An automated aquatic rack system for rearing marine invertebrates.

BACKGROUND: One hundred years ago, marine organisms were the dominant systems for the study of developmental biology. The challenges in rearing these organisms outside of a marine setting ultimately contributed to a shift towards work on a smaller number of so-called model systems. Those animals are typically non-marine organisms with advantages afforded by short life cycles, high fecundity, and relative ease in laboratory culture. However, a full understanding of biodiversity, evolution, and anthropogenic effects on biological systems requires a broader survey of development in the animal kingdom. To this day, marine organisms remain relatively understudied, particularly the members of the Lophotrochozoa (Spiralia), which include well over one third of the metazoan phyla (such as the annelids, mollusks, flatworms) and exhibit a tremendous diversity of body plans and developmental modes. To facilitate studies of this group, we have previously described the development and culture of one lophotrochozoan representative, the slipper snail Crepidula atrasolea, which is easy to rear in recirculating marine aquaria. Lab-based culture and rearing of larger populations of animals remain a general challenge for many marine organisms, particularly for inland laboratories. RESULTS: Here, we describe the development of an automated marine aquatic rack system for the high-density culture of marine species, which is particularly well suited for rearing filter-feeding animals. Based on existing freshwater recirculating aquatic rack systems, our system is specific to the needs of marine organisms and incorporates robust filtration measures to eliminate wastes, reducing the need for regular water changes. In addition, this system incorporates sensors and associated equipment for automated assessment and adjustment of water quality. An automated feeding system permits precise delivery of liquid food (e.g., phytoplankton) throughout the day, mimicking real-life feeding conditions that contribute to increased growth rates and fecundity. CONCLUSION: This automated system makes laboratory culture of marine animals feasible for both large and small research groups, significantly reducing the time, labor, and overall costs needed to rear these organisms.

PelagiCam: a novel underwater imaging system with computer vision for semi-automated monitoring of mobile marine fauna at offshore structures.

Engineered structures in the open ocean are becoming more frequent with the expansion of the marine renewable energy industry and offshore marine aquaculture. Floating engineered structures function as artificial patch reefs providing novel and relatively stable habitat structure not otherwise available in the pelagic water column. The enhanced physical structure can increase local biodiversity and benefit fisheries yet can also facilitate the spread of invasive species. Clear evidence of any ecological consequences will inform the design and placement of structures to either minimise negative impacts or enhance ecosystem restoration. The development of rapid, cost-effective and reliable remote underwater monitoring methods is crucial to supporting evidence-based decision-making by planning authorities and developers when assessing environmental risks and benefits of offshore structures. A novel, un-baited midwater video system, PelagiCam, with motion-detection software (MotionMeerkat) for semi-automated monitoring of mobile marine fauna, was developed and tested on the UK's largest offshore rope-cultured mussel farm in Lyme Bay, southwest England. PelagiCam recorded Atlantic horse mackerel (Trachurus trachurus), garfish (Belone belone) and two species of jellyfish (Chrysaora hysoscella and Rhizostoma pulmo) in open water close to the floating farm structure. The software successfully distinguished video frames where fishes were present versus absent. The PelagiCam system provides a cost-effective remote monitoring tool to streamline biological data acquisition in impact assessments of offshore floating structures. With the rise of sophisticated artificial intelligence for object recognition, the integration of computer vision techniques should receive more attention in marine ecology and has great potential to revolutionise marine biological monitoring.

On the impact of Citizen Science-derived data quality on deep learning based classification in marine images.

The evaluation of large amounts of digital image data is of growing importance for biology, including for the exploration and monitoring of marine habitats. However, only a tiny percentage of the image data collected is evaluated by marine biologists who manually interpret and annotate the image contents, which can be slow and laborious. In order to overcome the bottleneck in image annotation, two strategies are increasingly proposed: "citizen science" and "machine learning". In this study, we investigated how the combination of citizen science, to detect objects, and machine learning, to classify megafauna, could be used to automate annotation of underwater images. For this purpose, multiple large data sets of citizen science annotations with different degrees of common errors and inaccuracies observed in citizen science data were simulated by modifying "gold standard" annotations done by an experienced marine biologist. The parameters of the simulation were determined on the basis of two citizen science experiments. It allowed us to analyze the relationship between the outcome of a citizen science study and the quality of the classifications of a deep learning megafauna classifier. The results show great potential for combining citizen science with machine learning, provided that the participants are informed precisely about the annotation protocol. Inaccuracies in the position of the annotation had the most substantial influence on the classification accuracy, whereas the size of the marking and false positive detections had a smaller influence.

