Abstract
The seal picornavirus 1, species Aquamavirus A, is currently the only recognized member of the genus Aquamavirus within the family Picornaviridae. The bear picornavirus 1 was recently proposed as the second species in the genus under the name aquamavirus B. Herein, we determined the complete genomes of two novel pinniped picornaviruses, the harbor seal picornavirus (HsPV) and the ribbon seal picornavirus (RsPV). The HsPV and the RsPV were isolated in Vero.DogSLAMtag cells from samples collected from stranded harbor (Phoca vitulina) and ribbon (Histriophoca fasciata) seals. RsPV-infected Vero.DogSLAMtag cells displaying extensive cytopathic effects were processed for transmission electron microscopy and revealed non-enveloped viral particles aggregated into paracrystalline arrays in the cytoplasm. A next-generation sequencing approach was used to recover the complete genomes of the HsPV and the RsPV (6,709 and 6,683 bp, respectively). Phylogenetic and genetic analyses supported the HsPV and the RsPV as members of the Aquamavirus genus. Based on these results, RsPV represents a novel strain of Aquamavirus A, while the HsPV is a novel strain of the proposed species aquamavirus B. These discoveries provide information on the evolutionary relationships and ultrastructure of aquamaviruses and expands the known host range of those viruses. Our results underscore the importance of the application of classical virology and pathology techniques coupled with high-throughput sequencing technologies for the discovery and characterization of pathogens in wild marine mammals.
Highlights
The family Picornaviridae is a diverse assemblage of viruses that possess small spherical nucleocapsids and positive-sense RNA genomes ranging between 7 and 8.8 kb in size [1, 2]
The Vero.DogSLAMtag cultures inoculated with harbor seals spleen, mediastinal lymph node, and brain tissues developed cytopathic effects (CPE) after three blind passages
The flask inoculated with the ribbon seal rectal swab was selected for Transmission Electron Microscopy (TEM) and showed multiple vesicles within the cytoplasm of Vero.DogSLAMtag cells, along with virus particles aggregated into polygonal paracrystalline arrays (Figure 2)
Summary
The family Picornaviridae (order Picornavirales) is a diverse assemblage of viruses that possess small spherical nucleocapsids and positive-sense RNA genomes ranging between 7 and 8.8 kb in size [1, 2]. Picornaviruses may cause subclinical to severe infections in humans and animals, such as febrile illness and diseases of the heart, liver, respiratory tract, gastrointestinal tract, and central nervous system [3, 4]. They cause diseases of significance to public health including the common cold [most commonly caused by rhinovirus A, B, and C; [5]], poliomyelitis [caused by poliovirus; [6]], and hepatitis A [caused by hepatovirus A; [7]]. Enterovirus 76 is associated with diseases including aseptic meningitis, encephalitis, and myocarditis [9]
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