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Oral Administration of Bacterially Expressed VP28dsRNA to Protect Penaeus monodon from White Spot Syndrome Virus

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Abstract

We explored the possibility of protecting Penaeus monodon against white spot syndrome virus (WSSV) infection via interference RNA technology by oral administration of bacterially expressed WSSV VP28dsRNA. Shrimp were given dsRNA orally via two methods. In the first method, pellet feed was coated with inactivated bacteria containing overexpressed dsRNA of the WSSV VP28 gene, and in the second method, pellet feed was coated with VP28dsRNA–chitosan complex nanoparticles. The treated shrimp were orally challenged with WSSV by feeding WSSV-infected tissue. The experiment was conducted for 30 days. The dsRNA-treated shrimp challenged with WSSV showed higher survival compared to control shrimp. Sixty-eight percent survival was observed in shrimp fed with feed coated with inactivated bacteria containing dsRNA of the WSSV VP28 gene whereas 37% survival was observed in shrimp fed with VP28dsRNA–chitosan complex nanoparticle–coated feed. The WSSV caused 100% mortality in shrimp fed with pellet feed coated with inactivated bacteria with empty LITMUS38i vector. At the end of the experiment, the tissue samples prepared from randomly selected shrimp that survived were analyzed via reverse transcriptase-polymerase chain reaction and Western blot analysis for WSSV. The samples were negative for WSSV. Based on the present data and the advantages of dsRNA, we believe that oral administration of crude extract of bacterially expressed VP28dsRNA is a potential therapeutic agent against WSSV infection of shrimp.

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References

  • Balasubramanian G, Sarathi M, Rajesh Kumar S, Sahul Hameed AS (2007) Screening the antiviral activity of Indian medicinal plants against white spot syndrome virus in shrimp. Aquaculture 263, 15–19

    Article  Google Scholar 

  • Bernstein E, Caudy AA, Hammond SM, Hannon GJ (2001) Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409, 363–366

    Article  PubMed  CAS  Google Scholar 

  • Chang CF, Su MS, Chen HY, Liao IC (2003) Dietary-1,3-glucan effectively improves immunity and survival of Penaeus monodon challenged with white spot syndrome virus. Fish Shellfish Immunol 15, 297–310

    Article  PubMed  CAS  Google Scholar 

  • Dong Y, Friedrich M, Nympal (2005) RNAi mediated gene knockdown in juvenile grasshopper. BMC Biotechnol 5, 25

    Article  PubMed  CAS  Google Scholar 

  • Gao S, Chen J, Dong L, Ding Z, Yang Yh, Zhang J (2005) Targeting delivery of oligonucleotide and plasmid DNA to hepatocyte via galactosylated chitosan vector. Eur J Pharm Biopharm 60, 327–334

    Article  PubMed  CAS  Google Scholar 

  • George MR, Maharajan A, John KR, Prince Jeyaseelan MJ (2006) Shrimps survive white spot syndrome virus challenge following treatment with Vibrio bacterin. Indian J Exp Biol 44, 63–67

    PubMed  Google Scholar 

  • Huang R, Xie Y, Zhang J, Shi Z (2005) A novel envelope protein involved in White spot syndrome virus infection. J Gen Virol 86, 1357–1361

    Article  PubMed  CAS  Google Scholar 

  • Isobe R, Kojima K, Matsuyama T, Quan GX, Kanda T, Tamura T, Sahara K, Asano SI, Bando H (2004) Using of RNAi technology to confer enhanced resistance to BmNPV on transgenic silkworms. Arch Virol 149, 1913–1940

    Google Scholar 

  • Itami T, Asano M, Tokishige K, Kubono K, Nakagawa A, Takeno N, Nishimura H, Maeda M, Kondo M, Takahashi Y (1998) Enhancement of disease resistance of kuruma shrimp, Penaeus japonicus, after oral administration of peptidoglycan derived from Bifidobacterium thermophilum. Aquaculture 16, 277–288

