Patrícia Matos

Ultrastructural and phylogenetic data of Chloromyxum riorajum sp. nov. (Myxozoa), a parasite of the stingray Rioraja agassizii in Southern Brazil.

We describe a new myxozoan parasite found infecting the gall bladder of the cartilaginous fish Rioraja agassizii (Rajidae) from the South Atlantic coast of Brazil. Light microscopy, scanning and transmission electron microscopy and phylogenetic data were used. Numerous irregular polyspqric plasmodia externally covered by numerous microvilli containing different stages of sporogony, including free spores, were observed in bile. Ellipsoidal spores, on average 11.41 microm long, 8.48 microm wide and 7.32 microm thick, were formed by 2 equal-sized valves, each possessing 3 to 4 (rarely 5) elevated ridges which projected from the basal portion of the spore, and joined along a sinuous S-shaped sutural line. The basal portion of the valves bore a bundle of 33 to 37 extended tapering caudal filaments attached to the basal portion of the last ridge and basal portion of the sutural edge of the 2 valves. The caudal filaments, formed of material similar to the valves, were attached to the shell wall by a conical basis. The spores contained 4 equal-sized pyriform polar capsules (4.5 x 2.4 microm), located at the same level, each with a polar filament with 6 (rarely 7) coils. Binucleate sporoplasm was irregular in shape, with a granular matrix and dense bodies randomly distributed in a light area. Based on the shape and dimensions of the spore, on the number, position and arrangements of the surface ridges, caudal bundle of filaments, polar capsules and polar filament arrangements, as well as phylogenetic analyses using the small subunit ribosomal DNA (SSU rDNA) sequences, we propose the name Chloromyxum riorajum for this new myxozoan.

A New Species of Myxozoa, Henneguya rondoni n. sp. (Myxozoa), from the Peripheral Nervous System of the Amazonian Fish, Gymnorhamphichthys rondoni (Teleostei)

ABSTRACT. Henneguya rondoni n. sp. found in the peripheral lateral nerves located below the two lateral lines of the fish Gymnorhamphichthys rondoni (Teleostei, Rhamphichthyidae) from the Amazon river is described using light and electron microscopy. Spherical to ellipsoid cysts measuring up to 110 μm in length contained only immature and mature spores located in close contact with the myelin sheaths of the nervous fibres. Ellipsoidal spores measured 17.7 (16.9–18.1)‐μm long, 3.6 (3.0–3.9)‐μm wide, and 2.5 (2.2–2.8)‐μm (n=25) thick. The spore body measuring 7.0 (6.8–7.3)‐μm long was formed by two equal symmetric valves, each with an equal tapering tail 10.7 (10.3–11.0) μm in length. The tails were composed of an internal dense material surrounded by an external homogeneous sheath of hyaline substance. The valves surrounded two equal pyriform polar capsules measuring 2.5 (2.2–2.8)‐μm long and 0.85 (0.79–0.88)‐μm (n=25) wide and a binucleated sporoplasm cell containing globular sporoplasmosomes 0.38 (0.33–0.42) μm (n=25) in diam. with an internal eccentric dense structure with half‐crescent section. Each polar capsule contains an anisofilar polar filament with 6–7 turns obliquely to the long axis. The matrix of the polar capsule was dense and the wall filled with a hyaline substance. The spores differed from those of previously described species. Based on the ultrastructural morphology of the spore and specificity to the host species, we propose a new species name H. rondoni n. sp.

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).

Henneguya torpedo sp. nov. (Myxozoa), a parasite from the nervous system of the Amazonian teleost Brachyhypopomus pinnicaudatus (Hypopomidae)

A new species of Myxosporea, Henneguya torpedo sp. nov., is described from the brain and spinal cord of the Amazonian teleostean fish Brachyhypopomus pinnicaudatus collected from the Peixe Boi River, State of Park, Brazil. The spores were surrounded by a thick hyaline sheath that is homogeneous and electron translucent and consists of 2 layers of different densities. The total spore length is 48.62 +/- 0.51 microm (mean +/- SE), the ellipsoidal spore body length is 28.53 +/- 0.36 microm, the body width is 7.25 +/- 0.31 microm and the body thickness is 3.06 +/- 0.26 microm. Each of the 2 equal-sized valves presented a tapering tail (19.64 +/- 0.44 microm in length). The 2 equal-sized thin and smooth valves surrounded 2 equal-sized and elongated ellipsoidal polar capsules (6.41 +/- 0.26 x 1.84 +/- 0.19 microm) that contained 5 to 6 (rarely 7) polar filament coils. The binucleated sporoplasm contained numerous spherical sporoplasmosomes (-260 x -280 nm) with a laterally eccentric-dense structure containing a half-crescent section. The sporoplasmosomes are surrounded by a hyaline homogenous sheath. Based on the data obtained by light and electron microscopy and on the host specificity, the spores differed from the previously described Henneguya spp., mainly in the presence of a sheath surrounding the spores, the spore shape and size and the number and arrangement of the polar filament coils. Therefore, from this description we propose the establishment of a new species, which we have named Henneguya torpedo sp. nov.

