The influence of biopsy site and pregnancy on stable isotope ratios in humpback whale skin.

RATIONALE: Stable isotope analysis (SIA) of free-swimming mysticetes using biopsies is often limited in sample size and uses only one sample per individual, failing to capture both intra-individual variability and the influence of demographic and physiological factors on isotope ratios. METHODS: We applied SIA of δ13C and δ15N to humpback whale (Megaptera novaeangliae) biopsies taken during the foraging season along the western Antarctic Peninsula to quantify intra-individual variation from repeatedly sampled individuals, as well as to determine the effect of biopsy collection site, sex, and pregnancy on isotope ratios. RESULTS: There was substantial variability in δ13C from multiple biopsies taken from the same individuals, though δ15N was much more consistent. Side of the body (left versus right) and biopsy location (dorsal, anterior, ventral, and posterior) did marginally affect the isotopic composition of δ15N but not δ13C. Pregnancy had a significant effect on both δ13C and δ15N, where pregnant females were depleted in both when compared to non-pregnant females and males. CONCLUSIONS: These results indicate that isotopic signatures are influenced by multiple endogenous and exogenous factors and emphasize value in accounting for intra-individual variability and pregnancy status within a sampled population. Placed within an ecological context, the endogenous variability in δ13C observed here may be informative for future isotopic analyses.

Use of a behavioural response method to assess the risk of collision between migrating humpback whales and vessels.

With the substantial increase in many large whale populations, paired with the rise in global shipping and recreational vessel activity, it is not surprising that negative interactions between whales and vessels are increasing. Here, the collision risk between migrating groups of humpback whales (Megaptera novaeangliae) and vessels was assessed by determining if changes in their movement trajectories in response to an oncoming vessel translated to vessel avoidance. It was assumed groups would implement an escape response strategy, using cues such as the vessel speed, trajectory, proximity, and received level of noise to inform their response magnitude. However, many groups were unresponsive to an approaching vessel such that the vessel had to take evasive action. This study shows that humpback whales are not likely to take sufficient avoidance action when there is a potential for a vessel and whale to collide. Therefore, when developing a risk management strategy, mitigation measures that reduce the encounter rate between whales and vessels are likely to be the most effective.

Interactive bioacoustic playback as a tool for detecting and exploring nonhuman intelligence: "conversing" with an Alaskan humpback whale.

Here we report on a rare and opportunistic acoustic turn-taking with an adult female humpback whale, known as Twain, in Southeast Alaska. Post hoc acoustic and statistical analyses of a 20-min acoustic exchange between the broadcast of a recorded contact call, known as a 'whup/throp', with call responses by Twain revealed an intentional human-whale acoustic (and behavioral) interaction. Our results show that Twain participated both physically and acoustically in three phases of interaction (Phase 1: Engagement, Phase 2: Agitation, Phase 3: Disengagement), independently determined by blind observers reporting on surface behavior and respiratory activity of the interacting whale. A close examination of both changes to the latency between Twain's calls and the temporal matching to the latency of the exemplar across phases indicated that Twain was actively engaged in the exchange during Phase 1 (Engagement), less so during Phase 2 (Agitation), and disengaged during Phase 3 (Disengagement). These results, while preliminary, point to several key considerations for effective playback design, namely the importance of salient, dynamic and adaptive playbacks, that should be utilized in experimentation with whales and other interactive nonhuman species.

Novel Use of Cell Profiling Technology to Visualize Mitochondrial Responses of Humpback Whale Fibroblasts to Chemical Exposure.

