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At least 19 records

Establishing a process of irradiating small animal brain using a CyberKnife and a microCT scanner

Purpose: Establish and validate a process of accurately irradiating small animals using the CyberKnife G4 System (version 8.5) with treatment plans designed to irradiate a hemisphere of a mouse brain based on microCT scanner images. Methods: These experiments consisted of four parts: (1) building a mouse phantom for intensity modulated radiotherapy (IMRT) quality assurance (QA), (2) proving usability of a microCT for treatment planning, (3) fabricating a small animal positioning system for use with the CyberKnife's image guided radiotherapy (IGRT) system, and (4)in vivo verification of targeting accuracy. A set of solid water mouse phantoms was designed and fabricated, with radiochromic films (RCF) positioned in selected planes to measure delivered doses. After down-sampling for treatment planning compatibility, a CT image set of a phantom was imported into the CyberKnife treatment planning system—MultiPlan (ver. 3.5.2). A 0.5 cm diameter sphere was contoured within the phantom to represent a hemispherical section of a mouse brain. A nude mouse was scanned in an alpha cradle using a microCT scanner (cone-beam, 157 × 149 pixels slices, 0.2 mm longitudinal slice thickness). Based on the results of our positional accuracy study, a planning treatment volume (PTV) was created. A stereotactic body mold of the mouse was “printed” using a 3D printer laying UV curable acrylic plastic. Printer instructions were based on exported contours of the mouse's skin. Positional reproducibility in the mold was checked by measuring ten CT scans. To verify accurate dose delivery in vivo, six mice were irradiated in the mold with a 4 mm target contour and a 2 mm PTV margin to 3 Gy and sacrificed within 20 min to avoid DNA repair. The brain was sliced and stained for analysis. Results: For the IMRT QA using a set of phantoms, the planned dose (6 Gy to the calculation point) was compared to the delivered dose measured via film and analyzed using Gamma analysis (3% and 3 mm). A passing rate of 99% was measured in areas of above 40% of the prescription dose. The final inverse treatment plan was comprised of 43 beams ranging from 5 to 12.5 mm in diameter (2.5 mm size increments are available up to 15 mm in diameter collimation). Using the Xsight Spine Tracking module, the CyberKnife system could not reliably identify and track the tiny mouse spine; however, the CyberKnife system could identify and track the fiducial markers on the 3D mold.In vivo positional accuracy analysis using the 3D mold generated a mean error of 1.41 mm ± 0.73 mm when fiducial markers were used for position tracking. Analysis of the dissected brain confirmed the ability to target the correct brain volume. Conclusions: With the use of a stereotactic body mold with fiducial markers, microCT imaging, and resolution down-sampling, the CyberKnife system can successfully perform small-animal radiotherapy studies.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Transmitters for animals and methods for transmitting from animals

Animal transmitters are provided that can include: a transducer configured to transmit a signal; process circuitry coupled to the transducer; and an energy harvesting element coupled to the process circuitry. Animals having a transmitter coupled thereto are also provided with the transmitter including an energy harvesting element in operational alignment with the animal's musculoskeletal system. Methods for transmitting the location of an animal are also provided with the methods including: coupling a transmitter powered by an energy harvesting element to the animal; and monitoring the transmissions of the transmitter.

Deng, Z. Daniel↗

Unexpected competitiveness of Methanosaeta populations at elevated acetate concentrations in methanogenic treatment of animal wastewater

Acetoclastic methanogenesis is a key metabolic process in anaerobic digestion, a technology with broad applications in biogas production and waste treatment. Acetoclastic methanogenesis is known to be performed by two archaeal genera, Methanosaeta and Methanosarcina. The conventional model posits that Methanosaeta populations are more competitive at low acetate levels (<1 mM) than Methanosarcina and vice versa at higher acetate concentrations. While this model is supported by an extensive body of studies, reports of inconsistency have grown that Methanosaeta were observed to outnumber Methanosarcina at elevated acetate levels. In this study, monitoring of anaerobic digesters treating animal wastewater unexpectedly identified Methanosaeta as the dominant acetoclastic methanogen population at both low and high acetate levels during organic overloading. The surprising competitiveness of Methanosaeta at elevated acetate was further supported by the enrichment of Methanosaeta with high concentrations of acetate (20 mM). The dominance of Methanosaeta in the methanogen community could be reproduced in anaerobic digesters with the direct addition of acetate to above 20 mM, again supporting the competitiveness of Methanosaeta over Methanosarcina at elevated acetate levels. This study for the first time systematically demonstrated that the dominance of Methanosaeta populations in anaerobic digestion could be linked to the competitiveness of Methanosaeta at elevated acetate concentrations. Given the importance of acetoclastic methanogenesis in biological methane production, findings from this study could have major implications for developing strategies for more effective control of methanogenic treatment processes.

