Type II Grass Carp Reovirus Rapidly Invades Grass Carp ( Ctenopharyngodon idella ) via Nostril-Olfactory System-Brain Axis, Gill, and Skin on Head

Type II grass carp reovirus (GCRV-II) with high pathogenicity and infectivity causes severe hemorrhagic disease, which leads to extensive death in the grass carp and black carp aquaculture. However, the early invasion portal remains unclear. In this study, we explored the invasion portal, time, and pathway of GCRV-II by immersion infection in grass carp. Through the detection of the infected grass carp external body surface tissues, most of them could be detected to carry GCRV-II within 45 min except for the skin covered by scales. Further shortening the duration of infection, we proved that GCRV-II rapidly invades through the nostril (especially), gill, and skin on head at only 5 min post-immersion, rather than merely by adhesion. Subsequently, visual localization investigations of GCRV-II were conducted on the nostril, olfactory system (olfactory bulb and olfactory tract), and brain via immunofluorescence microscopy and transmission electron microscopy. We found that few viruses were located in the nostril at 5 min post-immersion infection, while a significantly increased quantity of viruses were distributed in all of the examined tissues at 45 min. Furthermore, the semi-qRT-PCR and Western blotting results of different infection times confirmed that GCRV-II invades grass carp via the nostril-olfactory system-brain axis and then viral replication unfolds. These results revealed the infection mechanism of GCRV-II in terms of the invasion portal, time, and pathway in grass carp. This study aims to understand the invasion mode of GCRV-II in grass carp, thus providing theoretical support for the prevention and control strategies of hemorrhagic disease.