Cetaceans are at elevated risk of accumulating persistent and lipophilic environmental contaminants due to their longevity and high proportion of body fat. Despite this, there is a paucity of taxa-specific chemical effect data, in part due to the ethical and logistical constraints in working with highly mobile aquatic species. Advances in cetacean cell culture have opened the door to the application of mainstream in vitro toxicological effect assessment approaches. Image-based cell profiling is a high-throughput, microscopy-based system commonly applied in drug development. It permits the analysis of the xenobiotic effect on multiple cell organelles simultaneously, hereby flagging its potential utility in the evaluation of chemical toxicodynamics. Here we exposed immortalized humpback whale skin fibroblasts (HuWaTERT) to six priority environmental contaminants known to accumulate in the Southern Ocean food web, in order to explore their subcellular organelle responses. Results revealed chemical-dependent modulation of mitochondrial texture, with the lowest observed effect concentrations for chlorpyrifos, dieldrin, trifluralin, and p,p'-dichlorodiphenyldichloroethane of 0.3, 4.1, 9.3, and 19.8 nM, respectively. By contrast, no significant changes were observed upon exposure to endosulfan and lindane. This study contributes the first fixed mitochondrial images of HuWaTERT and constitutes novel, taxa-specific chemical effect data in support of evidence-based conservation policy and management.

Antarctic sea-ice low resonates in the ecophysiology of humpback whales.

The past six years have been marked by some of the most dramatic climatic events observed in the Antarctic region in recent history, commencing with the 2017 sea-ice extreme low. The Humpback Whale Sentinel Programme is a circum-polar biomonitoring program for long term surveillance of the Antarctic sea-ice ecosystem. It has previously signalled the extreme La Niña event of 2010/11, and it was therefore of interest to assess the capacity of existing biomonitoring measures under the program to detect the impacts of 2017 anomalous climatic events. Six ecophysiological markers of population adiposity, diet, and fecundity were targeted, as well as calf and juvenile mortality via stranding records. All indicators, with the exception of bulk stable isotope dietary tracers, indicated a negative trend in 2017, whilst C and N bulk stable isotopes appeared to indicate a lag phase resulting from the anomalous year. The collation of multiple biochemical, chemical, and observational lines of evidence via a single biomonitoring platform provides comprehensive information for evidence-led policy in the Antarctic and Southern Ocean region.

Reproductive Steroid Hormone Patterns in Baleen of Two Pregnant Humpback Whales (Megaptera novaeangliae).

Understanding reproductive physiology in mysticetes has been slowed by the lack of repeated samples from individuals. Analysis of humpback whale baleen enables retrospective hormone analysis within individuals dating back 3-5 years before death. Using this method, we investigated differences in four steroid hormones involved in reproduction and mating during confirmed pregnant and non-pregnant periods in two female humpback whales (Megaptera novaeangliae) with known reproductive histories based on sightings and necropsy data. Cortisol, corticosterone, testosterone, and estradiol concentrations were determined via enzyme immunoassay using subsamples of each baleen plate at 2 cm intervals. There were no significant differences in cortisol or corticosterone during pregnancy when compared to non-pregnancy (inter-calving interval), but there were significant differences between the two whales in average glucocorticoid concentrations, with the younger whale showing higher values overall. For testosterone, levels for the younger female peaked at parturition in one pregnancy, but also had spikes during non-pregnancy. The older female had three large spikes in testosterone, one of which was associated with parturition. Estradiol had large fluctuations in both whales but had generally lower concentrations during non-pregnancy than during pregnancy. There were peaks in estradiol before each pregnancy, possibly coinciding with ovulation, and peaks coinciding with the month of parturition. Both estradiol and testosterone could be useful for determining ovulation or impending birth. Using baleen to investigate retrospective steroid hormone profiles can be used for elucidating long-term patterns of physiological change during gestation. LAY SUMMARY: Case studies of two pregnant humpback whales whose hormones were analyzed in baleen may illuminate when humpback whales ovulate, gestate, and give birth. These physiological metrics could assist in accurate population growth assessments and conservation of the species. This study shows that baleen hormone analysis can be a useful tool for understanding whale reproductive physiology.

Decadal-scale phenology and seasonal climate drivers of migratory baleen whales in a rapidly warming marine ecosystem.

