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Mass mortality occurred at an Anguilla japonica eel farm equipped with a recirculating aquaculture system in Gimcheon, Korea, from late spring to early summer 2015. The cumulative 3-month mortality was 16% (approximately 24,300-150,000 fish). The majority of affected fish displayed ulcerative lesions that progressed to petechial haemorrhages and small white granulomas in the major organs. A Gram-positive, acid-fast, nonmotile bacterium was isolated from internal organ lesions. Phylogenetic analysis of 16S rRNA identified the species as Nocardia seriolae and the strain was designated EM150506. Afterwards, naïve eels were injected with 1.8 × 10 colony-forming units per fish to confirm the strain's pathogenicity, which resulted in a 20% mortality rate within 4 weeks. However, surviving fish still exhibited white N. seriolae colonies in internal organs. To our knowledge, this is the first report of a N. seriolae infection in cultured eel.

BackgroundThermal denaturation of probe-target hybrid is highly reproducible, and which makes probe melting point analysis reliable in the detection of mutations, polymorphisms and epigenetic differences in DNA. To improve resolution of these detections, we used dual-labeled (quencher and fluorescence), full base of peptide nucleic acid (PNA) probe for fluorescence probe based melting point analysis. Because of their uncharged nature and peptide bond-linked backbone, PNA probes have more favorable hybridization properties, which make a large difference in the melting temperature between specific hybridization and partial hybridization.ResultsHere, we have shown that full base dual-labeled PNA is apt material for fluorescence probe-based melting point analysis with large difference in the melting temperature between full specific hybridization and that of partial hybridization, including insertion and deletion. In case of narrowly distributed mutations, PNA probe effectively detects three mutations in a single reaction tube with three probes. Moreover, we successfully diagnose virus analogues with amplification and melting temperature signal. Lastly, Melting temperature of PNA oligomer can be easily adjusted just by adding gamma-modified PNA probe.ConclusionsThe PNA probes offer advantage of improved flexibility in probe design, which could be used in various applications in mutation detection among a wide range of spectrums.Electronic supplementary materialThe online version of this article (doi:10.1186/s12575-015-0027-5) contains supplementary material, which is available to authorized users.

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