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PURPOSE: We describe two new Henneguya spp. (Myxobolidae) found parasitizing Cyphocharax modestus from Pardo river, Paraná river basin, municipality of Botucatu, São Paulo State, Brazil: Henneguya fastigata n. sp. from gill lamellae and Henneguya pardensis n. sp. from gill arches. We based the descriptions on myxospore morphology, histology, and small subunit ribosomal DNA sequences. METHODS: In this investigation, 50 specimens of Cyphocharax modestus were examined between July and December 2020 for myxozoan infections. Morphological characterization was based on the mature myxospores. The small subunit ribosomal DNA sequences were used for molecular and phylogenetic study. RESULTS: Phylogenetic analysis provided the evolutionary position of these new species with other myxobolids species. Myxospores of the two species had similar morphology and morphometrics, but differed in spore body width and length, and ssrDNA sequences differed by 7.2%. These data supported the diagnosis of the parasites as distinct and novel species. The phylogenetic analysis showed a well-supported subclade formed by species that parasitize curimatid fishes, with Henneguya gilbert as a sister species of Henneguya fastigata n. sp., and Henneguya pardensis n. sp. as a sister species of both species. CONCLUSION: Our analysis was consistent with previous studies suggesting that orders and families of the hosts are strongly correlated with phylogenetic signals in the Myxobolidae. These are the first species of myxozoans described in the Pardo river.

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Myxozoans are microscopic cnidarians that mainly parasitize fishes. The present study aimed to describe a new myxozoan parasite from the gills of Boulengerella cuvieri (Spix and Agassiz, 1829) by morphological and molecular analysis. The fish was collected in 2019 at the Pindaíba River, municipality of Cocalinho, Mato Grosso State, Brazil. Whitish and circular plasmodia were found in the primary gill filaments, occupying an intralamellar position, with an average of 0.5 mm in diameter. Henneguya Thélohan, 1892 myxospores found inside the plasmodia were elongated and ellipsoidal, consisting of two long and elliptical shell valves with two long, tapering caudal appendages. Morphometric measurements revealed a total spore length of 36.1 ± 2.0 µm; spore body length of 12.8 ± 0.5 µm; spore width of 4.9 ± 0.3 µm; tail length of 23.3 ± 1.6 µm; capsule length of 7.2 ± 0.4 µm; capsule width of 1.5 ± 0.2 µm; and 10 coils in the polar filament. Phylogenetic analysis showed that the isolates from this study were grouped into the main-clade of freshwater fishes, within a group of species parasitizing fishes from Brazil. Intergenotypic difference ranged from 23%-25.9% compared with other Brazilian myxozoan isolates. Using molecular and morphological characterization, this parasite was identified as a new species of the genus Henneguya.

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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.

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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.

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This study describes Henneguya sacacaensis n. sp. in specimens of the Osteichthyes Satanoperca jurupari (Heckel, 1840), collected in the Rio Curiaú Environmental Protection Area in the city of Macapá, state of Amapá Brazil. Using optical microscopy and molecular analysis, these cyst-shaped parasites were analyzed. The gills of 57.14% of the analyzed S. jurupari contained hundreds of spores. The cysts found on the gill lamellae were oval-shaped and whitish. The Henneguya spores had an average length of 46.5 (41.3-56.92) µm. The fusiform body of the Henneguya measured 16.5 (13.16-20.01) µm long and 5.1 (3.91-6.12) µm in width, the two polar capsules had a taper of 3.83 (3.4-4.32) µm and a width of 1.68 (1.4-1.99) µm, and the tail measured 30 (22.47-41.67) µm in length, containing a polar filament coiled seven to nine times. Morphogical and phylogenetic analysis allowed the preposition of a new species, Henneguya sacacaensis n. sp, that belongs to the family Myxobolidae and the genus Henneguya.(AU)