Species' response to rapid climate change can be measured through shifts in timing of recurring biological events, known as phenology. The Gulf of Maine is one of the most rapidly warming regions of the ocean, and thus an ideal system to study phenological and biological responses to climate change. A better understanding of climate-induced changes in phenology is needed to effectively and adaptively manage human-wildlife conflicts. Using data from a 20+ year marine mammal observation program, we tested the hypothesis that the phenology of large whale habitat use in Cape Cod Bay has changed and is related to regional-scale shifts in the thermal onset of spring. We used a multi-season occupancy model to measure phenological shifts and evaluate trends in the date of peak habitat use for North Atlantic right (Eubalaena glacialis), humpback (Megaptera novaeangliae), and fin (Balaenoptera physalus) whales. The date of peak habitat use shifted by +18.1 days (0.90 days/year) for right whales and +19.1 days (0.96 days/year) for humpback whales. We then evaluated interannual variability in peak habitat use relative to thermal spring transition dates (STD), and hypothesized that right whales, as planktivorous specialist feeders, would exhibit a stronger response to thermal phenology than fin and humpback whales, which are more generalist piscivorous feeders. There was a significant negative effect of western region STD on right whale habitat use, and a significant positive effect of eastern region STD on fin whale habitat use indicating differential responses to spatial seasonal conditions. Protections for threatened and endangered whales have been designed to align with expected phenology of habitat use. Our results show that whales are becoming mismatched with static seasonal management measures through shifts in their timing of habitat use, and they suggest that effective management strategies may need to alter protections as species adapt to climate change.

Individual and joint estimation of humpback whale migratory patterns and their environmental drivers in the Southwest Atlantic Ocean.

Humpback whales (Megaptera novaeangliae) perform seasonal migrations from high latitude feeding grounds to low latitude breeding and calving grounds. Feeding grounds at polar regions are currently experiencing major ecosystem modifications, therefore, quantitatively assessing species responses to habitat characteristics is crucial for understanding how whales might respond to such modifications. We analyzed satellite telemetry data from 22 individual humpback whales in the Southwest Atlantic Ocean (SWA). Tagging effort was divided in two periods, 2003-2012 and 2016-2019. Correlations between whale's movement parameters and environmental variables were used as proxy for inferring behavioral responses to environmental variation. Two versions of a covariate-driven continuous-time correlated random-walk state-space model, were fitted to the data: i) Population-level models (P-models), which assess correlation parameters pooling data across all individuals or groups, and ii) individual-level models (I-models), fitted independently for each tagged whale. Area of Restricted Search behavior (slower and less directionally persistent movement, ARS) was concentrated at cold waters south of the Polar Front (~ 50°S). The best model showed that ARS was expected to occur in coastal areas and over ridges and seamounts. Ice coverage during August of each year was a consistent predictor of ARS across models. Wind stress curl and sea surface temperature anomalies were also correlated with movement parameters but elicited larger inter-individual variation. I-models were consistent with P-models' predictions for the case of females accompanied by calves (mothers), while males and those of undetermined sex (males +) presented more variability as a group. Spatial predictions of humpback whale behavioral responses showed that feeding grounds for this population are concentrated in the complex system of islands, ridges, and rises of the Scotia Sea and the northern Weddell Ridge. More southernly incursions were observed in recent years, suggesting a potential response to increased temperature and large ice coverage reduction observed in the late 2010s. Although, small sample size and differences in tracking duration precluded appropriately testing predictions for such a distributional shift, our modelling framework showed the efficiency of borrowing statistical strength during data pooling, while pinpointing where more complexity should be added in the future as additional data become available.

Round-trip migration and energy budget of a breeding female humpback whale in the Northeast Atlantic.