    Article  Google Scholar 

  • Jha RK, Xu ZR, Shen J, Bai SJ, Sun JY, Wei FL (2006) The efficacy of recombinant vaccines against white spot syndrome virus in Procambarus clarkii. Immunol Lett 105, 68–76

    Article  PubMed  CAS  Google Scholar 

  • Jones L, Hamilton AJ, Voinnet O, Thomas CL, Maule AJ, Baulcombe DC (1999) RNA–DNA interactions and DNA methylation in posttranscriptional gene silencing. Plant Cell 11, 2291–2301

    Article  PubMed  CAS  Google Scholar 

  • Katas H, Alpar HO (2006) Development and characterization of chitosan nanoparticles for siRNA delivery. J Control Rel 115, 216–225

    Article  CAS  Google Scholar 

  • Kim DK, Jang IK, Seo HC, Shin SC, Yang SY, Kim JW (2004) Shrimp protected from WSSV disease treatment with egg yolk antibodies (IgY) against a truncated fusion protein derived from white spot syndrome virus. Aquaculture 237, 21–30

    Article  CAS  Google Scholar 

  • Kim CS, Kosuke Z, Nam YK, Kim SK, Kim KH (2007) Protection of shrimp (Penaeus chinensis) against white spot syndrome virus (WSSV) challenge by double-stranded RNA. Fish Shellfish Immunol 23, 242–246

    Article  PubMed  CAS  Google Scholar 

  • Lee MK, Chun SK, Choi WJ, Kim JK, Choi SH, Kim A, Oungbho K, Park JS, Ahn WS, Kim CK (2005) The use of chitosan as a condensing agent to enhance emulsion-mediated gene transfer. Biomaterials 26, 2147–2156

    Article  PubMed  CAS  Google Scholar 

  • Li HX, Meng XL, Xu JP, Lu W, Wang J (2005) Protection of crayfish, Cambarus clarkii, from white spot syndrome virus by polyclonal antibodies against a viral envelope fusion protein. J Fish Dis 28, 285–291

    Article  PubMed  CAS  Google Scholar 

  • Li LJ, Yuan JF, Cai CA, Gu WG, Shi ZL (2006) Multiple envelope proteins are involved in white spot syndrome virus (WSSV) infection in crayfish. Arch Virol 114, 1–9

    Google Scholar 

  • Lightner DV (Ed.) (1996) A Handbook of Shrimp Pathology and Diagnostic Procedures for Diseases of Cultured Penaeid Shrimp (Baton Rouge, LA:World Aquaculture Society), 305 pp

  • Liu F, Liu Y, Li F, Dong B, Xiang J (2005) Molecular cloning and expression profile of putative antilipopolysaccharide factor in Chinese shrimp (Fenneropenaeus chinensis). Mar Biotechnol 7, 600–608

    Article  PubMed  CAS  Google Scholar 

  • Lowry OH, Rosebrough NF, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol regent. J Biol Chem 193, 265–275

    PubMed  CAS  Google Scholar 

  • Namikoshi A, Wu JL, Yamashita T, Nishizawa T, Nishioka T, Arimoto M, Muroga K (2004) Vaccination trials with Penaeus japonicus to induce resistance to white spot syndrome virus. Aquaculture 229, 25–35

    Article  Google Scholar 

  • Newmark PA, Reddien PW, Cebria F, Alvarado AS (2003) Ingestion of bacterially expressed double-stranded RNA inhibits gene expression in planarians. PNAS 100, 11861–11865

    Article  PubMed  CAS  Google Scholar 

  • Perez F, Ortiz J, Zhinaula M, Gonzabay C, Calderon J, Volckaert FA (2005) Development of EST-SSR markers by data mining in three species of shrimp: Litopenaeus vannamei, Litopenaeus stylirostris, and Trachypenaeus birdy. Mar Biotechnol 7, 554f–569f

    Article  CAS  Google Scholar 

  • Ramos EA, Relucio JLV, Torres-Villanueva CAT (2005) Gene expression in tilapia following oral delivery of chitosan-encapsulated plasmid DNA incorporated into fish feeds. Mar Biotechnol 7, 89–94