Morphology and phylogeny of Henneguya jocu n. sp. (Myxosporea, Myxobolidae), infecting the gills of the marine fish Lutjanus jocu

Henneguya jocu n. sp. (Myxosporea, Myxobolidae) is described from the gill lamellae of the marine teleost fish Lutjanus jocu, with a focus on ultrastructural and molecular features. This myxosporean forms subspherical cysts up to ∼260 μm × 130 μm long, and develops asynchronously. Mature myxospores ellipsoidal with a bifurcated caudal process. Myxospore length 10.9 ± 0.4 μm (n=50); width, 8.2 ± 0.3 μm (n=50); and thickness, 2.9 ± 0.5 μm (n=50). Two equal caudal processes, 34.1±1.0 μm long (n=50); and total myxospore length, 45.2 ± 1.0 μm (n=50). Two symmetric valves surround two ellipsoidal polar capsules, 5.0 ± 0.3 × 1.4 ± 0.2 μm (n=20), each containing an isofilar polar filament forming 4-5 coils along the inner wall of these structures, as well as a binucleated sporoplasm presenting a spherical vacuole and several globular sporoplasmosomes. Both the morphological data and molecular analysis of the SSU rDNA gene identify this parasite as a new species of the genus Henneguya. Maximum Likelihood and Maximum Parsimony analyses further indicate that the parasite clusters within others marine Myxobolidae species, forming a group alongside other Henneguya species described from marine hosts.

Description of Triangulamyxa psittaca sp. nov. (Myxozoa: Myxosporea), a New Parasite in the Urinary Bladder of Colomesus psittacus (Teleostei) from the Amazon River, with Emphasis on the Ultrastructure of Plasmodial Stages

A fish-infecting myxosporean was found in the urinary bladder of the teleostean Colomesus psittacus, collected from the Amazon River, Brazil. Specimens were sampled in three different periods: May and June, with water temperature ranging from 18-23oC; August, with water temperature ranging from 24-28oC; and November and December, with water temperature ranging from 29-32oC. Upon observation, several fish displayed abnormal behaviour, consisting of erratic movements, and mortality was recorded among them. Necropsy of all sampled fishes revealed hypertrophy of the urinary bladder only among specimens previously displaying the irregular behaviour. Microscopic analysis of this organ confirmed the parasitic infection, resulting in the observation of spores floating free in the urine, and numerous plasmodia attached to the epithelium of the urinary bladder. Light and ultrastructural studies allowed recognition of the spores and plasmodia morphological characteristics. Coelozoic plasmodia were polysporic with varying organizational structure, according to the sampling period. Spores were equilaterally triangular with rounded ends in valvar view, measuring 8.8 ± 0.4 μm (n = 30) in length and 8.4 ± 0.5 μm (n = 30) in width, and displaying a ridge surface pattern. Two polar capsules were observed in the anterior end of the spores, measuring 3.1-3.2 μm in diameter. The spores were morphologically identified as belonging to the recently described genus Triangulamyxa. Further observation and comparison to the morphological features described for Triangulamyxa amazonica, the only other species within this genus, allowed us to conclude our parasite as a new species, herein named Triangulamyxa psittaca sp. nov. from the Amazon River, Brazil. Also, three different stages were distinguished in the plasmodium evolution, based on the observed morphological features at the three sampling periods. Fish sampled during May and June displayed small plasmodia (up to ~ 15-20 μm long), containing early stages of sporogenic development. Fish sampled during November and December presented larger plasmodia (up to ~ 850 μm long), which appeared flattened against and lining the urinary bladder epithelial cells and contained the later stages of sporogenic development, including some mature spores. Fish sampled during August presented plasmodia displaying intermediate morphological features between those observed in infected fish from the other sampling periods. Several immature and mature spores were among the different developmental stages. The parasite-host interface evolution is described throughout the different observed stages, with emphasis on the formation of septate junctions. Considering several previous reports, as well as the different environmental conditions during the sampling periods, the plasmodium development here described appears to be influenced by environmental factors, namely water temperature.

