Skip to main content
SpringerLink
Log in

Distribution of recombinant VP28 protein in tissues and its immunomodulatory effect against white spot syndrome virus in whiteleg shrimp, Litopenaeus vannamei (Boone, 1931)

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

The gene encoding VP28 protein of WSSV was cloned into pRSET B vector, and recombinant VP28 (r-VP28) protein was produced in E. coli GJ1158 by induction with NaCl. The r-VP28 protein was purified. The purified r-VP28 protein was injected intramuscularly in Whiteleg shrimp, Litopenaeus vannamei, to study its tissue distribution at different time intervals by ELISA using the antiserum raised against r-VP28, and the results showed the presence of r-VP28 protein in different organs such as heart, hepatopancreas, gill, muscle, and hemolymph. The r-VP28 protein was cleared from all the organs after 264 h post-injection (hpi). Various immunological parameters such as phenoloxidase activity (PO), superoxide anion production, activity of superoxide dismutase, and MDA content were assessed in normal and experimentally r-VP28 protein injected shrimp at different time intervals. The mRNA expression of ten immune-related genes was analyzed in hemocytes, hepatopancreas, and gill by quantitative real-time PCR in order to investigate their expression to r-VP28 injection in L. vannamei, and the results showed the upregulation of expression of immune genes such as PO, cMnSOD, lipopolysaccharides (LPs), BGBP, hemocyanin, crustin, lectin, toll receptor, lysozyme, and tumor necrosis factor receptor-associated factor 6 (TRAF6) in VP28-injected shrimp. The efficacy study revealed that 86.67% survival was observed in shrimp orally treated with r-VP28 protein after WSSV challenge without any clinical sign of WSSV infection till the end of experimental period.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Alabi A, Jones D, Latchford J (1999) The efficacy of immersion as opposed to oral vaccination of Penaeus indicus larvae against Vibrio harveyi. Aquaculture 178:1–11

    Article  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44(1):276–287

    Article  CAS  PubMed  Google Scholar 

  • Beuge JA, Aust AD (1978) Microsomal lipid peroxidation. Methods enzymology 52:302–310

    Article  Google Scholar 

  • Bhandari P, Gowrishankar J (1997) An Escherichia coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer. J Bacteriol 179(13):4403–4406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bright Singh IS, Manjusha M, Somnath Pai S, Rosamma P (2005) Fenneropenaeus indicus Is protected from white spot disease by oral administration of inactivated white spot syndrome virus. Dis Aquat Org 66(3):265–270

    Article  CAS  PubMed  Google Scholar 

  • Cai S, Huang T, Wang C, Song X (1995) Epidemiological studies on the explosive epidemic disease of prawn in 1993-94. J Fish China 19:112–117

    Google Scholar 

  • Caipang CMA, Verjan N, Ooi EL, Kondo H, Hirono I, Aoki T, Kiyono H, Yuki Y (2008) Enhanced survival of shrimp, Penaeus (marsupenaeus) japonicus from white spot syndrome disease after oral administration of recombinant VP28 expressed in Brevibacillus brevis. Fish Shellfish Immunol 25:315–320

    Article  CAS  PubMed  Google Scholar 

  • Chang PS, Chen HC, Wang YC (1998) Detection of white spot syndrome associated baculovirus (WSBV) in experimentally infected wild shrimp, crabs and lobsters by in situ hybridization. Aquaculture 164(1):233–242

    Article  CAS  Google Scholar 

  • Chang CF, Su MS, Chen HY (1999) A rapid method to quantify total haemocyte count of Penaeus monodon using ATP analysis. Fish Pathol 34(4):211–212

    Article  Google Scholar 

  • Chang CF, Su MS, Chen HY, Liao IC (2003) Dietary beta-1,3-glucan effectively improves immunity and survival of Penaeus monodon challenged with white spot syndrome virus. Fish Shellfish Immunol 15(4):297–310

