Occurrence of the host-parasite system Rhaphiodon vulpinus and Ceratomyxa barbata n. sp. in the two largest watersheds in South America.

While around world, species of the genus Ceratomyxa parasite majority marine hosts, growing diversity has been reported in South American freshwater fish. The present study reports Ceratomyxa barbata n. sp. parasitizing the gallbladder of the Rhaphiodon vulpinus fish from the Amazon and La Plata basins. Morphological (light and transmission electron microscopy), molecular (sequencing of small subunit ribosomal DNA - SSU rDNA), and phylogenetic analyses were used to characterize the new species. Worm-like plasmodia endowed with motility were found swimming freely in the bile. The myxospores were elongated, lightly arcuate, with rounded ends and had polar tubules with 3 coils in the polar capsules. Ultrastructural analysis revealed plasmodia composed of an outer cytoplasmic region, where elongated tubular mitochondria, a rough endoplasmic reticulum, sporogonic stages, and a large vacuole occupying the internal area were observed. Phylogenetic analysis, based on SSU rDNA, found that among all South America freshwater Ceratomyxa species, C. barbata n. sp. arises as an earlier divergent species. The present study reveals the occurrence of this host-parasite system (R. vulpinus/C. barbata n. sp.) in the two largest watersheds on the continent.

A novel Ceratomyxa species (Myxozoa: Cnidaria) infecting an Amazonian catfish.

Parasites are important organisms for the health of ecosystems. While the Amazon Basin is home to a great diversity of ichthyofauna, our knowledge of myxozoan diversity in the biome remains relatively limited. The present study describes a new myxozoan species, Ceratomyxa mandii n. sp., parasitizing the gallbladder of the Amazonian catfish Pimelodina flavipinnis (Pimelodidae) from the Solimões River, in the region of Manaus, Brazil. Light and electron microscopy,  small subunit ribosomal DNA (SSU rDNA) sequencing and phylogenetic analysis were performed. The new species exhibited worm-like plasmodia with undulatory motility. The SSU rDNA based phylogenetic analysis revealed it to be a sister taxon of C. gracillima, which also parasitizes an Amazonian pimelodid fish, possibly reflecting a host-parasite co-speciation process. This study contributes to our understanding of this little sampled group of organisms.

Two novel species of Myxobolus parasitizing the gills of Semaprochilodus insignis in the Brazilian Amazon.

The present study describes two new Myxobolus species infecting the gills of Semaprochilodus insignis, the most consumed freshwater fish species in the Brazilian Amazon. The fish specimens were caught in the Tapajós River, in the state of Pará, and the morphological, ultrastructural, small subunit ribosomal DNA (ssrDNA), and phylogenetic data of the myxosporean species were obtained. Two species of Myxobolus were found in the gills of S. insignis. Myxobolus maiai n. sp. developed in the gill filaments, and mature myxospores were round-shaped from the frontal view, measuring 12.5-14.8 (13.9 ± 0.5) µm in length, 11.4-13.8 (12.3 ± 0.5) µm in width, and have a thickness of 6.4-7.7 (6.9 ± 0.6) µm in the lateral view, with symmetric values. Its polar capsules were 4.4-6.6 (5.5 ± 0.5) µm in length and 2.3-3.7 (3.0 ± 0.3) µm in width, and the polar tubules had 4 - 5 coils. Myxobolus iarakiensis n. sp. was found infecting the gill arch. Mature myxospores were oval-shaped from the frontal view, and measured 6.7-8.6 (8.0 ± 0.4) µm in length, 4.5-6.3 (5.6 ± 0.4) µm in width, and had a thickness of 2.7-4.7 (3.8 ± 0.5) µm in the lateral view, with symmetric values. Its polar capsules were 2.1-3.7 (2.9 ± 0.3) µm in length and 1.1-2.0 (1.5 ± 0.2) µm in width, and its polar tubules had 4 - 5 coils. The ssrDNA based phylogeny showed these two novel species as grouping in a clade composed of parasite species of Prochilodontidae hosts.

Morphological, molecular, and histopathological characterization of a new species of Henneguya infecting farmed Astyanax lacustris in Brazil.

