Ultrastructure and phylogeny ofPhilasterides dicentrarchi(Ciliophora, Scuticociliatia) from farmed turbot in NW Spain

Abstract

Several species of opportunistic histophagous scuticociliates have been implicated in systemic infections of farmed fish. In turbot, scuticociliatosis is an emerging disease, and the identification of the parasite species involved is controversial. We have previously isolatedPhilasterides dicentrarchifrom farmed turbot scuticociliatosis outbreaks in northwest Spain. In the present study, we report detailed ultrastructural studies of this parasite, and investigate phylogenetic relations with other members of the order Philasterida on the basis of sequence comparison of the small-subunit rRNA (SSUrRNA) gene. Ultrastructural study indicates the presence of dikinetids in the anterior two-thirds of the body; micronucleus closely associated with the macronucleus, though not physically connected; numerous mitochondria located below the cell cortex, parallel to the surface; numerous spherical and fusiform extrusomes located close to the plasma membrane. We consider that these characteristics are useful for diagnosis of infections by this parasite. A nested 350-bp nucleotide sequence of the SSUrRNA gene of the turbotP. dicentrachiisolate showed high identity with previously reported SSUrRNA gene sequences from 2 scuticociliates isolated from olive flounderParalichthys olivaceusin Korea, namelyP. dicentrarchi(98%) andMiamiensis avidus(99%); conversely, ourP. dicentrarchisequence showed low identity (86%) with that ofUronema marinum, a scuticociliate that has also been implicated in scuticociliatosis outbreaks in turbot in Europe and olive flounder in Asia. Phylogenetic tree construction on the basis of the SSUrRNA gene sequences, using the neighbour-joining method, confirm that the differentP. dicentrarchiisolates andM. avidusare closely related and a possible synonymy between both ciliates species should be considered.