    Article  CAS  PubMed  Google Scholar 

  • Cheng W, Liu CH, Tsai CH, Chen JC (2005) Molecular cloning and characterization of a pattern recognition molecule, lipopolysaccharide - and β-1, 3 glucan binding protein (LGBP) from the white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol 18:297–310

  • Chou HY, Huang CY, Wang CH, Chiang HC, Lo CF (1995) Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan. Dis Aquat Org 23:165–173

    Article  Google Scholar 

  • Citarasu T, Sivaram V, Immanuel G, Rout N, Murugan V (2006) Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes. Fish Shellfish Immunol 21(4):372–384

    Article  PubMed  Google Scholar 

  • Dong C, Zhao J, Song L, Wang L, Qiu L, Zheng P, Li L, Gai Y, Yang G (2009) The immune responses in Chinese mitten crab Eriocheirsinensis challenged with double-stranded RNA. Fish shellfish. Immunol 26(3):438–442

    CAS  Google Scholar 

  • Downs C, Fauth JE, Woodley CM (2001) Assessing the health of grass shrimp (Palaemonetespugio) exposed to natural and anthropogenic stressors: a molecular biomarker system. Mar Biotechnol 3(4):380–397

    Article  CAS  PubMed  Google Scholar 

  • Du H-H, Hou CL, Wu XG, Xie RH, Wang YZ (2013) Antigenic and immunogenic properties of truncated vp28 protein of white spot syndrome virus in Procambarus clarkii. Fish shellfish immunol 34(1):332–338

    Article  CAS  PubMed  Google Scholar 

  • Flegel TW (1997) Major viral diseases of the black tiger prawn (Penaeus monodon) in Thailand. World J Microbiol Biotechnol 13:433–442

    Article  Google Scholar 

  • Fu LL, Shuai JB, Xu ZR, Li JR, Li WF (2010) Immune responses of Fenneropenaeus chinensis against white spot syndrome virus after oral delivery of vp28 using Bacillus Subtilis as vehicles. Fish Shellfish Immunol 28:49–55

    Article  CAS  PubMed  Google Scholar 

  • Gross PS, Bartlett TC, Browdy CL, Chapman RW, Warr GW (2001) Immune gene discovery by expressed sequence tag analysis of haemocytess and hepatopancreas in the Pacific white shrimp, Litopenaeus vannamei, and the Atlantic white shrimp, L. setiferus. Dev Comp Immunol 25(7):565–577

    Article  CAS  PubMed  Google Scholar 

  • Huang G, Zhang X, Lin Q, Xu X, Hu ZH, Hew CL (2002) Proteonic analysis of shrimp white syndrome viral proteins and characterization of a navel envelope protein VP466. Mol Cell Proteomics 1(3):223–231

    Article  CAS  PubMed  Google Scholar 

  • Itami T, Takahashi Y, Nakamura Y (1989) Efficacy of vaccination against vibriosis in cultured kuruma prawns Penaeus japonicus. J Aquat Anim Health 1:238–242

    Article  Google Scholar 

  • Itami T, Asano M, Tokushige 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 164:277–288

    Article  CAS  Google Scholar 

  • Jha RK, Roung XZ, Juan BS, Yu SJ, Fen LW, Shen J (2006a) Protection of Procambarus clarkii against white spot syndrome virus using vaccine expressed in Pichia pastoris. Fish Shellfish Immunol 72:1011–1019

    CAS  Google Scholar 

  • Jha RK, Roung XZ, Shen J, Juan BS, Yu SJ, Fen LW (2006b) The efficacy of recombinant vaccines against white spot syndrome virus in Procambarus clarkii. Immunol Lett 105(1):68–76

    Article  CAS  PubMed  Google Scholar 

  • Jha RK, Xu ZR, Bai SJ, Sun JY, Li WF, Shen J (2007) Protection of Procambarus clarkii against white spot syndrome virus using recombinant oral vaccine expressed in Pichia pastoris. Fish Shellfish Immunol 22(4):295–307

