Long-term persistence of West Nile virus (WNV) infection within vertebrate reservoir hosts is a potential mechanism for overwintering of this (and other) arbovirus(es) at temperate latitudes. The house sparrow (Passer domesticus), an established amplifying host for WNV and other arboviruses, was used as a model to confirm chronicity of WNV infection in passerine birds and to evaluate the feasibility of two overwintering mechanisms: blood-borne infection of arthropod vectors (recrudescence) and oral infection of vertebrate reservoir hosts (ingestion of infected tissues through predation). WNV-inoculated sparrows were monitored for persistent infection for up to 2 years. Infectious virus persisted in tissues through 43 days, but not in sera beyond 6 days. Viral RNA persisted in tissues through 65 days. Chronicity of WNV infection in some tissues, but not blood, supports the predation mechanism of WNV overwintering, but not recrudescence. RNA persistence impacts interpretation and etiologic determination of avian mortality.