59 BASIC BIOLOGICAL SCIENCES↗

Lighted display devices for producing static or animated visual displays, including animated facial features

An illuminated display device with a base member with a plurality of cavities therein. Illumination devices illuminate the cavities and emit light through an opening of the cavities in a pattern, and a speaker can emit sounds in synchronization with the pattern. A panel with translucent portions can overly the base member and the cavities. An animated talking character can have an animated mouth cavity complex with multiple predetermined mouth lighting configurations simulative of human utterances. The cavities can be open, or optical waveguide material or positive members can be disposed therein. Reflective material can enhance internal reflectance and light emission.

42 ENGINEERING↗

Using steam explosion or AFEX™ to produce animal feeds and biofuel feedstocks in a biorefinery based on sugarcane residues: Using Steam Explosion or AFEX™ to Produce Animal Feeds and Biofuel Feedstocks in a Biorefinery Based on Sugarcane Residues

Sustainable integration of biofuel (primarily bioethanol) and highly digestible livestock feed in production systems is a potential way to increase the economic returns to agriculture and simultaneously promote energy security, particularly in developing countries. In this work we evaluated the efficacy of steam explosion (StEx) and ammonia fiber expansion (AFEX™) as potential processes for improving the in vitro rumen digestibility, metabolizable energy, and ethanol yields from sugarcane crop residues (bagasse and cane leaf matter, CLM). Ammonia fiber expansion pretreatment enhanced the in vitro true digestibility and metabolizable energy content of sugarcane crop residues by 69% and 26%, respectively compared to untreated controls. On the other hand, StEx increased the true digestibility and metabolizable energy content of the sugarcane residues by 54% and 7%, respectively. Ammonia fiber expansion also increased the total nitrogen content of both sugarcane bagasse and CLM to more than 20.2 g kg -1 dry forage, a more than 230% improvement relative to untreated controls. High solid-loading enzymatic hydrolysis and fermentation of StEx- and AFEX™-pretreated sugarcane crop residues generated yields of up to 3368 and 4360 L of ethanol per hectare of sugarcane cultivated, respectively, at a biomass-degrading enzyme dosage of 20 mg protein per gram glucan. This research strongly suggests that the use of suitably pretreated sugarcane crop residues in integrated sugarcane biofuel-livestock production systems can increase the total per hectare agricultural output without increasing the area of sugarcane cultivated. In effect, this integrated approach promotes more sustainable biofuel production and increased food production while avoiding the potential for indirect land use change.

09 BIOMASS FUELS↗

MO-DE-BRA-06: 3D Image Acquisition and Reconstruction Explained with Online Animations

Purpose: Understanding the principles of 3D imaging and image reconstruction is fundamental to the field of medical imaging. Clinicians, technologists, physicists, patients, students, and inquisitive minds all stand to benefit from greater comprehension of the supporting technologies. To help explain the basic principles of 3D imaging, we developed multi-frame animations that convey the concepts of tomographic imaging. The series of free (gif) animations are accessible online, and provide a multimedia introduction to the main concepts of image reconstruction. Methods: Text and animations were created to convey the principles of analytic tomography in CT, PET, and SPECT. Specific topics covered included: principles of sinograms/image data storage, forward projection, principles of PET acquisitions, and filtered backprojection. A total of 8 animations were created and presented for CT, PET, and digital phantom formats. In addition, a free executable is also provided to allow users to create their own tomographic animations – providing an opportunity for interaction and personalization to help foster user interest. Results: Tutorial text and animations have been posted online, freely available to view or download. The animations are in first position in a google search of “image reconstruction animations”. The website currently receives approximately 200 hits/month, from all over the world, and the usage is growing. Positive feedback has been collected from users. Conclusion: We identified a need for improved teaching tools to help visualize the (temporally variant) concepts of image reconstruction, and have shown that animations can be a useful tool for this aspect of education. Furthermore, posting animations freely on the web has shown to be a good way to maximize their impact in the community. In future endeavors, we hope to expand this animated content, to cover principles of iterative reconstruction, as well as other phenomena relating to imaging.

