Phased nanopore assembly with Shasta and modular graph phasing with GFAse.

Reference-free genome phasing is vital for understanding allele inheritance and the impact of single-molecule DNA variation on phenotypes. To achieve thorough phasing across homozygous or repetitive regions of the genome, long-read sequencing technologies are often used to perform phased de novo assembly. As a step toward reducing the cost and complexity of this type of analysis, we describe new methods for accurately phasing Oxford Nanopore Technologies (ONT) sequence data with the Shasta genome assembler and a modular tool for extending phasing to the chromosome scale called GFAse. We test using new variants of ONT PromethION sequencing, including those using proximity ligation, and show that newer, higher accuracy ONT reads substantially improve assembly quality.

Observed transmissions and ocean-ice-acoustic coupled modelling in the Beaufort Sea.

An ocean-ice-acoustic coupled model is configured for the Beaufort Sea. The model uses outputs from a data assimilating global scale ice-ocean-atmosphere forecast to drive a bimodal roughness algorithm for generating a realistic ice canopy. The resulting range-dependent ice cover obeys observed roughness, keel number density, depth, and slope, and floe size statistics. The ice is inserted into a parabolic equation acoustic propagation model as a near-zero impedance fluid layer along with a model defined range-dependent sound speed profile. Year-long observations of transmissions at 35 Hz from the Coordinated Arctic Acoustic Thermometry Experiment and 925 Hz from the Arctic Mobile Observing System source were recorded over the winter of 2019-2020 on a free-drifting, eight-element vertical line array designed to vertically span the Beaufort duct. The ocean-ice-acoustic coupled model predicts receive levels that reasonably agree with the measurements over propagation ranges of 30-800 km. At 925 Hz, seasonal and sub-seasonal ocean and ice driven variations of propagation loss are captured in the data and reproduced in the model.

Fluid balance during physical work in the heat is not modified by the menstrual cycle when fluids are freely available.

We tested the hypothesis that women may be more at risk of becoming dehydrated during physical work in the heat in the early follicular phase (EF), compared with the late follicular (LF) and mid-luteal (ML) phases of the menstrual cycle when allowed free access to drink. Twelve healthy, eumenorrheic, unacclimated women (26 ± 5 yr) completed three trials (EF, LF, and ML phases) involving 4 h of exposure to 33.8 ± 0.8 °C, 54 ± 1% relative humidity. Each hour, participants walked on a treadmill for 30 min at a rate of metabolic heat production of 338 ± 9 W. Participants drank a cool, flavor-preferred non-caloric sport drink ad libitum. Nude body weight was measured pre- and post-exposure, and percent changes in body weight loss were interpreted as an index of changes in total body water. Total fluid intake and urine output were measured and sweat rate was estimated from changes in body mass corrected for fluid intake and urine output. Fluid intake was not different between phases (EF: 1,609 ± 919 mL; LF: 1,902 ± 799 mL; ML: 1,913 ± 671; P = 0.202). Total urine output (P = 0.543) nor sweat rate (P = 0.907) differed between phases. Percent changes in body mass were not different between phases (EF: -0.5 ± 0.9%; LF: -0.3 ± 0.9%; ML: -0.3 ± 0.7%; P = 0.417). This study demonstrates that the normal hormonal fluctuations that occur throughout the menstrual cycle do not alter fluid balance during physical work in the heat.NEW & NOTEWORTHY The effect of the menstrual cycle on fluid balance during physical work in the heat when fluids are freely available is unknown. This study demonstrates that fluid balance is not modified in women across three distinct phases of the menstrual cycle during physical work in the heat These results indicate that when women have free access to cool fluid during physical work in the heat, they respond similarly across all three phases to maintain fluid homeostasis across the menstrual cycle.

Phased nanopore assembly with Shasta and modular graph phasing with GFAse.

As a step towards simplifying and reducing the cost of haplotype resolved de novo assembly, we describe new methods for accurately phasing nanopore data with the Shasta genome assembler and a modular tool for extending phasing to the chromosome scale called GFAse. We test using new variants of Oxford Nanopore Technologies' (ONT) PromethION sequencing, including those using proximity ligation and show that newer, higher accuracy ONT reads substantially improve assembly quality.

Modeling the Evolution of Rates of Continuous Trait Evolution.

