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Diseases of Aquatic Organisms

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DAO 114:89-98 (2015)  -  DOI:

RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei 

Rubens Galdino Feijó1, Rodrigo Maggioni2, Pedro Carlos Cunha Martins2, Keuly Ladislau de Abreu2, João Mafaldo Oliveira-Neto2, Cristhiane Guertler3, Emily Bruna Justino3, Luciane Maria Perazzolo3, Luis Fernando Marins1,*

1Laboratório de Biologia Molecular, Instituto de Ciências Biológicas (ICB), Universidade Federal de Rio Grande (FURG), Av. Itália, Km 8, CEP 96203-900, Rio Grande, RS, Brazil
2Laboratório de Biologia Molecular, Centro de Diagnóstico de Enfermidades de Organismos Aquáticos (CEDECAM), Instituto de Ciências do Mar (LABOMAR), Universidade Federal do Ceará (UFC), Av. Abolição, 3207, Meireles, CEP 60165-081, Fortaleza, CE, Brazil
3Laboratório de Imunologia Aplicada à Aquicultura, Departamento de Biologia Celular, Embriologia e Genética (BEG), Universidade Federal de Santa Catarina (UFSC), CP 476, CEP 88040-900, Florianópolis, SC, Brazil
*Corresponding author:

ABSTRACT: Disease in Pacific white shrimp Litopenaeus vannamei caused by the infectious myonecrosis virus (IMNV) causes significant socioeconomic impacts in infection-prone shrimp aquaculture regions. The use of synthetic dsRNA to activate an RNA interference (RNAi) response is being explored as a means of disease prophylaxis in farmed shrimp. Here, survival was tracked in L. vannamei injected with long synthetic dsRNAs targeted to IMNV open reading frame (ORF) 1a, ORF1b, and ORF2 genome regions prior to injection challenge with IMNV, and real-time RT-PCR was used to track the progress of IMNV infection and mRNA expression levels of the host genes sid1, dicer2, and argonaute2. Injection of dsRNAs targeting the ORF1a and ORF1b genes but not the ORF2 gene strongly inhibited IMNV replication over a 3 wk period following IMNV challenge, and resulted in 90 and 83% shrimp survival, respectively. Host gene mRNA expression data indicated that the Sid1 protein, which forms a transmembrane channel involved in cellular import/export of dsRNA, increased in abundance most significantly in shrimp groups that were most highly protected by virus-specific dsRNA injection. Subclinical IMNV infections present in the experimental L. vannamei used increased markedly in the 2 d between injection of any of the 4 virus-specific or non-specific dsRNAs tested and IMNV challenge. While handling and injection stress are implicated in increasing IMNV replication levels, the underlying molecular factors that may have been involved remain to be elucidated.

KEY WORDS: IMNV · Litopenaeus vannamei · RNAi therapy · Gene expression · RT-qPCR

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Cite this article as: Feijó RG, Maggioni R, Martins PCC, de Abreu KL and others (2015) RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei . Dis Aquat Org 114:89-98.

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