Eudiplozoon nipponicum (Monogenea: Diplozoidae): Comparative study of transcriptome and secretome of fish parasite
2017
Ectoparasitic flatworms from the family Diplozoidae (Platyhelminthes: Monogenea) represent a serious hematophagous fish pathogens. Information related to the biochemical and molecular nature of the physiological processes is rather sporadic, as well as the knowledge of the molecules produced by monogeneans. Therefore, we performed complex transcriptomic and proteomic (secretomic) analysis of monogenean representative Eudiplozoon nipponicum Goto, 1891 (Polyopisthocotylea: Diplozoidae), which was realized by two sequencing strategies (Roche 454 and MiSeq Illumina) and mass spectrometry (HPLC MS/MS). Specific bioinformatics tools were used for filtering and removing of low quality bases as well as contaminants – reads related to host organisms (common carp). Processed reads were assembled into the form individual transcripts. 454/Roche and Illumina sequences were pooled into ~39,000 nucleotide sequences with open reading frame and several further bioinformatics approaches (especially searching for the closest homologous in non-redundant databases and prediction of proteins key properties) were used for annotation. We annotated ~19,000 nucleotide sequences and for ~13,000 of them a source organism from phylum Platyhelminthes was assigned. ~5,800 sequences were determined as peptidases and ~1,400 as their inhibitors. Most abundant peptidases belong to metallo (~1,800), serine (~1,500) and cysteine (~1,400) catalytic type. Some of these peptidases may be related to hematophagy, e.g. cathepsins C, D, H, K, L1, L3, S (40 unique entries), their inhibitors (e.g. kunitz inhibitors; 11 unique entries) and anticoagulant agents (e.g. heparan sulfate; 12 unique entries). For secretomic study, the living specimens of E. nipponicum were continuously collected from the gills of naturally infected and freshly netted carp, excretory-secretory products (ESP) were isolated from ~100 adults and characterized by mass spectrometric methods (LC MS/MS).
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