In the northern hemisphere, humpback whales (Megaptera novaeangliae) typically migrate between summer/autumn feeding grounds at high latitudes, and specific winter/spring breeding grounds at low latitudes. Northeast Atlantic (NEA) humpback whales for instance forage in the Barents Sea and breed either in the West Indies, or the Cape Verde Islands, undertaking the longest recorded mammalian migration (~ 9 000 km). However, in the past decade hundreds of individuals have been observed foraging on herring during the winter in fjord systems along the northern Norwegian coast, with unknown consequences to their migration phenology, breeding behavior and energy budgets. Here we present the first complete migration track (321 days, January 8th, 2019-December 6th, 2019) of a humpback whale, a pregnant female that was equipped with a satellite tag in northern Norway. We show that whales can use foraging grounds in the NEA (Barents Sea, coastal Norway, and Iceland) sequentially within the same migration cycle, foraging in the Barents Sea in summer/fall and in coastal Norway and Iceland in winter. The migration speed was fast (1.6 ms-1), likely to account for the long migration distance (18 300 km) and long foraging season, but varied throughout the migration, presumably in response to the calf's needs after its birth. The energetic cost of this migration was higher than for individuals belonging to other populations. Our results indicate that large whales can modulate their migration speed to balance foraging opportunities with migration phenology, even for the longest migrations and under the added constraint of reproduction.

The Southern Ocean Exchange: porous boundaries between humpback whale breeding populations in southern polar waters.

Humpback whales (Megaptera novaeangliae) are a cosmopolitan species and perform long annual migrations between low-latitude breeding areas and high-latitude feeding areas. Their breeding populations appear to be spatially and genetically segregated due to long-term, maternally inherited fidelity to natal breeding areas. In the Southern Hemisphere, some humpback whale breeding populations mix in Southern Ocean waters in summer, but very little movement between Pacific and Atlantic waters has been identified to date, suggesting these waters constituted an oceanic boundary between genetically distinct populations. Here, we present new evidence of summer co-occurrence in the West Antarctic Peninsula feeding area of two recovering humpback whale breeding populations from the Atlantic (Brazil) and Pacific (Central and South America). As humpback whale populations recover, observations like this point to the need to revise our perceptions of boundaries between stocks, particularly on high latitude feeding grounds. We suggest that this "Southern Ocean Exchange" may become more frequent as populations recover from commercial whaling and climate change modifies environmental dynamics and humpback whale prey availability.

Alternative method for assessment of southwestern Atlantic humpback whale population status.

The population of humpback whales (Megaptera novaeangliae) wintering off eastern South America was exploited by commercial whaling almost to the point of extinction in the mid-twentieth century. Since cessation of whaling in the 1970s it is recovering, but the timing and level of recovery is uncertain. We implemented a Bayesian population dynamics model describing the population's trajectory from 1901 and projecting it to 2040 to revise a previous population status assessment that used Sampling-Importance-Resampling in a Bayesian framework. Using our alternative method for model fitting (Markov chain Monte Carlo), which is more widely accessible to ecologists, we replicate a "base case scenario" to verify the effect on model results, and introduce additional data to update the status assessment. Our approach allowed us to widen the previous informative prior on carrying capacity to better reflect scientific uncertainty around historical population levels. The updated model provided more precise estimates for population sizes over the period considered (1901-2040) and suggests that carrying capacity (K: median 22,882, mean 22,948, 95% credible interval [CI] 22,711-23,545) and minimum population size (N1958: median 305, mean 319, 95% CI 271-444) might be lower than previously estimated (K: median 24,558, mean 25,110, 95% CI 22,791-31,118; N1958: median 503, mean 850, 95% CI 159-3,943). However, posterior 95% credible intervals of parameters in the updated model overlap those of the previous study. Our approach provides an accessible framework for investigating the status of depleted animal populations for which information is available on historical mortality (e.g., catches) and intermittent estimates of population size and/or trend.

Allopatric humpback whales of differing generations share call types between foraging and wintering grounds.

