Light and Electron Microscopy of the Spore of Myxobolus heckelii n. sp. (Myxozoa), Parasite from the Brazilian Fish Centromochlus heckelii (Teleostei, Auchenipteridae)

ABSTRACT. A myxosporean parasitizing the gill filaments of the freshwater teleost fish Centromochlus heckelii collected in the Tocantins River (Lower Amazonian Region, Brazil) is described using light and electron microscopy. This parasite produces spherical to ellipsoidal cyst-like plasmodia up to 250 μm in diameter, with a thick wall strengthened by several stratified juxtaposed crossed collagen layers, whose thickness varies according to the number of the layers. Several compressed fibroblasts are observed among the collagen fibrils. Deposits of spherical dense material are scattered at the internal periphery of the cysts. Plasmodia and different developmental stages, including immature and mature spores, filled the central region of the cysts. The spore body is ellipsoidal in valvar view and biconvex in sutural view. It is formed by two equal-sized and symmetric valves measuring 12.7 μm long (12.2–13.1) ( n =50), 6.6 μm wide (6.3–6.9) ( n =25), and 4.0 μm (3.7–4.4) ( n =20) thick. A thin layer formed by fine and anastomosed microfibrils is observed at the spore surface. Two equal, elongated pyriform polar capsules measure 2.9 μm (2.7–3.3) × 1.7 μm (1.4–2.0) ( n =25), each containing four or five oblique polar filament coils. The binucleated sporoplasm contains numerous spherical sporoplasmosomes, glycogen particles, and a large vacuole with fine granular matrix. Based on the morphological and ultrastructural differences and specificity of the host, we describe this isolate as a new myxosporidian, Myxobolus heckelii n. sp. (Myxozoa, Myxosporea).     

Myxobolus albi n. sp. (Myxozoa) from the Gills of the Common Goby Pomatoschistus microps Krøyer (Teleostei: Gobiidae)

ABSTRACT. A recent investigation into the myxozoan fauna of common gobies, Pomatoschistus microps , from the Forth Estuary in Scotland, revealed numerous myxosporean cysts within the gill cartilage. They were composed of polysporous plasmodia containing myxobolid spores that were morphologically different from the other known species of Myxobolus and from the myxosporeans previously recorded from this host (i.e. the ceratomyxid Ellipsomyxa gobii , infecting the gall bladder, and the kudoid Kudoa camarguensis , infecting the muscle tissues). Spores were ovoid, 9.4 × 9.1 μm with a thickness of 6.6 μm, with two pyriform polar capsules, the polar filaments of which had four to five turns. Molecular analysis of the parasite's small subunit rDNA region, based upon a contiguous sequence of 1,558 base pairs, discriminated it from other myxosporean species that have been characterized so far. A comparison of the spore morphology and the molecular sequences determined for this new isolate with other myxozoans described to date, confirmed its identity as a previously unknown myxobolid supporting the proposal that this isolate be elevated to the species level as a new species within the genus Myxobolus . A phylogenetic analysis places this new myxobolid, Myxobolus albi n. sp., in a basal position of a clade containing the majority of Henneguya spp. sequenced to date and various Myxobolus spp.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

Henneguya schizodon n. sp. (Myxozoa, Myxobolidae), a parasite of the Amazonian teleost fish Schizodon fasciatus (Characiformes, Anostomidae)

A new histozoic species of myxosporean (Henneguya schizodon n. sp.) is described from the Amazon River teleost fish Schizodon fasciatus Spix & Agassiz, 1892 (Characiformes, Anostomidae). The plasmodia, which showed asynchronous development, were located in the kidney of the host. The spore body was ellipsoidal and was 13.1 (12-14) micron long by 3.3 (3-4) micron wide. The total length of the spore was 28.9 (27-30) micron, and each value had a caudal process measuring 16.3 (15-17) micron. The polar capsules were 5.4 (5-6) micron long by 1.3 (1-1.5) micron wide, and each had a polar filament with 8-10 coils. The characteristics of the species were compared with nearly all the species described so far, including all the species reported from South American fishes. This comparison allows to consider the materials as a new species, and the name Henneguya schizodon n. sp. is proposed.     

