title: Evolution of Newcastle Disease Virus Quasispecies Diversity and Enhanced Virulence after Passage through Chicken Air Sacs.
abstract: It has been reported that lentogenic Newcastle disease virus (NDV) isolates have the potential to become velogenic after their transmission and circulation in chickens, but the underlying mechanism is unclear. In this study, a highly velogenic NDV variant, JS10-A10, was generated from the duck-origin lentogenic isolate JS10 through 10 consecutive passages in chicken air sacs. The velogenic properties of this selected variant were determined using mean death time (MDT) assays, intracerebral pathogenicity index (ICPI), the intravenous pathogenicity index (IVPI), histopathology, and the analysis of host tissue tropism. In contrast, JS10 remained lentogenic after 20 serial passages in chicken eggs (JS10-E20). The JS10, JS10-A10, and JS10-E20 genomes were sequenced and found to be nearly identical, suggesting that both JS10-A10 and JS10-E20 were directly generated from JS10. To investigate the mechanism for virulence enhancement, the partial genome covering the F0 cleavage site of JS10 and its variants were analyzed using ultradeep pyrosequencing (UDPS) and the proportions of virulence-related genomes in the quasispecies were calculated. Velogenic NDV genomes accumulated as a function of JS10 passaging through chicken air sacs. Our data suggest that lentogenic NDV strains circulating among poultry might be a risk factor to future potential velogenic NDV outbreaks in chickens.
  
  
  OBJECTIVE
  An avirulent isolate, JS10, was passaged through chicken air sacs and embryos, and the pathogenicity of the variants was assessed. A virulent variant, JS10-A10, was generated from consecutive passage in air sacs. We developed a deep-sequencing approach to detect low-frequency viral variants across the NDV genome. We observed that virulence enhancement of JS10 was due to the selective accumulation of velogenic quasispecies and the concomitant disappearance of lentogenic quasispecies. Our results suggest that because it is difficult to avoid contact between natural waterfowl reservoirs and sensitive poultry operations, circulating lentogenic NDV strains may represent a potential reservoir for emergent velogenic NDV strains that could cause outbreaks in chickens.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Chunchun
authors.initials: C
authors.lastname: Meng
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Xusheng
authors.initials: X
authors.lastname: Qiu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Shengqing
authors.initials: S
authors.lastname: Yu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Chuanfeng
authors.initials: C
authors.lastname: Li
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Zongyan
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Kaichun
authors.initials: K
authors.lastname: Liu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Xiangle
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Tan
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Cuiping
authors.initials: C
authors.lastname: Song
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China.
authors.forename: Guangqing
authors.initials: G
authors.lastname: Liu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, People's Republic of China shoveldeen@shvri.ac.cn.
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
mesh_terms: D000220:Adaptation, Biological; D000400:Air Sacs / Q000821:virology; D000818:Animals; D001921:Brain / Q000473:pathology; D002645:Chickens; D004372:Ducks; D014644:Genetic Variation; D016679:Genome, Viral; D006651:Histocytochemistry; D008969:Molecular Sequence Data; D009521:Newcastle Disease / Q000473:pathology* / Q000821:virology*; D009522:Newcastle disease virus / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D011201:Poultry Diseases / Q000473:pathology* / Q000821:virology*; D017422:Sequence Analysis, DNA; D012692:Serial Passage; D016019:Survival Analysis; D056189:Viral Tropism; D014774:Virulence