Henneguya sacacaensis n. sp. é descrito em espécimes do Osteichthyes Satanoperca jurupari (Heckel, 1840), coletados na área de Proteção Ambiental do rio Curiaú na cidade de Macapá no estado do Amapá, Brasil. Com auxílio de microscopia óptica e análises moleculares, esses parasitos foram analisados e observados nas brânquias em forma de cistos, contendo centenas de esporos e apresentaram a prevalência de 57,14%. Os cistos encontrados nas lamelas branquiais tinham formatos ovais e esbranquiçados. Seus esporos apresentaram um comprimento médio de 46,5 (41,3-56,92) µm, corpo fusiforme medindo 16,5 (13,16-20,01) µm de comprimento e 5,1 (3,91-6,12) µm de largura, suas duas cápsulas polares apresentam uma conicidade de 3,83 (3,4-4,32) µm e sua largura 1,68 µm (1,4-1,99), a cauda 30 (22,47-41,67) µm de comprimento, contento um filamento polar de 7 à 9 voltas. Análises morfológicas e filogenéticas permitiram a preposição de uma nova espécie, Henneguya sacacaensis n. sp, que pertence à família Myxobolidae e ao gênero Henneguya.(AU)

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Abstract This study describes Henneguya sacacaensis n. sp. in specimens of the Osteichthyes Satanoperca jurupari (Heckel, 1840), collected in the Rio Curiaú Environmental Protection Area in the city of Macapá, state of Amapá Brazil. Using optical microscopy and molecular analysis, these cyst-shaped parasites were analyzed. The gills of 57.14% of the analyzed S. jurupari contained hundreds of spores. The cysts found on the gill lamellae were oval-shaped and whitish. The Henneguya spores had an average length of 46.5 (41.3-56.92) µm. The fusiform body of the Henneguya measured 16.5 (13.16-20.01) µm long and 5.1 (3.91-6.12) µm in width, the two polar capsules had a taper of 3.83 (3.4-4.32) µm and a width of 1.68 (1.4-1.99) µm, and the tail measured 30 (22.47-41.67) µm in length, containing a polar filament coiled seven to nine times. Morphogical and phylogenetic analysis allowed the preposition of a new species, Henneguya sacacaensis n. sp, that belongs to the family Myxobolidae and the genus Henneguya.

Resumo Henneguya sacacaensis n. sp. é descrito em espécimes do Osteichthyes Satanoperca jurupari (Heckel, 1840), coletados na área de Proteção Ambiental do rio Curiaú na cidade de Macapá no estado do Amapá, Brasil. Com auxílio de microscopia óptica e análises moleculares, esses parasitos foram analisados e observados nas brânquias em forma de cistos, contendo centenas de esporos e apresentaram a prevalência de 57,14%. Os cistos encontrados nas lamelas branquiais tinham formatos ovais e esbranquiçados. Seus esporos apresentaram um comprimento médio de 46,5 (41,3-56,92) µm, corpo fusiforme medindo 16,5 (13,16-20,01) µm de comprimento e 5,1 (3,91-6,12) µm de largura, suas duas cápsulas polares apresentam uma conicidade de 3,83 (3,4-4,32) µm e sua largura 1,68 µm (1,4-1,99), a cauda 30 (22,47-41,67) µm de comprimento, contento um filamento polar de 7 à 9 voltas. Análises morfológicas e filogenéticas permitiram a preposição de uma nova espécie, Henneguya sacacaensis n. sp, que pertence à família Myxobolidae e ao gênero Henneguya.

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On the basis of morphological and molecular analyses, a new myxozoan parasite is described from the gills of the fish Hoplerythrinus unitaeniatus, collected in the municipality of Nova Xavantina, Mato Grosso State, Brazil. Plasmodia of Henneguya unitaeniata sp. nov. were oval and whitish and were found surrounded by collagen fibers forming plasmodia wall between gill filaments on the gill arch. The spores were ellipsoidal with two similar polar capsules. Morphometric analysis showed a total spore mean length of 23.8 ± 1.5 µm, spore body mean length of 14.5 ± 0.7 µm, caudal appendage mean length of 10.3 ± 1.4 µm, thickness mean length of 4.3 ± 0.3 µm, polar capsule mean length of 4.2 ± 0.5 µm, polar capsule mean width of 1.8 ± 0.3 µm, spore mean width of 4.8 ± 0.4 µm, and 4-5 polar filament coils. Phylogenetic analysis showed Henneguya unitaeniata sp. nov. as a basal species in a subclade formed by myxozoans that parasitize bryconid fishes.