Rates of phenotypic evolution vary markedly across the tree of life, from the accelerated evolution apparent in adaptive radiations to the remarkable evolutionary stasis exhibited by so-called "living fossils." Such rate variation has important consequences for large-scale evolutionary dynamics, generating vast disparities in phenotypic diversity across space, time, and taxa. Despite this, most methods for estimating trait evolution rates assume rates vary deterministically with respect to some variable of interest or change infrequently during a clade's history. These assumptions may cause underfitting of trait evolution models and mislead hypothesis testing. Here, we develop a new trait evolution model that allows rates to vary gradually and stochastically across a clade. Further, we extend this model to accommodate generally decreasing or increasing rates over time, allowing for flexible modeling of "early/late bursts" of trait evolution. We implement a Bayesian method, termed "evolving rates" (evorates for short), to efficiently fit this model to comparative data. Through simulation, we demonstrate that evorates can reliably infer both how and in which lineages trait evolution rates varied during a clade's history. We apply this method to body size evolution in cetaceans, recovering substantial support for an overall slowdown in body size evolution over time with recent bursts among some oceanic dolphins and relative stasis among beaked whales of the genus Mesoplodon. These results unify and expand on previous research, demonstrating the empirical utility of evorates. [cetacea; macroevolution; comparative methods; phenotypic diversity; disparity; early burst; late burst].

Minimal COVID-19 quieting measured in the deep offshore waters of the U.S. Outer Continental Shelf.

Using a 2-year time series (2019-2020) of 1-min sound pressure level averages from seven sites, the extension of COVID-related quieting documented in coastal soundscapes to deep (approximately 200-900 m) waters off the southeastern United States was assessed. Sites ranged in distance to the continental shelf break and shipping lanes. Sound level decreases in 2020 were observed at sites closest to the shelf break and shipping lanes but were inconsistent with the timing of shipping changes related to a COVID-19 slowdown. These observations are consistent with increased numbers of vessel tracks in 2020 compared to 2019 at a majority of sites.

Clinical impact of whole-genome sequencing in patients with early-onset dementia.

BACKGROUND: In the clinical setting, identification of the genetic cause in patients with early-onset dementia (EOD) is challenging due to multiple types of genetic tests required to arrive at a diagnosis. Whole-genome sequencing (WGS) has the potential to serve as a single diagnostic platform, due to its superior ability to detect common, rare and structural genetic variation. METHODS: WGS analysis was performed in 50 patients with EOD. Point mutations, small insertions/deletions, as well as structural variants (SVs) and short tandem repeats (STRs), were analysed. An Alzheimer's disease (AD)-related polygenic risk score (PRS) was calculated in patients with AD. RESULTS: Clinical genetic diagnosis was achieved in 7 of 50 (14%) of the patients, with a further 8 patients (16%) found to have established risk factors which may have contributed to their EOD. Two pathogenic variants were identified through SV analysis. No expanded STRs were found in this study cohort, but a blinded analysis with a positive control identified a C9orf72 expansion accurately. Approximately 37% (7 of 19) of patients with AD had a PRS equivalent to >90th percentile risk. DISCUSSION: WGS acts as a single genetic test to identify different types of clinically relevant genetic variations in patients with EOD. WGS, if used as a first-line clinical diagnostic test, has the potential to increase the diagnostic yield and reduce time to diagnosis for EOD.

High Intraindividual Variability in the Response of Serum Erythropoietin to Multiple Simulated Altitude Exposures.

Baranauskas, Marissa N., Timothy J. Fulton, Alyce D. Fly, Bruce J. Martin, Timothy D. Mickleborough, and Robert F. Chapman. High intraindividual variability in the response of serum erythropoietin to multiple simulated altitude exposures. High Alt Med Biol. 23:85-89, 2022. Purpose: To evaluate within-subject variability in the serum erythropoietin (EPO) response to multiple simulated altitude exposures. Methods: Seven physically active men and women (age 27 ± 3 years, body mass index = 24.6 ± 4.0 kg/m2) were exposed to normobaric hypoxia (fraction of inspired oxygen [FiO2] = 0.14) for 12 hours on three separate occasions. Serum EPO concentrations were measured before exposure (0 hour), after 6 hours, and after 12 hours in hypoxia. The EPO response to hypoxia was calculated as percent change from 0 to 12 hours (ΔEPO0-12). Results: Exposure time had a significant effect on EPO (p < 0.001) with concentrations increasing 3.2 ± 1.3 mIU/ml from 0 to 6 hours (p = 0.034) and 4.7 ± 1.2 mIU/ml from 0 to 12 hours (p = 0.001). Group mean ΔEPO0-12 remained unchanged (p = 0.688) between the three exposures; however, there was considerable intraindividual variability in EPO responses. The intrasubject coefficient of variation for ΔEPO0-12 was 61% ± 28% (range: 17%-103%) with intrasubject associations ranging r = 0.052 to r = 0.651 between repeated exposures. Conclusions: Athletes who routinely supplement training with simulated altitude methods (e.g., hypoxic tents) should expect inconsistent EPO responses to intermittent exposures lasting ≤12 hours.