60 APPLIED LIFE SCIENCES↗

Staufen1, Kinesin1 and microtubule function in cyclin B1 mRNA transport to the animal polar cytoplasm of zebrafish oocytes

Highlights: • Zebrafish cyclin B1 mRNA is transported to the animal polar cytoplasm of oocytes. • Microtubule, Kinesin1 and Stufen1 are involved in cyclin B1 mRNA transport. • The plus ends of microtubules accumulate to the animal pole of full-grown oocytes. • A common transport machinery is employed in zebrafish and Xenopus oocytes. • The direction of mRNA transport depends on microtubule organization in oocytes. In zebrafish oocytes, cyclin B1 mRNAs are transported to the animal polar cytoplasm. To elucidate the molecular basis of cyclin B1 mRNA transport, we analyzed zebrafish Staufen1, a protein known to play a central role in mRNA transport to the vegetal pole of Xenopus oocytes. Zebrafish Staufen1 interacts with cyclin B1 mRNA throughout oocyte growth. Both cyclin B1 mRNA and Staufen1 are evenly distributed in the cytoplasm of young oocytes but are co-localized to the animal polar cytoplasm in later stages. Real-time imaging showed that the plus ends of oocyte microtubules are free in the cytoplasm in early stages but anchored to the animal polar cytoplasm in later stages. Transport of cyclin B1 reporter mRNA to the animal polar cytoplasm was inhibited by disruption of microtubules and injection of antibodies against Staufen1 or Kinesin1, a plus-end-directed microtubule motor that interacts with Staufen1, indicating that the transport depends on movement along microtubules toward the plus ends. Reporter mRNAs with an element required for the vegetal localization of vg1 mRNA in Xenopus oocytes were localized to the animal polar cytoplasm in zebrafish oocytes, indicating that the element is functional for animal polar localization in zebrafish oocytes. Our findings suggest that cyclin B1 mRNA-Staufen1 protein complexes are transported toward the animal pole of zebrafish oocytes by the plus-end-directed motor protein Kinesin1 along microtubules and that a common mRNA transport machinery functions in zebrafish and Xenopus oocytes, although its transport direction is opposite due to different organizations of microtubules.

60 APPLIED LIFE SCIENCES↗

WE-H-206-02: Recent Advances in Multi-Modality Molecular Imaging of Small Animals

