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Protection of Shrimp Penaeus monodon from WSSV Infection Using Antisense Constructs

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Abstract

White spot syndrome caused by white spot syndrome virus (WSSV) is one of the most threatening diseases of shrimp culture industry. Previous studies have successfully demonstrated the use of DNA- and RNA-based vaccines to protect WSSV infection in shrimp. In the present study, we have explored the protective efficacy of antisense constructs directed against WSSV proteins, VP24, and VP28, thymidylate synthase (TS), and ribonucleotide reductase-2 (RR2) under the control of endogenous shrimp histone-3 (H3) or penaedin (Pn) promoter. Several antisense constructs were generated by inserting VP24 (pH3–VP24, pPn–VP24), VP28 (pH3–VP28, pPn–VP28), TS (pH3–TS, pPn–TS), and RR2 (pH3–RR2) in antisense orientation. These constructs were tested for their protective potential in WSSV infected cell cultures, and their effect on reduction of the viral load was assessed. A robust reduction in WSSV copy number was observed upon transfection of antisense constructs in hemocyte cultures derived from Penaeus monodon and Scylla serrata. When tested in vivo, antisense constructs offered a strong protection in WSSV challenged P. monodon. Constructs expressing antisense VP24 and VP28 provided the best protection (up to 90 % survivability) with a corresponding decrease in the viral load. Our work demonstrates that shrimp treated with antisense constructs present an efficient control strategy for combating WSSV infection in shrimp aquaculture.

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Acknowledgment

We are grateful to the Department of Fisheries, Government of Andhra Pradesh for participating and providing facilities and Mr. N.V. Ramakanth for his help in construction of vectors. Financial assistance of NAIP (ICAR) component-4 grant no. C4-30019 is greatly acknowledged. The authors also thank Dr. W.S. Lakra, Director, CIFE.

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Authors and Affiliations

  1. Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research (CSIR), E405-East Wing 3rd Floor, CCMB, Uppal Road, Hyderabad, 500007, India

    Sajad Ahanger, Madhu K. Mani, Ravinder Kondadhasula, Kshitish C. Majumdar & Rakesh K. Mishra

  2. State Institute of Fisheries Technology (SIFT), Kakinada, Andhra Pradesh, India

    Supriyanka Sandaka & Chandra Sekhar Reddy

  3. Central Institute for Fisheries Education (CIFE), Versova, 400061, Mumbai, India

    Deepika Ananad, Makesh Marappan & Rajendran K. Valappil

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  1. Sajad Ahanger
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  2. Supriyanka Sandaka
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  3. Deepika Ananad
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  4. Madhu K. Mani
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  5. Ravinder Kondadhasula
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  7. Makesh Marappan
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  8. Rajendran K. Valappil
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  9. Kshitish C. Majumdar
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  10. Rakesh K. Mishra
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Correspondence to Rakesh K. Mishra.

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S. A., S. S., D.A., and M.K.M contributed equally to this work.

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Ahanger, S., Sandaka, S., Ananad, D. et al. Protection of Shrimp Penaeus monodon from WSSV Infection Using Antisense Constructs. Mar Biotechnol 16, 63–73 (2014). https://doi.org/10.1007/s10126-013-9529-9

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