Isolation of Diverse Simian Arteriviruses Causing Hemorrhagic Disease

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S imarteriviruses (order Nidovirales, family Arteri- viridae, subfamily Simarterivirinae) are genetically diverse viruses that naturally infect cercopithecoid monkeys throughout sub-Saharan Africa.Many divergent simarteriviruses can infect macaques, although severity of disease ranges from subclinical to highly fatal in this nonnatural host (1,2).Simarteriviruses are not known to infect humans, but the close evolutionary history of simarterivirus hosts and humans, their interhost and intrahost diversity, and various molecular characteristics suggest that those viruses might pose a zoonotic threat (3-5).Simian hemorrhagic fever virus (SHFV), the best characterized simarterivirus, was identified as the cause of a highly fatal hemorrhagic fever epizootic at a National Institutes of Health quarantine facility in Bethesda, Maryland, USA, in 1964, affecting captive rhesus monkeys from Asia (Macaca mulatta), crab-eating macaques (Macaca fascicularis), and stump-tailed macaques (Macaca arctoides) (6-8).During the next 3 decades, several other epizootics in captive macaques were attributed to SHFV, but retrospective sequencing analyses indicated that other highly diverse simarteriviruses caused some of those outbreaks despite similar clinical manifestations (9).Healthy monkeys from Africa housed adjacent to the affected macaques from Asia were long suspected to be the natural simarterivirus hosts, but simarterivirus identification in healthy wild monkeys from Africa only began in 2011, when metagenomic sequencing was used (10-14).Arteriviruses in general, and simarteriviruses in particular, are difficult to isolate.SHFV will only replicate to high titers in the grivet-derived cell line MA-104 and its subclones (e.g., MARC-145) (5,8,15-19).Although Kibale red colobus virus 1 (KRCV-1) can replicate in MARC-145 cells, replication is inconsistent, and virus production is low.Thus, immortalized cell lines capable of sustaining robust replication of simarteriviruses other than SHFV do not exist.If a novel simarterivirus were to emerge, threatening human or animal health, the lack of in vitro systems for virus propagation and isolation would be a major hurdle in understanding basic virology and developing medical countermeasures.We describe a Isolation of Diverse Simian Arteriviruses Causing Hemorrhagic Disease

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