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.

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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.

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, 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.

A new myxosporean species, Ortholinea auratae n. sp., is described from the gilthead seabream, Sparus aurata Linnaeus, 1758 (Teleostei, Sparidae) from a fish farm in Algarve, Portugal. Plasmodia and spores were found in the urinary bladder and, less frequently, in the posterior kidney. Plasmodia were polymorphic, presenting an irregular cellular membrane due to the presence of several peripheral projections, which in turn were covered by a glycocalyx-like sheet. Mature spores were subspherical in valvular view and ellipsoidal in sutural view, measuring 9.0 ± 0.3 (8.2-10.1) μm in length, 8.3 ± 0.4 (7.5-9.1) μm in width, and 7.2 ± 0.5 (6.3-8.4) μm in thickness. The two valves comprising the spores displayed an intricate pattern of surface ridges and were also enveloped by a glycocalyx-like sheet. Two subspherical polar capsules, 3.2 ± 0.1 (2.9-3.6) μm long and 2.7 ± 0.1 (2.4-2.9) μm wide, were located at the anterior pole and displayed divergent orientation. The polar filament coiled in three to four turns. The comprehensive analysis of the parasite's ultrastructural observations and molecular data for the small subunit (SSU) ribosomal DNA (rDNA) gene identify O. auratae n. sp. as a new species, clustering together with other myxosporeans infecting the excretory system to form a subclade of the main freshwater clade.

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.

Known life cycles of myxosporean parasites have two hosts, but very few life cycles have been disclosed, especially in the marine environment. Sphaerospora dicentrarchi Sitjà-Bobadilla and Álvarez-Pellitero, 1992 is a systemic parasite from the European seabass, Dicentrarchus labrax (Linnaeus, 1758), a highly valuable commercial fish. It affects its health, leading to aquaculture production losses. During 2013 and 2014, an actinospore survey was conducted in a total of 5942 annelids collected from a fish farm in Algarve and from the Aveiro Estuary, in Portugal. A new tetractinomyxon actinospore was found in a capitellid polychaete, belonging to the genera Capitella collected at the fish farm. The tetractinomyxons were pyriform measuring 11·1 ± 0·7 µm in length and 7·2 ± 0·4 µm in width, and presented three rounded polar capsules measuring 2·4 ± 0·3 µm in diameter. The molecular analysis of the 18S rRNA gene sequences from the tetractinomyxons revealed a similarity of 100% with the DNA sequences deposited in the GenBank from S. dicentrarchi myxospores collected from the European seabass and the spotted seabass in the same fish farm and 99·9% similarity with the DNA sequence obtained from the myxospores found infecting the European seabass in the Aveiro Estuary. Therefore, the new tetractinomyxons are inferred to represent the actinospore phase of the S. dicentrarchi life cycle.