Associations between total mercury, trace minerals, and blood health markers in Northwest Atlantic white sharks (Carcharodon carcharias).

The ecology and life-histories of white sharks make this species susceptible to mercury bioaccumulation; however, the health consequences of mercury exposure are understudied. We measured muscle and plasma total mercury (THg), health markers, and trace minerals in Northwest Atlantic white sharks. THg in muscle tissue averaged 10.0 mg/kg dry weight, while THg in blood plasma averaged 533 µg/L. THg levels in plasma and muscle were positively correlated with shark precaudal length (153-419 cm), and THg was bioaccumulated proportionally in muscle and plasma. Nine sharks had selenium:mercury molar ratios in blood plasma >1.0, indicating that for certain individuals the potential protective effects of the trace mineral were diminished, whereas excess selenium may have protected other individuals. No relationships between plasma THg and any trace minerals or health markers were identified. Thus, we found no evidence of negative effects of Hg bioaccumulation, even in sharks with very high THg.

DNA metabarcoding of cloacal swabs provides insight into diets of highly migratory sharks in the Mid-Atlantic Bight.

The abundances of migratory shark species observed throughout the Mid-Atlantic Bight (MAB) during productive summer months suggest that this region provides critical habitat and prey resources to these taxa. However, the principal prey assemblages sustaining migratory shark biomass in this region are poorly defined. We applied high-throughput DNA metabarcoding to shark feces derived from cloacal swabs across nine species of Carcharhinid and Lamnid sharks to (1) quantify the contribution of broad taxa (e.g., invertebrates, fishes) supporting shark biomass during seasonal residency in the MAB and (2) determine whether the species displayed distinct dietary preference indicative of resource partitioning. DNA metabarcoding resulted in high taxonomic (species-level) resolution of shark diets with actinopterygian and elasmobranch fishes as the dominant prey categories across the species. DNA metabarcoding identified several key prey groups consistent across shark taxa that are likely integral for sustaining their biomass in this region, including Atlantic menhaden (Brevoortia tyrannus), Atlantic mackerel (Scomber scombrus), and benthic elasmobranchs, including skates. Our results are consistent with previously published stomach content data for the shark species of similar size range in the Northwest Atlantic Ocean, supporting the efficacy of cloacal swab DNA metabarcoding as a minimally invasive diet reconstruction technique. The high reliance of several shark species on Atlantic menhaden could imply wasp-waist food-web conditions during the summer months, whereby high abundances of forage fishes sustain a diverse suite of migratory sharks within a complex, seasonal food web.

An introduction to lesions and histology of scleractinian corals.

Stony corals (Scleractinia) are in the Phylum Cnidaria (cnidae referring to various types of stinging cells). They may be solitary or colonial, but all secrete an external, supporting aragonite skeleton. Large, colonial members of this phylum are responsible for the accretion of coral reefs in tropical and subtropical waters that form the foundations of the most biodiverse marine ecosystems. Coral reefs worldwide, but particularly in the Caribbean, are experiencing unprecedented levels of disease, resulting in reef degradation. Most coral diseases remain poorly described and lack clear case definitions, while the etiologies and pathogenesis are even more elusive. This introductory guide is focused on reef-building corals and describes basic gross and microscopic lesions in these corals in order to serve as an invitation to other veterinary pathologists to play a critical role in defining and advancing the field of coral pathology.

Tissue metal concentrations and antioxidant enzyme activity in western north Atlantic white sharks (Carcharodon carcharias).

Anthropogenic practices have increased metal contamination in marine ecosystems. Most sharks have long lifespans, occupy an important ecological position at the top of marine food webs, and can accumulate metals. However, reference levels of metal contaminants in the tissues of sharks, particularly, apex predators such as the white shark (Carcharodon carcharias), are lacking. In this study, concentrations of copper (Cu), cadmium (Cd), nickel (Ni), lead (Pb), silver (Ag), and zinc (Zn) were measured in the muscle tissue of white (n = 42) and tiger (Galeocerdo cuvier; n = 3) sharks. Metal exposure in various species, including sharks, has been correlated with increased oxidative stress. Therefore, the main objectives of this study were to assess metal accumulation and antioxidant enzyme activity (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)) in the muscle tissue of the population of white sharks and tiger sharks inhabiting the Western North Atlantic. The measured parameters were qualitatively compared between species. The small sample size of tiger sharks (collected from only one site) limited statistical analyses, therefore, white sharks were the primary focus of this study. Differences in tissue metal (Cu, Cd, Ni, and Zn) concentrations and antioxidant enzyme activities were detected based on collection site, with significant positive correlations between Cd and enzymes, SOD and CAT, and Zn and enzymes, SOD and GPx in C. carcharias. Differences in Ni concentration were detected based on sex, with females having higher Ni levels. Additionally, plasma osmolality was not correlated with tissue metal concentrations; however, osmolality decreased with increasing length in C. carcharias. This study is the first to report baseline levels of Cu, Zn, Cd, Ni, Ag, and Pb in muscle of North Atlantic white sharks and provides new insights into oxidative stress responses of these sensitive species to metal contaminants.

