Isolation and characterization of an astrovirus causing fatal visceral gout in domestic goslings

Zhang, Qingshui; Cao, Yanxin; Wang, Jun; Fu, Guanghua; Sun, Mengxu; Zhang, Lijiao; Meng, Li; Cui, Guolin; Huang, Yu; Hu, Xueying; Su, Jingliang

To isolate AstV, the kidney, spleen, and liver samples were homogenized with sterile phosphate buffered saline (PBS, pH 7.4) to a 20% suspension (w/v) and centrifuged at 8000×g, at 4°C for 10 min. The supernatant was filtered using a syringe-driven filter unit with a pore size of 0.2 μm and the filtrate was inoculated into five 9-day-old goose embryos (0.2 ml/egg) via the chorioallantoic membrane route. Embryos were incubated at 37°C and candled daily. Embryos that died beyond 24 h and those that survived until 5 day after inoculation were chilled to 4°C overnight. The allantoic fluids were collected for a hemagglutination (HA) activity test performed by a standard method using 1% chicken red blood cells as an indicator, and then were subjected to additional passage in goose embryos.