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Transcript Analysis of White spot syndrome virus Latency and Phagocytosis Activating Protein Genes in Infected Shrimp (Penaeus monodon)

Indian Journal of Virology volume 23, pages 333–343 (2012)Cite this article

Abstract

Viral latency has been recently observed to be associated with White spot syndrome virus (WSSV) infection in shrimp. In the present study, shrimp samples (Penaeus monodon) surviving WSSV infection were examined for presence of WSSV in latent phase. Virus latency was observed in shrimp which were either experimentally challenged with WSSV and survived the infection or those which survived the natural infection. Three viral transcripts (ORFs 427, 151, 366) associated with latency were analyzed by real-time PCR. The shrimp surviving the natural WSSV infection on estimation with RT-PCR were found to have low grade of WSSV infection (less than 56 copies of WSSV). All the shrimp samples were RT-PCR negative for structural protein genes of WSSV, VP24 and VP28, indicating that these samples were harboring latent phase virus. RT-PCR of all the shrimp samples which survived WSSV infection revealed amplification of phagocytosis activating protein (PAP) gene (435 bp) with higher gene expression levels in experimentally challenged shrimp when compared to naturally infected shrimp. The expression of PAP in WSSV infected shrimp samples indicates its possible role in host response for resistance against WSSV infection. PAP was cloned and expressed as recombinant protein for protection studies. Shrimp were injected with three doses (5, 15 and 20 μg g−1 body weight) of recombinant PAP. Relative percent survival of 10 % was observed in shrimp immunized with the dose of 15 μg g−1 body weight of recombinant PAP. The expression of both WSSV latency associated and PAP genes obtained from shrimp surviving the WSSV infection, indicates the possible role of these genes in host–pathogen interaction.

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References

  1. Amend DF. Potency testing of fish vaccines. In: Anderson DP, Hennessen W, editors. Fish biologics: serodiagnostic and vaccines. Basel: Karger; 1981. pp. 447–454.

  2. Argue BJ, Arce SM, Lotz JM, Moss SM. Selective breeding of Pacific white shrimp (Litopenaeus vannamei) for growth and resistance to Taura syndrome virus. Aquaculture. 2002;204:447–60.

    Article  Google Scholar 

  3. Bell TA, Lightner DV. IHHN virus: infectivity and pathogenicity studies in Penaeus stylirostris and Penaeus vannamei. Aquaculture. 1984;38:185–94.

    Article  Google Scholar 

  4. Bergallo M, Costa C, Baro S, Musso T, Balbo V, Merlino C, Cavallo R. Multiplex-nested RT-PCR to evaluate latent and lytic Epstein–Barr virus gene expression. J Biotechnol. 2007;128:462–76.

    PubMed  Article  CAS  Google Scholar 

  5. Brink AATP, Oudejans JJ, Jiwa M, Walboomers JMM, Meijer CJLM, Van den Brule AJC. Multiprimed cDNA synthesis followed by PCR is the most suitable method for Epstein–Barr virus transcript analysis in small lymphoma biopsies. Mol Cell Probes. 1997;11:39–47.

    PubMed  Article  CAS  Google Scholar 

  6. Chotigeat W, Deachamag P, Phongdara A. Identification of a protein binding to the phagocytosis activating protein (PAP) in immunized black tiger shrimp. Aquaculture. 2007;271:112–20.

    Article  CAS  Google Scholar 

  7. Cock J, Gitterle T, Salazar M, Rye M. Breeding for disease resistance of penaeid shrimps. Aquaculture. 2009;286:1–11.

    Article  Google Scholar 

  8. de la Rosa-Vélez J, Cedano-Thomas Y, Cid-Becerra J, Méndez-Payán JC, VegaPérez C, Zambrano-García J, Bonami JR. Presumptive detection of yellow head virus by reverse transcriptase-polymerase chain reaction and dot blot hybridization in Litopenaeus vannamei and L. stylirostris cultured on the Northwest coast of Mexico. J Fish Dis. 2006;29:717–28.

