摘要
In order to obtain a fowl avidenovirus type 4 strain with good immunogenicity,chicken liver tissues suspected of adenovirus infection in a chicken farm in Binzhou were treated and then inoculated to chicken liver hepatocellular carcinoma cells(LMH).The cell cultures were extracted for DNA,which was subjected to PCR identification and sequencing analysis,and animal regression test and immunogenicity test were also carried out.The results showed that one fowl avidenovirus strain was successfully isolated.The isolated strain was inoculated to LMH cells,and the first generation showed obvious cytopathic changes.The PCR identification result of the 8^(th)generation cell culture of the isolated virus strain on LMH cells was positive.The sequencing result and NCBI sequence alignment analysis showed that the isolated virus strain had the highest nucleotide similarity with fowl avidenovirus type 4,reaching 100%,indicating that the isolated strain was of fowl avidenovirus type 4.The strain could cause the death of 21-day-old SPF chickens,with a mortality rate of 100%,and could completely replicate the same symptoms as clinically infected chickens after being challenged.The three batches of oil vaccine prepared with the isolated strain had a protection rate of 100%,and the geometric mean values of serum agar expansion titers were 1∶4.6,1∶4.9,and 1∶4.6,respectively.It can be seen that the isolated virus is of fowl avidenovirus type 4 in group I,and has good immunogenicity.
基金
Supported by Poultry Innovation Team Project of Agriculture Research System in Shandong Province(SDAIT-11-16)
2017 Shandong Province Foreign Experts Double Hundred Plan Project(2017 Double Hundred Program for Chinese and British Foreign Experts)
Key Project of Natural Science Foundation of Shandong Province(ZR2020KC006)。