Unoccupied Aircraft Systems in Marine Science and Conservation.

The use of unoccupied aircraft systems (UASs, also known as drones) in science is growing rapidly. Recent advances in microelectronics and battery technology have resulted in the rapid development of low-cost UASs that are transforming many industries. Drones are poised to revolutionize marine science and conservation, as they provide essentially on-demand remote sensing capabilities at low cost and with reduced human risk. A variety of multirotor, fixed-wing, and transitional UAS platforms are capable of carrying various optical and physical sampling payloads and are being employed in almost every subdiscipline of marine science and conservation. This article provides an overview of the UAS platforms and sensors used in marine science and conservation missions along with example physical, biological, and natural resource management applications and typical analytical workflows. It concludes with details on potential effects of UASs on marine wildlife and a look to the future of UASs in marine science and conservation.

Windows into Microbial Seascapes: Advances in Nanoscale Imaging and Application to Marine Sciences.

Geochemical cycles of all nonconservative elements are mediated by microorganisms over nanometer spatial scales. The pelagic seascape is known to possess microstructure imposed by heterogeneous distributions of particles, polymeric gels, biologically important chemicals, and microbes. While indispensable, most traditional oceanographic observational approaches overlook this heterogeneity and ignore subtleties, such as activity hot spots, symbioses, niche partitioning, and intrapopulation phenotypic variations, that can provide a deeper mechanistic understanding of planktonic ecosystem function. As part of the movement toward cultivation-independent tools in microbial oceanography, techniques to examine the ecophysiology of individual populations and their role in chemical transformations at spatial scales relevant to microorganisms have been developed. This review presents technologies that enable geochemical and microbiological interrogations at spatial scales ranging from 0.02 to a few hundred micrometers, particularly focusing on atomic force microscopy, nanoscale secondary ion mass spectrometry, and confocal Raman microspectroscopy and introducing promising approaches for future applications in marine sciences.

A Dexterous, Glove-Based Teleoperable Low-Power Soft Robotic Arm for Delicate Deep-Sea Biological Exploration.

Modern marine biologists seeking to study or interact with deep-sea organisms are confronted with few options beyond industrial robotic arms, claws, and suction samplers. This limits biological interactions to a subset of "rugged" and mostly immotile fauna. As the deep sea is one of the most biologically diverse and least studied ecosystems on the planet, there is much room for innovation in facilitating delicate interactions with a multitude of organisms. The biodiversity and physiology of shallow marine systems, such as coral reefs, are common study targets due to the easier nature of access; SCUBA diving allows for in situ delicate human interactions. Beyond the range of technical SCUBA (~150 m), the ability to achieve the same level of human dexterity using robotic systems becomes critically important. The deep ocean is navigated primarily by manned submersibles or remotely operated vehicles, which currently offer few options for delicate manipulation. Here we present results in developing a soft robotic manipulator for deep-sea biological sampling. This low-power glove-controlled soft robot was designed with the future marine biologist in mind, where science can be conducted at a comparable or better means than via a human diver and at depths well beyond the limits of SCUBA. The technology relies on compliant materials that are matched with the soft and fragile nature of marine organisms, and uses seawater as the working fluid. Actuators are driven by a custom proportional-control hydraulic engine that requires less than 50 W of electrical power, making it suitable for battery-powered operation. A wearable glove master allows for intuitive control of the arm. The manipulator system has been successfully operated in depths exceeding 2300 m (3500 psi) and has been field-tested onboard a manned submersible and unmanned remotely operated vehicles. The design, development, testing, and field trials of the soft manipulator is placed in context with existing systems and we offer suggestions for future work based on these findings.

Early-career researchers: an interview with Kakani Katija.

Kakani Katija is a Principal Engineer at the Monterey Bay Aquarium Research Institute, USA, where she designs and builds instrumentation to study marine invertebrate ecomechanics. She received her Bachelor's degree in Aeronautics and Astronautics from the University of Washington, USA, before moving to the California Institute of Technology, USA, for her Master's degree in Aeronautics with Morteza Gharib and PhD in Bioengineering in the laboratory of John Dabiri, completed in 2010. Katija was recognised as a National Geographic Emerging Explorer in 2011 and has given presentations at TEDYouth and TEDWomen.

Buzz off! An evaluation of ultrasonic acoustic vibration for the disruption of marine micro-organisms on sensor-housing materials.