Light and ultrastructural description of Meglitschia mylei n. sp. (myxozoa) from Myleus rubripinnis (Teleostei: Serrasalmidae) in the Amazon River system

Meglitschia mylei n. sp. found in the gall bladder of the teleostean fish Myleus rubripinnis (Serrasalmidae) from the middle Amazonian region of Brazil is described using light and transmission electron microscopy. The spores observed in the bile averaged 24.6±0.8 μm long, 8.7±0.4 μm wide and 5.1±0.3 μm thick and were strongly furcate and arcuate ∩-shaped composed of two symmetric equal-sized valves, up to ～70 nm thick. Each valve possessed one opposed tapering appendage, 20.1±0.7 μm long, oriented parallel towards the basal tip of the appendages and joined along a right suture line forming a thick strand. The strand goes around the central part of the spore, which in turn surrounds two equal and symmetric spherical polar capsules (PC), 2.1±0.3 μm in diameter, located at the same level. Each capsule contains a polar filament with five (rarely six) coils. The binucleate sporoplasm was irregular in shape, contained several sporoplasmosomes, ～175 nm in diameter and filled all the space of the two caudal appendages. Based on the arc shape of the spore with two tapering caudal appendages oriented to the basis of spores, on the number and position of the PC and of the polar filament coils and arrangements, and on the host specificity, we propose the name M. mylei n. sp. for this new myxozoan. Accordingly, this is the second described species of this genus.     

Ultrastructural and molecular characterization of a new microsporidium parasite from the Amazonian fish, Gymnorhamphichthys rondoni (Rhamphichthyidae)

A new species of a microsporidium found in the freshwater teleost Gymnorhamphichthys rondoni, collected on the lower Amazon River, is described based on light, ultrastructural, and phylogenetic studies. This parasite develops in the skeletal muscle of the abdominal cavity, forming whitish cyst-like structures containing numerous spores. Mature spores, lightly pyriform to ellipsoidal with rounded ends and measuring 4.25 ± 0.38 × 2.37 ± 0.42 μm (n = 30), were observed. The spore wall, which measured about 102 nm, was composed of 2 layers with approximately the same thickness. The isofilar polar filament was coiled, with 9-10 (rarely 8) turns. The posterior vacuole appeared as a pale area, occupying about 1/3 of the spore length, and contained a spherical posterosome composed of granular material that was denser at the periphery. The myofibrils located near the spores appeared to be in advanced degradation. Molecular analysis of the rRNA genes, including the ITS region, and phylogenetic analyses using maximum parsimony, maximum likelihood, and Baysesian inference were performed. The ultrastructural characteristics of the spores and the phylogenetic data strongly suggested that it is a new species related to Kabatana, Microgemma, Potaspora, Spraguea, and Tetramicra. We named this new microsporidian from Amazonian fauna as Kabatana rondoni n. sp.     

Ultrastructural data on the spore of Myxobolus maculatus n. sp. (phylum Myxozoa), parasite from the Amazonian fish Metynnis maculatus (Teleostei)

Light and electron microscopy studies of a myxosporean, parasitic in the intertubular interstitial tissue of the kidney of the freshwater teleost fish Metynnis maculatus Kner, 1860 (Characidae) from the lower Amazon River (Brazil), are described. We observed polysporic histozoic plasmodia delimited by a double membrane and with several pinocytic channels and containing several life cycle stages, including mature spores. The spore body was of pyriform shape and was 21.0 microm long, 8.9 microm wide and 7.5 microm thick. Elongated-pyriform polar capsules were of equal size (12.7 x 3.2 microm) and contained a polar filament with 14 or 15 coils. The spore features fit those of the genus Myxobolus. Densification of the capsular primordium matrix, which increased in density from the inner core outwards, differentiating at the periphery into small microfilaments measuring 45 nm each, and tubuli arranged in aggregates and dispersed within the capsular matrix of the mature spores, are described. Based on the morphological differences and specificity of the host, we propose the creation of a new species named Myxobolus maculatus n. sp.     

