Genomic recombination between infectious laryngotracheitis vaccine strains occurs under a broad range of infection conditions in vitro and in ovo

Gallid alphaherpesvirus 1 causes infectious laryngotracheitis (ILT) in farmed poultry worldwide. Intertypic recombination between vaccine strains of this virus has generated novel and virulent isolates in field conditions. In this study, in vitro and in ovo systems were co-infected and superinfected under different conditions with two genomically distinct and commonly used ILTV vaccines. The progeny virus populations were examined for the frequency and pattern of recombination events using multi-locus high-resolution melting curve analysis of polymerase chain reaction products. A varied level of recombination (0 to 58.9%) was detected, depending on the infection system (in ovo or in vitro), viral load, the composition of the inoculum mixture, and the timing and order of infection. Full genome analysis of selected recombinants with different in vitro phenotypes identified alterations in coding and non-coding regions. The ability of ILTV vaccines to maintain their capacity to recombine under such varied conditions highlights the significance of recombination in the evolution of this virus and demonstrates the capacity of ILTV vaccines to play a role in the emergence of recombinant viruses.