The present study describes the morphological, histopathological and SSU rDNA data of a new myxosporean species infecting farmed Astyanax lacustris fish from the state of São Paulo, Brazil. Henneguya lambariensis sp. nov. was found infecting the gills, and the plasmodial development resulted in displacement, blood congestion, compression, deformation and lamellar fusion, stretching of the epithelia, hyperplasia of the epithelial cells, edema, and mild infiltration of the mast cells and lymphocytes. The SSU rDNA sequencing resulted in the sequencing of 1804 nucleotides that did not correspond to any myxosporean sequences deposited in GenBank. The closest phylogenetic affinity of the new species was to the South American Henneguya loreotoensis and Henneguya guanduensis, which also parasite the gills. The present study suggests this new myxosporean species has considerable pathogenic potential, and health monitoring should be implemented in A. lacustris fish farms to ensure production.

The Amazonian fish Colossoma macropomum harbors high myxosporean diversity: A description of three novel species from a fish farm.

The farming of Colossoma macropomum has intensified in recent years, leading to an increased need for research into the health of the fish. We therefore investigated the diversity of myxosporeans (Cnidaria: Myxozoa) infecting C. macropomum in a breeding system in the municipality of Rio Branco, in the state of Acre, Brazil. Twenty-four fish specimens were examined from June to August 2018. Our results revealed a high prevalence of infection, with 23 specimens (95.8%) exhibiting myxosporean plasmodia. Morphological analysis, based on light and electron microscopies, and molecular analysis (small subunit ribosomal DNA [SSU rDNA] sequencing) revealed the occurrence of three novel species of the genus Myxobolus. Plasmodia of Myxobolus guttae n. sp. were found in the fins of 75% of the specimens, and the myxospores were pear-shaped, measuring 12.3 ± 0.6 (10.3-13.5) µm in length, 8.1 ± 0.3 (7.1-8.6) µm in width, and 5.1 ± 0.6 (4.5-6.5) µm in thickness. The polar capsules were elongated and equal in size, measuring 6.8 ± 0.5 (5.8-7.6) µm in length and 2.5 ± 0.3 (1.8-3.1) µm in width, exhibiting polar tubules with 9-10 coils. The plasmodia of Myxobolus longus n. sp. were found in the gills of 45.8% of the C. macropomum specimens, and the myxospores were fusiform, measuring 16.4 ± 0.6 (14.7-17.3) µm in length, 7.1 ± 0.2 (6.8-7.7) µm in width and 5.5 ± 0.6 (4.4-6.6) µm in thickness. The polar capsules occupied more than half of the myxospore, exhibiting different sizes, the largest measuring 9.2 ± 0.5 (7.3-10.1) µm in length and 2.5 ± 0.3 (2.0-3.1) µm in width, while the smallest measured 8.5 ± 0.4 (7.1-9.1) µm in length and 2.4 ± 0.2 (1.9-3.0) µm in width. Both polar capsules contained polar tubules with 10-12 coils. For Myxobolus tambaquiensis n. sp., plasmodia were found in the opercular cavity of 41.7% of the fish specimens, and the myxospores had an oval shape, measuring 10.0 ± 0.4 (9.5-11.3) µm in length, 6.5 ± 0.2 (6.1-7.1) µm in width, and 4.6 ± 0.4 (3.9-5.6) µm in thickness. The polar capsules were elongated and equal in size, measuring 4.9 ± 0.2 (4.4-5.3) µm in length and 1.9 ± 0.2 (1.5-2.2) µm in width, closing with 8-9 coils of the polar tubule. The morphological and sequencing data of the SSU rDNA showed that the three species studied herein remain unknown to science, increasing the diversity of myxosporeans infecting C. macropomum, an iconic fish in South American freshwater fish farming. The SSU rDNA based phylogenetic analysis revealed that Myxobolus spp. parasites of C. macropomum did not have a monophyletic origin, identifying different times and pathways of the acquisition of parasites by this host species.

Broad but restricted detection of malacosporeans in a Neotropical cradle of diversification.