Perspective: Opportunities for advancing aquatic invertebrate welfare.

Welfare considerations and regulations for invertebrates have lagged behind those for vertebrates, despite invertebrates comprising more than 95% of earth's species. Humans interact with and use aquatic invertebrates for exhibition in zoos and aquaria, as pets, research subjects, and important food sources. Recent research has indicated that aquatic invertebrates, in particular cephalopod mollusks and decapod crustaceans, experience stress and may be able to feel pain. With this article, we present results of a survey on attitudes of aquatic animal health professionals toward aquatic invertebrate welfare and provide practical recommendations for advancing aquatic invertebrate welfare across four areas of opportunity: use of anesthesia, analgesia, and euthanasia; development of less invasive diagnostic and research sampling methods based on 3R principles; use of humane slaughter methods for aquatic invertebrates; and reducing impacts of invasive procedures in aquaculture and fisheries. We encourage consideration of these opportunities to achieve far-reaching improvements in aquatic invertebrate welfare.

A Review of Asteroid Biology in the Context of Sea Star Wasting: Possible Causes and Consequences.

AbstractSea star wasting-marked in a variety of sea star species as varying degrees of skin lesions followed by disintegration-recently caused one of the largest marine die-offs ever recorded on the west coast of North America, killing billions of sea stars. Despite the important ramifications this mortality had for coastal benthic ecosystems, such as increased abundance of prey, little is known about the causes of the disease or the mechanisms of its progression. Although there have been studies indicating a range of causal mechanisms, including viruses and environmental effects, the broad spatial and depth range of affected populations leaves many questions remaining about either infectious or non-infectious mechanisms. Wasting appears to start with degradation of mutable connective tissue in the body wall, leading to disintegration of the epidermis. Here, we briefly review basic sea star biology in the context of sea star wasting and present our current knowledge and hypotheses related to the symptoms, the microbiome, the viruses, and the associated environmental stressors. We also highlight throughout the article knowledge gaps and the data needed to better understand sea star wasting mechanistically, its causes, and potential management.

Energetic consequences of resource use diversity in a marine carnivore.

Understanding how intraspecific variation in the use of prey resources impacts energy metabolism has strong implications for predicting long-term fitness and is critical for predicting population-to-community level responses to environmental change. Here, we examine the energetic consequences of variable prey resource use in a widely distributed marine carnivore, juvenile sand tiger sharks (Carcharias taurus). We used carbon and nitrogen isotope analysis to identify three primary prey resource pools-demersal omnivores, pelagic forage, and benthic detritivores and estimated the proportional assimilation of each resource using Bayesian mixing models. We then quantified how the utilization of these resource pools impacted the concentrations of six plasma lipids and how this varied by ontogeny. Sharks exhibited variable reliance on two of three predominant prey resource pools: demersal omnivores and pelagic forage. Resource use variation was a strong predictor of energetic condition, whereby individuals more reliant upon pelagic forage exhibited higher blood plasma concentrations of very low-density lipoproteins, cholesterol, and triglycerides. These findings underscore how intraspecific variation in resource use may impact the energy metabolism of animals, and more broadly, that natural and anthropogenically driven fluctuations in prey resources could have longer term energetic consequences.

Microbiome structure in large pelagic sharks with distinct feeding ecologies.