Biofouling is a process of ecological succession which begins with the attachment and colonization of micro-organisms to a submerged surface. For marine sensors and their housings, biofouling can be one of the principle limitations to long-term deployment and reliability. Conventional antibiofouling strategies using biocides can be hazardous to the environment, and therefore alternative chemical-free methods are preferred. In this study, custom-made testing assemblies were used to evaluate ultrasonic vibration as an antibiofouling process for marine sensor-housing materials over a 28-day time course. Microbial biofouling was measured based on (i) surface coverage, using fluorescence microscopy and (ii) bacterial 16S rDNA gene copies, using Quantitative polymerase chain reaction (PCR). Ultrasonic vibrations (20 KHz, 200 ms pulses at 2-s intervals; total power 16·08 W) significantly reduced the surface coverage on two plastics, poly(methyl methacrylate) and polyvinyl chloride (PVC) for up to 28 days. Bacterial gene copy number was similarly reduced, but the results were only statistically significant for PVC, which displayed the greatest overall resistance to biofouling, regardless of whether ultrasonic vibration was applied. Copper sheet, which has intrinsic biocidal properties was resistant to biofouling during the early stages of the experiment, but inhibited measurements made by PCR and generated inconsistent results later on. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, ultrasonic acoustic vibration is presented as a chemical-free, ecologically friendly alternative to conventional methods for the perturbation of microbial attachment to submerged surfaces. The results indicate the potential of an ultrasonic antibiofouling method for the disruption of microbial biofilms on marine sensor housings, which is typically a principle limiting factor in their long-term operation in the oceans. With increasing deployment of scientific apparatus in aquatic environments, including further offshore and for longer duration, the identification and evaluation of novel antifouling strategies that do not employ hazardous chemicals are widely sought.

New Ulvaceae (Ulvophyceae, Chlorophyta) from mesophotic ecosystems across the Hawaiian Archipelago.

Ulvalean algae (Chlorophyta) are most commonly described from intertidal and shallow subtidal marine environments worldwide, but are less well known from mesophotic environments. Their morphological simplicity and phenotypic plasticity make accurate species determinations difficult, even at the generic level. Here, we describe the mesophotic Ulvales species composition from 13 locations across 2,300 km of the Hawaiian Archipelago. Twenty-eight representative Ulvales specimens from 64 to 125 m depths were collected using technical diving, submersibles, and remotely operated vehicles. Morphological and molecular characters suggest that mesophotic Ulvales in Hawaiian waters form unique communities comprising four species within the genera Ulva and Umbraulva, each with discrete geographic and/or depth-related distributional patterns. Three genetically distinct taxa are supported by both plastid (rbcL and tufA) and nuclear (ITS1) markers, and are presented here as new species: Umbraulva kaloakulau, Ulva ohiohilulu, and Ulva iliohaha. We also propose a new Umbraulva species (Umbraulva kuaweuweu), which is closely related to subtidal records from New Zealand and Australia, but not formally described. To our knowledge, these are the first marine species descriptions from Hawai'i resulting from the collaboration of traditional Hawaiian nomenclature specialists, cultural practitioners and scientists. The difficulty of finding reliable diagnostic morphological characters for these species reflects a common problem worldwide of achieving accurate identification of ulvalean taxa using solely morphological criteria. Mesophotic Ulvales appear to be distinct from shallow-water populations in Hawai'i, but their degree of similarity to mesophotic floras in other locations in the Pacific remains unknown.

Automated detection of Antarctic blue whale calls.

This paper addresses the problem of automated detection of Z-calls emitted by Antarctic blue whales (B. m. intermedia). The proposed solution is based on a subspace detector of sigmoidal-frequency signals with unknown time-varying amplitude. This detection strategy takes into account frequency variations of blue whale calls as well as the presence of other transient sounds that can interfere with Z-calls (such as airguns or other whale calls). The proposed method has been tested on more than 105 h of acoustic data containing about 2200 Z-calls (as found by an experienced human operator). This method is shown to have a correct-detection rate of up to more than 15% better than the extensible bioacoustic tool package, a spectrogram-based correlation detector commonly used to study blue whales. Because the proposed method relies on subspace detection, it does not suffer from some drawbacks of correlation-based detectors. In particular, it does not require the choice of an a priori fixed and subjective template. The analytic expression of the detection performance is also derived, which provides crucial information for higher level analyses such as animal density estimation from acoustic data. Finally, the detection threshold automatically adapts to the soundscape in order not to violate a user-specified false alarm rate.

Three-dimensional tracking of Cuvier's beaked whales' echolocation sounds using nested hydrophone arrays.

