Morphometric and molecular characterization of Kudoa encrasicoli n. sp. (Myxozoa: Myxosporea) from the European anchovy, Engraulis encrasicolus (L.) (Clupeiformes: Engraulidae).

The European anchovy represents the main fisheries for countries in the Mediterranean and Black Sea basins. The skeletal muscle of 13 of 48 (27.1%) Engraulis encrasicolus (L.) specimens from North East Atlantic waters (FAO 27.8.c) was found infected with interfibrillar elongated plasmodia (130-980 µm in length) containing mature myxospores belonging to the genus Kudoa Meglitsch, 1947. No flesh softening was found associated with infection. Fresh myxospores were 10.8 ± 0.7 (9.1-12.3) µm in width 1, 11.3 ± 0.9 (9.5-13.4) µm in width 2, 6.7 ± 0.4 (5.8-7.4) µm in thickness, and 6.9 ± 0.5 (5.8-7.5) µm in length. They were almost stellate in apical view having three pointed-edged shell valves bearing three small polar capsules equal in size 5.0 ± 0.3 (4.4-5.4) µm long and 2.4 ± 0.2 (2.0-3.0) µm wide, and one rounded- to rarely bluntly pointed-edged shell valve bearing a large and particularly wide polar capsule 6.8 ± 0.4 (5.9-7.6) µm long and 4.1 ± 0.2 (3.6-4.4) µm wide. Morphological and morphometrical comparisons between these myxospores and those of Kudoa thyrsites (Gilchrist, 1923) from the clupeid Sardina pilchardus (Walbaum) (North East Atlantic waters, FAO 27.9.a), with which exhibited a similarity of 98.9% and 96.2% using SSU and LSU rDNA sequences, respectively, support the creation of Kudoa encrasicoli n. sp. Morphometrical analysis of the polar capsules of flattened myxospores is suggested as a useful approach to differentiate phylogenetically related kudoids with stellate or almost stellate myxospores bearing four polar capsules.

Morphological and molecular analysis of Henneguya tietensis n. sp. (Cnidaria: Myxosporea), parasitizing the gill filaments of Prochilodus lineatus (Valenciennes, 1837) from Brazil.

The present study describes Henneguya tietensis n. sp. parasitizing Prochilodus lineatus (Valenciennes, 1837) from Brazil, through morphological and molecular analysis. Myxospores were found in the gill tissue of P. lineatus with a prevalence of 10%. The myxospores consisted of two elongate, elliptical shell valves each with a long, tapering caudal appendage. The morphology was consistent with Henneguya and the myxospore measured as follows (mean ± SD): total length 55.5 ± 2.1 µm, body length 16.2 ± 1.1 µm, body width 5.5 ± 0.1 µm, caudal appendages length 39.0 ± 2.0 µm. The polar capsules were 7.3 ± 0.2 µm long, 1.7 ± 0.2 µm wide and contained a polar filament coiled 11 to 13 turns. Histological analysis showed the plasmodia developing in the middle region of each gill filament and caused a focally extensive distension by forming a space-occupying mass within the gill filament. The ultrastructural analysis allowed the observation of the presence of mature myxospores throughout the plasmodium. Phylogenetic analysis with the SSU rDNA gene showed Henneguya tietensis n. sp. as a sister species of the subclade formed by Henneguya piaractus and Henneguya brachypomus that infect fishes of the genus Piaractus. The genetically closest species was H. piaractus, which showed a similarity of 82.4%. Using molecular and morphological characterization, the myxozoan parasite described herein represents a new species of the genus Henneguya.

A new species, Henneguya lacustris n. sp. (Cnidaria: Myxosporea), infecting the gills of Astyanax lacustris from Brazil.

The present study describes Henneguya lacustris n. sp. parasitizing Astyanax lacustris (Lütken 1875), from the Tietê River, State of São Paulo, Brazil, through morphological and molecular analysis. Myxospores of the parasites were found in the gills of A. lacustris with 13% prevalence of infection. The myxospores were oval and presented as measures (mean ± standard deviation): total length 18.3 ± 2.2 µm, body length 10.4 ± 1.6 µm, body width 4.9 ± 0.9 µm, tail length 7.2 ± 2.5 µm. The polar capsule was 4.8 ± 0.3 µm long and 1.5 ± 0.2 µm wide. The polar tubules, present inside the polar capsules, had 6 to 7 turns. The ultrastructural analysis allowed the observation of the asynchronous development of the myxospores. Phylogenetic analysis showed H. lacustris n. sp. as a sister species of H. chydadea Barassa and Cordeiro 2003 in a subclade formed by species that parasitize the gills of Characiform fish from Brazil. Using molecular and morphological characterization, this parasite was identified as a new species of the genus Henneguya Thélohan 1892.

The potential role of the sphaeractinomyxon collective group (Cnidaria, Myxozoa) in the life cycle of mugiliform-infecting myxobolids, with the morphological and molecular description of three new types from the oligochaete Tubificoides insularis.