This study undertook the first investigation of malacosporean infections in Neotropical fish. We used polymerase chain reaction detection with a primer set generally targeting known malacosporeans to assay for infection in the kidney of 146 fish in 21 species belonging to 12 families collected from two areas in the Amazon Basin. Infections were found in 13 fish variously belonging to seven species in six families and included the first identification of a malacosporean infection in cartilaginous fish (a freshwater stingray). Based on ssrDNA, all infections represented a single Buddenbrockia species (Buddenbrockia sp. E) that demonstrates an exceptionally broad range of fish species infected, and countered our expectations of high Neotropical malacosporean diversity. Infections were characterized at varying and often high prevalences in fish species but sample sizes were small. Ascertaining whether highly divergent malacosporeans have not been detected by current primers, and more comprehensive sampling may reveal whether malacosporeans are truly as species poor in the Amazon Basin as present data suggest. Our results prompt speculations about evolutionary scenarios including introduction via marine incursions and patterns of host use over time.

PCR-RFLP: a targeted method to reveal host specific malacosporean infection profiles (Cnidaria: Myxozoa: Malacosporea).

Malacosporeans are a group of endoparasitic cnidarians (Myxozoa) that use freshwater bryozoans and fish as final and intermediate hosts, respectively. The malacosporean Tetracapsuloides bryosalmonae causes proliferative kidney disease (PKD), an emerging disease in aquaculture and wild fish populations, including threatened salmonids in Europe and the USA. Mixed infections of malacosporeans are often encountered, and a monitoring tool for screening of multiple malacosporean species in either their fish or bryozoan hosts is therefore desirable. We describe an inexpensive method that combines PCR amplification of the partial 18S rRNA gene (~260 bp) and a single-step restriction fragment length polymorphism (RFLP) method for identification of 10 malacosporean lineages and species. We demonstrate and test this methodology on a set of DNA extracted from malacosporeans infecting fish kidney and tissues sampled from bryozoan colonies and compare the results with Sanger sequencing of the same parasite DNA isolates. The PCR-RFLP and Sanger sequencing methods agreed in 100% of cases. The PCR-RFLP method offers a number of opportunities, including screening large panels of host tissue samples to gain insights into infection patterns, characterizing mixed infections, and confirming highly pathogenic T. bryosalmonae infections. The method can also be further refined as new sequence data become available for malacosporeans.

Increasing the known biodiversity of cnidarian parasites of bryconid fishes from South America: two novel Myxobolus species with ultrastructure and ssrDNA-based phylogeny.

This study increases the known biodiversity of cnidarian parasites in neotropical bryconid fishes. Two novel Myxobolus species are described based on morphology, ultrastructure and small subunit ribosomal DNA (ssrDNA) sequencing: Myxobolus vetuschicanus n. sp. infecting fins of Salminus franciscanus and Myxobolus mineirus n. sp. infecting the mesentery of Brycon orthotaenia from the São Francisco River basin, Minas Gerais State, Brazil. Ultrastructural analysis of the two species revealed an asynchronous sporogenesis process, with germinative cells and young developmental stages of myxospores in the periphery of the plasmodia. In M. vetuschicanus n. sp., the plasmodia were surrounded by a layer of fibroblasts and in M. mineirus n. sp., the plasmodial membrane had direct contact with the host tissue. The phylogenetic analysis based on the ssrDNA of Henneguya/Myxobolus species showed that the two novel Myxobolus species grouped in subclades together with other parasite species of bryconid fishes.

Novel myxosporean species parasitizing an economically important fish from the Amazon basin.

This paper provides morphological and phylogenetic analyses of two new myxobolid species found infecting Piaractus brachypomus from the Amazon basin. The fish were caught in the Tapajós River, in the municipality of Santarém, in the state of Pará, Brazil. The plasmodial development of Henneguya brachypomus n. sp. occurred in the gill lamellae while Myxobolus pirapitingae n. sp. developed in the pyloric cecum. Morphological analyses did not identify inflammatory infiltrate for either species, but H. brachypomus n. sp. induced stretching of the epithelium, compression of the adjacent tissues, and displacement and deformation of the neighboring lamellae. The mature myxospores of H. brachypomus n. sp. were ellipsoid, with a length of 11.7-13.8 µm, a width of 4.0-4.6 µm, and a thickness of 3.5-4.3 µm. The polar capsules were elongated, with a length of 5.6-7.3 µm and a width of 1.3-2.0 µm, and each contained a polar filament with 8-9 coils. The caudal process was 40.5-48.1 µm long and the total length of the myxospore was 52.4-61.6 µm. Myxobolus pirapitingae n. sp. exhibited rounded mature myxospores measuring 10.0-11.1 µm in length, 7.0-7.6 µm in width, and 5.4-6.3 µm in thickness. The polar capsules were of equal size and occupied less than half the myxospore, measuring 3.5-4.0 µm in length and 2.0-2.6 µm in width, with each containing a polar filament with 6-7 coils. Phylogenetic analysis based on partial small subunit ribosomal DNA (ssrDNA) sequences showed that H. brachypomus n. sp. clustered as a sister species of Henneguya piaractus, while M. pirapitingae n. sp. was grouped in a sub-clade together with Myxobolus matosi and Myxobolus colossomatis.