    Article  PubMed  CAS  Google Scholar 

  • Robalino J, Browdy CL, Prior S, Metz A, Parnell P, Gross P, Warr G (2004) Induction of antiviral immunity by double-stranded RNA in a marine invertebrate. J Virol 78, 10442–10448

    Article  PubMed  CAS  Google Scholar 

  • Robalino J, Bartlett T, Shepard E, Prior S, Jaramillo G, Scura E, Chapman RW, Gross PS, Browdy CL, Warr GW (2005) Double-stranded RNA induces sequence-specific antiviral silencing in addition to nonspecific immunity in a marine shrimp: convergence of RNA interference and innate immunity in the invertebrate antiviral response? J Virol 79, 13561–13571

    Article  PubMed  CAS  Google Scholar 

  • Rosenberry B (2002) World shrimp farming 2002. (San Diego: Shrimp News International)

    Google Scholar 

  • Sarathi M, Simon MC, Ishaq Ahmed VP, Rajesh Kumar S, Sahul Hameed AS (2007) Silencing VP28 gene of white spot syndrome virus of shrimp by bacterially expressed dsRNA. Mar Biotechnol DOI 10.1007/s10126-007-9052-y

  • Song YL, Liu JJ, Chan LC, Sung HH (1997) Glucan-induced disease resistance in tiger shrimp (Penaeus monodon). Fish Vaccinol 90, 413–421

    CAS  Google Scholar 

  • Syed Musthaq S, Yoganandhan K, Sudhakaran R, Rajesh Kumar S, Sahul Hameed AS (2006) Neutralization of white spot syndrome virus of shrimp by antiserum raised against recombinant VP28. Aquaculture 253, 98–104

    Article  Google Scholar 

  • Takahashi Y, Itami T, Maeda M, Fujii R, Tomonaga S, Supamattaya K, Boonyaratpalin S (1994) Electron microscopic evidence of bacilliform virus infection in kuruma shrimp Penaeus japonicus. Fish Pathol 29, 121–125

    Google Scholar 

  • Takahashi Y, Kondo M, Itami T, Honda T, Inagawa H, Niahizawa T (2000) Enhancement of disease resistance against penaeid acute viraemia and induction of virus-inactivating activity in haemolymph of kuruma shrimp, Penaeus japonicus by oral administration of Pantoea agglomerans lipopolysaccharide (LPS). Fish Shellfish Immunol 10, 555–558

    Article  PubMed  CAS  Google Scholar 

  • Tan LT, Soon S, Lee KL, Shariff M, Hassan MD, Omar AR (2001) Quantitative analysis of an experimental white spot syndrome virus (WSSV) infection in Penaeus monodon Fabricius with competitive polymerase chain reaction. J Fish Dis 24, 315–323

    Article  CAS  Google Scholar 

  • Timmons L, Fire A (1998) Specific interference by ingested dsRNA. Nature 395, 854

    Article  PubMed  CAS  Google Scholar 

  • Tirasophon W, Roshorm Y, Panyim S (2005) Silencing of yellow head virus replication in penaeid shrimp cell by dsRNA. Biochem Biophys Res Commun 334, 102–107

    Article  PubMed  CAS  Google Scholar 

  • Tsutsui N, Ohira T, Kawazoe I, Takahashi A, Wilder MN (2007) Purification of sinus gland peptides having vitellogenesis-inhibiting activity from the whiteleg shrimp litopenaeus vannamei. Mar Biotechnol 9, 360–369

    Article  PubMed  CAS  Google Scholar 

  • Van Hulten MCW, Witteveldt J, Snippe M, Vlak JM (2001) White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. Virology 285, 228–233

    Article  PubMed  CAS  Google Scholar 

  • Venegas CA, Nonaka L, Mushiake K, Nishizawa T, Muroga K (2000) Quasi-immune response of Penaeus japonicus to penaeid rod-shaped DNA virus (PRDV). Dis Aquat Organ 42, 83–89