Cuvier's beaked whales (Ziphius cavirostris) were tracked using two volumetric small-aperture (∼1 m element spacing) hydrophone arrays, embedded into a large-aperture (∼1 km element spacing) seafloor hydrophone array of five nodes. This array design can reduce the minimum number of nodes that are needed to record the arrival of a strongly directional echolocation sound from 5 to 2, while providing enough time-differences of arrivals for a three-dimensional localization without depending on any additional information such as multipath arrivals. To illustrate the capabilities of this technique, six encounters of up to three Cuvier's beaked whales were tracked over a two-month recording period within an area of 20 km(2) in the Southern California Bight. Encounter periods ranged from 11 min to 33 min. Cuvier's beaked whales were found to reduce the time interval between echolocation clicks while alternating between two inter-click-interval regimes during their descent towards the seafloor. Maximum peak-to-peak source levels of 179 and 224 dB re 1 µPa @ 1 m were estimated for buzz sounds and on-axis echolocation clicks (directivity index = 30 dB), respectively. Source energy spectra of the on-axis clicks show significant frequency components between 70 and 90 kHz, in addition to their typically noted FM upsweep at 40-60 kHz.

Thermal reference points as an index for monitoring body temperature in marine mammals.

BACKGROUND: Monitoring body temperature is essential in veterinary care as minor variations may indicate dysfunction. Rectal temperature is widely used as a proxy for body temperature, but measuring it requires special equipment, training or restraining, and it potentially stresses animals. Infrared thermography is an alternative that reduces handling stress, is safer for technicians and works well for untrained animals. This study analysed thermal reference points in five marine mammal species: bottlenose dolphin (Tursiops truncatus); beluga whale (Delphinapterus leucas); Patagonian sea lion (Otaria flavescens); harbour seal (Phoca vitulina); and Pacific walrus (Odobenus rosmarus divergens). RESULTS: The thermogram analysis revealed that the internal blowhole mucosa temperature is the most reliable indicator of body temperature in cetaceans. The temperatures taken during voluntary breathing with a camera held perpendicularly were practically identical to the rectal temperature in bottlenose dolphins and were only 1 °C lower than the rectal temperature in beluga whales. In pinnipeds, eye temperature appears the best parameter for temperature control. In these animals, the average times required for temperatures to stabilise after hauling out, and the average steady-state temperature values, differed according to species: Patagonian sea lions, 10 min, 31.13 °C; harbour seals, 10 min, 32.27 °C; Pacific walruses, 5 min, 29.93 °C. CONCLUSIONS: The best thermographic and most stable reference points for monitoring body temperature in marine mammals are open blowhole in cetaceans and eyes in pinnipeds.

A single-station method for the detection, classification and location of fin whale calls using ocean-bottom seismic stations.

Passive seismic monitoring in the oceans uses long-term deployments of Ocean Bottom Seismometers (OBSs). An OBS usually records the three components of ground motion and pressure, typically at 100 Hz. This makes the OBS an ideal tool to investigate fin and blue whales that vocalize at frequencies below 45 Hz. Previous applications of OBS data to locate whale calls have relied on single channel analyses that disregard the information that is conveyed by the horizontal seismic channels. Recently, Harris, Matias, Thomas, Harwood, and Geissler [J. Acoust. Soc. Am. 134, 3522-3535 (2013)] presented a method that used all four channels recorded by one OBS to derive the range and azimuth of fin whale calls. In this work, the detection, classification, and ranging of calls using this four-channel method were further investigated, focusing on methods to increase the accuracy of range estimates to direct path arrivals. Corrections to account for the influences of the sound speed in the water layer and the velocity structure in the top strata of the seabed were considered. The single station method discussed here is best implemented when OBSs have been deployed in deep water on top of seabed strata with low P-wave velocity. These conditions maximize the ability to detect and estimate ranges to fin whale calls.

Dissolved Oxygen Sensor in Animal-Borne Instruments: An Innovation for Monitoring the Health of Oceans and Investigating the Functioning of Marine Ecosystems.

