Heard off Heard: Acoustic detections of sperm whales (Physeter macrocephalus) and other cetaceans off Heard Island.

An underwater acoustic recorder was moored off Heard Island from September 2017 through March 2018 to listen for marine mammals. Analysis of data was initially conducted by visual inspection of long-term spectral averages to reveal sounds from sperm whales, Antarctic and pygmy blue whales, fin whales, minke whales, odontocete whistles, and noise from nearby ships. Automated detection of sperm whale clicks revealed they were seldom detected from September through January (n = 35 h) but were detected nearly every day of February and March (n = 684 h). Additional analysis of these detections revealed further diel and demographic patterns.

Diel patterns of fin whale 20 Hz acoustic presence in Eastern Antarctic waters.

This study presents evidence of diel patterns in fin whale (Balaenoptera physalus) 20 Hz acoustic presence in Eastern Antarctic waters. Passive acoustic recordings were collected at four sites in Eastern Antarctica from 2013 to 2019. A generalized linear model fitted by a generalized estimating equation was used to test the hypothesis that fin whale 20 Hz acoustic presence shows significant variation between light regimes dawn, day, dusk and night. In the Indian sector of Antarctica, at the Prydz and Southern Kerguelen Plateau sites, fin whale acoustic presence was significantly more common during the night and dawn before declining during the day and dusk periods. A different diel pattern was observed in the Pacific sector, at the Dumont d'Urville site: fin whale acoustic presence was significantly more common during the day than dusk and night periods. No diel pattern was identified at the Casey site. The identified diel patterns in the Indian sector of Eastern Antarctica correlate with previously identified diel patterns of the fin whales' prey. We suggest an indirect association between fin whale acoustic presence and foraging, with the animals more likely to produce the 20 Hz pulse during the night when not foraging and less likely to vocalize when foraging during the day.

Satellite tag derived data from two Antarctic blue whales (Balaenopteramusculusintermedia) tagged in the east Antarctic sector of the Southern Ocean.

Background: Satellite tags were deployed on two Antarctic blue whales (Balaenopteramusculusintermedia) in the east Antarctic sector of the Southern Ocean as part of the International Whaling Commission's Southern Ocean Research Partnership initiative. The satellite tracks generated are the first and currently, the only, satellite telemetry data that exist for this critically endangered species. These data provide valuable insights into the movements of Antarctic blue whales on their Antarctic feeding ground. The data were collected between February and April 2013 and span a 110° longitudinal range. New information: This dataset is the first and only detailed movement data that exist for this critically endangered species. As such, this dataset provides the first measures of movement rates (distances travelled, speeds) and movement behaviour (distinguishing transit behaviour from area restricted search behaviour) within the Southern Ocean. These movement-based measures are critical to the ongoing management of Antarctic blue whales as they recover from commercial whaling as they provide insight into foraging behaviour, habitat use, population structure and overlap with anthropogenic threats.

Acoustic analysis of crabeater seal (Lobodon carcinophaga) vocalizations in the Southern Kerguelen Plateau region of East Antarctica.

Crabeater seals are circumpolar, ice-dependent seals that produce distinct vocalizations during the breeding season. This study provides the first description of the acoustic repertoire of the crabeater seal in East Antarctica, using data from a stationary hydrophone at 1.8 km depth in the Southern Kerguelen Plateau region in 2014-2015. Two call types were identified in the data set: the low and the high moan calls. Of the 8821 calls detected, 92.5% were classified as low moan calls and 7.5% were high moan calls. The mean duration of the two call types was similar (2.3 and 2.8 s, respectively), however, the high moan calls had a higher frequency range (1020-4525 Hz, n = 11) than the low moan calls (360-2753 Hz, n = 120). The calls were primarily detected in the austral spring. Diel analysis showed that the low moan calls were mostly made at nighttime or proximal to dusk and dawn. The results of this study could aid in the development of automated detectors for crabeater seal vocalizations. This would facilitate comparisons of the distribution and abundance of the species using extant acoustic data and could increase knowledge on the breeding behavior of crabeater seals.

An open access dataset for developing automated detectors of Antarctic baleen whale sounds and performance evaluation of two commonly used detectors.

