased from 23 to 120 kHz in a nearly linear fashion (on a logarithm plot). Target strength variation with frequency was similar to a previous backscatter measurement on a bottlenose dolphin over a comparable frequency range (23-80 kHz). The porpoise seems to be a stealth body with low backscatter properties. The target strength of the porpoise was also about 15-16 dB lower than that of the bottlenose dolphin. The difference in lung volume of the two species when expressed in dB was also approximately 15 dB.

Drivers of the dive response in trained harbour porpoises (Phocoena phocoena).

Pronounced dive responses through peripheral vasoconstriction and bradycardia enable prolonged apnoea in marine mammals. For most vertebrates, the dive response is initiated upon face immersion, but little is known about the physical drivers of diving and surfacing heart rate in cetaceans whose faces are always mostly submerged. Using two trained harbour porpoises instrumented with an ECG-measuring sound-and-movement tag (DTAG-3), we investigated the initiation and progression of bradycardia and tachycardia during apnoea and eupnoea for varying levels of immersion. We show that paranasal wetting drives bradycardia initiation and progression, whereas apnoea leads to dive-level bradycardia eventually, but not instantly. At the end of dives, heart rate accelerates independently of lung expansion, perhaps in anticipation of surfacing; however, full tachycardia is only engaged upon inhalation. We conclude that breathing drives surface tachycardia, whereas blowhole wetting is an important driver of bradycardia; however, anticipatory/volitional modulation can overrule such responses to sensory inputs.

Context-dependent biosonar adjustments during active target approaches in echolocating harbour porpoises.

Echolocating mammals generally target individual prey items by transitioning through the biosonar phases of search (slow-rate, high-amplitude outputs), approach (gradually increasing rate and decreasing output amplitude) and buzzing (high-rate, low-amplitude outputs). The range to the main target of interest is often considered the key or sole driver of such biosonar adjustments of acoustic gaze. However, the actively generated auditory scene of an echolocator invariably comprises a large number of other reflectors and noise sources that likely also impact the biosonar strategies and source parameters implemented by an echolocating animal in time and space. In toothed whales, the importance of context on biosonar adjustments is largely unknown. To address this, we trained two harbour porpoises to actively approach the same sound recording target over the same approach distance in two highly different environments: a PVC-lined pool and a semi-natural net pen in a harbour, while blind-folded and wearing a sound recording tag (DTAG-4). We show that the approaching porpoises used considerably shorter interclick intervals (ICIs) in the pool than in the net pen, except during the buzz phase, where slightly longer ICIs were used in the pool. We further show that average click source levels were 4-7 dB higher in the net pen. Because of the very low-level in-band ambient noise in both environments, we posit that the porpoises adapted their echolocation strategy to the different reverberation levels between the two settings. We demonstrate that harbour porpoises use different echolocation strategies and biosonar parameters in two different environments for solving an otherwise identical target approach task and thus highlight that biosonar adjustments are both range and context dependent.

Porpoise click classifier (PorCC): A high-accuracy classifier to study harbour porpoises (Phocoena phocoena) in the wild.

Harbour porpoises are well-suited for passive acoustic monitoring (PAM) as they produce highly stereotyped narrow-band high-frequency (NBHF) echolocation clicks. PAM systems must be coupled with a classification algorithm to identify the signals of interest. Here, the authors present a harbour porpoise click classifier (PorCC) developed in matlab, which uses the coefficients of two logistic regression models in a decision-making pathway to assign candidate signals to one of three categories: high-quality clicks (HQ), low-quality clicks (LQ), or high-frequency noise. The receiver operating characteristics of PorCC was compared to that of PAMGuard's Porpoise Click Detector/Classifier Module. PorCC outperformed PAMGuard's classifier achieving higher hit rates (correctly classified clicks) and lower false alarm levels (noise classified as HQ or LQ clicks). Additionally, the detectability index (d') for HQ clicks for PAMGuard was 2.2 (overall d' = 2.0) versus 4.1 for PorCC (overall d' = 3.4). PorCC classification algorithm is a rapid and highly accurate method to classify NBHF clicks, which could be applied for real time monitoring, as well as to study harbour porpoises, and potentially other NBHF species, throughout their distribution range from data collected using towed hydrophones or static recorders. Moreover, PorCC is suitable for studies of acoustic communication of porpoises.

Effect of impact pile driving noise on marine mammals: A comparison of different noise exposure criteria.

Regulators in Europe and in the United States have developed sound exposure criteria. Criteria range from broadband levels to frequency weighted received sound levels. The associated differences in impact assessment results are, however, not yet understood. This uncertainty makes environmental management of transboundary anthropogenic noise challenging and causes confusion for regulators who need to choose appropriate exposure criteria. In the present study, three established exposure criteria frameworks from Germany, Denmark, and the US were used to analyse the effect of impact pile driving at a location in the Baltic Sea on harbor porpoise and harbor seal hearing. The acoustic modeling using MIKE showed that an unmitigated scenario would lead to auditory injury for all three criteria. Despite readily apparent variances in impact ranges among the applied approaches, it was also evident that noise mitigation measures could reduce underwater sound to levels where auditory injuries would be unlikely in most cases. It was concluded that each of the frameworks has its own advantages and disadvantages. Single noise exposure criteria follow the precautionary principle and can be enforced relatively easily, whereas criteria that consider hearing capabilities and animal response movement can improve the accuracy of the assessment if data are available.

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.

Cytokine expression and lymphocyte proliferative capacity in diseased harbor porpoises (Phocoena phocoena) - Biomarkers for health assessment in wildlife cetaceans.

Harbor porpoises (Phocoena phocoena) in the North and Baltic Seas are exposed to anthropogenic influences including acoustic stress and environmental contaminants. In order to evaluate immune responses in healthy and diseased harbor porpoise cells, cytokine expression analyses and lymphocyte proliferation assays, together with toxicological analyses were performed in stranded and bycaught animals as well as in animals kept in permanent human care. Severely diseased harbor porpoises showed a reduced proliferative capacity of peripheral blood lymphocytes together with diminished transcription of transforming growth factor-ß and tumor necrosis factor-α compared to healthy controls. Toxicological analyses revealed accumulation of polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (DDE), and dichlorodiphenyltrichloroethane (DDT) in harbor porpoise blood samples. Correlation analyses between blood organochlorine levels and immune parameters revealed no direct effects of xenobiotics upon lymphocyte proliferation or cytokine transcription, respectively. Results reveal an impaired function of peripheral blood leukocytes in severely diseased harbor porpoises, indicating immune exhaustion and increased disease susceptibility.