Taxonomy, phylogeny and host-parasite interaction of two novel Myxobolus species infecting Brycon orthotaenia from the São Francisco River, Brazil.

Two new Myxobolus species were described infecting Brycon orthotaenia from the São Francisco River, in the state of Minas Gerais, Brazil. From a total of 39 B. orthotaenia collected, two specimens (5.1%) exhibited infection of the ovary and 12 specimens (30.8%) displayed infection of the liver. The plasmodia of both Myxobolus species were white and spherical measuring around 1 mm in length. The plasmodium found in the ovary showed mature myxospores, which were oval shaped from the frontal view and measured 9.2-11.0 (9.8 ± 0.4) µm in length, 5.9-6.9 (6.5 ± 0.3) µm in width and 4.6-5 (4.9 ± 0.1) µm in diameter. The two polar capsules were the same size and measured 3.9-6.2 (4.7 ± 0.5) µm in length and 1.8-2.4 (2.1 ± 0.2) µm in width. The polar tubules had 9 coils. The plasmodium found in the liver showed mature myxospores which were ellipsoidal in shape from the frontal view and measured 10.0-11.4 (10.7 ± 0.5) µm in length, 7.3-8.6 (8.1 ± 0.4) µm in width and 5.3-7.0 (6.8 ± 0.4) µm in diameter. The two polar capsules were the same size and measured 4.2-5.4 (4.9 ± 0.3) µm in length and 1.9-2.9 (2.7 ± 0.3) µm in width. The polar tubules had 8 coils. Ultrastructural analysis revealed an asynchronous sporogenesis process, with young developmental myxospore stages more often found in the periphery of the plasmodium and mature myxospores in the centre of the plasmodium. The plasmodial wall was formed by a single membrane which was not surrounded by a layer of host tissue. A thick layer of fibrous material was found in the peripheral ectoplasm close to the plasmodial wall of the plasmodium found in the ovary. Phylogenetic analysis based on the small-subunit ribosomal DNA - ssrDNA sequences and using the closest myxozoan sequences to each one of the species studied here based on previous GenBank data and Henneguya/Myxobolus/Thelohanellus species parasitizing fish from South American, revealed that the new species are grouped in a subclade together with other Myxobolus species parasitizing bryconid hosts.

Malacosporean myxozoans exploit a diversity of fish hosts.

Myxozoans are widespread and common endoparasites of fish with complex life cycles, infecting vertebrate and invertebrate hosts. There are two classes: Myxosporea and Malacosporea. To date about 2500 myxosporean species have been described. By comparison, there are only five described malacosporean species. Malacosporean development in the invertebrate hosts (freshwater bryozoans) has been relatively well studied but is poorly known in fish hosts. Our aim was to investigate the presence and development of malacosporeans infecting a diversity of fish from Brazil, Europe and the USA. We examined kidney from 256 fish belonging variously to the Salmonidae, Cyprinidae, Nemacheilidae, Esocidae, Percidae, Polyodontidae, Serrasalmidae, Cichlidae and Pimelodidae. Malacosporean infections were detected and identified by polymerase chain reaction and small subunit ribosomal DNA sequencing, and the presence of sporogonic stages was evaluated by ultrastructural examination. We found five malacosporean infections in populations of seven European fish species (brown trout, rainbow trout, white fish, dace, roach, gudgeon and stone loach). Ultrastructural analyses revealed sporogonic stages in kidney tubules of three fish species (brown trout, roach and stone loach), providing evidence that fish belonging to at least three families are true hosts. These results expand the range of fish hosts exploited by malacosporeans to complete their life cycle.