Since 2001, hundreds of thousands of hours of underwater acoustic recordings have been made throughout the Southern Ocean south of 60° S. Detailed analysis of the occurrence of marine mammal sounds in these circumpolar recordings could provide novel insights into their ecology, but manual inspection of the entirety of all recordings would be prohibitively time consuming and expensive. Automated signal processing methods have now developed to the point that they can be applied to these data in a cost-effective manner. However training and evaluating the efficacy of these automated signal processing methods still requires a representative annotated library of sounds to identify the true presence and absence of different sound types. This work presents such a library of annotated recordings for the purpose of training and evaluating automated detectors of Antarctic blue and fin whale calls. Creation of the library has focused on the annotation of a representative sample of recordings to ensure that automated algorithms can be developed and tested across a broad range of instruments, locations, environmental conditions, and years. To demonstrate the utility of the library, we characterise the performance of two automated detection algorithms that have been commonly used to detect stereotyped calls of blue and fin whales. The availability of this library will facilitate development of improved detectors for the acoustic presence of Southern Ocean blue and fin whales. It can also be expanded upon to facilitate standardization of subsequent analysis of spatiotemporal trends in call-density of these circumpolar species.

Southern right whale vocalizations on foraging grounds in South Georgia.

Southern right whale vocalizations were recorded concurrently with visual observations off the sub-Antarctic Island of South Georgia, and the characteristics of these calls were described. Calls were also compared to those of humpback whales at South Georgia, to determine how the two species might reliably be distinguished acoustically. The southern right whale calls measured (which were all upcalls) had lower frequency with peak energy and were mostly shorter in duration than the calls measured from humpback whales. The frequency upsweep and the lack of harmonics of southern right whale calls were also diagnostic characteristics.

The seasonal occupancy and diel behaviour of Antarctic sperm whales revealed by acoustic monitoring.

The seasonal occupancy and diel behaviour of sperm whales (Physeter macrocephalus) was investigated using data from long-term acoustic recorders deployed off east Antarctica. An automated method for investigating acoustic presence of sperm whales was developed, characterised, and applied to multi-year acoustic datasets at three locations. Instead of focusing on the acoustic properties of detected clicks, the method relied solely on the inter-click-interval (ICI) for determining presence within an hour-long recording. Parameters for our classifier were informed by knowledge of typical vocal behaviour of sperm whales. Sperm whales were detected predominantly from Dec-Feb, occasionally in Nov, Mar, Apr, and May, but never in the Austral winter or early spring months. Ice cover was found to have a statistically significant negative effect on sperm whale presence. In ice-free months sperm whales were detected more often during daylight hours and were seldom detected at night, and this effect was also statistically significant. Seasonal presence at the three east Antarctic recording sites were in accord with what has been inferred from 20th century whale catches off western Antarctica and from stomach contents of whales caught off South Africa.

Estimating drift of directional sonobuoys from acoustic bearings.

A maximum likelihood method is presented for estimating drift direction and speed of a directional sonobuoy given the deployment location and a time series of acoustic bearings to a sound source at known position. The viability of this method is demonstrated by applying it to two real-world scenarios: (1) during a calibration trial where buoys were independently tracked via satellite, and (2) by applying the technique to sonobuoy recordings of a vocalising Antarctic blue whale that was simultaneously tracked by photogrammetric methods. In both test cases, correcting for sonobuoy drift substantially increased the accuracy of acoustic locations.

Software for real-time localization of baleen whale calls using directional sonobuoys: A case study on Antarctic blue whales.

Directional frequency analysis and recording (DIFAR) sonobuoys can allow real-time acoustic localization of baleen whales for underwater tracking and remote sensing, but limited availability of hardware and software has prevented wider usage. These software limitations were addressed by developing a module in the open-source software PAMGuard. A case study is presented demonstrating that this software provides greater efficiency and accessibility than previous methods for detecting, localizing, and tracking Antarctic blue whales in real time. Additionally, this software can easily be extended to track other low and mid frequency sounds including those from other cetaceans, pinnipeds, icebergs, shipping, and seismic airguns.

Red shift, blue shift: investigating Doppler shifts, blubber thickness, and migration as explanations of seasonal variation in the tonality of Antarctic blue whale song.