    Article  CAS  PubMed  Google Scholar 

  • Jing D, Yan-Bo W, Ling-Lin F (2013) Immune-related genes expression and phagocytosis against white spot syndrome virus after oral delivery of VP28 using Bacillus Subtilis as vehicles in Litopenaeus vannamei. Acta Hydrobiol Sin 37(4):705–711

    Google Scholar 

  • Johnson PT (1987) A review of fixed phayocytic and pinocytotic cell of decapods crustaceans, with remarks on haemocytess. Dev Comp Immunol 11(4):679–704

    Article  CAS  PubMed  Google Scholar 

  • Lee SY, Soderhall K (2002) Early events in crustacean innate immunity. Fish Shellfish Immunol 12:421–437

    Article  CAS  PubMed  Google Scholar 

  • Leonard C, Ratcliffe NA, Rowley AF (1985) The role of prophenoloxidase activation in non-self recognition and phagocytosis by insect blood cells. J Insect Physiol 31(10):789–799

    Article  CAS  Google Scholar 

  • Lightner DV (1996) A handbook of shrimp pathology and diagnostic procedures for diseases of cultured penaeid shrimp, vol. 80. World Aquaculture Society, Baton Rouge, pp 3021–3029

    Google Scholar 

  • Lightner DV (2003) Exclusion of specific pathogens for disease prevention in a penaeid shrimp biosecurity program. In: Lee CS, O’Bryen PJ (eds) Biosecurity in aquaculture production systems: exclusion of pathogens and other undesirables. The World Aquaculture Society, Baton Rouge, pp 81–116

    Google Scholar 

  • Lo CF, Ho CH, Peng SE, Chen CH, Hsu HC, Chiu YL (1996) White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp, crabs and other arthropods. Dis Aquat Org 27:215–225

    Article  Google Scholar 

  • Mantel LH, Farmer LL (1983) Osmotic and ionic regulation. In: Farmer LL (ed) Internal anatomy and physiological regulation. Academic Press, New York, pp 153–161

    Google Scholar 

  • Mavichak R, Kondo H, Hirono I, Aoki T, Kiyono H, Yuki Y (2009) Protection of Pacific white shrimp, Litopenaeus vannamei against white spot virus following administration of N-terminus truncated recombinant VP28 protein expressed in gram-positive bacteria, Brevibacillus choshinensis. Aquacult Sci 57:83–90

    CAS  Google Scholar 

  • Mu Y, Lan JF, Zhang XW, Wang XW, Zhao XF, Wang JX (2012) A vector that expresses VP28 of WSSV can protect red swamp crayfish from white spot disease. Dev Comp Immunol 36(2):442–449

    Article  CAS  PubMed  Google Scholar 

  • Munoz M, Cedeno R, Rodriquez J, Van der Knap W, Mialhe E, Bacher E (2000) Measurement of reactive oxygen intermediate production in haemocytes of Penaeid shrimp, PenaeusVannamei. Aquaculture 191:89–107

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Ongvarrasopone C, Chanasakulniyom M, Sritunyalucksana K, Panyim S (2008) Suppression of PmRap7 by dsRNA inhibits WSSV or YHV infection in shrimp. Mar Biotechnol 10:374–381

    Article  CAS  PubMed  Google Scholar 

  • Pequeux A (1995) Osmotic regulation in crustaceans. J Crust Biol 5:1–60

    Article  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):45

    Article  Google Scholar 

  • Rajesh Kumar S, Ishaq Ahamed VP, Sarathi M, Nazeer Basha A, Sahul Hameed AS (2008) Immunological responses of Penaeus monodon to DNA vaccine and its efficacy to protect shrimp against white spot syndrome virus (WSSV). Fish Shellfish Immunol 24(4):467–478

    Article  CAS  PubMed  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(21):13561–13571

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rout N, Kumar S, Jaganmohan S, Murugan V (2007) DNA vaccines encoding viral envelope proteins confer protective immunity against WSSV in black tiger shrimp. Vaccine 25(15):2778–2786