BACKGROUND: Sharks play essential roles in ocean food webs and human culture, but also face population declines worldwide due to human activity. The relationship between sharks and the microbes on and in the shark body is unclear, despite research on other animals showing the microbiome as intertwined with host physiology, immunity, and ecology. Research on shark-microbe interactions faces the significant challenge of sampling the largest and most elusive shark species. We leveraged a unique sampling infrastructure to compare the microbiomes of two apex predators, the white (Carcharodon carcharias) and tiger shark (Galeocerdo cuvier), to those of the filter-feeding whale shark (Rhincodon typus), allowing us to explore the effects of feeding mode on intestinal microbiome diversity and metabolic function, and environmental exposure on the diversity of microbes external to the body (on the skin, gill). RESULTS: The fecal microbiomes of white and whale sharks were highly similar in taxonomic and gene category composition despite differences in host feeding mode and diet. Fecal microbiomes from these species were also taxon-poor compared to those of many other vertebrates and were more similar to those of predatory teleost fishes and toothed whales than to those of filter-feeding baleen whales. In contrast, microbiomes of external body niches were taxon-rich and significantly influenced by diversity in the water column microbiome. CONCLUSIONS: These results suggest complex roles for host identity, diet, and environmental exposure in structuring the shark microbiome and identify a small, but conserved, number of intestinal microbial taxa as potential contributors to shark physiology.

Elasmobranch microbiomes: emerging patterns and implications for host health and ecology.

Elasmobranchs (sharks, skates and rays) are of broad ecological, economic, and societal value. These globally important fishes are experiencing sharp population declines as a result of human activity in the oceans. Research to understand elasmobranch ecology and conservation is critical and has now begun to explore the role of body-associated microbiomes in shaping elasmobranch health. Here, we review the burgeoning efforts to understand elasmobranch microbiomes, highlighting microbiome variation among gastrointestinal, oral, skin, and blood-associated niches. We identify major bacterial lineages in the microbiome, challenges to the field, key unanswered questions, and avenues for future work. We argue for prioritizing research to determine how microbiomes interact mechanistically with the unique physiology of elasmobranchs, potentially identifying roles in host immunity, disease, nutrition, and waste processing. Understanding elasmobranch-microbiome interactions is critical for predicting how sharks and rays respond to a changing ocean and for managing healthy populations in managed care.

PHARMACOKINETICS AND CLINICAL SAFETY OF A SUSTAINED-RELEASE FORMULATION OF CEFTIOFUR CRYSTALLINE FREE ACID IN RINGNECK DOVES (STREPTOPELIA RISORIA) AFTER A SINGLE INTRAMUSCULAR INJECTION.

Ceftiofur crystalline free acid (CCFA) is a third-generation, oil-based, cephalosporin antimicrobial marketed as a once weekly treatment in cattle and swine, and as a two-time dose with 10-day duration in horses. Because handling and restraint times can be reduced, long-acting antibiotic preparations are particularly useful for treatment of nondomestic species. This study evaluated the pharmacokinetics of CCFA in ringneck doves (Streptopelia risoria). A single intramuscular (IM) injection of CCFA at 50 mg/kg was administered to each of 30 doves, and blood was collected from subsamples of 6 birds at predetermined sampling times (i.e., with a postinjection range of 0.5 to 192 hr). All ringneck doves were scheduled for euthanasia because of reasons unrelated to the study; this was performed at the conclusion of the study; and complete postmortem and histopathologic examinations were performed. Plasma concentrations of CCFA remained above the minimum inhibitory concentration (1.0 µg/ml; observed for most avian pathogenic bacteria) for 108 hr. No abnormalities were identified on individual birds before and after clinical pathology results (i.e., hematocrits and plasma biochemistry profiles), and only minimal gross and histopathologic changes such as mild tissue inflammation at the injection site were observed. Based on these results, one IM injection of CCFA at 50 mg/kg seems to be a potential option for treatment of ringneck doves.

Infection With a Novel Rickettsiella Species in Emperor Scorpions (Pandinus imperator).

