Biological Characterization and Evolutionary Dynamics of Pigeon Paramyxovirus Type 1 in China

Pigeon paramyxovirus type 1 (PPMV-1) is considered as an antigenic variant of Newcastle disease virus (NDV) which has an obvious host preference for pigeons and has caused significant economic losses to the global poultry industry. The evolutionary dynamics of PPMV-1 in China, however, are poorly understood. In this study, we characterized seven PPMV-1 isolates from diseased pigeons collected in Jiangsu, Anhui, and Henan provinces during 2020. Phylogenetic analysis revealed that seven isolates belonged to sub-genotype VI.2.1.1.2.2. Biological characterization indicated that seven isolates were mesogenic based on the mean death time (69.6–91.2 h) and intracerebral pathogenicity index (1.19–1.40) and had similar growth kinetics in chicken embryos and CEFs. Furthermore, the four representative viruses (AH/01/20/Pi, JS/06/20/Pi, HN/01/20/Pi, and HN/02/20/Pi) could result in marked cytopathic effects (CPE) in CEFs and induced syncytium formation in Vero cells. Our Bayesian phylogenetic analysis showed that PPMV-1 might first emerge in East China in 1974 and East China had the highest genotypic diversity of PPMV-1. Besides, phylogeographic analysis indicated that East China and South China were probably the major epicenters of dissemination of PPMV-1 in China. Selection pressure analysis and amino acid substitutions analysis revealed that the viral replication complex (NP, P, and L proteins) was likely related with the host preference of PPMV-1. Collectively, this study uncovered the epidemiology and evolutionary dynamics of PPMV-1 circulating in China, emphasizing the importance of strengthening the monitoring of PPMV-1 in East China and South China and providing significant clues for further studies on the molecular mechanism underlying host preference of PPMV-1.

Evolutionary Dynamics and Age-Dependent Pathogenesis of Sub-Genotype VI.2.1.1.2.2 PPMV-1 in Pigeons

Pigeon paramyxovirus type 1 (PPMV-1) infection causes high morbidity in pigeons, resulting in a significant burden to the poultry industry. In this study, we isolated three PPMV-1 strains from diseased pigeons collected in Guangdong Province, South China, from June 2017 to April 2019. Genetic analysis revealed that these three PPMV-1 strains and most of the PPMV-1 strains isolated from China after 2011 were clustered into sub-genotype VI.2.1.1.2.2. Our Bayesian analysis revealed that the VI.2.1.1.2.2 viruses might have originated in Europe. Phylogeographic analyses revealed that East and South China might have played a key role in seeding the VI.2.1.1.2.2 PPMV-1 epidemic in China. To characterize the effect of age at infection on the outcome of PPMV-1 infection in pigeons, we investigated the pathogenesis and transmission of the pigeon/Guangdong/GZ08/2017 (GZ08) virus in 3-, 6-, and 12-week-old pigeons. Two of six 12-week-old pigeons inoculated with GZ08 survived, and all of the 3- and 6-week-pigeons inoculated with GZ08 died. Moreover, the GZ08 virus could be transmitted to 3-, 6-, and 12-week-old naïve contact pigeons. The lethality of the GZ08 virus through contact with 3-, 6-, and 12-week-old pigeons was 100%, 66.7%, and 0%, respectively, suggesting that the transmissibility of the GZ08 virus was stronger in young pigeons. These findings demonstrated that East and South China was the epicenter for dissemination of VI.2.1.1.2.2 PPMV-1, and age at infection has an impact on the outcome of PPMV-1 infection in pigeons.

Genetic changes in pigeon paramyxovirus type-1 induced by serial passages in chickens and microscopic lesions caused by the virus in various avian hosts

Introduction: Genotype VI of avian avulavirus 1 (AAvV-1) has pigeons and doves as its reservoir and is often termed pigeon paramyxovirus type-1 (PPMV-1). The pathogenesis of PPMV-1 infections in poultry is largely obscure. It is known that PPMV-1 requires a series of passages in chickens before it becomes adapted to gallinaceous poultry.Material and Methods: Changes in the genome of PPMV-1 were analysed after serial passages in specific pathogen free (SPF) chickens, using high-throughput sequencing. Additionally, histopathological lesions induced by PPMV-1 in experimentally inoculated pigeons, chickens, and turkeys were evaluated.Results: Following six passages of PPMV-1 in chickens, 10 nonsynonymous substitutions were found including one (in the NP protein) which dominated the genetic pool of viral quasispecies. Histopathological changes induced by the post-passage PPMV-1 strain were more prominent than changes wrought by the pre-passaged PPMV-1 strain and the lesions were most intense in pigeons followed by chickens and turkeys.Conclusion: PPMV-1 is highly adapted to pigeons and passaging through chickens results in the acquisition of novel amino acids in the polymerase complex, which may alter the pathogenic potential of the virus.