Estrutura fina do espermatozóide de Acestrorhynchus falcatus Bloch (Teleostei, Characidae) da região norte do Brasil

Spermatozoa of amazon fish, Acestrorhynchus falcatus Bloch, 1794 were investigated by optical microscopy (DIC) and electron microscopy (TEM). Spermatozoa have no acrosome, the head is ovoid, and the midpiece region cylindrical, contains spherical mitochondria. The centriolar complex is located at the lateral side of the nucleus, formed by nine peripheral pairs of microtubules and arranged in a right angle to each other. The flagellum consists of nine pairs of peripheral and two central microtubules.

Light and electron microscopy of Myxobolus sciades n. sp. (Myxozoa), a parasite of the gills of the Brazilian fish Sciades herzbergii (Block, 1794) (Teleostei: Ariidae)

A myxosporean parasite in the gill lamellae of the freshwater teleost fish, Sciades herzbergii (Ariidae) (Block, 1794), from the Poti River (Northeast of Brazil) was described by light and electron microscopy studies. Polysporic histozoic cyst-like plasmodia containing several life-cycle stages, including mature spores, were observed. The spores were pyriform and uninucleate, measuring 9.15 +/- 0.39 microm (n = 50) long, 4.36 +/- 0.23 microm (n = 25) wide and 2.61 +/- 0.31 microm (n = 25) thick. Elongated pyriform polar capsules (PC) were of equal size (4.44 +/- 0.41 microm long and 1.41 +/- 0.42 microm in diameter) and each contained a polar filament with 9-10 coils obliquely arranged in relation to the axis of PC. The PC wall was composed of two layers of different electron densities. Histological analysis revealed the close contact of the cyst-like plasmodia with the basal portion of the epithelial gill layer, which exhibited some alterations in the capillary vessels. Based on the morphological and ultrastructural differences, the similarity of the spore features to those of the genus Myxobolus and the specificity of this host to previously described species, we describe a new species named Myxobolus sciades n. sp. in this study.

Morphological data of Calyptospora spinosa n. sp. (Apicomplexa, Calyptosporidae) parasite of Crenicichla lepidota HECKEL, 1840 (Teleostei) from Amazon river

A new coccidian species, Calyptospora spinosa (fam. Calytosporidae) is described from liver, testis and ovary tissue of a freshwater fish, Crenicichla lepidota, from the lower Amazon river near Belém, Brazil, where it causes great mortality. Each oocyst contains four ellipsoidal sporocysts, each one surrounded by a membranous veil. Each sporocyst, containing two sporozoites, has an apical sporopodium possessing several lateral sporopodium projections. These sporocysts differ from those of the four other species of the genus Calyptospora in size and shape, in the insertion of the lateral sporopodium projections and in the identity of the host. This paper describes the morphology and ultrastructural characteristics of the oocyst and sporocyst of this coccidium and draws morphological comparisons with the other four species of the same genus.

Fine structure of Chloromyxum menticirrhi n. sp. (Myxozoa) infecting the urinary bladder of the marine teleost Menticirrhus americanus (Sciaenidae) in Southern Brazil.

A myxosporidian was found in the urinary bladder of the teleost Menticirrhus americanus Linnaeus, 1758 (Sciaenidae) collected from the South Atlantic coast of Brazil. Polysporic amoeboid plasmodia containing sporoblasts, developing pansporoblasts and spores were free in the bladder lumen. The prevalence of infection was 17.64% (15/85). Unfixed spores were spherical to subspherical, on average 10.5microm long, 9.8microm wide and 10.1microm thick (n=25), and fixed spores measured 10.1 x 9.5 x 9.7microm. The two spore valves were of equal size and each possessed prominent sutural lines and about 41 (37-45) surface ridges aligned parallel with the suture line. These ridges gave transverse sections a cog-wheel-like outline. The spores contained four pyriform polar capsules of equal size (3.20 x 2.0microm) (n=25) (fixed), each with a polar filament having 3-4 (rarely 5) coils. The binucleate sporoplasm was irregular in shape, with granular matrix and randomly distributed dense bodies. The shape and dimensions of the spore, as well as the number, position and arrangement of the surface ridges, polar capsules and polar filament indicate that this is a new species, herein designated Chloromyxum menticirrhi. The gill, liver, gall bladder and intestine of the host showed no abnormalities.