The song of Antarctic blue whales (Balaenoptera musculus intermedia) comprises repeated, stereotyped, low-frequency calls. Measurements of these calls from recordings spanning many years have revealed a long-term linear decline as well as an intra-annual pattern in tonal frequency. While a number of hypotheses for this long-term decline have been investigated, including changes in population structure, changes in the physical environment, and changes in the behaviour of the whales, there have been relatively few attempts to explain the intra-annual pattern. An additional hypothesis that has not yet been investigated is that differences in the observed frequency from each call are due to the Doppler effect. The assumptions and implications of the Doppler effect on whale song are investigated using 1) vessel-based acoustic recordings of Antarctic blue whales with simultaneous observation of whale movement and 2) long-term acoustic recordings from both the subtropics and Antarctic. Results from vessel-based recordings of Antarctic blue whales indicate that variation in peak-frequency between calls produced by an individual whale was greater than would be expected by the movement of the whale alone. Furthermore, analysis of intra-annual frequency shift at Antarctic recording stations indicates that the Doppler effect is unlikely to fully explain the observations of intra-annual pattern in the frequency of Antarctic blue whale song. However, data do show cyclical changes in frequency in conjunction with season, thus suggesting that there might be a relationship among tonal frequency, body condition, and migration to and from Antarctic feeding grounds.

A simulation study of acoustic-assisted tracking of whales for mark-recapture surveys.

Collecting enough data to obtain reasonable abundance estimates of whales is often difficult, particularly when studying rare species. Passive acoustics can be used to detect whale sounds and are increasingly used to estimate whale abundance. Much of the existing effort centres on the use of acoustics to estimate abundance directly, e.g. analysing detections in a distance sampling framework. Here, we focus on acoustics as a tool incorporated within mark-recapture surveys. In this context, acoustic tools are used to detect and track whales, which are then photographed or biopsied to provide data for mark-recapture analyses. The purpose of incorporating acoustics is to increase the encounter rate beyond using visual searching only. While this general approach is not new, its utility is rarely quantified. This paper predicts the "acoustically-assisted" encounter rate using a discrete-time individual-based simulation of whales and survey vessel. We validate the simulation framework using existing data from studies of sperm whales. We then use the framework to predict potential encounter rates in a study of Antarctic blue whales. We also investigate the effects of a number of the key parameters on encounter rate. Mean encounter rates from the simulation of sperm whales matched well with empirical data. Variance of encounter rate, however, was underestimated. The simulation of Antarctic blue whales found that passive acoustics should provide a 1.7-3.0 fold increase in encounter rate over visual-only methods. Encounter rate was most sensitive to acoustic detection range, followed by vocalisation rate. During survey planning and design, some indication of the relationship between expected sample size and effort is paramount; this simulation framework can be used to predict encounter rates and establish this relationship. For a case in point, the simulation framework indicates unequivocally that real-time acoustic tracking should be considered for quantifying the abundance of Antarctic blue whales via mark-recapture methods.

Blue whale vocalizations recorded around New Zealand: 1964-2013.

Previous underwater recordings made in New Zealand have identified a complex sequence of low frequency sounds that have been attributed to blue whales based on similarity to blue whale songs in other areas. Recordings of sounds with these characteristics were made opportunistically during the Southern Ocean Research Partnership's recent Antarctic Blue Whale Voyage. Detections of these sounds occurred all around the South Island of New Zealand during the voyage transits from Nelson, New Zealand to the Antarctic and return. By following acoustic bearings from directional sonobuoys, blue whales were visually detected and confirmed as the source of these sounds. These recordings, together with the historical recordings made northeast of New Zealand, indicate song types that persist over several decades and are indicative of the year-round presence of a population of blue whales that inhabits the waters around New Zealand. Measurements of the four-part vocalizations reveal that blue whale song in this region has changed slowly, but consistently over the past 50 years. The most intense units of these calls were detected as far south as 53°S, which represents a considerable range extension compared to the limited prior data on the spatial distribution of this population.

Acoustically derived growth rates of sperm whales (Physeter macrocephalus) in Kaikoura, New Zealand.

A non-invasive acoustic method for measuring the growth of sperm whales was developed based on estimating the length of individuals by measuring the inter-pulse interval (IPI) of their clicks. Most prior knowledge of growth in male sperm whales has come from from fitting growth curves to length data gained from whaling. Recordings made at Kaikoura, New Zealand, were used to estimate the length and growth of 32 photographically identified, resident whales that have been recorded repeatedly between 1991 and 2009. All whales recorded more than six months apart (n = 30) showed an increase in IPI. Using established relationships between IPI and total length, it was found that the average growth rate in the Kaikoura population is lower, especially for smaller whales (13-14.5 m), than that derived from historical whaling data from other populations. This difference may be due to ecological differences among populations but might also reflect upward bias in measurements gained in whaling. The ability to track growth of individuals through time is only possible via non-lethal means and offers a fundamentally different kind of data because differences among individuals can be measured.