Three new types of sphaeractinomyxon (Cnidaria, Myxozoa) are described from the coelomic cavity of the marine oligochaete Tubificoides insularis, collected from the Alvor estuary, Algarve, Portugal. Another known type is also registered from this location and host: Sphaeractinomyxon type 10 of Rangel et al. (2016), which was originally described from the marine oligochaete Tubificoides pseudogaster in the Aveiro estuary, Portugal. Phylogenetic analysis revealed the case isolates and all other available SSU rRNA sequences of sphaeractinomyxon clustering within a clade containing Myxobolus spp. that infect mullets, thus suggesting that this collective group plays a role in the life cycle of mugiliform-infecting myxobolids. Also clustering within this clade were all types of tetraspora and endocapsa, calling into question the distinctiveness of these collective groups. Acknowledging a previous work showing that the pansporocysts of sphaeractinomyxon produce a variable number of actinospores, we suggest that the tetraspora collective group be deemed invalid and its types transferred to sphaeractinomyxon. In turn, endocapsa requires validation through the description of new types truly differentiating them from sphaeractinomyxon.

Ultrastructure and phylogeny of Ceratomyxa diplodae (Myxosporea: Ceratomyxidae), from gall bladder of European seabass Dicentrarchus labrax.

The myxosporean parasite Ceratomyxa diplodae Lubat et al. 1989 sensu Sitjà-Bobadilla & Álvarez-Pellitero, 1993, originally described from the annular seabream Diplodus annularis in the Adriatic Sea, has subsequently been reported from several other sparid hosts, and also a moronid fish, the European seabass Dicentrarchus labrax from the Mediterranean Sea. Here, molecular identity and additional morphological data are given for this parasite infecting the gall bladder of D. labrax in a southern Portuguese fish farm. In the bile, disporic plasmodia were spherical to subspherical with a smooth surface membrane. Most myxospores were crescent-shaped, 5.1 ± 0.5 (4.8-6.7) µm long (mean ± SD) and 21.9 ± 1.0 (20.4-23.9) µm thick; a few were more arcuate, 5.7 ± 0.4 (5.3-6.3) µm long and 17.3 ± 1.0 (16.3-19.1) µm thick. The wall consisted of 2 symmetrical valves united along a slightly curved suture line, with moderately tapering to rounded ends. Two spherical polar capsules, measuring 2.9 ± 0.3 (2.5-3.4) µm in diameter, contained a polar filament forming 8 to 9 coils organized in 2 rows. Host species, tissue tropism, and myxospore morphology supported species identification. Phylogenetic analyses of the small subunit ribosomal RNA sequence positioned the parasite among most sparid-infecting Ceratomyxa spp., suggesting the existence of a common ancestor for these species. The acquisition of molecular data from infections of C. diplodae in its original host and in other sparids is essential in order to ascertain if the morphological and biological variations found among reports of this parasite are intra- or inter-specific.

Body size-dependent effects on the distribution patterns of phoretic mite species assemblages on Rhynchophorus ferrugineus (Olivier, 1790).

Phoretic mites attach to different body parts of the red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790), to disperse. However, the question of how the patterns of attachment sites are formed remains intriguing. Here, we examined RPW-associated phoretic mites in four districts in Northern Portugal (macrohabitat), and investigated the patterns of mite distribution on six body parts of RPW (microhabitat). At the macrohabitat level, we detected seven phoretic mite taxa using the RPW host in each of the four studied districts, all documented for the first time in association with this invasive exotic species in Portugal. However, their relative abundance (species evenness) varied between districts, as did species diversity. All examined weevils carried mites, and the prevalence of the different taxa did not differ between districts or sex of weevils. Measured by mean abundance and degree of aggregation, Centrouropoda sp. proved to be the dominant taxon, while Acarus sp. and Curculanoetus rhynchophorus were considered common subordinate taxa and Uroovobella sp., Mesostigmata, Nenteria extremica and Dendrolaelaps sp. sparse taxa. At the microhabitat level, all taxa were present on all body parts of the RPW; the highest abundance was in a region encompassing the inner surface of the elytra and the membranous hind wings (subelytral space). Analysis of niche overlap revealed that the distribution patterns of phoretic mite taxa on the RPW were not randomly structured. In the subelytral space, interspecific coexistence of mites increased as a function of body size difference with the dominant Centrouropoda sp. We found that in the subelytral space the large dominant species Centrouropoda sp. displaced the larger species Uroobovella sp. and the similarly sized species Nenteria extremica, but coexisted with smaller taxa.

Ultrastructural aspects of Ellipsomyxa mugilis (Myxozoa: Ceratomyxidae) spores and developmental stages in Nereis diversicolor (Polychaeta: Nereidae).

The ultrastructure of the spores and developmental stages of Ellipsomyxa mugilis in Nereis diversicolor were studied by transmission electron microscopy. The ultrastructure features and the developmental stages show many similarities with the general pattern described for other actinospores. However, several new features are definitely worth noting. For example, tetranucleated cells precede the formation of the initial pansporocyst, which preserves the 2 original enveloping cells until the end of sporogony. In the initial stages of sporogony, the future sporoplasm cell acquires the first secondary cell by an engulfment process. In the final stage of sporogony, spores are formed by a sporoplasm with 2 secondary cells and 1 somatic nucleus, and the polar capsule has a polar filament with a helicoidal arrangement possessing 7-8 coils.