Animal Industry Report Animal Industry Report

AS 658 ASL R2670

2012

Injection of Double Stranded RNA Enhances Survival of Litopenaeus vannamei Challenged with White Spot Syndrome Virus

Duan S. Loy Iowa State University

Lyric Bartholomay Iowa State University

D.L. Hank Harris Iowa State University

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Recommended Citation Loy, Duan S.; Bartholomay, Lyric; and Harris, D.L. Hank (2012) "Injection of Double Stranded RNA Enhances Survival of Litopenaeus vannamei Challenged with White Spot Syndrome Virus," Animal Industry Report: AS 658, ASL R2670. DOI: https://doi.org/10.31274/ans_air-180814-1051 Available at: https://lib.dr.iastate.edu/ans_air/vol658/iss1/2

This Animal Health is brought to you for free and open access by the Animal Science Research Reports at Iowa State University Digital Repository. It has been accepted for inclusion in Animal Industry Report by an authorized editor of Iowa State University Digital Repository. For more information, please contact [email protected]. Iowa State University Animal Industry Report 2012

Injection of Double Stranded RNA Enhances Survival of Litopenaeus vannamei Challenged with White Spot Syndrome Virus

A.S. Leaflet R2670 Materials and Methods Specific Pathogen Free (SPF) juvenile L. vannamei Duan S. Loy, graduate student, Department of Animal weighing 3-5 grams were acquired from Shrimp Science; Lyric Bartholomay, associate professor, Improvement Systems (SIS) and were acclimated into 200L Department of Entomology; tanks containing synthetic seawater. Animals were divided D. L. (Hank) Harris, professor, Department of Animal into 5 groups with 3 replicates per group and 10 animals per Science tank. Shrimp were injected with 5 micrograms of Summary and Implications dsRNAVP19, dsRNAVP28, a non-specific dsRNA, or an Double stranded RNA was synthesized in vitro with equivalent volume of 2% sterile saline. Shrimp were sequences corresponding to portions of the WSSV genome challenged 3 days later with a lethal dose of WSSV by and were injected into shrimp 3 days prior to challenge with intramuscular injection into the third abdominal segment a lethal dose of WSSV. using a previously clarified and diluted tissue homogenate positive for WSSV by Real Time Quantitative PCR. Introduction Mortalities were recorded for 17 days. White Spot Syndrome Virus (WSSV) can cause 100 % mortality within 7-10 days resulting in the huge economic Results and Discussion loss in shrimp farming. The aim of this study was to test After 17 days post challenge, VP19 dsRNA, VP28 novel means of protecting the Pacific white shrimp, dsRNA and non-specific dsRNA showed a percent survival Litopenaeus vannamei, against disease caused by WSSV. of 100%, 80% and 23.33%, respectively. The positive Double-stranded RNA (dsRNA) is a hallmark of viral control group showed 2% survival (Figure 1). Data were infection which responds to the immune system of shrimp analyzed using one-way ANOVA (SPSS software). VP19 by RNA interference (RNAi) pathway. Mechanistically this dsRNA and VP28 dsRNA groups were significantly was done by testing RNAi through exposure to double- different from nonspecific dsRNA, positive control group stranded RNA targeting virus protein VP19, VP28. (p<0.05). The data suggests that virus-specific dsRNAs are capable of enhancing the antiviral mechanism.

100

80 VP19 dsRNA VP28 dsRNA 60 Nonspecific 40 dsRNA Positive control

20 Negative control Percent survival (%) survival Percent 0 0 5 10 15 20 Days post infection (days)

Figure 1. Precent survival of L.vannamei after WSSV challenge.