Lihong V. Wang: Photoacoustic tomography (PAT), combining non-ionizing optical and ultrasonic waves via the photoacoustic effect, provides in vivo multiscale functional, metabolic, and molecular imaging. Broad applications include imaging of the breast, brain, skin, esophagus, colon, vascular system, and lymphatic system in humans or animals. Light offers rich contrast but does not penetrate biological tissue in straight paths as x-rays do. Consequently, high-resolution pure optical imaging (e.g., confocal microscopy, two-photon microscopy, and optical coherence tomography) is limited to penetration within the optical diffusion limit (∼1 mm in the skin). Ultrasonic imaging, on the contrary, provides fine spatial resolution but suffers from both poor contrast in early-stage tumors and strong speckle artifacts. In PAT, pulsed laser light penetrates tissue and generates a small but rapid temperature rise, which induces emission of ultrasonic waves due to thermoelastic expansion. The ultrasonic waves, orders of magnitude less scattering than optical waves, are then detected to form high-resolution images of optical absorption at depths up to 7 cm, conquering the optical diffusion limit. PAT is the only modality capable of imaging across the length scales of organelles, cells, tissues, and organs (up to whole-body small animals) with consistent contrast. This rapidly growing technology promises to enable multiscale biological research and accelerate translation from microscopic laboratory discoveries to macroscopic clinical practice. PAT may also hold the key to label-free early detection of cancer by in vivo quantification of hypermetabolism, the quintessential hallmark of malignancy. Learning Objectives: To understand the contrast mechanism of PAT To understand the multiscale applications of PAT Benjamin M. W. Tsui: Multi-modality molecular imaging instrumentation and techniques have been major developments in small animal imaging that has contributed significantly to biomedical research during the past decade. The initial development was an extension of clinical PET/CT and SPECT/CT from human to small animals and combine the unique functional information obtained from PET and SPECT with anatomical information provided by the CT in registered multi-modality images. The requirements to image a mouse whose size is an order of magnitude smaller than that of a human have spurred advances in new radiation detector technologies, novel imaging system designs and special image reconstruction and processing techniques. Examples are new detector materials and designs with high intrinsic resolution, multi-pinhole (MPH) collimator design for much improved resolution and detection efficiency compared to the conventional collimator designs in SPECT, 3D high-resolution and artifact-free MPH and sparse-view image reconstruction techniques, and iterative image reconstruction methods with system response modeling for resolution recovery and image noise reduction for much improved image quality. The spatial resolution of PET and SPECT has improved from ∼6–12 mm to ∼1 mm a few years ago to sub-millimeter today. A recent commercial small animal SPECT system has achieved a resolution of ∼0.25 mm which surpasses that of a state-of-art PET system whose resolution is limited by the positron range. More recently, multimodality SA PET/MRI and SPECT/MRI systems have been developed in research laboratories. Also, multi-modality SA imaging systems that include other imaging modalities such as optical and ultrasound are being actively pursued. In this presentation, we will provide a review of the development, recent advances and future outlook of multi-modality molecular imaging of small animals. Learning Objectives: To learn about the two major multi-modality molecular imaging techniques of small animals. To learn about the spatial resolution achievable by the molecular imaging systems for small animal today. To learn about the new multi-modality imaging instrumentation and techniques that are being developed. Sang Hyun Cho; X-ray fluorescence (XRF) imaging, such as x-ray fluorescence computed tomography (XFCT), offers unique capabilities for accurate identification and quantification of metals within the imaging objects. As a result, it has emerged as a promising quantitative imaging modality in recent years, especially in conjunction with metal-based imaging probes. This talk will familiarize the audience with the basic principles of XRF/XFCT imaging. It will also cover the latest development of benchtop XFCT technology. Additionally, the use of metallic nanoparticles such as gold nanoparticles, in conjunction with benchtop XFCT, will be discussed within the context of preclinical multimodal multiplexed molecular imaging. Learning Objectives: To learn the basic principles of XRF/XFCT imaging To learn the latest advances in benchtop XFCT development for preclinical imaging Funding support received from NIH and DOD; Funding support received from GE Healthcare; Funding support received from Siemens AX; Patent royalties received from GE Healthcare; L. Wang, Funding Support: NIH; COI: Microphotoacoustics; S. Cho, Yes: ;NIH/NCI grant R01CA155446 DOD/PCRP grant W81XWH-12-1-0198.

60 APPLIED LIFE SCIENCES↗

Improving the accessibility and transferability of machine learning algorithms for identification of animals in camera trap images: MLWIC2

Motion-activated wildlife cameras (or “camera traps”) are frequently used to remotely and noninvasively observe animals. The vast number of images collected from camera trap projects has prompted some biologists to employ machine learning algorithms to automatically recognize species in these images, or at least filter-out images that do not contain animals. These approaches are often limited by model transferability, as a model trained to recognize species from one location might not work as well for the same species in different locations. Furthermore, these methods often require advanced computational skills, making them inaccessible to many biologists. We used 3 million camera trap images from 18 studies in 10 states across the United States of America to train two deep neural networks, one that recognizes 58 species, the “species model,” and one that determines if an image is empty or if it contains an animal, the “empty-animal model.” Our species model and empty-animal model had accuracies of 96.8% and 97.3%, respectively. Furthermore, the models performed well on some out-of-sample datasets, as the species model had 91% accuracy on species from Canada (accuracy range 36%–91% across all out-of-sample datasets) and the empty-animal model achieved an accuracy of 91%–94% on out-of-sample datasets from different continents. Our software addresses some of the limitations of using machine learning to classify images from camera traps. By including many species from several locations, our species model is potentially applicable to many camera trap studies in North America. We also found that our empty-animal model can facilitate removal of images without animals globally. We provide the trained models in an R package (MLWIC2: Machine Learning for Wildlife Image Classification in R), which contains Shiny Applications that allow scientists with minimal programming experience to use trained models and train new models in six neural network architectures with varying depths.