    Article  CAS  PubMed  Google Scholar 

  • Sahul Hameed AS, Yoganandhan K, Sathish S, Murugan V, Rasheed M, Jayaraman K (2001) White spot syndrome virus (WSSV) in two species of freshwater crabs (Paratelphusa hydrodomous and P. pulvinata). Aquaculture 201(3–4):179–186

    Article  Google Scholar 

  • Sahul Hameed AS, Balasubramanian G, Syed Musthaq S, Yoganandhan K (2003) Experimental infection of twenty species of Indian marine crabs with white spot syndrome virus (WSSV). Dis Aquat Org 57(1–2):157–161

    Article  Google Scholar 

  • Schurrs AHWM, van Weemen BK (1977) Enzyme immunoassay. Clin Chim Acta 81:1–40

    Article  Google Scholar 

  • Soderhall K, Thornqvist PO (1997) Crustacean immunity--a short review. Dev Biol Stand 90:45–51

    CAS  PubMed  Google Scholar 

  • Song YL, Hsieh YT (1994) Immunostimulation of tiger shrimp Penaeus monodon haemocytes for generation of microbicidal substances: analysis of reactive oxygen species. Dev Comp Immunol 18(3):201–209

    Article  CAS  PubMed  Google Scholar 

  • 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, Kwang J (2011) Oral vaccination of Baculovirus-expressed VP28 displays enhanced protection against white spot syndrome virus in Penaeus monodon. PLoS One 6(11):e26428

    Article  CAS  PubMed  Google Scholar 

  • Syed Musthaq S, Sudhakaran R, Ishaq Ahmed VP, Balasubramanian G, Sahul Hameed AS (2006) Variability in the tandem repetitive DNA sequences of white spot syndrome virus (WSSV) genome and suitability of VP28 gene to detect different isolates of WSSV from India. Aquaculture 256(1–4):34–41

    Article  Google Scholar 

  • Taju G, Madan N, Abdul Majeed S, Raj Kumar T, Thamizvanan S, Otta SK, Sahul Hameed AS (2014) Immune responses of white leg shrimp, Litopenaeus vannamei (Boone, 1931) to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus. J Fish Dis 38:451–465. doi:10.1111/jfd.12256

    Article  PubMed  Google Scholar 

  • Takahashi Y, Itami T, Maeda M, Suzuki N, Kasornchandra J, Supamattaya K, Khongpradit R, Boonyaratpalin S, Kondo M, Kawai K, Kusda R, Hirono I, Aoki T (1996) Polymerase chain reaction PCR amplification of bacilliform virus RV–PJ DNA Inpenaeus japonicus bate and systemic ectodermal and mesodermal baculovirus SEMBV DNA in Penaeus monodon Fabricius. J Fish Dis 19(5):399–403

    Article  CAS  Google Scholar 

  • Takahashi Y, Kondo M, Itami T, Honda T, Inagawa H, Nishizawa T, Soma GI, Yokomizo Y (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(6):555–558

    Article  CAS  PubMed  Google Scholar 

  • Teunissen OSP, Faber R, Booms GHR, Latscha T, Boon JH (1998) Influence of vaccination on vibriosis resistance of the giant black tiger shrimp Penaeus monodon (Fabricius). Aquaculture 164:359–366

    Article  Google Scholar 

  • van Hulten MC, Westenberg M, Goodall SD, Vlak JM (2000) Identification of two major virion protein genes of white spot syndrome virus of shrimp. Virology 266(2):227–236

    Article  PubMed  Google Scholar 

  • van Hulten MCW, Witteveldt J, Peters S, Kloosterboer N, Tarchini R, Fiers M, Sand Brink H, Klein Lankhorst R, Vlak JM (2001a) The white spot syndrome virus DNA genome sequence. Virology 286(1):7–22

    Article  PubMed  Google Scholar 

  • van Hulten MC, Witteveldt J, Snippe M, Vlak JM (2001b) White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. Virology 285(2):228–233

