Isolation, Identification, and Characterization of Novel Arenaviruses, the Etiological Agents of Boid Inclusion Body Disease

Abstract

g Boid inclusion body disease (BIBD) is a progressive, usually fatal disease of constrictor snakes, characterized by cytoplasmic inclusion bodies (IB) in a wide range of cell types. To identify the causative agent of the disease, we established cell cultures from BIBD-positive and -negative boa constrictors. The IB phenotype was maintained in cultured cells of affected animals, and supernatants from these cultures caused the phenotype in cultures originating from BIBD-negative snakes. Viruses were purified from the supernatants by ultracentrifugation and subsequently identified as arenaviruses. Purified virus also induced the IB phenotype in naive cells, which fulfilled Koch's postulates in vitro. One isolate, tentatively designated University of Helsinki virus (UHV), was studied in depth. Sequencing confirmed that UHV is a novel arenavirus species that is distinct from other known arenaviruses including those recently identified in snakes with BIBD. The morphology of UHV was established by cryoelectron tomography and subtomographic averaging, revealing the trimeric arenavirus spike structure at 3.2-nm resolution. Immunofluorescence, immunohistochemistry, and immunoblotting with a polyclonal rabbit antiserum against UHV and reverse transcription-PCR (RT-PCR) revealed the presence of genetically diverse arenaviruses in a large cohort of snakes with BIBD, confirming the causative role of arenaviruses. Some snakes were also found to carry arenavirus antibodies. Furthermore, mammalian cells (Vero E6) were productively infected with UHV, demonstrating the potential of arenaviruses to cross species barriers. In conclusion, we propose the newly identified lineage of arenaviruses associated with BIBD as a novel taxonomic entity, boid inclusion body disease-associated arenaviruses (BIBDAV), in the family Arenaviridae.

Doença do corpúsculo de inclusão e espondilite por Salmonella sp. em uma Boa constrictor constrictor

“…Doença do corpúsculo de inclusão (Inclusion body disease -IBD) é uma enfermidade que afeta serpentes, principalmente Pythonidae e Boidae (Schumacher et al 1994, Schumacher 2006, Chang & Jacobson 2010, Stenglein et al 2012, Hetzel et al 2013, Chang et al 2013, Turchetti et al 2013. Foi descrita pela primeira vez há quatro décadas e têm sido uma preocupação mundial devido à alta morbidade e mortalidade (Chang & Jacobson 2010, Stenglein et al 2012, Hetzel et al 2013, Chang et al 2013.…”

“…Foi descrita pela primeira vez há quatro décadas e têm sido uma preocupação mundial devido à alta morbidade e mortalidade (Chang & Jacobson 2010, Stenglein et al 2012, Hetzel et al 2013, Chang et al 2013. Entretanto, até o momento, só há um relato de caso de IBD no Brasil (Turchetti et al 2013).…”

“…A doença se caracteriza por corpúsculos intracitoplasmáticos em diversos tecidos (Schumacher et al 1994, Schumacher 2006, Chang & Jacobson 2010, Stenglein et al 2012, Chang et al 2013, Hetzel et al 2013, Turchetti et al 2013) e os sinais clínicos são consequência das inclusões no sistema nervoso central, levando a opistótono, desequilíbrio, paralisia flácida e inabilidade de voltar à posição quando colocados em decúbito dorsal (Chang & Jacobson 2010, Stenglein et al 2012, anisocoria e tremores da cabeça (Hetzel et al 2013). Além dos sinais neurológicos, sinais clínicos inespecíficos como anorexia, apatia (Hetzel et al 2013, Turchetti et al 2013) e regurgitação já foram associados à enfermidade (Schumacher et al 1994, Chang & Jacobson 2010, Stenglein et al 2012, Hetzel et al 2013, Turchetti et al 2013.…”

“…Além dos sinais neurológicos, sinais clínicos inespecíficos como anorexia, apatia (Hetzel et al 2013, Turchetti et al 2013) e regurgitação já foram associados à enfermidade (Schumacher et al 1994, Chang & Jacobson 2010, Stenglein et al 2012, Hetzel et al 2013, Turchetti et al 2013.…”

“…Recentemente, identificou-se que um Arenavirus modificado é o responsável pela doença (Stenglein et al 2012, Bodewes et al 2013, Hetzel et al 2013). Acredita-se que a transmissão ocorra por contato direto entre animais infectados e por via transovariana.…”