    Article  PubMed  CAS  Google Scholar 

  • Wang B, Li F, Dong B, Zhang X, Zhang C, Xiang J (2006) Discovery of the genes in response to white spot syndrome virus (WSSV) Infection in fenneropenaeus chinensis through cDNA microarray. Mar Biotechnol 8, 491–500

    Article  PubMed  CAS  Google Scholar 

  • Warren AA, Meola DM, Wang Y, Guo X, Zhou L, Xiang J, Moss S, Arce S, Warren W, Xu Z, Bell K (2006) Isolation and mapping of telomeric pentanucleotide (TAACC)n repeats of the Pacific whiteleg shrimp, Penaeus vannamei, using fluorescence in situ hybridization. Mar Biotechnol 8, 467–480

    Article  PubMed  CAS  Google Scholar 

  • Wei KQ, Xu ZR (2005) Effect of white spot syndrome virus envelope protein VP28 expressed in silkworm (Bombyx mori) pupae on disease resistance in Procambarus clarkii. Shi Yan Sheng Wu Xue Bao 38, 190–198

    PubMed  Google Scholar 

  • Westenberg M, Heinhuis B, Zuidema D, Vlak JM (2005) siRNA injection induces sequence independent protection in Penaeus monodon against white spot syndrome virus (WSSV). Virus Res 114, 133–139

    Article  PubMed  CAS  Google Scholar 

  • Witteveldt J, Cifuentes CC, Vlak JM, van Hulten MC (2004) Protection of Penaeus monodon against white spot syndrome virus by oral vaccination. J Virol 78, 2057–2061

    Article  PubMed  CAS  Google Scholar 

  • Xu J, Han F, Zhang X (2007) Silencing shrimp white spot syndrome virus (WSSV) genes by siRNA. Antiviral Res 73, 126–131

    Article  PubMed  CAS  Google Scholar 

  • Yi G, Wang Z, Qi Y, Yao L, Qian J, Hu L (2004) VP28 of shrimp white spot syndrome virus is involved in the attachment and penetration into shrimp cells. J Biochem Mol Biol 37, 726–734

    PubMed  CAS  Google Scholar 

  • Yodmuang S, Tirasophon W, Roshorm Y, Chinnirunvong W, Panyim S (2006) YHV-protease dsRNA inhibits YHV replication in Penaeus monodon and prevents mortality. Biochem Biophys Res Commun 341, 351–356

    Article  PubMed  CAS  Google Scholar 

  • Yoganandhan K, Sathish S, Narayanan RB, Sahul Hameed AS (2003) Rapid non-enzymatic method of DNA extraction for PCR detection of white spot syndrome virus in shrimp. Aquacult Res 34, 1–5

    Article  Google Scholar 

  • Zamore PD, Tuschl T, Sharp PA, Bartel DP (2000) RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell 101, 25–33

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Huang C, Xu X, Hew CL (2002) Identification and localization of a prawn white spot syndrome virus gene that encodes an envelope protein. J Gen Virol 83, 1063–1074

    Google Scholar 

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Acknowledgments

M. Sarathi is a recipient of a Senior Research Fellowship award from the Council of Scientific and Industrial Research (CSIR), New Delhi. This study was funded by the Department of Biotechnology, Government of India, New Delhi. The authors thank the Management of C. Abdul Hakeem College, Melvisharam for providing the facilities to carry out this work.

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  1. Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam, –632 509, Vellore Dist., Tamil Nadu, India

    M. Sarathi, C. Venkatesan & A. S. Sahul Hameed

  2. Department of Biology, University of Kaiserslautern, Erwin-Schrödinger-Street, Kaiserslautern, –67663, Germany

    Martin C. Simon

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  1. M. Sarathi
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  4. A. S. Sahul Hameed
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Correspondence to A. S. Sahul Hameed.

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Sarathi, M., Simon, M.C., Venkatesan, C. et al. Oral Administration of Bacterially Expressed VP28dsRNA to Protect Penaeus monodon from White Spot Syndrome Virus. Mar Biotechnol 10, 242–249 (2008). https://doi.org/10.1007/s10126-007-9057-6

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  • DOI: https://doi.org/10.1007/s10126-007-9057-6

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