RESUMO
While farming of the Pacific white shrimp Litopenaeus vannamei is well established in North and South America, the industry has more recently been introduced to Asia, and the Pacific white shrimp is now the most commonly farmed species in Thailand. However, outbreaks of yellow head virus (YHV) disease in the Pacific white shrimp have caused severe economic losses and currently there is no effective prevention or treatment of YHV infections. The YHV-protease double-stranded RNA (YHV-Pro dsRNA) can act as both a prophylactic agent and as a treatment to inhibit YHV replication in infected black tiger shrimp Penaeus monodon. The utility of this methodology to other shrimp species has not, however, been established. The purpose of this study was to determine whether YHV-Pro dsRNA can be applied to the Pacific white shrimp. To assess prophylactic efficiency, YHV-Pro dsRNA was injected into juvenile shrimp 24 h prior to challenge with YHV. Subsequent YHV replication was inhibited by YHV-Pro dsRNA as compared with injection of an unrelated dsRNA. For therapeutic treatment of YHV-infected shrimp, shrimp were challenged with YHV before dsRNA injection. Injection of YHV-Pro dsRNA up to 6 h post-infection resulted in the almost complete elimination of YHV replication. These results suggest that YHV-Pro dsRNA can also be broadly applied as a prophylactic agent to inhibit YHV replication and therapeutic treatment of YHV-infected Pacific white shrimp.
Assuntos
Penaeidae/virologia , Peptídeo Hidrolases , RNA de Cadeia Dupla , RNA Viral , Roniviridae , Proteínas Virais/genética , Animais , Replicação ViralRESUMO
Yellow head virus (YHV) is an invertebrate nidovirus which causes a severe mortality in cultured Penaeus monodon. The mortality may be prevented by prior treatment of shrimps with YHV-protease dsRNA. Whether the YHV infected shrimp might be cured by the dsRNA remains to be investigated. P. monodon injected with 10(-6) YHV showed a high virus replication and mortality within 2 days. Injection of 25 microg YHV-protease dsRNA at 3, 6, 12 or 24 h post YHV infection showed a strong inhibition of YHV replication up to 12 h. Unrelated dsRNA-GFP showed no inhibition, indicating that the inhibition was nucleic acid sequence specific through RNAi pathway. Shrimp mortality could be prevented at 3h post YHV infection by the dsRNA, but not at 24 h. These results demonstrate that YHV-protease dsRNA gives therapeutic effect and pave the way to develop a cure for YHV-infected shrimps.
Assuntos
Antivirais/farmacologia , Penaeidae/virologia , RNA de Cadeia Dupla/farmacologia , RNA Viral/antagonistas & inibidores , Roniviridae/efeitos dos fármacos , Proteínas Virais/biossíntese , Animais , Modelos Animais de Doenças , Peptídeo Hidrolases/biossíntese , Peptídeo Hidrolases/genética , Interferência de RNA , RNA de Cadeia Dupla/genética , RNA Viral/genética , Roniviridae/genética , Roniviridae/fisiologia , Proteínas Virais/genética , Replicação Viral/efeitos dos fármacosRESUMO
Yellow head virus infects cultured shrimps and causes severe mortality resulting in a great economic loss. Haemolymph injection of dsRNA(pro) corresponding to the protease motif of YHV genome resulted in a complete inhibition of YHV replication. The effect of dsRNA lasted for at least 5 days. Injecting sequence-unrelated dsRNA(gfp) or dsRNA(TSV-pol) also resulted in an inhibition of YHV replication but at a comparatively much less extent. Shrimp mortality was monitored for 10 days when more than 90% shrimps receiving no dsRNA died within 8 dpi. However, those receiving dsRNA(pro) showed no mortality. A partial mortality was observed among the shrimps receiving dsRNA(gfp) or dsRNA(TSV-pol). Thus, Penaeus monodon possesses the sequence-specific protection to YHV infection, most likely through the RNAi pathway, in addition to sequence-independent protection. It gives a new notion that dsRNA induction of antiviral immunity in shrimp goes through two pathways, sequence-independent and sequence-dependent.
