This paper sums up the results of light microscopical, ultrastructural and molecular studies of five strains of amoeboid organisms isolated as endocommensals from coelomic fluid of sea urchins, Sphaerechinus granularis (Lamarck), collected in the Adriatic Sea. The organisms are reported as Didymium-like myxogastrids. Of the life-cycle stages, the attached amoeboids, flagellated trophozoites, cysts and biflagellated swarmers are described. Formation of fruiting bodies was not observed. Although phylogenetic analyses of SSU rDNA sequences indicated a close relationship with Hyperamoeba dachnaya, our sea-urchin strains have not been assigned to the genus Hyperamoeba Alexeieff, 1923. The presence of either one or two flagella reported in phylogenetically closely related organisms and mutually distant phylogenetic positions of strains declared as representatives of the genus Hyperamoeba justify our approach. Data obtained in this study may be useful in future analyses of relationships of the genera Didymium, Hyperamoeba, Physarum and Pseudodidymium as well as in higher-order phylogeny of Myxogastrea.
Effort was made to identify Naegleria strains isolated from organs of fish, using phylogenetic analyses of SSU rDNA and ITS sequences. Eighteen fish-isolated strains studied enlarged substantially the so far available set of Naegleria strains characterized by both molecular markers. The phylogenetic analyses of separate and concatenated SSU rDNA and ITS sequences revealed phylogenetic relationships of strains under study; however, they failed to solve classification of fish-isolated strains into species. The sequence similarity of strain-representatives of Naegleria species as well as data obtained on intragenomic variation of ITS sequences discouraged the authors from the definition of new species. The results of the present study provide evidence of a need to re-evaluate the current practice of setting boundaries between species of the genus Naegleria. Sequences obtained in this study have been deposited in GenBank with accession numbers DQ768714-DQ768743.
Based on morphological and molecular characterisation, four amoeba strains isolated from organs of freshwater fish were identified as Hartmannella vermiformis Page, 1967. Small subunit rRNA gene sequences of these strains expand the set of corresponding complete and almost complete sequences of this species to twelve. A new species-specific oligonucleotide probe inferred from recently available SSU rRNA gene sequences was designed and successfully tested in tissue lesions produced by one strain of H. vermiformis in experimentally infected fish.
A new myxosporean species, Henneguya cynoscioni sp. n., is described from the spotted seatrout, Cynoscion nebulosus (Cuvier) (Sciaenidae) as a causative agent of cardiac henneguyosis. This new myxosporean species is characterized by the morphology of spores and the sequence of SSU rDNA. Examination of 227 spotted seatrout from four South Carolina estuaries in 2008-2010 revealed a 33.5% total prevalence of H. cynoscioni. Henneguya cynoscioni produces lesions in the bulbus arteriosus, its specific site of infection. The severity of lesions and their impact on the bulbus arteriosus is proportional to the number of plasmodial stages developing in this segment of the heart, being most pronounced in host reaction directed against spores liberated from plasmodia.
A new multivalvulid myxosporean species, Kudoa dianae sp. n., is described from bullseye puffer, Sphoeroides annulatus (Jenyns) (Tetraodontiformes: Tetraodontidae). Plasmodia develop in extramuscular sites, in the wall of oesophagus and less frequently on mesenteries. Mature spores can reach lumen of the digestive tract directly by disruption of plasmodial wall or via macrophage transport to the oesophageal epithelium. New species is characterised by morphology of spores and by the complete sequence of SSU rRNA gene that differs from all hitherto known sequences of Kudoa species. Spore morphology (moderate-sized, simple non-ornate spores, quadrate in apical view) clusters with that of Kudoa scienae, K. cerebralis, K. chilkaensis, K. leiostomi, K. funduli, K. cascasia and K. ovivora. Analysis of phylogenetic relationships (using SSU rRNA gene sequences) among five Kudoa species, the molecular data of which are available thus far, revealed that K. dianae is distinguishable from these five species and that its closest relation is with K. miniauriculata.
