The morphology of Zschokkella mugilis Sitjà-Bobadilla and Alvarez-Pellitero, 1993 (Myxosporea, Bivalvulida) in Nereis diversicolor O. F. Müller, 1776 is described for the first time. The molecular data show that the actinospore has 100% similarity to the myxospore of Z. mugilis. Fully mature actinospores are tri-radiate, the spore body has a small process, and the sporoplasm has 2 inner daughter cells. In the polychaete, the spores of the parasite develop in groups of 8 inside pansporocysts. The schizogony phase takes place in the intestinal epithelium, while gametogony and sporogony occur in the coelom of the polychaete. Observations indicate that mature spores are released only during the polychaete reproductive season. Infection was detected only in the winter and spring. In the Aveiro estuary (Portugal), the overall prevalence of infection of the polychaete was 0.5%.

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Ten new types of sphaeractinomyxon actinospores are morphologically and molecularly described from the coelomic cavity of two marine oligochaete hosts, Hrabě, 1967 and Tubificoides pseudogaster (Dahl, 1960), from Aveiro 2 estuary, Portugal. The smallest sphaeractinomyxon type measured 17 µm (length) × 19 µm (width) × 19 µm (apical diameter), whereas the largest type measured 61 µm × 76 µm × 80 µm. While considering the ten types of sphaeractinomyxon, it was found that the number of spores developing inside pansporocysts varied between one, two, four and eight. The total prevalence of infection was of 19% for the two host species, with a maximum recorded for spring and summer (25-26%). While considering each type of sphaeractinomyxon individually, it was found that the prevalence values ranged between 0.3 and 1.7%. All described sphaeractinomyxons were most similar to mugilids infecting Myxobolus species. Limnodriloides agnesThe validity of the tetraspora and endocapsa collective group names is discussed.

This work reports a new type of actinospore, Unicapsulactinomyxon, which exhibits a unique morphological characteristic in that it has a single and large polar capsule (9·3×4·1 μm) (which possesses a longitudinally-folded polar filament) instead of the 3 polar capsules previously described for actinosporeans. The spore has a binucleated sporoplasm and 3 valves, each of which forms a long process. The spore has a total length of 241·3 μm. This parasite develops in groups of 8 inside pansporocysts in the coelomic cavity of the polychaete host. Molecular investigations on the SSU rDNA show that the new actinospore type is most closely related to Enteromyxum species (81-84% similarity). A survey of actinospore infections of the marine polychaete Diopatra neapolitana in 2007 and 2009, in the Aveiro Estuary (Portugal), showed an annual prevalence of 1·0% and 0·3%, respectively.

Actinospores released from the marine oligochaete Limnodriloides agnes inhabiting a Southern Portuguese fish farm are molecularly recognized as developmental stages of the life cycle of Ortholinea auratae, a myxosporean parasite that infects the urinary bladder of Sparus aurata. The molecular analysis of the 18S rRNA gene reveals a similarity of 99.9 to 100 % of the actinospores analyzed to the myxospores of O. auratae. The actinospores belong to the triactinomyxon morphotype and occur in groups of eight within pansporocysts that develop in the intestinal epithelium of the oligochaete host. This is the first record of a myxosporean using an oligochaete as its invertebrate host in the marine environment.

A new myxosporean, Zschokkella auratis sp. nov., infecting the gall bladder of the gilthead seabream Sparus aurata in a southern Portuguese fish farm, is described using microscopic and molecular procedures. Plasmodia and mature spores were observed floating free in the bile. Plasmodia, containing immature and mature spores, were characterized by the formation of branched glycostyles, apparently due to the release of segregated material contained within numerous cytoplasmic vesicles. Mature spores were ellipsoidal in sutural view and slightly semicircular in valvular view, with rounded ends, measuring 9.5 ± 0.3 SD (8.7−10.3) µm in length and 7.1 ± 0.4 (6.5−8.0) µm in width/thickness. The spore wall was composed of 2 symmetrical valves united along a slightly curved suture line, each displaying 10 to 11 elevated surface ridges. Two equal subspherical polar capsules, 3.7 ± 0.3 (3.0−4.1) µm long and 3.0 ± 0.2 (2.6−3.2) µm wide, were located separately at the spore's extremities. Each polar capsule contained a polar filament forming 4 to 5 coils. The sporoplasm was binucleate and contained numerous sporoplasmosomes. Morphological data, tissue tropism, and molecular analysis of the small subunit rDNA gene identified this parasite as a new species of Zschokkella. Maximum parsimony, neighbor-joining, and maximum likelihood inferences clustered the parasite in a subclade containing other Zschokkella species parasitizing the gall bladder of brackish and marine fish hosts, located within the coelozoic clade of the major freshwater clade; this supports the existence of a marine subclade within the 'freshwater' clade, as well as the existence of a correlation between tissue tropism and myxosporean phylogeny.

Ortholinea labracis n. sp. is described and its life cycle is inferred from a Southern Portuguese fish farm, with basis on microscopic and molecular procedures. This myxosporean parasite infects the urinary bladder of the European seabass Dicentrarchus labrax and the intestinal epithelium of a marine oligochaete of the genus Tectidrilus. Myxospores subspherical in valvular view and ellipsoidal in sutural view measuring 7.6 ± 0.3 (6.8-8.7) μm in length, 7.2 ± 0.2 (6.7-7.7) μm in width and 6.5 ± 0.4 (5.8-7.7) μm in thickness. Two polar capsules, 3.0 ± 0.2 (2.6-3.4) μm long and 2.4 ± 0.1 (2.0-2.9) μm wide, located at the same level, but with divergent orientation and opening to opposite sides of the suture line. Sequencing of the SSU rRNA gene revealed a similarity of 100% between the analysed myxospores and triactinomyxon actinospores. The phylogenetic setting of O. labracis n. sp. shows subgrouping in correlation with tissue tropism, but identifies this parasite as another exception to the main division of Myxosporea into the main freshwater and marine lineages.

A new myxosporean parasite is described from the gall bladder of the gilthead seabream Sparus aurata in a Southern Portuguese fish farm, with basis on light and transmission electron microscopy, as well as in molecular procedures. In the bile, young and mature mono- to disporic plasmodia were elliptical and presented smooth surface membranes. Crescent-shaped myxospores measured 6.7±0.7 (5.3-7.6) μm in length and 27.0±3.0 (19.7-31.2) μm in thickness. The myxospore wall was constituted by two symmetrical valves united along a slightly curved suture line, each presenting a lateral projection with a rounded end. Two equal-sized subspherical polar capsules, measuring 3.6±0.2 (2.9-3.8) μm in length and 3.5±0.3 (2.9-3.8) μm in width, were located at the same level, each displaying a polar filament coiled in 5 turns. Molecular analysis of the SSU rRNA gene confirmed the parasite as a new member of the genus Ceratomyxa, making this the fourth report of Ceratomyxa from the gall bladder of S. aurata in the Iberian Peninsula. This reinforces the assumption that species richness of ceratomyxids in South European sparids is high, but the phylogenetic analysis performed disagrees with the existence of a common ancestor for Ceratomyxa species infecting sparid hosts, as well as their clustering according to geographical location. The main Ceratomyxa clade is not monophyletic due to the inclusion of Palliatus indecorus and Pseudoalatospora kovalevae; a situation that will probably be resolved by the taxonomic revision of these genera.