    Article  PubMed  Google Scholar 

  • van Hulten MCW, Reijns M, Vermeesch AMG, Zandbergen F, Vlak JM (2002) Identification of VP19 and VP15 of white spot syndrome virus (WSSV) and glycosylation status of the WSSV major structural proteins. J Gen Virol 83:257–265

    Article  PubMed  Google Scholar 

  • Vaseeharan B, Prem Anand T, Murugan T, Chen JC (2006) Shrimp vaccination trials with the VP292 protein of white spot syndrome virus. Lett Appl Microbiol 43(2):137–142

    Article  CAS  PubMed  Google Scholar 

  • Vogt G (1996) Cytopathology of bay of Piran shrimp virus (BPSV), a new crustacean virus from the Mediterranean Sea. J Inverteb Pathol 68(3):239–245

    Article  CAS  Google Scholar 

  • Wang CH, Lo CF, Leu JH, Chou CM, Yeh PY, Chou HY, Tung MC, Chang CF, Su MS, Kou GH (1995) Purification and genomic analysis of baculovirus associated with white spot syndrome (WSBV) of Penaeus monodon. Dis Aquat Org 23:239–242

    Article  Google Scholar 

  • Wang YC, Lo CF, Chang PS, Kou GH (1998) Experimental infection of white spot baculovirus in some cultured and wild decapods in Taiwan. Aquaculture 164:221–231

    Article  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Witteveldt J, Vlak JM, van Hulten MCW (2004b) Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine. Fish Shellfish Immunol 16(5):571–579

    Article  CAS  PubMed  Google Scholar 

  • Witteveldt J, Vlak JM, van Hulten MCW (2006) Increased tolerance of Litopenaeus vannamei to white spot syndrome virus (WSSV) infection after oral application of the viral envelope protein VP28. Dis Aquat Org 70(1–2):167–170

    Article  PubMed  Google Scholar 

  • Xu Z, Du H, Xu Y, Sun J, Shen J (2006) Crayfish Procambarus clarkii protected against white spot syndrome virus by oral administration of viral proteins expressed in silkworms. Aquaculture 253(1–4):178–183

    Google Scholar 

  • Yang FJ, He X, Lin Q, Li D, Pan X, Zhang X, Xu (2001) Complete genome sequence of the shrimp white spot bacilliform virus. J Virol 75(23):11811–11820

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yoganandhan K, Syed Musthaq S, Sahul Hameed AS (2004) Production of polyclonal antiserum against recombinant VP28 protein and its application for the white spot syndrome virus in crustaceans. J Fish Dis 27(9):517–522

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Management of C. Abdul Hakeem College, Melvisharam, India, for providing the facilities to carry out this work. This work was funded by the Department of Biotechnology, Government of India, New Delhi, India, under the scheme of Tata Innovation Fellowship awarded to Dr. A. S. Sahul Hameed.

Author information

Authors and Affiliations

  1. Aquatic Animal Health Laboratory (OIE Reference for WTD), PG and Research Department of Zoology, C. Abdul Hakeem College, Vellore District, Melvisharam, Tamilnadu, 632 509, India

    G. Taju, N. Madan, S. Abdul Majeed, T. Rajkumar, M. A. Farook, A. Nazeer Basha & A. S. Sahul Hameed

  2. Department of Bio-Medical Sciences, School of Bio Science and Technology, VIT University, Vellore, Tamilnadu, India

    S. Vimal

Authors
  1. G. Taju
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Madan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Abdul Majeed
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Rajkumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. A. Farook
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S. Vimal
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. Nazeer Basha
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A. S. Sahul Hameed
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. S. Sahul Hameed.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Taju, G., Madan, N., Abdul Majeed, S. et al. Distribution of recombinant VP28 protein in tissues and its immunomodulatory effect against white spot syndrome virus in whiteleg shrimp, Litopenaeus vannamei (Boone, 1931). Aquacult Int 25, 1761–1776 (2017). https://doi.org/10.1007/s10499-017-0155-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-017-0155-9

Keywords

Search

Navigation