Light and ultrastructural data on Henneguya testicularis n. sp. (Myxozoa,Myxobolidae), a parasite from the testis of the Amazonian fish Moenkhausia oligolepis

Light and electron microscopical data on a myxosporidian found in the testis of Moenkhausia oligolepis Gunther (Teleostei, Characidae, Tetragonopterinae) from the lower Amazon River near Belém, Brazil, are described. Based on spore morphology, we conclude that this species belongs to the family Myxobolidae and genus Henneguya. Mature spores (total length 27.5 (27.0–28.5) µm) were observed at the periphery of the testis. The ellipsoidal spore body consists of two unequal shell valves adhering together along the suture lines. The spore body is ellipsoidal, 14.0 µm long and 6.5 µm wide. Each valve tapers to a single caudal projection, forming a 13.5 µm long tail (13.0–14.5 µm). The spore is surrounded by a homogenous dense sheath. The polar capsules, measuring 9.0 × 2.0 µm, contain 12–13 coils of the polar filaments. Morphological differences between this material and other species of the genus Henneguya indicated the erection of a new species, which was named H. testicularis. The taxonomic affinities of this parasite are discussed.     

Two new species of Myxozoa, Myxobolus inaequus sp. n. and Henneguya theca sp. n. from the brain of a South American knife fish, Eigemannia virescens (V.)

Two new species of Myxozoa from the brain of the green knife fish Eigemannia virescens are described: Myxobolus inaequus sp. n. has an unusually large spore body and extremely unequal polar capsules, and Henneguya theca sp. n. has an attenuated spore encased in a sheath not previously described in other Myxozoa . Only spores of the two species were observed, and infections caused no obvious pathological changes in the brain.     

Ultrastructural studies of Henneguya rhamdia n. sp. (Myxozoa) a parasite from the Amazon teleost fish, Rhamdia quelen (Pimelodidae)

Henneguya rhamdia n. sp. is described in the gill filaments of the teleost fish Rhamdia quelen, collected from the Peixe Boi River, State of Pará, Brazil. This myxosporean produced spherical to ellipsoidal plasmodia, up to 300 microm in diameter, which contained developmental stages, including spores. Several dense bodies up to 2 microm in diameter were observed among the spores. The spore body was ellipsoidal (13.1 microm in length, 5.2 microm in width, and 2.5 microm in thickness) and each of the two valves presented a tapering tail (36.9 microm in length). These valves surrounded the binucleated sporoplasm cell and two equal ellipsoidal polar capsules (4.7 x 1.1 microm), which contained 10-11 (rarely 12) polar filament coils. The sporoplasm contained sporoplasmosomes with a laterally eccentric dense structure with a half-crescent section. Based on the data obtained by electron microscopy and on the host specificity, the spores differed from previously described Henneguya species, mainly in their shape and size, number and arrangement of the polar filament coils, and sporoplasmosome morphology.     

Ultrastructural aspects of the myxosporean Henneguya astyanax n. sp. (Myxozoa: Myxobolidae), a parasite of the Amazonian teleost Astyanax keithi (Characidae)

This study reports light and electron microscopical aspects of a myxosporean found in the gills of the freshwater teleost Astyanax keithi Géry, Planquete & Le Bail, 1996 (family Characidae), collected from the estuarine region of the Amazon River, near Belém, Brazil. The prevalence of infection was 23%. In interlamellar spaces of the gills, ellipsoidal whitish cyst-like plasmodia structures were present, which contained spores. The spores had a spermatozoa-like appearance (47.8 +/- 0.71 microm in total length) with a fusiform body (15.2 +/- 0.77 pm in length, 5.7 +/- 0.71 microm in width and 4.2 +/- 0.31 microm in thickness), and each of the 2 valves presented a tapering tail (32.6 +/- 1.11 microm in length). The valves surrounded a binucleate sporoplasm cell and 2 polar capsules (5.0 +/- 0.13 microm in length, 1.5 +/- 0.07 microm in width) that contained 8 to 9 coils of the polar filament. In the sporoplasm, several unique sporoplasmosomes were visible. A synoptic table of spore measurements of known Brazilian Henneguya species is presented. The spores differed from those of previously described species. Based on spore morphology, it is concluded that this species belongs to the family Myxobolidae, genus Henneguya, and that it constitutes a new species: H. astyanax n. sp.     