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Plasmodia containing spores of a new species of myxozoan, Henneguya jundiai n. sp., were found in the gill arches of Rhamdia quelen catfish. The present study describes H. jundiai n. sp. based on morphological and molecular analysis. The mature spores were elongated and measured 26.9 ± 1.9 (22.9-29.2) µm in total spore length, 9.5 ± 0.4 (8.8-10.0) µm in spore body length, 4.6 ± 0.4 (4.1-5.5) µm in spore body width, 17.3 ± 1.8 (14.1-19.8) µm in tail length, 4.9 ± 0.3 (4.6-5.5) µm in polar capsule length and 1.4 ± 0.2 (1.2-1.7) µm in polar capsule width. The polar filaments had 6-7 coils. Histological analysis revealed the formation of a large plasmodium connected to the gill arch through the epithelial tissue. Ultrastructural analysis revealed the presence of a binucleated sporoplasm, as well as the polar capsules and the coils of the polar filament. Phylogenetic analysis found that H. jundiai n. sp. is located within a subclade formed by species of Henneguya which parasitize siluriform fish from Brazil. Henneguya jundiai n. sp. appears as a sister species of H. quelen. The results of the analyzes clearly indicated the existence of a new species, H. jundiai.

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Henneguya guanduensis was originally described as a gill parasite of the Hoplosternum litoralle, a siluriform fish belonging to the Callichthyidae family. A study was undertaken with 58 specimens of H. litoralle taken from Batalha River in the state of São Paulo. The fish were collected and examined searching for lesions and/or myxosporean plasmodia. The prevalence of infection was 9.31% in the gills and 5.17% in the intestine. The mature spores had elongated bodies with polar capsules of inequal size and a caudal length greater than body length. Morphological characteristics identified the parasite as H. guanduensis. Molecular analysis of the SSU rDNA partials sequences resulted in a 1796 bp and 1712 bp for gills and intestine repectively, demonstrating significant genetic differences with previously described species of Henneguya and 99.7% similarity to each other when aligned. Phylogenetic analysis comparing the SSU rDNA sequence of H. guanduensis with closest species as indicated by BLASTn Max Score showed H. guanduensis as sister species of H. loretoensis in a subclade compose by species that parasites fishes from Amazon basin. This is the first report of the finding of Henneguya guanduensis spores in the intestine of Hoplosternum littorale.

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A new myxozoan was found parasitizing the freshwater catfish, Rhamdia quelen (Quoy Gaimard), in the Marajó island, Amazon region, Brazil. The new species is described based on the results of morphological and molecular analyses. The parasite is approximately 1.5 mm in diameter and develops in the musculature of the host in the form of spherical, whitish cysts, which are visible macroscopically between the epaxial and hypaxial layers. When ruptured, these cysts produced ellipsoidal spores with a mean length of 11.4 µm (10.7-12.6) and width of 7.2 µm (6.4-7.9). Anomalous spores with a caudal elongation, vesicles in the peripheral portion of the spore and ornamentation of the valves were also observed. The results of the phylogenetic analysis of sequences of the 18S rDNA gene using Bayesian Inference indicated clear differences among the Myxobolus species that reinforced the taxonomic position of the parasite, confirming its status as a new species, denominated Myxobolus arariensis n. sp.

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We describe two novel myxosporean parasites from Brachyplatystoma rousseauxii, an economically important freshwater catfish from the Amazon basin, Brazil. Myxobolus tapajosi n. sp., was found in the gill filaments of 23.5% of 17 fish, with myxospores round to oval in frontal view and biconvex in lateral view: length 15 (13.5-17) µm and width 10.7 (9.6-11.4) µm; polar capsules equal, length 5.8 (4.6-7.1) µm and width 3 (2.3-3.8) µm containing polar tubules with 6-7 turns. Ellipsomyxa amazonensis n. sp. myxospores were found floating freely or inside plasmodia in the gall bladder of 23.5% of fish. The myxospores were ellipsoidal with rounded extremities: length 12.8 (12.3-13.6) µm and width 7.6 (6.7-8.7) µm; with two equal, slightly pyriform polar capsules, length 3.8 (3.8-4.0) µm and width 3.1 (2.5-3.4) µm, containing polar tubules with 2-3 turns. We combined spore morphometry, small-subunit ribosomal DNA data, specific host, and phylogenetic analyses, to identify both of these parasites as new myxozoan species. Maximum likelihood and Bayesian analyses showed that Myxobolus tapajosi n. sp. clustered in a basal branch in a subclade of parasites from exclusively South American pimelodid fishes. Ellipsomyxa amazonensis n. sp. clustered within the marine Ellipsomyxa lineage, but we suspect that although the parasite was collected in freshwater, its hosts perform a large migration throughout the Amazon basin and may have become infected from a brackish/marine polychaete host during the estuary phase of its life.