North Sea soundscapes from a fish perspective: Directional patterns in particle motion and masking potential from anthropogenic noise.

The aquatic world of animals is an acoustic world as sound is the most prominent sensory capacity to extract information about the environment for many aquatic species. Fish can hear particle motion, and a swim bladder potentially adds the additional capacity to sense sound pressure. Combining these capacities allows them to sense direction, distance, spectral content, and detailed temporal patterns. Both sound pressure and particle motion were recorded in a shallow part of the North Sea before and during exposure to a full-scale airgun array from an experimental seismic survey. Distinct amplitude fluctuations and directional patterns in the ambient noise were found to be fluctuating in phase with the tidal cycles and coming from distinct directions. It was speculated that the patterns may be determined by distant sources associated with large rivers and nearby beaches. Sounds of the experimental seismic survey were above the ambient conditions for particle acceleration up to 10 km from the source, at least as detectable for the measurement device, and up to 31 km for the sound pressure. These results and discussion provide a fresh perspective on the auditory world of fishes and a shift in the understanding about potential ranges over which they may have access to biologically relevant cues and be masked by anthropogenic noise.

Challenges of antithrombotic therapy in the management of cardiovascular disease in patients with inherited bleeding disorders: A single-centre experience.

INTRODUCTION: Cardiovascular events in patients with inherited bleeding disorders are challenging to manage. The risk of bleeding secondary to antithrombotic treatment must be balanced against the risk of thrombosis secondary to haemostatic therapy. METHODS: Patients with inherited bleeding disorders with coronary artery bypass grafting (CABG), percutaneous coronary intervention (PCI) or atrial fibrillation (AF) from a single centre (2010-2018) are included. RESULTS: A total of 11 patients undergoing CABG (n = 3), PCI (n = 5) or with AF (n = 3) and a diagnosis of haemophilia A (n = 8), haemophilia B (n = 1), factor XI deficiency (n = 1) and von Willebrand disease (n = 1) managed by a multidisciplinary team are reported. In patients undergoing CABG, factor levels were normalized for 7-10 days with trough levels of 70-80% with severe patients continuing high-dose factor prophylaxis (trough 20-30%) three weeks post-operatively with daily aspirin. In a patient with mild haemophilia A and an inhibitor, recombinant factor VIIa dosing was monitored with thromboelastometry. For PCI, a 3rd-generation drug-eluting stent with one month of dual antiplatelet therapy in addition to high-dose prophylaxis as needed was preferred. Patients with AF and severe haemophilia did not receive antithrombotic treatment, and a thrombin generation assay was used to guide heparin dosing in mild haemophilia. CONCLUSION: Our experience demonstrates the importance of interdisciplinary communication to identify strategies that decrease the risk of bleeding and thrombosis. The use of extended, increased intensity prophylaxis facilitated antiplatelet therapy. Global assays may help balance the intensity of haemostatic and antithrombotic treatment.

Effects of a seismic survey on movement of free-ranging Atlantic cod.

