Name: Fei Fan Ng

Email: fng@mail.usf.edu

Author: Terry Fei Fan Ng 1, Charles Manire 2, Wm. Kirk Suedmeyer 3, Shankar Alavandi 4, Kelly Borrowman 5, Tammy Langer 6, Elizabeth Wheeler7 , Frances Gulland 7, Llewellyn Ehrhart 5, and Mya Breitbart 1

Author affiliation: 1) University of South Florida, Saint Petersburg, FL, USA; 2) Atlantis, Paradise Island, Bahamas; 3) Kansas City Zoo, Kansas City, MO, USA; 4) Central Institute of Brackishwater Aquaculture, Chennai, India; 5)University of Central Florida, Orlando, FL, USA; 6) Volusia Marine Science Center, Ponce Inlet, FL, USA; 7) Marine Mammal Center, CA, USA

Abstract title: Discovering Novel Marine Animal Viruses Using Viral Metagenomics

Absstract:

New diseases in marine animals are emerging at an increasing rate and the causative agents of these diseases are mostly unknown. Currently, diagnosing novel viral infections is difficult due to limitations of virus discovery methods. PCR, immunological assays, and pan-viral microarrays are only useful in identifying viruses with high levels of similarity to known viruses. Viral metagenomics, which involves viral particle purification and shotgun sequencing, has proven to be useful for describing novel viruses, including those with limited homology to previously described viral families. Viral metagenomics is useful for investigating unknown etiological agents in mortality events, characterizing viruses in known disease, and assessing existing viral pathogens (both known and unknown) in endangered species. We have successfully performed viral metagenomics directly from animal tissues, resulting in the discovery of two novel single-stranded DNA viruses. Sea turtle tornovirus 1 (STTV1) and California sea lion anellovirus (ZcAV) both share only limited sequence similarity to previously described viral genomes (1, 2). STTV1 was discovered from the fibropapilloma of a Florida green sea turtle. The single-stranded, circular genome of STTV1 was approximately 1800 nucleotides in length. STTV1 only had weak amino acid level identities (25%) to Chicken Anemia Virus in short regions of its genome, hence STTV1 may represent the first member of a novel virus family. ZcAV was an anellovirus discovered from the lung of a California sea lion that died in a respiratory-related mortality event. The single-stranded circular genome of ZcAV was 2140 nucleotides in length, with only 35% amino acid identity to feline anelloviruses in the ORF1 region. Our studies have shown that viral metagenomics can be used to discover viruses directly from many tissue types in a wide range of animals. Therefore, this approach has the potential to significantly enhance the fields of virus discovery and infectious disease monitoring. LITERATURE CITED 1. Ng, T. F. F., C. Manire, K. Borrowman, T. Langer, L. Ehrhart, and M. Breitbart. 2009. Discovery of a novel single-stranded DNA virus from a sea turtle fibropapilloma using viral metagenomics. J. Virol. 83:2500-2509. 2. Ng, T. F. F., W. K. Suedmeyer, F. Gulland, E. Wheeler, and M. Breitbart. 2009. Novel anellovirus discovered from a mortality event of captive California sea lions. J. Gen. Virol. 90:1256 -1261.