Macrobrachium rosenbergii nodavirus infection in a giant freshwater prawn hatchery in Indonesia

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Abstract

A pathogen of giant freshwater prawn, Macrobrachium rosenbergii, was recently recorded in a hatchery in Yogyakarta. The clinical symptom in post-larvae (PL) was a whitish appearance of the muscles in the tail. Histological examination revealed myonecrosis with massive infiltration of myonuclei and hemocytes. RT-PCR products of 850 bp were obtained when using RNA from diseased PL as a template. The clinical signs and RT-PCR amplicon were reproduced in M. rosenbergii inoculated with bacteria-free inocula. Electron microscopy demonstrated that the M. rosenbergii nodavirus (MrNV) was icosahedral in shape and 28.12 ± 2.31 nm in diameter. RT-PCR products of the RNA-dependent RNA polymerase gene (RNA-1) and capsid protein gene (RNA-2) of MrNV were obtained using designed primer pairs, cloned into pBluescript-KS, and sequenced. The 1312 nucleotide (nt) sequence of MrNV RNA-1 revealed 98.0 % identity with isolates from China and India. Additionally, the 1112 nt sequence of MrNV RNA-2 displayed 98.0 % identity with isolates from China and Taiwan. Disease control efforts involving disinfection of PL, broodstocks, water media, tanks, equipment and ponds successfully eradicated white tail disease from the hatchery. This study is the first report on white tail disease and the isolation and characterization of MrNV in Indonesia.

Figures

  • Fig. 1 Post-larvae of Macrobrachium rosenbergii with different levels of whitish tails
  • Fig. 4 Agarose gel showing reverse transcription-polymerase chain reaction products of Macrobrachium rosenbergii nodavirus (MrNV) amplified using the described primer pairs. An 850 bp amplified product was obtained from sample 1 (S1) and sample 2 (S2) from diseased PL but was not obtained from the non-template control (S-). M: marker DNA with indicated size
  • Fig. 2 Infiltration of hematocytes in the tail of Macrobrachium rosenbergii post-larvae (scale bar 250 µm)
  • Fig. 3 Cumulative percent mortality of post-larvae of Macrobrachium rosenbergii infected by feeding with infected PL, immersion and feeding with inoculum bioencapsulated-artemia. The control PL were reared without inoculum immersion and fed with untreated artemia
  • Fig. 5 Transmission electron micrographs of Macrobrachium rosenbergii nodavirus (MrNV). (2 % PTA, scale bar 100 nm)
  • Fig. 6 Phylograms of the sequence of the Indonesia isolate of Macrobrachium rosenbergii nodavirus (MrNV) compared with other isolates of MrNV using both the 1312 nt sequences from the RdRP gene (a) and the 1112 nt sequences of the capsid protein gene (b)

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CITATION STYLE

APA

Murwantoko, M., Bimantara, A., Roosmanto, R., & Kawaichi, M. (2016). Macrobrachium rosenbergii nodavirus infection in a giant freshwater prawn hatchery in Indonesia. SpringerPlus, 5(1). https://doi.org/10.1186/s40064-016-3127-z

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