Humpback whales (Megaptera novaeangliae) are a cosmopolitan baleen whale species with geographically isolated lineages. Despite last sharing an ancestor ~ 2-3 million years ago, Atlantic and Pacific foraging populations share five call types. Whether these call types are also shared between allopatric breeding and foraging populations is unclear, but would provide further evidence that some call types are ubiquitous and fixed. We investigated whether these five call types were present on a contemporary foraging ground (Newfoundland, 2015-2016) and a historic breeding ground (Hawaii, 1981-1982). Calls were classified using aural/visual (AV) characteristics; 16 relevant acoustic variables were measured and a Principal Component Analysis (PCA) was used to examine within-call and between-population variation. To assess whether between-population variation influenced classification, all 16 variables were included in classification and regression tree (CART) and random forest analyses (RF). All five call types were identified in both populations. Between-population variation in combined acoustic variables (PC1, PC2, PC3) was lower within call types than among call types, and high agreement between AV and quantitative classification (CART: 83% agreement; RF: 77% agreement) suggested that acoustic characteristics were more similar within than among call types. Findings indicate that these five call types are shared across allopatric populations, generations, and behavioural contexts.

Steroid hormone profiles and body conditions of migrating male humpback whales (Megaptera novaeangliae).

Simultaneous analysis of multiple steroid hormones from remotely obtained blubber biopsies has the potential to concurrently provide information regarding stress and reproductive status from free-swimming cetaceans, while also investigating correlations between hormone concentrations and other health biomarkers. In this study we measured blubber concentration profiles of eight reproductive and adrenal steroid hormones (17α-hydroxy-progesterone, testosterone, androstenedione, progesterone, cortisol, 11-deoxy-corticosterone, oestrone, and oestradiol) together with body condition, as determined by the inverse Adipocyte Index, of 101 male humpback whales. Whales were sampled randomly at two time points, while migrating to and from their northeast Australian breeding grounds, allowing for intra- and inter-seasonal profile analysis. Testosterone, progesterone and cortisol together with androstenedione 17α-hydroxyprogesterone, and oestrone concentrations (the latter quantified for the first time in live biopsied male humpback whales) decreased between the northward and southward migrations. Decreasing testosterone levels during the height of humpback whale conceptions suggests asynchronicity between blubber testosterone levels and the expected peak of male fertility. Statistically significant relationships between levels of certain steroid analytes were observed and appeared to change between the early and late breeding seasons. During the northward migration, testosterone, progesterone, androstenedione, oestrone and 17α-hydroxyprogesterone levels were positively correlated. Cortisol concentrations correlated positively with those of testosterone during the northward migration, but negatively during the southward migration. Androstenedione and testosterone were positively correlated with adiposity during the late breeding season. These hormone-hormone and hormone-adiposity correlations may be reflective of the activation of certain steroid hormone synthesis pathways, or alternatively, of concomitant physiological stimuli. As steroid hormones work in concert, information on multiple steroid hormones is needed to interpret endocrinological status and understand the relationships between these compounds and ancillary health markers. This study provides steroid hormone profiles of wild male humpback whales, as well as the first insight into seasonal male endocrinology as a function of adiposity.

Social survival: Humpback whales (Megaptera novaeangliae) use social structure to partition ecological niches within proposed critical habitat.

Animal culture and social bonds are relevant to wildlife conservation because they influence patterns of geography, behavior, and strategies of survival. Numerous examples of socially-driven habitat partitioning and ecological-niche specialization can be found among vertebrates, including toothed whales. But such social-ecological dynamics, described here as 'social niche partitioning', are not known among baleen whales, whose societies-particularly on foraging grounds-are largely perceived as unstructured and incidental to matters of habitat use and conservation. However, through 16 years of behavioral observations and photo-identifications of humpback whales (Megaptera novaeangliae) feeding within a fjord system in the Canadian Pacific (primarily within Gitga'at First Nation waters), we have documented long-term pair bonds (up to 12 years) as well as a complex societal structure, which corresponds closely to persistent patterns in feeding strategy, long-term site fidelity (extended occupancy and annual rate of return up to 75%), specific geographic preferences within the fjord system, and other forms of habitat use. Randomization tests of network congruency and clustering algorithms were used to test for overlap in patterns of social structure and habitat use, which confirmed the occurrence of social niche partitioning on the feeding grounds of this baleen whale species. In addition, we document the extensive practice of group bubble net feeding in Pacific Canada. This coordinated feeding behavior was found to strongly mediate the social structure and habitat use within this humpback whale society. Additionally, during our 2004-2019 study, we observed a shift in social network structure in 2010-2012, which corresponded with environmental and demographic shifts including a sudden decline in the population's calving rate. Our findings indicate that the social lives of humpback whales, and perhaps baleen whales generally, are more complex than previously supposed and should be a primary consideration in the assessment of potential impacts to important habitat.