The current decline in dissolved oxygen concentration within the oceans is a sensitive indicator of the effect of climate change on marine environment. However the impact of its declining on marine life and ecosystems' health is still quite unclear because of the difficulty in obtaining in situ data, especially in remote areas, like the Southern Ocean (SO). Southern elephant seals (Mirounga leonina) proved to be a relevant alternative to the traditional oceanographic platforms to measure physical and biogeochemical structure of oceanic regions rarely observed. In this study, we use a new stage of development in biologging technology to draw a picture of dissolved oxygen concentration in the SO. We present the first results obtained from a dissolved oxygen sensor added to Argos CTD-SRDL tags and deployed on 5 female elephant seals at Kerguelen. From October 2010 and October 2011, 742 oxygen profiles associated with temperature and salinity measurements were recorded. Whether a part of the data must be considered cautiously, especially because of offsets and temporal drifts of the sensors, the range of values recorded was consistent with a concomitant survey conducted from a research vessel (Keops-2 project). Once again, elephant seals reinforced the relationship between marine ecology and oceanography, delivering essential information about the water masses properties and the biological status of the Southern Ocean. But more than the presentation of a new stage of development in animal-borne instrumentation, this pilot study opens a new field of investigation in marine ecology and could be enlarged in a near future to other key marine predators, especially large fish species like swordfish, tuna or sharks, for which dissolved oxygen is expected to play a crucial role in distribution and behaviour.

An assessment of the effectiveness of high definition cameras as remote monitoring tools for dolphin ecology studies.

Research involving marine mammals often requires costly field programs. This paper assessed whether the benefits of using cameras outweighs the implications of having personnel performing marine mammal detection in the field. The efficacy of video and still cameras to detect Indo-Pacific bottlenose dolphins (Tursiops aduncus) in the Fremantle Harbour (Western Australia) was evaluated, with consideration on how environmental conditions affect detectability. The cameras were set on a tower in the Fremantle Port channel and videos were perused at 1.75 times the normal speed. Images from the cameras were used to estimate position of dolphins at the water's surface. Dolphin detections ranged from 5.6 m to 463.3 m for the video camera, and from 10.8 m to 347.8 m for the still camera. Detection range showed to be satisfactory when compared to distances at which dolphins would be detected by field observers. The relative effect of environmental conditions on detectability was considered by fitting a Generalised Estimation Equations (GEEs) model with Beaufort, level of glare and their interactions as predictors and a temporal auto-correlation structure. The best fit model indicated level of glare had an effect, with more intense periods of glare corresponding to lower occurrences of observed dolphins. However this effect was not large (-0.264) and the parameter estimate was associated with a large standard error (0.113). The limited field of view was the main restraint in that cameras can be only applied to detections of animals observed rather than counts of individuals. However, the use of cameras was effective for long term monitoring of occurrence of dolphins, outweighing the costs and reducing the health and safety risks to field personal. This study showed that cameras could be effectively implemented onshore for research such as studying changes in habitat use in response to development and construction activities.

Cross-correlation, triangulation, and curved-wavefront focusing of coral reef sound using a bi-linear hydrophone array.

A seven element, bi-linear hydrophone array was deployed over a coral reef in the Papahãnaumokuãkea Marine National Monument, Northwest Hawaiian Islands, in order to investigate the spatial, temporal, and spectral properties of biological sound in an environment free of anthropogenic influences. Local biological sound sources, including snapping shrimp and other organisms, produced curved-wavefront acoustic arrivals at the array, allowing source location via focusing to be performed over an area of 1600 m(2). Initially, however, a rough estimate of source location was obtained from triangulation of pair-wise cross-correlations of the sound. Refinements to these initial source locations, and source frequency information, were then obtained using two techniques, conventional and adaptive focusing. It was found that most of the sources were situated on or inside the reef structure itself, rather than over adjacent sandy areas. Snapping-shrimp-like sounds, all with similar spectral characteristics, originated from individual sources predominantly in one area to the east of the array. To the west, the spectral and spatial distributions of the sources were more varied, suggesting the presence of a multitude of heterogeneous biological processes. In addition to the biological sounds, some low-frequency noise due to distant breaking waves was received from end-fire north of the array.

Design and application of new low-cost instruments for marine environmental research.

The development of low-cost instrumentation plays a key role in marine environmental studies and represents one of the most innovative aspects of current oceanographic research. These kinds of devices can be used for several applications, ranging from vertical profilers to stand-alone systems, and can be installed on different platforms (buoys, Voluntary Observing Ships, underwater vehicles, etc.). The availability of low-cost technologies enables the realization of extended observatory networks for the study of marine physical and biological processes through an integrated approach merging in situ observations, forecasting models and remotely sensed data. We present new low-cost sensors and probes developed to measure marine temperature, conductivity, chlorophyll a and Chromophoric Dissolved Organic Matter fluorescence, focusing on sensing strategies, general architecture, laboratory trials, in situ tests and comparison with standard instruments. Furthermore, we report the expendable (New T-FLaP), vertical profiler (T-FLaPpro) and stand-alone (Spectra) applications of these technological developments that were tested during several oceanographic surveys in the Mediterranean Sea.