Morphological, ultrastructural, and phylogenetic analysis of two novel Myxobolus species (Cnidaria: Myxosporea) parasitizing bryconid fish from São Francisco River, Brazil.

Twelve Myxobolus species have been previously described to parasitize Bryconidae fish in South America. Here, we describe two novel myxosporean species that parasitize economically important Bryconidae from the São Francisco River basin in Brazil. Myxospores morphometry, morphology, small-subunit ribosomal DNA - ssrDNA sequences, and other biological traits were used in the taxonomic analysis. Phylogenetic analysis was performed to assess the position of the new Myxobolus species among the closest Myxobolus/Henneguya. Myxobolus iecoris n. sp. was found infecting the liver of Salminus franciscanus (dourado). Myxospores were oval with the anterior region aculiform in frontal view and biconvex in lateral view and measured 11.4-14.2 (12.8 ±â€¯0.8) µm long, 7.7-9.9 (8.7 ±â€¯0.6) µm wide, 6.5-7.5 (6.9 ±â€¯0.4) µm thick. Two pyriform and equal-sized polar capsules measuring 4.9-7.4 (5.9 ±â€¯0.5) µm long and 2.3-3.5 (3.0 ±â€¯0.2) µm wide contained polar tubules with 8-9 turns. Myxobolus lienis n. sp. was found infecting the spleen of Brycon orthotaenia (matrinxã). Myxospores were round to oval in frontal view and biconvex in lateral view and measured 10.3-13.8 (12 ±â€¯0.6) µm long, 6.8-9.3 (8.3 ±â€¯0.5) µm wide, and 6.9-7.0 (7.0 ±â€¯0.6) µm thick. Two oval and equal-sized polar capsules measured 3.9-5.8 (4.6 ±â€¯0.5) µm long and 2.0-3.5 (2.8 ±â€¯0.3) µm wide contained polar tubules with 5-6 turns. Ultrastructural analysis revealed asynchronous sporogenesis with germinative cells and young sporogonic stages in the periphery of the plasmodia. A connective tissue capsule was observed surrounding Myxobolus lienis n. sp., but it was absent for Myxobolus iecoris n. sp. Maximum likelihood and Bayesian inferences showed the two novel species clustering in a well-supported subclade composed by Myxobolus spp. of bryconids. Myxobolus iecoris n. sp. appeared as a sister species of M. aureus and Myxobolus lienis n. sp. as sister to M. umidus.

Extensive Uncharted Biodiversity: The Parasite Dimension.

Parasites are often hidden in their hosts and exhibit patchy spatial distributions. This makes them relatively difficult to detect and sample. Consequently we have poor knowledge of parasite diversities, distributions, and extinction. We evaluate our general understanding of parasite diversity and highlight the enormous bias in research on parasites such as helminths and arthropods that infect vertebrate hosts. We then focus on Myxozoa as an exemplary case for demonstrating uncharted parasite diversity. Myxozoans are a poorly recognized but speciose clade of endoparasitic cnidarians with complex life cycles that have radiated to exploit freshwater, marine, and terrestrial hosts by adopting strategies convergent to those of parasitic protists. Myxozoans are estimated to represent some 20% of described cnidarian species-greatly outnumbering the combined species richness of scyphozoans, cubozoans, and staurozoans. We summarize limited understanding of myxozoan diversification and geographical distributions, and highlight gaps in knowledge and approaches for measuring myxozoan diversity. We close by reviewing methods and problems in estimating parasite extinction and concerns about extinction risks in view of the fundamental roles parasites play in ecosystem dynamics and in driving host evolutionary trajectories.

New myxosporeans parasitizing Phractocephalus hemioliopterus from Brazil: morphology, ultrastructure and SSU-rDNA sequencing.

