Open Access Research article

Persistence of Penaeus stylirostris densovirus delays mortality caused by white spot syndrome virus infection in black tiger shrimp (Penaeus monodon)

Sudkhate Molthathong12, Sarocha Jitrakorn23, Yutthana Joyjinda2, Chuenchit Boonchird1, Boonsirm Witchayachamnarnkul24, Pattira Pongtippatee5, Timothy Flegel123 and Vanvimon Saksmerprome23*

Author Affiliations

1 Department of Biotechnology, Faculty of Science, Mahidol University, 10400, Bangkok, Thailand

2 Centex Shrimp, Faculty of Science, Mahidol University, 10400, Bangkok, Thailand

3 National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Thailand Science Park, 12120, Pathum Thani, Thailand

4 Shrimp Genetics Improvement Center, Surat Thani, Thailand

5 Aquatic Animal Biotechnology Research Center, Surat Thani Campus, 84100, Surat Thani, Thailand

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BMC Veterinary Research 2013, 9:33  doi:10.1186/1746-6148-9-33

Published: 15 February 2013

Abstract

Background

Persistent infection of Penaeus stylirostris densovirus (PstDNV) (also called IHHNV) and its non-infectious inserts in the black tiger shrimp, Penaeus monodon (P. monodon) genome are commonly found without apparent disease. Here, we introduced the method of multiplex PCR in order to differentiate shrimp with viral inserts from ones with the infectious virus. The method allowed us to study the effect of pre-infection of IHHNV, in comparison to IHHNV inserts, on WSSV resistance in P. monodon.

Results

A multiplex PCR system was developed to amplify the entire IHHNV genome, ensuring the accurate diagnosis. Field samples containing IHHNV DNA templates as low as 20 pg or equivalent 150 viral copies can be detected by this method. By challenging the two groups of diagnosed shrimp with WSSV, we found that shrimp with IHHNV infection and those with viral inserts responded to WSSV differently. Considering cumulative mortality, average time to death of shrimp in IHHNV-infected group (day 14) was significantly delayed relative to that (day 10) of IHHNV-inserted group. Real-time PCR analysis of WSSV copy number indicated the lower amount of WSSV in the IHHNV-infected group than the virus-inserted group. The ratio of IHHNV: WSSV copy number in all determined IHHNV-infected samples ranged from approximately 4 to 300-fold.

Conclusion

The multiplex PCR assay developed herein proved optimal for convenient differentiation of shrimp specimens with real IHHNV infection and those with insert types. Diagnosed shrimp were also found to exhibit different WSSV tolerance. After exposed to WSSV, the naturally pre-infected IHHNV P. monodon were less susceptible to WSSV and, consequently, survived longer than the IHHNV-inserted shrimp.

Keywords:
PstDNV; IHHNV; Shrimp; WSSV; Real-time PCR; Multiplex PCR