Ultrastructure of Myxobolus brycon n. sp. (Phylum Myxozoa), parasite of the piraputanga fish Brycon hilarii (Teleostei) from Pantanal (Brazil)

Light and electron microscopy studies of a myxosporean, parasitizing the gill filaments of the freshwater fish Brycon hilarii (Valenciennes, 1850) (Characidae) collected in the Paraguay River (18°49'S, 57°39'W) (Pantanal), Brazil, is described. This parasite produces spherical to ellipsoidal polysporic histozoic plasmodia (Pmd) (up to ～180 μm in diameter) delimited by a double membrane and with several pinocytic channels. The plasmodial cyst contained the youngest developmental stages at the cortical periphery and immature and mature spores more internally. The Pmd developed near the cartilaginous structure of the gill filament, forming a prominent deformation where the gill lamellae disappear. Pyriform spores measured 6.9±0.6 (range 6.5-7.2) μm long, 4.2±0.5 (range 3.9-4.8) μm wide, and 2.5±0.7 (range 1.9-2.8) μm thick. The spores composed of two equal shell valves (～70 nm thick), adhering together along the straight suture line, surrounded two equal symmetric and elongated to pyriform polar capsules (PC) 4.2±0.6 (range 3.8-4.7) × 1.9±0.6 (1.7-2.5) μm; each PC contained a coiled polar filament with eight or nine (rarely 10) turns and a binucleated sporoplasm cell. Dense irregular masses were observed among the polar filaments coils. An intercapsular appendix was not observed. The sporoplasm contained several globular sporoplasmosomes randomly distributed among an extensive rough endoplasmic reticulum system with numerous vesicles and cisternae. Based on the morphological and ultrastructural differences and specificity of the host, we establish the new species, Myxobolus brycon n. sp.     

Myxobolus insignis sp. n. (Myxozoa, Myxosporea, Myxobolidae), a parasite of the Amazonian teleost fish Semaprochilodus insignis (Osteichthyes, Prochilodontidae)

A new myxosporean species is described from the fish Semaprochilodus insignis captured from the Amazon River, near Manaus. Myxobolus insignis sp. n. was located in the gills of the host forming plasmodia inside the secondary gill lamellae. The spores had a thick wall (1.5-2 microm) all around their body, and the valves were symmetrical and smooth. The spores were a little longer than wide, with rounded extremities, in frontal view, and oval in lateral view. They were 14.5 (14-15) microm long by 11.3 (11-12) microm wide and 7.8 (7-8) microm thick. Some spores showed the presence of a triangular thickening of the internal face of the wall near the posterior end of the polar capsules. This thickening could occur in one of the sides of the spore or in both sides. The polar capsules were large and equal in size surpassing the midlength of the spore. They were oval with the posterior extremity rounded, and converging anteriorly with tapered ends. They were 7.6 (7-8) microm long by 4.2 (3-5) microm wide, and the polar filament formed 6 coils slightly obliquely to the axis of the polar capsule. An intercapsular appendix was present. There was no mucous envelope or distinct iodinophilous vacuole.     

A new species of Remijia (Rubiaceae) from Amazonian Brazil

Remijia hubbardiorum B. M. Boom, endemic to white sandcampinas orerra firme forest in the Rio Uatumã region in the State of Amazonas of north central Brazil, is described and illustrated, and its morphological differentiation with respect toR. asperula Standley is discussed.     