Geophysical exploration of the seabed is typically done through seismic surveys, using airgun arrays that produce intense, low-frequency-sound pulses1 that can be heard over hundreds of square kilometers, 24/7.2,3 Little is known about the effects of these sounds on free-ranging fish behavior.4-6 Effects reported range from subtle individual change in activity and swimming depth for captive fish7,8 to potential avoidance9 and changes in swimming velocity and diurnal activity patterns for free-swimming animals.10 However, the extent and duration of behavioral responses to seismic surveys remain largely unexplored for most fish species.4 In this study, we investigated the effect of a full-scale seismic survey on the movement behavior of free-swimming Atlantic cod (Gadus morhua). We found that cod did not leave the detection area more than expected during the experimental survey but that they left more quickly from 2 days to 2 weeks after the survey. Furthermore, during the exposure, cod decreased their activity, with time spent being "locally active" (moving small distances, showing high body acceleration) becoming shorter, and time spent being "inactive" (moving small distances, having low body acceleration) becoming longer. Additionally, diurnal activity cycles were disrupted with lower locally active peaks at dusk and dawn, periods when cod are known to actively feed.11,12 The combined effects of delayed deterrence and activity disruption indicate the potential for seismic surveys to affect energy budgets and to ultimately lead to population-level consequences.13.

Influence of Zinc on the Acute Changes in Erythropoietin and Proinflammatory Cytokines with Hypoxia.

Baranauskas, Marissa N., Joseph Powell, Alyce D. Fly, Bruce J. Martin, Timothy D. Mickleborough, Hunter L. Paris, and Robert F. Chapman. Influence of zinc on the acute changes in erythropoietin and proinflammatory cytokines with hypoxia. High Alt Med Biol. 22: 148-156, 2021. Background: Considerable, unexplained, interindividual variability characterizes the erythropoietin (EPO) response to hypoxia, which can impact hematological acclimatization for individuals sojourning to altitude. Zinc supplementation has the potential to alter EPO by attenuating increases in inflammation and oxidative stress. Yet, the application of such an intervention has not been evaluated in humans. In this proof-of-concept study, we aimed to evaluate the EPO and inflammatory responses to acute hypoxia in human participants following chronic zinc supplementation. Methods: Nine physically active participants (men n = 5, women n = 4, age 28 ± 4 years, height 176 ± 11 cm, mass 77 ± 21 kg) were exposed to 12 hours of normobaric hypoxia simulating an altitude of 3,000 m (FiO2 = 0.14) before and after 8 weeks of supplementation with 40 mg/day of elemental zinc from picolinate. Blood samples for subsequent analysis of serum zinc, EPO, superoxide dismutase (extracellular superoxide dismutase [EC-SOD]), C-reactive protein (CRP), and proinflammatory cytokines were obtained pre- and postsupplementation and exposure to hypoxia. Results: After zinc supplementation, EPO increased by 64.9 ± 36.0% (mean ± standard deviation) following 12 hours of hypoxia, but this response was not different from presupplementation (70.8 ± 46.1%). Considerable interindividual (range: -1% to +208%) variability was apparent in the acute EPO response. While most markers of inflammation did not change with hypoxia, interleukin-6 concentrations increased from 1.17 ± 0.05 to 1.97 ± 0.32 pg/ml during the final 6 hours. The acute EPO response at 12 hours was not related to changes in serum zinc, EC-SOD, CRP, or proinflammatory cytokines. Conclusions: Zinc supplementation does not influence the acute EPO or inflammatory response with short-term exposure to moderate levels of normobaric hypoxia (3,000 m) in apparently healthy young adults.

Ventilatory Responsiveness during Exercise and Performance Impairment in Acute Hypoxia.

INTRODUCTION: An adequate increase in minute ventilation to defend arterial oxyhemoglobin saturation (SpO2) during hypoxic exercise is commonly viewed as an important factor contributing to large inter-individual variations in the degree of exercise performance impairment in hypoxia. Although the hypoxic ventilatory response (HVR) could provide insight into the underpinnings of such impairments, it is typically measured at rest under isocapnic conditions. Thus, we aimed to determine whether 1) HVR at rest and during exercise are similar and 2) exercise HVR is related to the degree of impairment in cycling time trial (TT) performance from normoxia to acute hypoxia (∆TT). METHODS: Sixteen endurance-trained men (V˙O2peak, 62.5 ± 5.8 mL·kg-1·min-1) performed two poikilocapnic HVR tests: one during seated rest (HVRREST) and another during submaximal cycling (HVREX). On two separate visits, subjects (n = 12) performed a 10-km cycling TT while breathing either room air (FiO2 = 0.21) or hypoxic gas mixture (FiO2 = 0.16) in a randomized order. RESULTS: HVREX was significantly (P < 0.001) greater than HVRREST (1.52 ± 0.47 and 0.22 ± 0.13 L·min-1·%SpO2-1, respectively), and these measures were not correlated (r = -0.16, P = 0.57). ∆TT was not correlated with HVRREST (P = 0.70) or HVREX (P = 0.54), but differences in ventilation and end-tidal CO2 between hypoxic and normoxic TT and the ventilatory equivalent for CO2 during normoxic TT explained ~85% of the variance in performance impairment in acute hypoxia (P < 0.01). CONCLUSION: We conclude that 1) HVR is not an appropriate measure to predict the exercise ventilatory response or performance impairments in acute hypoxia and 2) an adequate and metabolically matched increase in exercise ventilation, but not the gain in the ventilatory response to hypoxia, is essential for mitigating hypoxia-induced impairments in endurance cycling performance.