Impact of prefractionation using Gradiflow on two-dimensional gel electrophoresis and protein identification by matrix assisted laser desorption/ionization-time of flight-mass spectrometry.

Prefractionation of protein samples prior to two-dimensional electrophoresis (2-DE) has the potential to increase the dynamic detection range for proteomic analysis. We evaluated a membrane-based electrophoretic separation technique (Gradiflow) for its ability to fractionate an exoproteome sample from the filamentous fungus Trichoderma reesei. The sample was separated on the basis of size and charge. Buffer optimization was found to be necessary for successful size fractionation. Fractionation by charge was used to resolve the sample into four fractions that were subjected to analysis by two-dimensional electrophoresis (2-DE). Enhanced detection of low-abundance proteins with selective removal of high-abundance species was achieved. Fractionated and unfractionated samples were examined for differences in the ability to identify proteins following 2-DE using trypsin in-gel digestion followed by peptide mass fingerprinting using matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Fractionated samples showed marked improvement in protein identification ability and sequence coverage. This study demonstrates the utility of the Gradiflow for fractionation, resulting in an enhancement of resolution and characterization of a moderately complex proteome.

Differential proteomic analysis of Bacillus subtilis nitrate respiration and fermentation in defined medium.

A comparative investigation of protein expression by two-dimensional gel electrophoresis was conducted between Bacillus subtilis cultures grown in defined medium under aerobic, anaerobic nitrate respiration, or fermentation conditions. Defined medium specific for either nitrate respiration or fermentation allowed distinction between proteins induced by each individual growth process. Our differential protein profiling analysis between aerobic and anaerobic conditions showed that anaerobic fermentation induced at least 44 proteins and nitrate respiration induced at least 19 proteins compared to aerobic controls. Certain proteins were specifically induced during nitrate respiration or fermentation, while others were induced by both anaerobic processes. Eleven proteins induced by nitrate respiration and/or fermentation were identified by peptide mass matching using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Proteins encoded by feuA, hmp, and ytkD were induced by nitrate respiration. Proteins encoded by pyrR, sucD, trpC, and ywjH were induced by fermentation. Proteins encoded by acuB, pdhC, ydjL, and yvyD were induced by nitrate respiration and fermentation. This proteomic analysis has provided a more complete characterization of B. subtilis anaerobic growth and increased our understanding of its metabolic pathways of nitrate respiration and fermentation.

Comparative growth analysis of the facultative anaerobes Bacillus subtilis, Bacillus licheniformis, and Escherichia coli.

Bacillus subtilis anaerobic respiration and fermentative growth capabilities were compared to two other facultative anaerobes, Bacillus licheniformis and Escherichia coli. In glycerol defined medium, B. subtilis grew with nitrate, but not nitrite or fumarate, while B. licheniformis grew with nitrate or fumarate, but not nitrite. Growth of E. coli occurred in glycerol defined medium with either nitrate, nitrite, or fumarate. In order to grow by fermentation, B. subtilis required both glucose and pyruvate, while B. licheniformis and E. coli were capable of using either glucose or pyruvate.

Impact of deglycosylation methods on two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry for proteomic analysis.

Glycosylation is a common post-translational modification that can add complexity to the proteome of many cell types. We used enzymatic and chemical methods of deglycosylation to treat a heavily glycosylated exoproteome sample from the filamentous fungus Trichoderma reesei. Deglycosylated samples were resolved on one-dimensional (1-D) and two-dimensional (2-D) gels in order to determine the effect of deglycosylation on the electrophoresis patterns and on the ability to identify proteins by peptide mass matching using matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis of in-gel tryptic digests. We found that deglycosylation of the protein sample resulted in different protein patterns on 1-D and 2-D gels, reduced the complexity of gel patterns, and enhanced the protein identification of some proteins via MALDI-TOF-MS. Deglycosylation with trifluoromethanesulfonic acid (TFMS) was found to be more effective than enzymatic treatments. These deglycosylation techniques may be employed in whole proteome analysis to locate glycosylated proteins and assist in their identification by MS.