59 BASIC BIOLOGICAL SCIENCES↗

Small animal PET: a review of what we have done and where we are going

Abstract Small animal research is an essential tool in studying both pharmaceutical biodistribution and disease progression over time. Furthermore, through the rapid development of in vivo imaging technology over the last few decades, small animal imaging (also referred to as preclinical imaging) has become a mainstay for all fields of biologic research and a center point for most preclinical cancer research. Preclinical imaging modalities include optical, MRI and MRS, microCT, small animal PET, ultrasound, and photoacoustic, each with their individual strengths. The strong points of small animal PET are its translatability to the clinic; its quantitative imaging capabilities; its whole-body imaging ability to dynamically trace functional/biochemical processes; its ability to provide useful images with only nano- to pico- molar concentrations of administered compounds; and its ability to study animals serially over time. This review paper gives an overview of the development and evolution of small animal PET imaging. It provides an overview of detector designs; system configurations; multimodality PET imaging systems; image reconstruction and analysis tools; and an overview of research and commercially available small animal PET systems. It concludes with a look toward developing technologies/methodologies that will further enhance the impact of small animal PET imaging on medical research in the future.

Engineering↗

Envisioning the Future of Aquatic Animal Tracking: Technology, Science, and Application

Electronic tags have proven to be extremely useful for broadening our understanding of aquatic animals by answering diverse questions about their behaviours, physiologies, and life histories fundamental to ecology. Simultaneously, many applied conservation and management efforts are informed by animals tagged with electronic tags. In spite of the many advances in tracking software and hardware, an uncertain future in the world’s aquatic ecosystems portends great challenges for science. Aquatic animal tracking with electronic tags represents both the present and future of integrative biology and ecology in aquatic ecosystems. Here we identify what we regard as the future of aquatic animal tracking in a horizon scanning exercise. We submit that the future of aquatic animal tracking will include opportunities for multi-platform tracking systems for simultaneously monitoring position, activity, physiology, and microhabitat of animals, improved data collection and accessibility with new infrastructure (e.g. tags, receivers) and cyberinfrastructure, and integrated tagging information with animal traits derived from biopsy during tagging. We discuss parallel needs and opportunities in areas related to the application of animal tracking in the future such as knowledge mobilization and governance.

biologging↗

A global database of nitrogen and phosphorus excretion rates of aquatic animals

Though their importance varies greatly among species and ecosystems, animals can be important in modulating ecosystem-level nutrient cycling. Nutrient cycling rates of individual animals represent valuable data for testing the predictions of important frameworks such as the Metabolic Theory of Ecology (MTE) and ecological stoichiometry (ES). They also represent an important set of functional traits that may reflect both environmental and phylogenetic influences. Over the past two decades, studies of animal-mediated nutrient cycling have increased dramatically, especially in aquatic ecosystems. Here we present a global compilation of aquatic animal nutrient excretion rates. The dataset includes 10,534 observations from freshwater and marine animals of N and/or P excretion rates. Furthermore, these observations represent 491 species, including most aquatic phyla. Coverage varies greatly among phyla and other taxonomic levels. The dataset includes information on animal body size, ambient temperature, taxonomic affiliations, and animal body N:P. We used this data set to test predictions of MTE and ES, as described in Vanni and McIntyre (2016; Ecology DOI: 10.1002/ecy.1582).

54 ENVIRONMENTAL SCIENCES↗

Allometric-kinetic model predictions of concentration ratios for anthropogenic radionuclides across animal classes and food selection

Protection of the environment from radiation fundamentally relies on dose assessments for non-human biota. These dose assessments use measured or predicted concentrations of radionuclides in soil or water combined with Concentration Ratios (CRs) to estimate tissue concentrations in animals and plants, yet there is a paucity of CR data relative to the vast number of potential taxa and radioactive contaminants in the environment and their taxon-specific ecosystems. Because there are many taxa each having very different behaviors and biology and many possible bioavailable radionuclides, CRs have the potential to vary by orders-of-magnitude, as often seen in published data. Given the diversity of taxa, the International Commission on Radiological Protection (ICRP) has categorized non-human biota into 12 representative organisms (ROs), while the U.S. Department of Energy (DOE) uses the non-taxon specific categories of terrestrial, riparian, and aquatic animals. The question we examine here is: do these categorizations adequately “represent” the broad diversity of animals? To explore this question, here we analyze an Allometric-Kinetic (A-K) model to calculate radionuclide-specific CRs for common animal classes which are then further subcategorized into herbivores, omnivores, carnivores, and invertebrate detritivores. Comparisons in CRs among animal classes exhibited only small differences, but there was order of magnitude differences between herbivores, carnivores, and especially detritivores, for many radionuclides of interest. These findings suggest that the ICRP ROs and the DOE categories are reasonable, but their accuracy could be improved by including sub-categories related to animal dietary ecology. Finally, comparisons of A-K model predicted CR values to published CRs show order-of-magnitude variations, providing justification for additional studies of assimilation across radionuclides and animal classes.