A new myxosporean species, Kudoa inornata sp. n. (Myxosporea: Multivalvulida), is described from skeletal muscles of the spotted seatrout Cynoscion nebulosus (Cuvier), collected in estuarine waters along the coast of South Carolina, USA. Light microscopic and ultrastructural characters rank this species to the group of Kudoa species with simple-shaped spores. The uniqueness of the SSU and LSU rDNA sequences justifies its status of a new species with sister relationship to Kudoa paniformis. The 100% prevalence in seatrout from four out of five localities sampled and pathogenicity of K. inornata recognized in this study should motivate further screening for infections in its host, which is considered a commercially important game fish with a wide distribution in the Western North Atlantic.
A new highly pathogenic muscle-infecting species of the genus Myxobolus Bütschli, 1882 is described from the Prussian carp, Carassius gibelio (Bloch, 1782) using spore morphology and SSU rDNA sequence data. Phylogenetic analyses elucidated relationship of the newly described Myxobolus lentisuturalis to other Myxobolus species and supported its position of an independent species.
Myxobolus pseudodispar Gorbunova, 1936 (Myxozoa) was originally described as a parasite of common roach, Rutilus rutilus (Linnaeus), with developing stages in muscles and spores disseminated in macrophage centres of different organs and tissues. Later, this parasite was described from several other cyprinids, but with relatively large intraspecific differences based on SSU rDNA gene sequences. Within our long-term study on myxozoan biodiversity, we performed a broad microscopic and molecular screening of various freshwater fish species (over 450 specimens, 36 species) from different localities. We investigated the cryptic species status of M. pseudodispar. Our analysis revealed four new unique SSU rDNA sequences of M. pseudodispar as well as an infection in new fish host species. Myxobolus pseudodispar sequence analysis showed clear phylogenetic grouping according to fish host criterion forming 13 well-recognised clades. Using 1% SSU rDNA-based genetic distance criterion, at least ten new species of Myxobolus Bütschli, 1882 may be recognised in the group of M. pseudodispar sequences. Our analysis showed the paraphyletic character of M. pseudodispar sequences and the statistical tests rejected hypothetical tree topology with the monophyletic status of the M. pseudodispar group. Myxobolus pseudodispar represents a species complex and it is a typical example of myxozoan hidden diversity phenomenon confirming myxozoans as an evolutionary very successful group of parasites with a great ability to adapt to a new hosts with subsequent speciation events.
The original description of Myxobolus longisporus Nie et Li, 1992, the species infecting gills of Cyprinus carpio haematopterus L., is supplemented with new data on the spore morphology and pathogenicity. Spores are elongate pyriform with pointed anterior end, 15.7 (15.5-16.5) µm long, 6.7 (6-8) µm wide and 5.5 µm thick. Sutural ridge is straight and narrow. Mucus envelope is lacking. Two equal-sized elongate pyriform polar capsules are 8.5 µm long and 2.5 µm wide with convergent long axes. Polar filament coiled perpendicularly to the long axis of the capsule makes 9 (8-10) turns. Posterior end of polar capsules exceeds mid-spore by 15-20%. Cyst-like plasmodia are localised in the gill secondary lamellae. The infection is described in adult big host specimens. Gross lesions manifested as dark red colouration of gill tissues were restricted to the ventral part of the first gill arches. Remarkable site specificity (apical part of secondary lamellae) was observed in the course of development of microscopic lesions. M. longisporus is characterised also on the molecular level using sequences of SSU rRNA gene. Phylogenetic analysis based on these sequences has allowed clearer phylogenetic relationships to be established with other species of the genus Myxobolus sequenced to date.
A myxosporean species found to develop in the liver of 10 out of 24 common shrews, Sorex araneus L., caught in South Bohemia, Czech Republic, was identified as Soricimyxum fegati Prunescu, Prunescu, Pucek et Lom, 2007, the unique representative of the genus and the first myxosporean species known to develop from plasmodia to spores in a terrestrial mammal. The original description of this species, based on fixed material, is supplemented with new data based on fresh material and with partial sequence of SSU rDNA (GenBank Acc. No. EU232760). Phylogenetic analysis of SSU rDNA revealed that S. fegati is closely related to myxosporeans infecting gall bladders of freshwater fish.