Rickettsiella infection was diagnosed in 4 adult emperor scorpions (Pandinus imperator) from 2 different collections over a 3-year period. One case had a 2-day history of weakness, failure to lift the tail, or respond to stimulation, with rapid progression to death. The other 3 cases were found dead. There were no gross lesions, but histologically the hemolymphatic vasculature and sinuses, presumed hematopoietic organ, heart, midgut and midgut diverticula, nerves, and skeletal muscle were infiltrated with phagocytic and granular hemocytes with necrosis. Phagocytic hemocytes contained abundant intracellular microorganisms that were Fite's acid-fast-positive, Macchiavello-positive, variably gram-positive or gram-negative, and Grocott's methenamine silver-negative. By transmission electron microscopy, hemocytes contained numerous phagocytic vacuoles with small dense bacterial forms (mean 0.603 × 0.163 µm) interspersed with large bacterial forms (mean 1.265 × 0.505 µm) and few intermediary forms with electron-dense nucleoids and membrane-bound crystalline arrays (average 4.72 µm). Transmission electron microscopy findings were consistent with bacteria of the family Coxiellaceae. Based on sequencing the 16S ribosomal RNA gene, the identity was confirmed as Rickettsiella, and phylogenetic analysis of protein-coding genes gidA, rspA, and sucB genes suggested the emperor scorpion pathogen as a new species. This study identifies a novel Rickettsiella causing infection in emperor scorpions and characterizes the unique pathological findings of this disease. We suggest this organism be provisionally named Rickettsiella scorpionisepticum.

Coelomic Fluid Evaluation in Pisaster ochraceus Affected by Sea Star Wasting Syndrome: Evidence of Osmodysregulation, Calcium Homeostasis Derangement, and Coelomocyte Responses.

Sea Star Wasting Syndrome (SSWS) is one of the largest marine wildlife die-offs ever recorded, killing millions of sea stars from more than 20 Asteroid species from Alaska to Mexico from 2013 to 2015 from yet undetermined cause(s). Coelomic fluid surrounds the sea star's organs, playing critical roles in numerous systemic processes, including nutrient transportation and immune functions. Coelomocytes, which are cellular components of coelomic fluid and considered functionally equivalent to vertebrate leukocytes, are responsible for innate cell-mediated immunity. The objectives of this study were to (1) evaluate changes in coelomic fluid chemistry, coelomocyte counts, and cytology from ochre sea stars (Pisaster ochraceus) (n = 55) with clinical signs consistent with SSWS at varying intensity (SSWS score 1: n = 4, score 2: n = 2, score 3: n = 3, score 4: n = 18, score 5: n = 26) in comparison to coelomic fluid from clinically normal sea stars (n = 26) and to (2) correlate SSWS score with cellular and biochemical analytes. SSWS-affected sea stars had wider ranges of all electrolytes, except calcium; statistically significantly higher chloride, osmolality, and total protein; lower calcium; and higher coelomocyte counts when compared to clinically normal sea stars maintained under identical environmental conditions. Free and/or phagocytized bacteria were noted in 29% (16 of 55) coelomic fluid samples from SSWS-affected sea stars but were absent in clinically normal sea stars. SSWS score correlated significantly with increasing chloride concentration, osmolality, and coelomocyte counts. These chemistry and cytological findings in coelomic fluid of SSWS-affected sea stars provide insight into the pathophysiology of SSWS as these results suggest osmo- and calcium dysregulation, coelomocyte responses, and presumptive opportunistic bacterial infection in SSWS-affected sea stars. This information provides potential future research applications for the development of treatment strategies for sea stars in managed care and for understanding the complexity of various biochemical and cellular pathophysiological mechanisms involved in sea star wasting.

Coelomic Fluid Evaluation in Clinically Normal Ochre Sea Stars Pisaster ochraceus: Cell Counts, Cytology, and Biochemistry Reference Intervals.

Coelomic fluid sampling is a noninvasive technique that is used to access the body fluid of sea stars for diagnostics and research. Given recent mortality events including sea star wasting disease, which has killed millions of sea stars along the Pacific coast since 2013, there is a need for validated diagnostic tests to evaluate sea star health. The objectives of this study were to establish coelomic fluid reference intervals for clinically normal ochre sea stars Pisaster ochraceus in an open system aquarium, to describe the cytologic findings, and to compare the chemistries of coelomic fluid with open system tank water. Coelomic fluid from 26 clinically normal sea stars was sampled for coelomocyte counts, cytologic evaluation, and biochemical analysis including magnesium, sodium, potassium, chloride, calcium, and total protein. The number of coelomocytes and total protein did not fit normal distribution and were excluded from analyses. Reference intervals were established for other chemistry analytes. There was no statistical difference in biochemistries between sea star coelomic fluid and water from five open system tanks, which supports previous evidence that sea stars are osmoconformers. Very low numbers of coelomocytes were observed cytologically. These results provide a useful baseline and diagnostic tool for health assessments of sea stars.

Invertebrate Oncology: Diseases, Diagnostics, and Treatment.

