34Monodelphis domestica, also known as the laboratory opossum, is a marsupial native to South 35 America. At birth, these animals are developmentally equivalent to human embryos at 36 approximately 5 weeks of gestation which, when coupled with other characteristics including the 37 size of the animals, the development of a robust immune system during juvenile development, 38 and the relative ease of experimental manipulation, have made M. domestica a valuable model in 39 many areas of biomedical research. However, their suitability as models for infectious diseases, 40 especially diseases caused by viruses such as Zika virus (ZIKV), is currently unknown. Here, we 41 describe the replicative effects of ZIKV using a fetal intra-cerebral model of inoculation. Using 42 immunohistochemistry and in situ hybridization, we found that opossum embryos and fetuses are 43 susceptible to infection by ZIKV administered intra-cerebrally, that the infection persists long 44 term, and that the infection and viral replication consistently results in neural pathology and may 45 occasionally result in global growth restriction. These results demonstrate the utility of M. 46 domestica as a new animal model for investigating ZIKV infection in vivo. This new model will 47facilitate further inquiry into viral pathogenesis, particularly for those viruses that are 48 neurotropic, that may require a host with the ability to support sustained viral infection, and/or 49 that may require intra-cerebral inoculations of large numbers of embryos or fetuses.
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AUTHOR SUMMARY
51Here we show that the laboratory opossum (Monodelphis domestica) is a valuable new model for 52 studying Zika virus pathogenesis. Newborns are at the developmental stage of 5-week human 53 embryos. Zika virus inoculated on a single occasion into the brains of pups at the human 54 developmental stages of 8-20 weeks post conception replicated in neuronal cells and persisted as 55 a chronic infection until the experimental endpoint at 74-days post infection. In addition, we 3 56 observed global growth restriction in one of 16 inoculated animals; global growth restriction has 57 been observed in humans and other animal models infected with Zika virus. The results illustrate 58 great potential for this new animal model for high throughput research on the neurological 59 effects of Zika virus infection of embryos and fetuses. 60 INTRODUCTION 61 Zika virus (ZIKV) is a small, enveloped positive-sense RNA virus from the family 62 Flaviviridae. Typically transmitted in a zoonotic cycle that alternates between a vertebrate host 63 and an invertebrate vector, ZIKV gained notoriety following the 2015 outbreak in Brazil, which 64 saw a dramatic increase in the number of neurological abnormalities in infants born to ZIKV-65 infected mothers [1]. Significant increases in Guillain-Barre syndrome and microcephaly during 66 this outbreak were also observed when compared to previous years [2], perhaps fulfilling the 67 theory posited by Hayes when he declared ZIKV to be neurovirulent [3...