Myxozoans are a diverse group of parasitic cnidarians, with some species recognized as serious pathogens to their hosts. The present study describes 2 new myxobolid species (Myxobolus figueirae sp. nov. and Henneguya santarenensis sp. nov.) infecting skin and gill filaments of the Amazonian pimelodid fish Phractocephalus hemioliopterus, based on ultrastructural, histology and phylogenetic analysis. The fish were caught in the Amazon River, Pará, Brazil. The plasmodial development of M. figueirae sp. nov. was in the dermis and those of H. santarenensis sp. nov. were of the intralamellar type. For both species, the plasmodia were surrounded by a connective tissue layer, but there was no inflammatory infiltrate. For M. figueirae sp. nov., mature spores were ovoid measuring 9.1 to 10 (9.5 ± 0.3) µm in length, 5.8 to 6.9 (6.4 ± 0.3) µm in width and 4.4 to 4.5 (4.5 ± 0.1) µm in thickness. Two polar capsules were elongated and of unequal size. For H. santarenensis sp. nov., mature spores were ellipsoidal in the frontal view, measuring 26.3 to 36.1 (31.9 ± 3) µm in total length, 9.6 to 11.9 (10.8 ± 0.5) µm in body length, 3.7 to 4.9 (4.3 ± 0.3) µm in width and 16.6 to 25.6 (21 ± 3.1) µm in caudal process. The polar capsules were elongated and of equal size. Phylogenetic analysis, based on partial small subunit ribosomal DNA (SSU rDNA) sequences and using the closest myxozoan sequences to each one of the species studied here based on previous GenBank data, showed M. figueirae sp. nov. and H. santarenensis sp. nov. clustering in distinct lineages. While H. santarenensis sp. nov. clustered in a well-supported subclade composed of Henneguya species that infect gills of South American pimelodid hosts, M. figueirae sp. nov. clustered in a weakly supported subclade containing parasite species of bryconid hosts.

Morphology and small subunit rDNA-based phylogeny of a new Henneguya species, infecting the ornamental fish Corydoras leucomelas from the Peruvian Amazon.

A new species of Myxosporea, Henneguya loreotoensis n. sp. is described parasitizing the gill filaments from 17 of 35 specimens (48.5%) of Corydoras leucomelas (Siluriformes: Callichthyidae) caught in the Nanay River, near village Ninarumi, in the Loreto state, Peru. Mature spores were ellipsoidal in shape from the frontal view, measuring 36.2±0.1µm (36.1-36.3) in total length, 14.3±0.1µm (14.2-14.4) in body length, 5.1±0.1µm (4.9-5.3) in width and 21.9±0.1µm (21.8-22.0) in the caudal process. The two polar capsules were symmetrical and elongated, measuring 5.1±0.1µm (4.9-5.3) in length and 2.4±0.2µm (2.1-2.7) in width, containing a polar filament with five coils arranged obliquely to the longitudinal axis. The sporoplasm was binucleate. Partial sequencing of the ssu-rDNA of H. loretoensis n. sp. resulted in a total of 1676 nucleotides, and this sequence did not match any of the myxozoan available in the GenBank. The phylogenetic analysis shows H. loretoensis n. sp. as a sister species of Henneguya paraensis, another amazonian myxozoan parasite of Cichla temensis (Perciformes: Cichlidae).

Morphology and small subunit rDNA-based phylogeny of Ceratomyxa amazonensis n. sp. parasite of Symphysodon discus, an ornamental freshwater fish from Amazon.

The specious genus Ceratomyxa Thélodan, 1892, infect mainly gallbladder of marine fishes, with only five species reported infecting species from freshwater environment. This study performed morphological and phylogenetic analyses involving a new Ceratomyxa species (Ceratomyxa amazonensis n. sp.) found in gallbladder of Symphysodon discus Heckel, 1840 (Perciformes: Cichlidae), an important ornamental fish endemic to Amazon basin. Mature spores were strongly arcuate shaped and measured 7.0 ± 0.3 (6.2-7.6) µm in length, 15.8 ± 0.4 (15.0-16.7) µm in thickness, and polar capsules 3.22 ± 0.34 (2.4-3.6) µm in length and 2.63 ± 0.17 (2.4-2.9) µm in width. This was the first small subunit ribosomal DNA (SS rDNA) sequencing performed to Ceratomyxa species parasite of freshwater fish, and the phylogenetic analysis showed C. amazonensis n. sp. clustering in the early diverging subclade of the ceratomyxids, together with species of parasites of amphidromous/estuaries fishes, suggesting some role of the transition of the fishes between marine/freshwater environments in the evolutionary history of these parasites.

Host-parasite and phylogenetic relationships of Myxobolus filamentum sp. n. (Myxozoa: Myxosporea), a parasite of Brycon orthotaenia (Characiformes: Bryconidae) in Brazil.