Neoplasia is a documented occurrence across invertebrate taxa, but challenges remain with regard to tumor diagnosis and treatment. Literature reports of neoplasia are frequent in mollusks and insects, infrequent in Cnidaria and crustaceans, and are yet to be documented in Porifera and echinoderms. A significant contribution could be made by veterinary practitioners documenting and treating neoplasms in invertebrates. Traditional methods of veterinary diagnosis are encouraged, but the anatomy and tissue biology of each invertebrate species need to be considered. Most neoplasms described in the invertebrate literature have been considered benign, making external lesions potentially amenable to surgical resection.

MANAGEMENT OF OSTEOMYELITIS CAUSED BY SALMONELLA ENTERICA SUBSP. HOUTENAE IN A TAYLOR'S CANTIL (AGKISTRODON BILINEATUS TAYLORI) USING AMIKACIN DELIVERED VIA OSMOTIC PUMP.

An adult female Taylor's cantil (Agkistrodon bilineatus taylori) presented with marked spinal and mandibular osteomyelitis that cultured positive for Salmonella enterica subsp. houtenae, serovar IV 43:z4,z32:-. Progression of osteomyelitis was arrested by treatment using amikacin (0.026 mg/kg per hour) delivered via subcutaneous osmotic pump for 10 mo, replacing the pump every 4 wk. No adverse effects on renal function were appreciated throughout the course of therapy. Amikacin therapy was discontinued after improvement with treatment, but 5 mo later, bony lesions worsened, and an additional abscess formed at the previous pump site. The animal's condition declined and euthanasia was elected. Postmortem examination confirmed marked osteomyelitis with Salmonella infection of same serovar as the initial biopsy. This report highlights the pathogenicity of the S. enterica subsp. houtenae serovar and the ability to deliver effective amikacin dosage via osmotic pump to arrest osteomyelitis due to salmonellosis in a venomous snake.

Emydid herpesvirus 1 infection in northern map turtles (Graptemys geographica) and painted turtles (Chrysemys picta).

A captive, juvenile, female northern map turtle (Graptemys geographica) was found dead following a brief period of weakness and nasal discharge. Postmortem examination identified pneumonia with necrosis and numerous epithelial, intranuclear viral inclusion bodies, consistent with herpesviral pneumonia. Similar intranuclear inclusions were also associated with foci of hepatocellular and splenic necrosis. Polymerase chain reaction (PCR) screening of fresh, frozen liver for the herpesviral DNA-dependent DNA polymerase gene yielded an amplicon with 99.2% similarity to recently described emydid herpesvirus 1 (EmyHV-1). Molecular screening of turtles housed in enclosures that shared a common circulation system with the affected map turtle identified 4 asymptomatic, EmyHV-1 PCR-positive painted turtles (Chrysemys picta) and 1 asymptomatic northern map turtle. Herpesvirus transmission between painted and map turtles has been previously suggested, and our report provides the molecular characterization of a herpesvirus in asymptomatic painted turtles that can cause fatal herpesvirus-associated disease in northern map turtles.

Three novel herpesviruses of endangered Clemmys and Glyptemys turtles.

The rich diversity of the world's reptiles is at risk due to significant population declines of broad taxonomic and geographic scope. Significant factors attributed to these declines include habitat loss, pollution, unsustainable collection and infectious disease. To investigate the presence and significance of a potential pathogen on populations of critically endangered bog turtles (Glyptemys muhlenbergii) as well sympatric endangered wood (G. insculpta) and endangered spotted (Clemmys guttata) turtles in the northeastern United States, choanal and cloacal swabs collected from 230 turtles from 19 sites in 5 states were screened for herpesvirus by polymerase chain reaction. We found a high incidence of herpesvirus infection in bog turtles (51.5%; 105/204) and smaller numbers of positive wood (5) and spotted (1) turtles. Sequence and phylogenetic analysis revealed three previously uncharacterized alphaherpesviruses. Glyptemys herpesvirus 1 was the predominant herpesvirus detected and was found exclusively in bog turtles in all states sampled. Glyptemys herpesvirus 2 was found only in wood turtles. Emydid herpesvirus 2 was found in a small number of bog turtles and a single spotted turtle from one state. Based on these findings, Glyptemys herpesvirus 1 appears to be a common infection in the study population, whereas Glyptemys herpesvirus 2 and Emydid herpesvirus 2 were not as frequently detected. Emydid herpesvirus 2 was the only virus detected in more than one species. Herpesviruses are most often associated with subclinical or mild infections in their natural hosts, and no sampled turtles showed overt signs of disease at sampling. However, infection of host-adapted viruses in closely related species can result in significant disease. The pathogenic potential of these viruses, particularly Emydid herpesvirus 2, in sympatric chelonians warrants additional study in order to better understand the relationship of these viruses with their endangered hosts.

