Turkey Hemorrhagic Enteritis Virus Can Be Titrated but Not Propagated in Chicken Embryos

Abstract

This study aimed to investigate the feasibility of propagating and titrating hemorrhagic enteritis virus (HEV) in chicken embryos. A total of 308 embryonated eggs were used. At 10 days of embryonic age, eggs were inoculated via allantoic sac or chorioallantoic membrane routes with non–heat-treated (live) HEV or heat-treated (dead) HEV or served as negative controls. Allantoic fluid retrieved at 0, 1, 3, 5, and 7 days postinoculation (dpi) was tested for HEV by quantitative PCR. Inoculation with HEV did not cause visible growth impairment or lesions in the chicken embryos. Overall, there was no difference in postinoculation mortality rates among groups sham-inoculated (6/30, 20.0%) or inoculated with live (34/252, 13.4%) or dead (3/26, 6.9%) HEV (𝑃 = 0.58). The amount of HEV DNA detected in allantoic fluid at 7 dpi in eggs inoculated with live virus was similar to the inoculated dose, indicating that virus propagation in chicken embryos is not efficient. No HEV DNA was detected after 3 dpi in eggs inoculated with dead virus. Inoculation of chicken embryos combined with qualitative PCR can be used for titration of HEV virus stocks and presents a high correlation with in vivo titration using chickens (R² 0.98, 𝑃 = 0.007). This method may be relevant in countries in which specific-pathogen-free turkeys are unavailable and in which the importation of RP19 cells, the only cell that supports effective propagation of HEV, is not permitted.