Differential host passaging alters pathogenicity and induces genomic variation in white spot syndrome virus

To understand the pathological effects of passaging WSSV through different animals, we passaged WSSV through crabs ( Portunus sanguinolentus and P. pelagicus), prawn ( Macrobrachium rosenbergii) and artemia ( Artemia sp.). Among the three geographically distinct WSSV isolates (A166, G9 and G27) that caused over 95% mortalities in experimental infections in P. monodon, only two isolates (A166 and G9) caused mortality (5%) in M. rosenbergii. On subsequent infection of P. monodon with the passage one virus (P1V) from M. rosenbergii, only P1V of A166 caused 5% mortality while P1V of G9 failed to cause any mortality, though the infected shrimp were positive for WSSV by nested PCR. Crabs showed differential susceptibility to WSSV isolates. While passaging WSSV (G9) through P. sanguinolentus retained pathogenicity, passaging through P. pelagicus completely attenuated the two strains (A117 and G27). Passaging through M. rosenbergii changed pathogenicity of isolates (A166 and G9) from 95% to 5% cumulative mortality in experimental infections. WSSV was unable to infect Artemia and could not transmit the disease to P. monodon. WSSV (A117) passaged through P. monodon retained its virulence. An examination of tandem repeats (TR) in ORF 94 of the WSSV genome revealed variations upon passaging through different hosts. One isolate (A117), with 12 TR yielded 8 TR when passaged through P. pelagicus and 18 TR when passaged through P. monodon. Two other isolates (G9 and G27) with 9 TR, retained them when passaged through P. monodon and P. sanguinolentus but yielded 8 and 12 TR when passaged through P. pelagicus and M. rosenbergii, respectively. Thus, differences in TR number appeared to result from host selection rather than geographical isolation.