Myxobolus filamentum sp. n. was found infecting gill filaments of three of 39 Brycon orthotaenia Günther specimens examined (8%), which were taken from the river São Francisco in Minas Gerais state, Brazil. Plasmodia of the parasite were white and long, measuring 5 mm in lenght. Mature spores of M. filamentum sp. n. were oval from the frontal view and biconvex from the lateral view, measuring 7.5-9.7 µm (9.0 ± 0.3 µm) in length and 5.2-7.3 µm (6.2 ± 0.4 µm) in width. The polar capsules were elongated and equal in size, measuring 3.8-5.5 µm (4.7 ± 0.3 µm) in length and 1.3-2.2 µm (1.7 ± 0.1 µm) in width. The development of the parasite led to compression of the adjacent tissues and inflammatory infiltrate with granulocytic cells. Ultrastructural observation revealed that the plasmodia were delimited by two membranes, which had numerous and extensive pinocytotic channels extending into the wide ectoplasm zone. The plasmodial wall exhibited abundant villi-like projections and a thin layer of granular material prevented direct contact between the plasmodial wall and the host tissue. Phylogenetic analysis, based on 18S rDNA, showed M. filamentum sp. n. as a sister species of Myxobolus oliveirai Milanin, Eiras, Arana, Maia, Alves, Silva, Carriero, Ceccarelli et Adriano, 2010, a parasite of other fish species of the genus Brycon Müller et Troschel from South America.

Morphology, ultrastructure and phylogeny of Myxobolus curimatae n. sp. (Myxozoa: Myxosporea) a parasite of Prochilodus costatus (Teleostei: Prochilodontidae) from the São Francisco River, Brazil.

Myxobolus curimatae n. sp. has been found infecting the gill filaments of Prochilodus costatus (Prochilodontidae) from the São Francisco River in the state of Minas Gerais, Brazil. The prevalence of the species was 18.7%. Mature spores were rounded from a frontal view, with elongated polar capsules of equal size, and had polar filaments with 9-10 turns. Ultrastructural analysis revealed that sporogenesis patterns followed those of other Myxobolus species. The plasmodium walls had numerous invaginations and protrusions, and few pinocytic channels. Numerous mitochondria, generative cells and young pansporoblasts were observed in the peripherical areas of the plasmodia, and mature spores were found in deeper layers. A layer of collagenic fibrils surrounded the plasmodia. The morphological data and molecular analysis of the 18S rDNA identified this parasite as a new species. The maximum likelihood phylogenetic tree showed M. curimatae n. sp., as a sister species of Thelohanellus marginatus, in a basal branch of the subclade composed by parasites with tropism to different organs and host families.

Henneguya cuniculator sp. nov., a parasite of spotted sorubim Pseudoplatystoma corruscans in the São Francisco Basin, Brazil.

Henneguya cuniculator sp. nov. was found infecting spotted sorubim catfish Pseudoplatystoma corruscans from the São Francisco River, Minas Gerais, Brazil. The parasites form elongated plasmodia of up to 1 cm in length in the gill filaments. Mature spores were ellipsoidal from the frontal view, with total length of 29.4 ± 2.4 (mean ± SD, range 23.3-32.4) µm, body length of 12.1 ± 1.0 (10.0-14.7) µm, width of 4.8 ± 0.4 (4.0-5.9) µm, and tail length of 16.7 ± 2.0 (12.3-19.4) µm. From the lateral view, spores were biconvex, with thickness of 4.2 ± 0.7 (3.9-4.9) µm. The polar capsules were elongated and equal in size, 6.2 ± 0.3 (5.2-6.2) µm in length, and 1.8 ± 0.1 (1.4-1.9) µm in width. Ultrastructural analysis showed that the plasmodial wall had delicate projections towards the host tissue and a thin layer that prevented contact between the host cells and the parasite. In the ectoplasm, few mitochondria were observed, while generative cells, early stages of sporogenesis, and advanced spore development occurred in the plasmodial periphery, and more mature spores in internal regions. Histopathological analysis showed that plasmodia developed in the sub-epithelial connective tissue of gill filaments, causing compression of the adjacent tissues, deformation of gill filaments, and lamellar fusion. Phylogenetic analysis, based on 18S rDNA genes and using only Henneguya/Myxobolus species parasites of siluriform fish, showed grouping according to the fish family.