    PubMed  Article  Google Scholar 

  9. Deachamag P, Intaraphad U, Phongdara A, Chotigeat W. Expression of a phagocytosis activating protein (PAP) gene in immunized black tiger shrimp. Aquaculture. 2006;255:165–72.

    Article  CAS  Google Scholar 

  10. Flegel TW, Nielsen L, Thamavit V, Kongtim S, Pasharawipas T. Presence of multiple viruses in non-diseased, cultivated shrimp at harvest. Aquaculture. 2004;240:55–68.

    Article  Google Scholar 

  11. Gopikrishna G, Gopal C, Shekhar MS, Vinaya Kumar K, Kannappan S, Ponniah AG. Growth performance and white spot syndrome virus resistance in families of kuruma shrimp (Marsupenaeus japonicus). Indian J Geo-mar Sci. 2012;41:56–60.

    Google Scholar 

  12. Hasson KW, Lightner DV, Mohney LL, Redman RM, Poulos BT, White BL. Taura syndrome virus (TSV) lesion development and the disease cycle in the Pacific white shrimp Penaeus vannamei. Dis Aquat Org. 1999;36:81–93.

    Article  Google Scholar 

  13. He F, Kwang J. Identification and characterization of a new E3 ubiquitin ligase in white spot syndrome virus involved in virus latency. Virol J. 2008;5:151.

    PubMed  Article  Google Scholar 

  14. Khadijah S, Ying Neo S, Hossain MS, Miller LD, Mathavan S, Kwang J. Identification of white spot syndrome virus latency-related genes in specific-pathogen-free shrimps by use of a microarray. J Virol. 2003;77:10162–7.

    PubMed  Article  CAS  Google Scholar 

  15. Kubat NJ, Amelio AL, Giordani NV, Bloom DC. The herpes simplex virus type 1 latency-associated transcript (LAT) enhancer/rcr is hyperacetylated during latency independently of LAT transcription. J Virol. 2004;78:12508–18.

    PubMed  Article  CAS  Google Scholar 

  16. Leight ER, Sugden B. EBNA-1: a protein pivotal to latent infection by Epstein–Barr virus. Rev Med Virol. 2000;10:83–100.

    PubMed  Article  CAS  Google Scholar 

  17. Lu L, Kwang J. Identification of a novel shrimp protein phosphatase and its association with latency-related ORF427 of white spot syndrome virus. FEBS Lett. 2004;577:141–6.

    PubMed  Article  CAS  Google Scholar 

  18. Lu L, Wang H, Manopo I, Yu L, Kwang J. Baculovirus-mediated promoter assay and transcriptional analysis of white spot syndrome virus orf427 gene. Virol J. 2005;2:71.

    PubMed  Article  Google Scholar 

  19. Luo T, Zhang X, Shao Z, Xu X. PmAV, a novel gene involved in virus resistance of shrimp Penaeus monodon. FEBS Lett. 2003;551:53–7.

    PubMed  Article  CAS  Google Scholar 

  20. Magbanua FO, Natividad KT, Migo VP, Alfafara CG, Peña F, Miranda RO, Albaladejo JD, Nadala ECB Jr, Loh PC, Tapay LM. White spot syndrome virus (WSSV) in cultured Penaeus monodon in the Philippines. Dis Aquat Org. 2000;42:77–82.

    PubMed  Article  CAS  Google Scholar 

  21. Ramakrishnan R, Levine M, Fink DJ. PCR-based analysis of herpes simplex virus type 1 latency in the rat trigeminal ganglion established with a ribonucleotide reductase-deficient mutant. J Virol. 1994;68:7083–91.

    PubMed  CAS  Google Scholar 

  22. Reed LJ, Muench H. A simple method of estimating fifty percent endpoints. Am J Hyg. 1938;27:493–7.

    Google Scholar 

  23. Reeves M, Sinclair J. Aspects of human cytomegalovirus latency and reactivation. Curr Top Microbiol Immunol. 2008;325:297–313.