Assessing auditory masking for management of underwater anthropogenic noise.

Masking is often assessed by quantifying changes, due to increasing noise, to an animal's communication or listening range. While the methods used to measure communication or listening ranges are functionally similar if used for vocalizations, they differ in their approaches: communication range is focused on the sender's call, while the listening range is centered on the listener's ability to perceive any signal. How these two methods differ in their use and output is important for management recommendations. Therefore it was investigated how these two methods may alter the conclusions of masking assessments based on Atlantic cod calls in the presence of a commercial air gun array. The two methods diverged with increasing distance from the masking noise source with maximum effects lasting longer between air gun pulses in terms of communication range than listening range. Reductions in the cod's communication ranges were sensitive to fluctuations in the call's source level. That instability was not observed for the listening range. Overall, changes to the cod's communication range were more conservative but very sensitive to the call source level. A high level of confidence in the call is therefore required, while confidence in the receiver's audiogram and soundscape is required for the listening range method.

Exploring movement patterns and changing distributions of baleen whales in the western North Atlantic using a decade of passive acoustic data.

Six baleen whale species are found in the temperate western North Atlantic Ocean, with limited information existing on the distribution and movement patterns for most. There is mounting evidence of distributional shifts in many species, including marine mammals, likely because of climate-driven changes in ocean temperature and circulation. Previous acoustic studies examined the occurrence of minke (Balaenoptera acutorostrata) and North Atlantic right whales (NARW; Eubalaena glacialis). This study assesses the acoustic presence of humpback (Megaptera novaeangliae), sei (B. borealis), fin (B. physalus), and blue whales (B. musculus) over a decade, based on daily detections of their vocalizations. Data collected from 2004 to 2014 on 281 bottom-mounted recorders, totaling 35,033 days, were processed using automated detection software and screened for each species' presence. A published study on NARW acoustics revealed significant changes in occurrence patterns between the periods of 2004-2010 and 2011-2014; therefore, these same time periods were examined here. All four species were present from the Southeast United States to Greenland; humpback whales were also present in the Caribbean. All species occurred throughout all regions in the winter, suggesting that baleen whales are widely distributed during these months. Each of the species showed significant changes in acoustic occurrence after 2010. Similar to NARWs, sei whales had higher acoustic occurrence in mid-Atlantic regions after 2010. Fin, blue, and sei whales were more frequently detected in the northern latitudes of the study area after 2010. Despite this general northward shift, all four species were detected less on the Scotian Shelf area after 2010, matching documented shifts in prey availability in this region. A decade of acoustic observations have shown important distributional changes over the range of baleen whales, mirroring known climatic shifts and identifying new habitats that will require further protection from anthropogenic threats like fixed fishing gear, shipping, and noise pollution.

Displaying bioacoustic directional information from sonobuoys using "azigrams".

The AN/SSQ-53 Directional Frequency Analysis and Recording (DIFAR) sonobuoy is an expendable device that can derive acoustic particle velocity along two orthogonal horizontal axes, along with acoustic pressure. This information enables computation of azimuths of low-frequency acoustic sources from a single compact sensor. The standard approach for estimating azimuth from these sensors is by conventional beamforming (i.e., adding weighted time series), but the resulting "cardioid" beampattern is imprecise, computationally expensive, and vulnerable to directional noise contamination for weak signals. Demonstrated here is an alternative multiplicative processing scheme that computes the "active intensity" of an acoustic signal to obtain the dominant directionality of a noise field as a function of time and frequency. This information is conveniently displayed as an "azigram," which is analogous to a spectrogram, but uses color to indicate azimuth instead of intensity. Data from several locations demonstrate this approach, which can be computed without demultiplexing the raw signal. Azigrams have been used to help diagnose sonobuoy issues, improve detectability, and estimate bearings of low signal-to-noise ratio signals. Azigrams may also enhance the detection and potential classification of signals embedded in directional noise fields.