A new species of Henneguya (Myxozoa) in the big-eyed scad (Selar crumenophthalmus) from Hawaii

We describe a new myxozoan, Henneguya akule n. sp., infecting the carangid fish Selar crumenophthalmus in Hawaii. Spores were found only in the aortic bulb, characterized by elliptical capsule with 2 tails, and pyriform polar capsules that angled toward the anterior end of the spore. Polar filaments had 3-4 coils. Parasites were present in apparently healthy fishes and caused no evident gross pathology. On microscopy, parasites evinced a mild inflammatory response in the host characterized by accumulations of eosinophilic fibrillar material around spores and a mononuclear infiltrate in the adventitia of the bulbus arteriosus. Overall prevalence was 20%, and prevalence between 2001 and 2006 ranged from 12 to 27%, but did not differ significantly between years. In contrast, prevalence of infection was highest in south-central Oahu. There was no relationship between infection status and body condition or gender of fish, and infection was absent in the smallest and largest fishes. Phylogenetically, H. akule n. sp. is most closely related to other Henneguya species infecting the heart of marine fishes based on ribosomal DNA analysis. This is the first documentation of a myxozoan parasite in marine fishes from Hawaii.     

Parvicapsula bicornis n. sp. and P. limandae n. sp. (Myxozoa, Parvicapsulidae) in Pleuronectidae (Teleostei, Heterosomata) from Denmark

Two species of Parvicapsula were found in the kidney tubules and the urinary bladder of 2 pleuronectid fish from the northern Oresund, Denmark. The coelozoic, spherical, disporic trophozoites of both species are 10 to 12 pm in diameter. The myxospores of both species are elongate, asymmetrical and slightly curved, and have spherical polar capsules. Parvicapsula bicornis n. sp. (6-8 x 5-6 microm, polar capsule 2.5 microm in diameter) occurs in Pleuronectes platessa. The polar capsules of P. bicornis are arranged symmetrically on either side of the longitudinal axis and its spores differ from other species of Parvicapsula in having two 2-3 microm long posterior processes of different length. Parvicapsula limandae n. sp. (8-11 x 4-5 pm, polar capsule 1.6 microm in diameter) is found in Limanda limanda. The polar capsules are arranged along the longitudinal axis. It differs from Parvicapsula unicornis Kabata, 1962, recorded from L. limanda, in the arrangement of the polar capsules and in the absence of a posterior horn-like projection. The phylogenetic relationship between P. bicornis n. sp., P. limandae n. sp. and other Parvicapsula spp. was examined with their partial small subunit rDNA (SSU rDNA) sequences. P. limandae n. sp. and P. asymmetrica appear to be closely related, while P. bicornis n. sp. and P. minibicornis are the most divergent members of the genus.     

A new genus Gadimyxa with three new species (Myxozoa, Parvicapsulidae) parasitic in marine fish (Gadidae) and the two-host life cycle of Gadimyxa atlantica n. sp

The myxozoans Gadimyxa atlantica n. sp. and G. sphaerica n. sp., and G. arctica n. sp. (Myxozoa, Parvicapsulidae), are described from Gadus morhua L. and Arctogadus glacialis (Peters) (Gadidae), respectively. They develop coelozoic in bisporic plasmodia in the urinary systems. Two morphological forms of spores were found in all 3 species, i.e., wide and (sub)spherical forms. Both spore types are bilaterally symmetrical along the suture line. The wide spores, semicircular in frontal view and elliptical in apical view, have 2 spherical polar capsules, which open in the sutural or median plane mid on the flat side of the spore. Mean widths of the wide spores of G. atlantica, G. sphaerica, and G. arctica are 7.5, 10.0, and 10.0 microm, respectively. The older, more thick-walled, (sub)spherical spores with binucleate sporoplasm are 8.0, 5.3, and 7.3 microm in mean width, respectively. The mean diameters of the polar capsules of (sub)spherical spores are 2.4, 1.7, and 2.2 microm, respectively. The (sub)spherical forms of Gadimyxa are most similar to Ortholinea within the Ortholineidae, but they differ in the development of the spores and in the arrangement of the polar capsules. The polychaetes Spirorbis spp. (Spirorbidae) act as invertebrate hosts of G. atlantica. The previously described actinospores of the tetractinomyxon type develop to myxospores in Gadus morhua within 8 wk. This is the second known myxozoan 2-host life cycle in the marine environment. Phylogenetic analyses based on partial small subunit rDNA sequences places Gadimyxa spp. among Parvicapsula spp. in the Parvicapsulidae.