Uropygial gland squamous cell carcinoma in chinstrap (Pygoscelis antarcticus) and gentoo (Pygoscelis papua) penguins at the Wildlife Conservation Society's Central Park Zoo.

Uropygial, or preen, glands are found in a variety of avian species including penguins. These glands have a multitude of functions and can develop a variety of conditions including impaction, rupture, adenitis, squamous metaplasia, and neoplasia of various types, with squamous cell carcinoma the most commonly reported. A case series of uropygial gland squamous cell carcinoma in five penguins at the Wildlife Conservation Society's Central Park Zoo is described. Most birds were aged (>10 yr) with a history of chronic, recurrent uropygial gland problems including impaction, rupture, abscess formation, or a combination of conditions. Before and after neoplasia diagnosis, these cases were managed conservatively, and palliative care was provided. Because many of these cases were preceded by chronic inflammation, it is possible this inflammation predisposed the uropygial gland to neoplastic transformation, and more aggressive treatment early in the disease process may therefore be warranted.

A Mycoplasma species of Emydidae turtles in the northeastern USA.

Mycoplasma infections can cause significant morbidity and mortality in captive and wild chelonians. As part of a health assessment of endangered bog turtles (Glyptemys muhlenbergii) in the northeastern US, choanal and cloacal swabs from these and other sympatric species, including spotted turtles (Clemmys guttata), eastern box turtles (Terrapene carolina carolina), wood turtles (Glyptemys insculpta), and common snapping turtles (Chelydra serpentina) from 10 sampling sites in the states (US) of Delaware, New Jersey, and Pennsylvania, were tested by PCR for Mycoplasma. Of 108 turtles tested, 63 (58.3%) were PCR positive for Mycoplasma including 58 of 83 bog turtles (70%), three of three (100%) eastern box turtles, and two of 11 (18%) spotted turtles; all snapping turtles (n = 7) and wood turtles (n = 4) were negative. Sequence analysis of portions of the 16S-23S intergenic spacer region and the 16S ribosomal RNA gene revealed a single, unclassified species of Mycoplasma that has been previously reported in eastern box turtles, ornate box turtles (Terrapene ornata ornata), western pond turtles (Emys marmorata), and red-eared sliders (Trachemys scripta elegans). We document a high incidence of Mycoplasma, in the absence of clinical disease, in wild emydid turtles. These findings, along with wide distribution of the identified Mycoplasma sp. across a broad geographic region, suggest this bacterium is likely a commensal inhabitant of bog turtles, and possibly other species of emydid turtles, in the northeastern US.

Densovirus associated with sea-star wasting disease and mass mortality.

Populations of at least 20 asteroid species on the Northeast Pacific Coast have recently experienced an extensive outbreak of sea-star (asteroid) wasting disease (SSWD). The disease leads to behavioral changes, lesions, loss of turgor, limb autotomy, and death characterized by rapid degradation ("melting"). Here, we present evidence from experimental challenge studies and field observations that link the mass mortalities to a densovirus (Parvoviridae). Virus-sized material (i.e., <0.2 µm) from symptomatic tissues that was inoculated into asymptomatic asteroids consistently resulted in SSWD signs whereas animals receiving heat-killed (i.e., control) virus-sized inoculum remained asymptomatic. Viral metagenomic investigations revealed the sea star-associated densovirus (SSaDV) as the most likely candidate virus associated with tissues from symptomatic asteroids. Quantification of SSaDV during transmission trials indicated that progression of SSWD paralleled increased SSaDV load. In field surveys, SSaDV loads were more abundant in symptomatic than in asymptomatic asteroids. SSaDV could be detected in plankton, sediments and in nonasteroid echinoderms, providing a possible mechanism for viral spread. SSaDV was detected in museum specimens of asteroids from 1942, suggesting that it has been present on the North American Pacific Coast for at least 72 y. SSaDV is therefore the most promising candidate disease agent responsible for asteroid mass mortality.