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗

SU-E-T-299: Dosimetric Characterization of Small Field in Small Animal Irradiator with Radiochromic Films

Purpose: The small animal irradiator has been used with small animals to optimize new radiation therapy as preclinical studies. The small animal was irradiated by whole- or partial-body exposure. In this study, the dosimetric characterizations of small animal irradiator were carried out in small field using Radiochromic films Material & Methods: The study was performed in commercial animal irradiator (XRAD-320, Precision x-ray Inc, North Brantford) with Radiochromic films (EBT2, Ashland Inc, Covington). The calibration curve was generated between delivery dose and optical density (red channel) and the films were scanned by and Epson 1000XL scanner (Epson America Inc., Long Beach, CA).We evaluated dosimetric characterization of irradiator using various filter supported by manufacturer in 260 kV. The various filters were F1 (2.0mm Aluminum (HVL = about 1.0mm Cu) and F2 (0.75mm Tin + 0.25mm Copper + 1.5mm Aluminum (HVL = about 3.7mm Cu). According to collimator size (3, 5, 7, 10 mm, we calculated percentage depth dose (PDD) and the surface –source distance(SSD) was 17.3 cm considering dose rate. Results: The films were irradiated in 260 kV, 10mA and we increased exposure time 5sec. intervals from 5sec. to 120sec. The calibration curve of films was fitted with cubic function. The correlation between optical density and dose was Y=0.1405 X{sup 3}−2.916 X{sup 2}+25.566 x+2.238 (R{sup 2}=0.994). Based on the calibration curve, we calculated PDD in various filters depending on collimator size. When compared PDD of specific depth (3mm) considering animal size, the difference by collimator size was 4.50% in free filter and F1 was 1.53% and F2 was within 2.17%. Conclusion: We calculated PDD curve in small animal irradiator depending on the collimator size and the kind of filter using the radiochromic films. The various PDD curve was acquired and it was possible to irradiate various dose using these curve.

60 APPLIED LIFE SCIENCES↗

Animal displacement from marine energy development: Mechanisms and consequences

For marine wave and tidal energy to successfully contribute to global renewable energy goals and climate change mitigation, marine energy projects need to expand beyond small deployments to large-scale arrays. However, with large-scale projects come potential environmental effects not observed at the scales of single devices and small arrays. One of these effects is the risk of displacing marine animals from their preferred habitats or their migration routes, which may increase with the size of arrays and location. Many marine animals may be susceptible to some level of displacement once large marine energy arrays are increasingly integrated into the seascape, including large migratory animals, non-migratory pelagic animals with large home ranges, and benthic and demersal mobile organisms with more limited ranges, among many others. Yet, research around the mechanisms and effects of displacement have been hindered by the lack of clarity within the international marine energy community regarding the definition of displacement, how it occurs, its consequences, species of concern, and methods to investigate the outcomes. This review paper leveraged lessons learned from other industries, such as offshore development, to establish a definition of displacement in the marine energy context, explore which functional groups of marine animals may be affected and in what way, and identify pathways for investigating displacement through modeling and monitoring. In the marine energy context, we defined displacement as the outcome of one of three mechanisms (i.e., attraction, avoidance, and exclusion) triggered by an animal's response to one or more stressors acting as a disturbance, with various consequences at the individual through population levels. The knowledge gaps highlighted in this study will help the regulatory and scientific communities prepare for mitigating, observing, measuring, and characterizing displacement of various animals around marine energy arrays in order to prevent irreversible consequences.

16 TIDAL AND WAVE POWER↗