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Plasmodia containing myxospores belonging to the genus Myxobolus Bütschli, 1882 were found in the ovaries and liver of Salminus hilarii. Despite its economic value, this fish host has no previous reports of myxozoan infections. Herein, we describe Myxobolus batalhensis n. sp. using morphological and ultrastructural data, as well as histological and SSU rDNA molecular data. The mature myxospores were elongated, measuring in average 15.2 ± 0.8 µm in length, 8.4 ± 0.4 µm in width, and 5.1 ± 0.2 µm in thickness. Polar capsules were elongated and measured 5.3 ± 0.3 µm in length and 2.8 ± 0.3 µm in width. Polar filaments had 6-9 coils. Histopathological analysis showed coagulation necrosis associated with cell lysis as a response of the host cell to the parasite in the ovaries. No inflammatory reaction was observed in the liver, although the presence of the plasmodia caused changes in tissue structure. The phylogenetic analysis of South American myxobolid species showed M. batalhensis n. sp. as sister species of Myxobolus aureus. This is the first report of a myxozoan species parasitizing S. hilarii and the first myxozoan species described in the Batalha river.

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Myxosporean Myxobolus saladensis sp. nov. in the gills of Mugil liza Valenciennes, 1836 from Samborombón Bay was described by light and electron microscopy studies. Spores were pyriform and binucleated, measuring 10.63±0.36 µm (n=20) long, 9.24±0.50 µm (n=20) wide and 4.13±0.36 µm (n=20) thick, included in polysporic cyst-like plasmodia. Elongated pyriform polar capsules were of equal size (3.84±0.27 µm long and 2.30±0.12 µm wide). The sporoplasm contained some sporoplasmosomes. Each PC contained a polar filament with 4-5 coils obliquely arranged in relation to the polar capsules axis. The PC wall was composed of two layers of different electron densities. Based on the morphological and ultrastructure differences of the spore to those of previously described species of Myxobolus, we describe a new species, Myxobolus saladensis sp. nov.

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ABSTRACT Myxosporean Myxobolus saladensis sp. nov. in the gills of Mugil liza Valenciennes, 1836 from Samborombón Bay was described by light and electron microscopy studies. Spores were pyriform and binucleated, measuring 10.63±0.36 µm (n=20) long, 9.24±0.50 µm (n=20) wide and 4.13±0.36 µm (n=20) thick, included in polysporic cyst-like plasmodia. Elongated pyriform polar capsules were of equal size (3.84±0.27 µm long and 2.30±0.12 µm wide). The sporoplasm contained some sporoplasmosomes. Each PC contained a polar filament with 4-5 coils obliquely arranged in relation to the polar capsules axis. The PC wall was composed of two layers of different electron densities. Based on the morphological and ultrastructure differences of the spore to those of previously described species of Myxobolus, we describe a new species, Myxobolus saladensis sp. nov.

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Myxosporean Myxobolus saladensis sp. nov. in the gills of Mugil liza Valenciennes, 1836 from Samborombón Bay was described by light and electron microscopy studies. Spores were pyriform and binucleated, measuring 10.63±0.36 µm (n=20) long, 9.24±0.50 µm (n=20) wide and 4.13±0.36 µm (n=20) thick, included in polysporic cyst-like plasmodia. Elongated pyriform polar capsules were of equal size (3.84±0.27 µm long and 2.30±0.12 µm wide). The sporoplasm contained some sporoplasmosomes. Each PC contained a polar filament with 4-5 coils obliquely arranged in relation to the polar capsules axis. The PC wall was composed of two layers of different electron densities. Based on the morphological and ultrastructure differences of the spore to those of previously described species of Myxobolus, we describe a new species, Myxobolus saladensis sp. nov.(AU)

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