Assessing the effects of lipid extraction and lipid correction on stable isotope values (δ13 C and δ15 N) of blubber and skin from southern hemisphere humpback whales.

RATIONALE: The coupled analysis of δ13 C and δ15 N stable isotope values of blubber and skin biopsy samples is widely used to study the diet of free-ranging cetaceans. Differences in the lipid content of these tissues can affect isotopic variability because lipids are depleted in 13 C, reducing the bulk tissue 13 C/12 C. This variability in carbon isotope values can be accounted for either by chemically extracting lipids from the tissue or by using mathematical lipid normalisation models. METHODS: This study examines (a) the effects of chemical lipid extraction on δ13 C and δ15 N values in blubber and skin of southern hemisphere humpback whales, (b) whether chemical lipid extraction is more favourable than mathematical lipid correction and (c) which of the two tissues is more appropriate for dietary studies. Strategic comparisons were made between chemical lipid extraction and mathematical lipid correction and between blubber and skin tissue δ13 C and δ15 N values, as well as C:N ratios. Six existing mathematical normalisation models were tested for their efficacy in estimating lipid-free δ13 C for skin. RESULTS: Both δ13 C and δ15 N values of lipid-extracted skin (δ13 C: -25.57‰, δ15 N: 6.83‰) were significantly higher than those of bulk skin (δ13 C: -26.97‰, δ15 N: 6.15‰). Five of the six tested lipid normalisation models had small error terms for predicting lipid-free δ13 C values. The average C:N ratio of lipid-extracted skin was within the lipid-free range reported in other studies, whereas the average C:N ratio of blubber was higher than previously reported. CONCLUSIONS: These results highlight the need to account for lipids when analysing δ13 C and δ15 N values from the same sample. For optimised dietary assessments using parallel isotope analysis from a single sample, we recommend the use of unextracted skin tissue. δ15 N values should be obtained from unextracted skin, whereas δ13 C values may be adequately lipid corrected by a mathematical correction.

Modeling changes in baleen whale seasonal abundance, timing of migration, and environmental variables to explain the sudden rise in entanglements in California.

We document changes in the number of sightings and timing of humpback (Megaptera novaeangliae), blue (Balaenoptera musculus), and gray (Eschrichtius robustus) whale migratory phases in the vicinity of the Farallon Islands, California. We hypothesized that changes in the timing of migration off central California were driven by local oceanography, regional upwelling, and basin-scale climate conditions. Using 24 years of daily whale counts collected from Southeast Farallon Island, we developed negative binomial regression models to evaluate trends in local whale sightings over time. We then used linear models to assess trends in the timing of migration, and to identify potential environmental drivers. These drivers included local, regional and basin-scale patterns; the latter included the El Niño Southern Oscillation, the Pacific Decadal Oscillation, and the North Pacific Gyre Oscillation, which influence, wind-driven upwelling, and overall productivity in the California Current System. We then created a forecast model to predict the timing of migration. Humpback whale sightings significantly increased over the study period, but blue and gray whale counts did not, though there was variability across the time series. Date of breeding migration (departure) for all species showed little to no change, whereas date of migration towards feeding areas (arrival) occurred earlier for humpback and blue whales. Timing was significantly influenced by a mix of local oceanography, regional, and basin-scale climate variables. Earlier arrival time without concomitant earlier departure time results in longer periods when blue and humpback whales are at risk of entanglement in the Gulf of the Farallones. We maintain that these changes have increased whale exposure to pot and trap fishery gear off the central California coast during the spring, elevating the risk of entanglements. Humpback entanglement rates were significantly associated with increased counts and early arrival in central California. Actions to decrease the temporal overlap between whales and pot/trap fishing gear, particularly when whales arrive earlier in warm water years, would likely decrease the risk of entanglements.