Are rural residents missing out on teaching? A comparison of teaching opportunities for urban and rural family medicine residents at the University of Manitoba.

BACKGROUND: The scholar competency of the CanMEDS-FM framework requires residents to develop teaching skills, and with the number of rural residency positions tripling over the last decade, it is essential for residency programs to provide rural residents with teaching opportunities. The purpose of this study was to investigate the differences in teaching opportunities offered to urban and rural family medicine residents at the University of Manitoba. METHODS: The 117 urban and rural family medicine residents were surveyed to quantify their interactions with medical students. Specific respondents were interviewed to provide context to the survey. RESULTS: On family medicine, only 20% of rural residents reported frequent opportunities to informally teach, compared to 57% of urban residents. Similarly, 86% of urban residents reported organized teaching opportunities compared to only 5% of rural residents. Residents placed high value in receiving additional teaching opportunities. CONCLUSION: This study suggests that there are fewer teaching opportunities for rural family medicine residents compared to urban residents at the University of Manitoba. Given the small sample size, a larger study could determine whether this trend exists across the country. It will be incumbent on residency programs to ensure rural residents have the opportunities to become competent educators.

Signals from the deep: Spatial and temporal acoustic occurrence of beaked whales off western Ireland.

Little is known of the spatio-temporal occurrence of beaked whales off western Ireland, limiting the ability of Regulators to implement appropriate management and conservation measures. To address this knowledge gap, static acoustic monitoring was carried out using eight fixed bottom-mounted autonomous acoustic recorders: four from May to December 2015 on Ireland's northern slope and four from March to November 2016 on the western and southern slopes. Recorders ran for 205 to 230 days, resulting in 4.09 TB of data sampled at 250 kHz which could capture beaked whale acoustic signals. Zero-crossing-based automated detectors identified beaked whale clicks. A sample of detections was manually validated to evaluate and optimize detector performance. Analysis confirmed the occurrence of Sowerby's and Cuvier's beaked whales and Northern bottlenose whales. Northern bottlenose whale clicks occurred in late summer and autumn, but were too few to allow further analysis. Cuvier's and Sowerby's clicks occurred at all stations throughout the monitoring period. There was a significant effect of month and station (latitude) on the mean daily number of click detections for both species. Cuvier's clicks were more abundant at lower latitudes while Sowerby's were greater at higher latitudes, particularly in the spring, suggesting a spatial segregation between species, possibly driven by prey preference. Cuvier's occurrence increased in late autumn 2015 off northwest Porcupine Bank, a region of higher relative occurrence for each species. Seismic airgun shots, with daily sound exposure levels as high as 175 dB re 1 µPa2·s, did not appear to impact the mean daily number of Cuvier's or Sowerby's beaked whale click detections. This work provides insight into the significance of Irish waters for beaked whales and highlights the importance of using acoustics for beaked whale monitoring.

Long-term passive acoustic recordings track the changing distribution of North Atlantic right whales (Eubalaena glacialis) from 2004 to 2014.

Given new distribution patterns of the endangered North Atlantic right whale (NARW; Eubalaena glacialis) population in recent years, an improved understanding of spatio-temporal movements are imperative for the conservation of this species. While so far visual data have provided most information on NARW movements, passive acoustic monitoring (PAM) was used in this study in order to better capture year-round NARW presence. This project used PAM data from 2004 to 2014 collected by 19 organizations throughout the western North Atlantic Ocean. Overall, data from 324 recorders (35,600 days) were processed and analyzed using a classification and detection system. Results highlight almost year-round habitat use of the western North Atlantic Ocean, with a decrease in detections in waters off Cape Hatteras, North Carolina in summer and fall. Data collected post 2010 showed an increased NARW presence in the mid-Atlantic region and a simultaneous decrease in the northern Gulf of Maine. In addition, NARWs were widely distributed across most regions throughout winter months. This study demonstrates that a large-scale analysis of PAM data provides significant value to understanding and tracking shifts in large whale movements over long time scales.