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White spot syndrome virus (WSSV) infection is commonly detected by vp 28-qPCR assay in wild crayfish, Procambarus clarkii , a widespread crustacean species in the aquatic environment in China. The virions of crayfish WSSV have been isolated and purified. Based on TEM observation, they exhibited morphological structures that are identical to known WSSV. In addition, the WSSV major envelope protein VP28 was observed based on Western blot analysis of the total structural proteins of crayfish WSSV. PCR amplification and sequencing analyses of variable regions of ORF14/15, ORF23/24 and ORF94, along with viral genomic sequencing and phylogenomic analysis, indicated that the crayfish WSSV, named WSSV-CN-Pc, represents a new WSSV genotype. Intramuscular injection bioassay revealed that WSSV-CN-Pc was as virulent as the WSSV Taiwan strain. The WSSV-CN-Pc exhibited characteristics of a dominant genotype, with high infection load (10 7 -10 8  WSSV/mg) and high prevalence (91.7%, 110 of 120 crayfish samples) observed in the surveyed wild crayfish. WSSV-CN-Pc was also detected, with similar infection pattern as observed in crayfish, in farmed Litopenaeus vannamei shrimp that shared similar ecological niches with the sampled crayfish. Our results indicated that there was horizontal transmission of WSSV-CN-Pc between crayfish and shrimp in nature. Our findings also implicated that crayfish and shrimp farming should be integrated and managed with cautions in order to reduce the risk of spread and circulation of WSSV in the aquatic environment.

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Characterization and prevalence of a novel white spot syndrome viral genotype in naturally infected wild crayfish, Procambarus clarkii, in Shanghai, China