Interannual variability in the lipid and fatty acid profiles of east Australia-migrating humpback whales (Megaptera novaeangliae) across a 10-year timeline.

Southern hemisphere humpback whales are classified as high-fidelity Antarctic krill consumers and as such are vulnerable to variability and long-term changes in krill biomass. Evidence of heterogeneous feeding patterns of east coast of Australia migrating humpback whales has been observed, warranting a comprehensive assessment of interannual variability in their diet. We examined the lipid and fatty acid profiles of individuals of the east coast of Australia migrating stock sampled between 2008 and 2018. The use of live-sampled blubber biopsies showed that fatty acid profiles varied significantly among all years. The two trophic indicator fatty acids for Antarctic krill, 20:5ω3 and 22:6ω3 remained largely unchanged across the 10-year period, suggesting that Antarctic krill is the principal prey item. A distance-based linear model showed that 33% of the total variation in fatty acid profiles was explained by environmental variables and climate indices. Most of the variation was explained by the Southern Annular Mode (23.7%). The high degree of variability observed in this study was unexpected for a species that is thought to feed primarily on one prey item. We propose that the observed variability likely arises from changes in the diet of Antarctic krill rather than changes in the whale's diet.

Modeling predator and prey hotspots: Management implications of baleen whale co-occurrence with krill in Central California.

As global ocean-bound commerce increases, managing human activities has become important in reducing conflict with threatened wildlife. This study investigates environmental factors determining abundance and distribution of blue whales (Balaenoptera musculus), humpback whales (Megaptera novaeangliae) and their prey (Euphausia pacifica and Thysanoessa spinifera) in central California. We provide insights into environmental drivers of the ecology and distribution of these species, model whale distributions and determine coincident hotspots of whales and their prey that will help decrease human threats to whales and protect critical feeding habitat. We developed separate predictive models of whale abundances (using negative binomial regression on count data) and krill abundance (using a two-part hurdlemodel combining logistic and negative binomial regressions) over a 14 year period (2004-2017). Variables included in situ surface and midwater oceanographic measures (temperature, salinity, and fluorescence), basin-scale climate indices, and bathymetric- and distance-related data. Predictions were applied to 1 km2 cells spanning the study area for May, June, July, and September during each of the 14 years of surveys to identify persistent distribution patterns. Both whales and krill were found to consistently use the northeast region of Cordell Bank, the Farallon Escarpment, and the shelf-break waters. The main identified blue whale hotspots were also krill hotspots, while co-occurrence was more limited and varied seasonally for humpback whales and krill. These results are valuable in identifying patterns in important areas of ecological interaction to assist management of whales. Areas north of Cordell Bank are of particular management concern since they overlap with the end of the San Francisco Bay northern shipping lane. Our findings can help decrease threats to whales, particularly in important foraging areas, by supporting implementation of vessel management and informing potential conflicts with other human uses.

Corticosterone in central North Pacific male humpback whales (Megaptera novaeangliae): Pairing sighting histories with endocrine markers to assess stress.