    PubMed  Article  CAS  Google Scholar 

  24. Rowe DT. Epstein–Barr virus immortalization and latency. Front Biosci. 1999;4:D346–71.

    PubMed  Article  CAS  Google Scholar 

  25. Shekhar MS, Sahoo PK, Dillikumar M, Abhilipsa D. Cloning, expression and sequence analysis of Macrobrachium rosenbergii nodavirus genes: Indian isolate. Aquacult Res. 2011;42:1778–88.

    Article  CAS  Google Scholar 

  26. Sinclair J, Sissons P. Latency and reactivation of human cytomegalovirus. J Gen Virol. 2006;87:1763–79.

    PubMed  Article  CAS  Google Scholar 

  27. Slobedman B, Mocarski ES. Quantitative analysis of latent human cytomegalovirus. J Virol. 1999;73:4806–12.

    PubMed  CAS  Google Scholar 

  28. Spivack JG, Fraser NW. Detection of herpes simplex virus type 1 transcripts during latent infection in mice. J Virol. 1987;61:3841–7.

    PubMed  CAS  Google Scholar 

  29. Stevens JG, Wagner EK, Devi-Rao GB, Cook ML, Feldman LT. RNA complementary to a herpesvirus alpha gene mRNA is prominent in latently infected neurons. Science. 1987;235:1056–9.

    PubMed  Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  31. Tang KFJ, Durand SV, White BL, Redman RM, Pantoja CR, Lightner DV. Postlarvae and juveniles of a selected line of Penaeus stylirostris are resistant to infectious hypodermal and hematopoietic necrosis virus infection. Aquaculture. 2000;190:203–10.

    Article  Google Scholar 

  32. Tsai M, Kou G, Liu H, Liu K, Chang C, Peng S, Hsu H, Wang C, Lo C. Long-term presence of white spot syndrome virus (WSSV) in a cultivated shrimp population without disease outbreaks. Dis Aquat Org. 1999;38:107–14.

    Article  Google Scholar 

  33. Van Hulten MC, Witteveldt J, Peters S, Kloosterboer N, Tarchini R, Fiers M, Sandbrink H, Lankhorst RK, Valk JM. The white spot syndrome virus DNA genome sequence. Virology. 2001;286:7–22.

    PubMed  Article  Google Scholar 

  34. Watanabe T. Isolation of cDNA encoding a homologue of ribosomal protein L26 in the decapod crustacean Penaeus japonicus. Mol Mar Biol Biotechnol. 1998;7:259–62.

    PubMed  CAS  Google Scholar 

  35. Withyachumnarnkul B. Results from black tiger shrimp Penaeus monodon culture ponds stocked with postlarvae PCR-positive or-negative for white-spot syndrome virus (WSSV). Dis Aquat Org. 1999;39:21–7.

    PubMed  Article  CAS  Google Scholar 

  36. Yang F, He J, Lin X, Li Q, Pan D, Zhang X, Xu X. Complete genome sequence of the shrimp white spot bacilliform virus. J Virol. 2001;75:11811–20.

    PubMed  Article  CAS  Google Scholar 

  37. You Z, Nadala EC, Yang J, Loh PC. Conservation of the DNA sequences encoding the major structural viral proteins of WSSV. Dis Aquat Org. 2004;61:159–63.

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

The authors are thankful to Mr. S. Santhana Krishnan for providing the shrimp samples from the farms which had survived the WSSV natural infection for this study.

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Affiliations

  1. Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, 75, Santhome High Road, R.A. Puram, Chennai, Tamil Nadu, India

    M. S. Shekhar, M. DilliKumar, K. Vinaya Kumar, G. Gopikrishna, S. Rajesh, J. Kiruthika & A. G. Ponniah

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  1. M. S. Shekhar
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  2. M. DilliKumar
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Correspondence to M. S. Shekhar.

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Shekhar, M.S., DilliKumar, M., Vinaya Kumar, K. et al. Transcript Analysis of White spot syndrome virus Latency and Phagocytosis Activating Protein Genes in Infected Shrimp (Penaeus monodon). Indian J. Virol. 23, 333–343 (2012). https://doi.org/10.1007/s13337-012-0119-7

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Keywords

  • White spot syndrome virus
  • Penaeus monodon
  • Latency
  • Phagocytosis activating protein