Isolation effects of GoAstV JL01 using different avian embryos and inoculation routes
Goose astrovirus(GoAstV)can cause gout and even death in goslings aged between 1 to 15 days.In order to explore effective methods for isolating GoAstV,the homogenized samples of gosling liver and kidney identified as GoAstV positive by RT-PCR were inoculated into SPF chicken embryos,SPF duck embryos and non-SPF goose astrovirus by the embryo yolk sac(YA),chorioallan-toic membrane(CAM)and chorioallantoic antrum(CAA)routes.The optimal method for isolating GoAstV JL01 on different avian embryos would be determined by comprehensive analyzing the results of the pathological changes in the embryos,mortality rates,and the detection of nucleic acid and viral loads after multiple passages.The results indicated that inoculation through the CAA route to non-SPF goose astrovirus and SPF chicken embryos,the CAM route to non-SPF goose astrovirus,SPF chicken embryos,and SPF duck embryos,and the YA route to SPF duck embryos did not lead to significant pathological changes or regular mortality in the embryos.RT-PCR and RT-qPCR tests revealed that the virus could not effectively replicate in the embryos,allantoic fluid or other tissues.However,when SPF chicken embryos were inoculated through the YA route and passed through 10 generations,all exhibited characteristic lesions such as retardation and hemorrhaging.All the generations except for the F1 died.The yolk sac and embryo tissues contained 104.5-106.0 copies of goose astrovirus per 1 μL of tissue cDNA.Therefore,inoculation via the YA route in 8-day-old SPF chicken embryos was identified as the optimal method for isolating GoAstV JL01.This study provided a reference pathway for the isolation of GoAstV.
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