Developing a better understanding of the stress response is critical to ensuring the health and sustainability of marine mammal populations. However, accurately measuring and interpreting a stress response in free-ranging, large cetaceans is a nascent field. Here, an enzyme immunoassay for corticosterone was validated for use in biopsy samples from male humpback whales (Megaptera novaeangliae). Analyses were conducted on 247 male North Pacific humpback whale blubber samples, including 238 non-calves and 9 calves that were collected on the Hawaiian breeding and Southeast Alaskan feeding grounds from 2004 to 2006. Significant relationships were found when corticosterone concentrations were examined by year, age class and distribution between locations. When examined by year, corticosterone concentrations for male humpback whales were higher in Hawaii in 2004 than in 2005 and 2006 (p < 0.05). Corticosterone concentration also varied by age class with initially high concentrations at birth which subsequently tapered off and remained relatively low until sexual maturity was reached around age 8-10 years. Corticosterone concentrations appeared to peak in male humpback whales around 15-25 years of age. Blubber biopsies from Alaska and Hawaii had similar mean corticosterone concentrations, yet the variability in these samples was much greater for whales located in Hawaii. It is clear that much work remains to be done in order to accurately define or monitor a stress response in male humpback whales and that specific attention is required when looking at age, sex, and yearly trends. Our results suggest that a stress response may be most impacted by age and yearly oceanographic conditions and needs to be initially examined at the individual level.

Vessel noise levels drive behavioural responses of humpback whales with implications for whale-watching.

Disturbance from whale-watching can cause significant behavioural changes with fitness consequences for targeted whale populations. However, the sensory stimuli triggering these responses are unknown, preventing effective mitigation. Here, we test the hypothesis that vessel noise level is a driver of disturbance, using humpback whales (Megaptera novaeangliae) as a model species. We conducted controlled exposure experiments (n = 42) on resting mother-calf pairs on a resting ground off Australia, by simulating whale-watch scenarios with a research vessel (range 100 m, speed 1.5 knts) playing back vessel noise at control/low (124/148 dB), medium (160 dB) or high (172 dB) low frequency-weighted source levels (re 1 µPa RMS@1 m). Compared to control/low treatments, during high noise playbacks the mother's proportion of time resting decreased by 30%, respiration rate doubled and swim speed increased by 37%. We therefore conclude that vessel noise is an adequate driver of behavioural disturbance in whales and that regulations to mitigate the impact of whale-watching should include noise emission standards.

Whale-watching is a multi-billion-dollar industry that is growing around the world. Typically, tour operators use boats to transport tourists into coastal waters to see groups of whales, dolphins or porpoises. There is, however, accumulating evidence that boat-based whale-watching negatively affects the way these animals behave and so many countries have put guidelines in place to mitigate activities that may disturb the animals. These guidelines generally stipulate the boat's angle of approach, how close the boat can get and the speed at which it can pass by the animals. In general, these guidelines are based on the assumption that the animals are disturbed by the closeness of the whale-watching boats. However, whales, dolphins and porpoises have very sensitive hearing, and only have a short range of vision underwater. Therefore, it seems plausible that the animals hear whale-watching boats long before they see them and so the loudness of underwater noise from the boats may be enough to disturb these animals' behaviour. To test this hypothesis, Sprogis et al. performed experiments where they simulated a whale-watching vessel approaching humpback whale mothers and calves who were resting off the northwest coast of Australia. A small motorised research boat travelling at a low speed passed different mother-calf pairs at a target distance of 100 meters, which is a common whale-watching distance guideline in many countries. The boat had an underwater speaker that played recordings of the boat noise at different volumes, while a drone with a video camera flew overhead to record the whales' behaviours in detail and to identify individual animals. These "controlled exposure experiments" showed that the quiet boat noise did not appear to disturb the mothers and calves. However, compared to when the quiet boat passed the animals the louder boat noise decreased how long the mother whale rested on the surface by 30%, made her swim 37% faster, and doubled the number of breaths she took per minute. If there are many disturbances from humans, then it can negatively impact the energy the mother and calf have available for nursing, fending off males and predators, and migrating back to their feeding ground nearer the Earth's poles. Based on these findings, it is shown that the loudness of the underwater noise from boats can explain why whales may be disturbed during whale-watching activities. To help reduce this disturbance, Sprogis et al. recommend that noise emission standards should be added to the current whale-watching regulations such that boats should be as quiet as possible and ideally around the volume of the ambient background noise. This would allow operators to approach the animals in a responsible, sustainable manner and offer tourists a view of undisturbed wildlife.