NOVEL POXVIRUS INFECTION IN NORTHERN AND SOUTHERN SEA OTTERS (ENHYDRA LUTRIS KENYONI AND ENHYDRA LUTRIS NEIRIS), ALASKA AND CALIFORNIA, USA

Abstract

Small superficially ulcerated skin lesions were observed between October 2009 and September 2011 during captive care of two orphaned sea otter pups: one northern (Enhydra lutris kenyoni) in Alaska and one southern (Enhydra lutris nereis) in California. Inclusions consistent with poxviral infection were diagnosed by histopathology in both cases. Virions consistent with poxvirus virions were seen on electron microscopy in the northern sea otter, and the virus was successfully propagated in cell culture. DNA extraction, pan-chordopoxviral PCR amplification, and sequencing of the DNA-dependent DNA polymerase gene revealed that both cases were caused by a novel AT-rich poxvirus. Bayesian and maximum likelihood phylogenetic analyses found that the virus is divergent from other known poxviruses at a level consistent with a novel genus. These cases were self-limiting and did not appear to be associated with systemic illness. To our knowledge, this is the first report of a poxvirus in a mustelid species. The source of this virus, mode of transmission, zoonotic potential, and biological significance are undetermined.

Alphaherpesvirus: Isolation, Identification, Partial Characterisation, Associated Pathologic Findings and Epidemiology in Beluga Whales (Delphinapterus leucas) in Alaska and Arctic Canada

“…Briefly, frozen ampoules were rapidly thawed and 350 μL aliquot samples were aseptically inoculated onto 25 cm 2 tissue culture flasks (Corning Inc., Corning, New York, USA) containing 80% confluent cultures of BWK cells grown in Dulbecco's Minimal Essential Medium/Ham's F-12 (1:1) containing HEPES buffer (to control the pH), supplemented with 10% foetal bovine serum (HyClone Laboratories Inc., Logan, Utah, USA) containing antibiotics (penicillin 200 units mL −1 , streptomycin 200 μg mL −1 , and gentamycin 50 μg mL −1 ) as previously described (Tuomi et al 2014). After adsorption for 1 h at 37°C, the inoculum was removed and 5.0 mL of fresh media were added to each flask.…”

Emerging viruses in marine mammals.

“…These cells have abundant oeosinophilic granular cytoplasm often with a single large round cytoplasmic inclusion body, a round vesicular nucleus and prominent nucleolus. Sea otter cutaneous pox lesions have epidermal hyperplasia with ulceration, and rete peg formation projecting into the dermis [153]. Intracytoplasmic inclusions consistent with Bollinger bodies may be present which ultrastructurally are consistent with pox inclusions.…”

“…A new species of poxvirus related to an orthopoxvirus was associated with raised, ulcerated cutaneous lesions in Steller sea lion (Eumetopias jubatus) pups [147,152]. A novel poxvirus was also described in two orphaned northern and southern sea otters in California [153].…”

Genomic characterization of a novel poxvirus from a flying fox: evidence for a new genus?

“…For poxvirus screening, DNA from lesions was extracted using a DNEasy blood and tissue kit (Qiagen), according to the manufacturer's instructions and the primers Poxpol_556F (5¢-GAYTAYAAYWSNYTNTAYCC NAAYGTITG-3¢) and Poxpol_673R (5¢-RAANCCCATNARNCC RTAIAC-3¢) designed to amplify a segment of approximately 374 bp of the DNA polymerase for all Chordopoxvirinae species (Tuomi et al, 2014) were used. Reactions were performed using GoTaq Green Master Mix (Promega), 1 µM of each primer and 2 µl of DNA template in a On: Thu, 10 May 2018 11:23:21 20 µl reaction with the following reaction conditions: 94 C for 2 min followed by 40 cycles of 94 C for 30 s, 49 C for 45 s and 72 C for 30 s, and a final extension of 72 C for 7 min.…”

“…Due to the gross pathology representing a pox-type lesion, DNA was extracted from the wing lesions and tested for poxviruses by PCR with degenerate primers targeting the viral DNA polymerase. The Poxpol_556F/Poxpol_673R primer set (Tuomi et al, 2014) amplified a product of approximately 350 bp (data not shown). Direct Sanger sequencing of the purified gel band resulted in a 325 bp sequence after trimming off primer sequences.ABLASTNsearchwiththissequencereturnedmultiple hits to the DNA polymerase gene from a very diverse set of poxviruses, all with very similar scores; however, the best match was to the sea otter poxvirus DNA polymerase gene (GenBank Accession KF425534.1).…”