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The family Togaviridae comprises several significant human and veterinary mosquito-borne pathogens. Two togaviruses (genus Alphavirus) have been previously identified in association with marine mammals, the southern elephant seal virus (SESV) and Eastern equine encephalitis virus (EEEV) from a fatal captive harbor seal infection. Herein we report the ultrastructural and phylogenomic characterization of a novel marine togavirus, the first isolated from a cetacean, an Alaskan harbor porpoise (Phocoena phocoena) displaying ulcerative dermatitis. A skin sample was processed for virus isolation on Vero.DogSLAMtag cells and cytopathic effects (CPE) were observed on primary isolation approximately 20 days post-infection. Transmission electron microscopy of the infected Vero.DogSLAMtag cells revealed typical alphavirus particles budding from both plasma and vacuolar membranes of infected cells. A next-generation sequencing approach was used to determine the near complete genome of the Alaskan harbor porpoise alphavirus (AHPV). Phylogenetic analysis supported the AHPV as the sister species to the SESV, forming a marine mammal alphavirus clade separate from the recognized alphavirus antigenic complexes. Genetic comparison of the protein coding sequence of the AHPV to other alphaviruses demonstrated amino acid identities ranging from 42.1-67.1%, with the highest identity to the SESV. Based on its genetic divergence, we propose the AHPV represents a novel alphavirus species, pending formal proposal to and ratification by the International Committee on Taxonomy of Viruses. The ecological and genetic characteristics of the AHPV and the SESV also suggest they represent a novel antigenic complex within the genus Alphavirus, which we propose to be named the Marine Mammal Virus Complex. The role of the AHPV in the associated harbor porpoise cutaneous pathology, if any, remains unclear. Further research is needed to determine AHPV's route(s) of transmission and potential vectors, host range, prevalence, and pathogenicity in cetaceans including harbour porpoises.

Summary Background In May, 2013, an outbreak of symptomatic hepatitis A virus infections occurred in the USA. Federal, state, and local public health officials investigated the cause of the outbreak and instituted actions to control its spread. We investigated the source of the outbreak and assessed the public health measures used. Methods We interviewed patients, obtained their shopping information, and did genetic analysis of hepatitis A virus recovered from patients' serum and stool samples. We tested products for the virus and traced supply chains. Findings Of 165 patients identified from ten states, 69 (42%) were admitted to hospital, two developed fulminant hepatitis, and one needed a liver transplant; none died. Illness onset occurred from March 31 to Aug 12, 2013. The median age of patients was 47 years (IQR 35–58) and 91 (55%) were women. 153 patients (93%) reported consuming product B from retailer A. 40 patients (24%) had product B in their freezers, and 113 (68%) bought it according to data from retailer A. Hepatitis A virus genotype IB, uncommon in the Americas, was recovered from specimens from 117 people with hepatitis A virus illness. Pomegranate arils that were imported from Turkey—where genotype IB is common—were identified in product B. No hepatitis A virus was detected in product B. Interpretation Imported frozen pomegranate arils were identified as the vehicle early in the investigation by combining epidemiology—with data from several sources—genetic analysis of patient samples, and product tracing. Product B was removed from store shelves, the public were warned not to eat product B, product recalls took place, and postexposure prophylaxis with both hepatitis A virus vaccine and immunoglobulin was provided. Our findings show that modern public health actions can help rapidly detect and control hepatitis A virus illness caused by imported food. Our findings show that postexposure prophylaxis can successfully prevent hepatitis A illness when a specific product is identified. Imported food products combined with waning immunity in some adult populations might make this type of intervention necessary in the future. Funding US Centers for Disease Control and Prevention, US Food and Drug Administration, and US state and local public health departments.

We report here the first documented case of a cetacean fatality from entanglement in recreational Spectra® fishing twine. Spectra® twine is a relatively new microfilament braided twine that is marketed to replace nylon monofilament twine in rod and reel fisheries. Following the case of this entangled bottlenose dolphin (Tursiops truncatus), we conducted tests with Spectra® and comparable monofilament twines on Tursiops tissue from stranded animals to compare the abrasion properties of the twines. We found that Spectra® twine was significantly more abrasive on bottlenose dolphin fluke tissue than a similar strength and diameter monofilament. With the same forces applied, the Spectra® twine cut deeper than the monofilament, exhibiting a linear relationship with force applied where the monofilament appeared to reach a maximum depth of penetration of approximately 2 mm. These tests may explain why this bottlenose dolphin was so severely debilitated from carrying a relatively light load of twine over a short period of time (20 days). Future public and corporate outreach will be essential to minimize the effect that this increasingly popular fishing twine will have on non-target species.

The neurotoxin domoic acid (DA) was detected in urine and fecal samples recovered from pygmy sperm whales (Kogia breviceps) and dwarf sperm whales (Kogia sima) stranding along the U.S. Atlantic coast from 1997 to 2008. Of the 41 animals analyzed from Virginia, North Carolina, South Carolina and Florida, 24 (59%) tested positive for DA at concentrations of 0.4–1.8 ng/mL in urine and 12–13,566 ng/g in feces as determined by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Feces appeared to be the best indicator of DA exposure in Kogia spp., with 87% of all fecal samples analyzed testing positive for this toxin. Additional stranded animals (n = 40) representing 11 other cetacean species were recovered from the same region between 2006 and 2008 and analyzed by LC–MS/MS, however DA was not detected in any of these individuals. DA is produced naturally by diatoms in the genus Pseudo-nitzschia. Although blooms of DA-producing Pseudo-nitzschia have been associated with repeated large-scale marine mammal mortalities on the west coast of the U.S., there is no documented history of similar blooms on the southeast U.S. coast, and there were no observed Pseudo-nitzschia blooms in the region associated with any of these strandings. The feeding habits of Kogia spp. are poorly documented; thus, the vector(s) for DA exposure to these deep-diving species remains to be identified. Toxin accumulation in these pelagic whale species may be an indication of cryptic harmful algal bloom activity in offshore areas not currently being monitored. This study highlights the need for a better understanding of the role of toxigenic algae in marine mammal morbidity and mortality globally. Published by Elsevier B.V.

Toxicologic sampling in cases of suspected toxic exposures, be they accidental or intentional, involves not only animals, living or dead, but also the environment. Diagnosis of a toxic exposure is not just laboratory driven, but also involves history, clinical signs, diagnostic tests, and necropsy findings including gross and histologic results. Although the list of toxic agents and tests available is extensive, the samples to be collected are more focused. The purpose of this techniques article is to familiarize practitioners with the components of a toxicologic investigation and to enhance sampling techniques.

Field necropsies of individual or multiple animals can be fraught with difficulty and frustration. Planning and preparation for these events, including development of required supplies and equipment checklists, necropsy forms, designated test sites, and a personnel pool, can alleviate many problems. Flexibility is paramount in field situations as is the need for individuals from multiple disciplines to collaboratively collect, record, disseminate, and interpret findings.