Sarah Arana

Henneguya pseudoplatystoma n. sp. causing reduction in epithelial area of gills in the farmed pintado, a South American catfish: Histopathology and ultrastructure

The present study is part of an ongoing investigation into the characteristics of Myxozoan parasites of Brazilian freshwater fish and was carried out using morphology, histopathology and electron microscopy analysis. A new Myxosporea species (Henneguya pseudoplatystoma) is described causing an important reduction in gill function in the farmed pintado (a hybrid fish from a cross between Pseudoplatystoma corruscans and Pseudoplatystoma fasciatum), which is a commercially important South American catfish. From a total of 98 pintado juveniles from fish farms in the states of São Paulo and Mato Grosso do Sul (Brazil), 36 samples (36.7%) exhibited infection of the gill filaments. Infection was intense, with several plasmodia occurring on a same gill filament. The plasmodia were white and measured up to 0.5mm in length; mature spores were ellipsoidal in the frontal view, measuring 33.2+/-1.9 microm in total length, 10.4+/-0.6 microm in body length, 3.4+/-0.4 microm in width and 22.7+/-1.7 microm in the caudal process. The polar capsules were elongated, measuring 3.3+/-0.4 microm in length and 1.0+/-0.1 microm in width and the polar filaments had six to seven turns. Histopathological analysis revealed the parasite in the connective tissue of the gill filaments and lamella. No inflammatory process was observed, but the development of the plasmodia reduced the area of functional epithelium. Ultrastructural analyses revealed a single plasmodial wall, which was in direct contact with the host cells and had numerous projections in direction of the host cells as well as extensive pinocytotic canals. A thick layer (2-6 microm) of fibrous material and numerous mitochondria were found in the ectoplasm. Generative cells and the earliest stage of sporogenesis were seen more internally. Advanced spore developmental stages and mature spores were found in the central portion of the plasmodia.

Myxobolus cordeiroi n. sp., a parasite of Zungaro jahu (Siluriformes: Pimelodiade) from Brazilian Pantanal: Morphology, phylogeny and histopathology☆

This work is part of an ongoing investigation into the characteristics of Myxozoan parasites of freshwater fish in Brazil and was carried out using morphology, histopathology and molecular analysis. A new Myxosporea species (Myxobolus cordeiroi) is described infecting the jaú catfish (Zungaro jahu). Fifty jaú specimens were examined and 78% exhibited plasmodia of the parasite. The plasmodia were white and round, measuring 0.3-2.0mm in diameter and the development occurred in the gill arch, skin, serosa of the body cavity, urinary bladder and eye. The spores had an oval body and the spore wall was smooth. Partial sequencing of the 18S rDNA gene resulted in a total of 505bp and the alignment of the sequences obtained from samples in different organs revealed 100% identity. In the phylogenetic analysis, the Myxobolus species clustered into two clades-one primarily parasites of freshwater fish and the other primarily parasites of marine fish. M. cordeiroi n. sp. was clustered in a basal position in the freshwater fish species clade. The histological analysis revealed the parasite in the connective tissue of the different infected sites, thereby exhibiting affinity to this tissue. The plasmodium was surrounded by an outer collagen capsule of fibers with distinct orientation from the adjacent connective tissue and an inner layer composed of delicate collagen fibrils-more precisely reticular fibers. The development of the parasite in the cornea and urinary bladder caused considerable stretching of the epithelium.

A new species of Henneguya, a gill parasite of Astyanax altiparanae (Pisces: Characidae) from Brazil, with comments on histopathology and seasonality

A new species of Myxosporea, Henneguya chydadea, is described parasitizing the gills of Astyanax altiparanae collected from a lake on Rio das Pedras farm near Campinas, state of São Paulo, Brazil. Of the fish examined, 88.3% had gills parasitized by myxosporeans. The prevalence of the parasite ranged from 80% in the spring and fall, 93% in the summer and 100% in the winter. The parasite induced the formation of white, oval-shaped cysts measuring 40-64 microm x 64-80 microm which deformed the gill lamellae, compressed the capillaries, and caused retraction of the neighboring lamellae. The mature spores were elongated and had two identical, parallel elongate polar capsules. Each capsule contained a polar filament with 9-10 turns. There was no mucous envelope or iodinophilous vacuole. Morphometric differences between this parasite and other species of the genus Henneguya indicated, that the parasite observed in A. altiparanae is a new species. This is the first report of a myxosporeanparasitizing A. altiparanae.

Histophatology and ultrastructure of Henneguya caudalongula sp. n. infecting Prochilodus lineatus (Pisces: Prochilodontidae) cultivated in the state of São Paulo, Brazil

The histological and ultrastructural characteristics of a new species of Henneguya and the host reactions to infection by this species are reported. Henneguya caudalongula sp. n. was found in the inter and intralamellar regions of the gills of Prochilodus lineatus (Valenciennes, 1836) cultivated at Center for the Research and Management of Continental Fishing Resources located in the municipality of Pirassununga, state of São Paulo, Brazil. The plasmodia were white and round or ellipsoidal and measured 0.2 to 1 mm in length. The development of the parasite was asynchronous and the mature spores were fusiform, with a total length 71 +/- 1.4 microm, body length of 16.6 +/- 0.54 microm and width 4.6 +/- 0.2 microm. The caudal process was 52.6 +/- 1.5 microm long. The polar capsules were elongate (length 6.1 +/- 0.19 microm, width 1.6 +/- 0.15 microm) and of equal size. The polar filament was coiled in 10-11 turns. The prevalence of the parasite was 48.3% and did not vary significantly with the season or host size.

Phylogenetic and host-parasite relationship analysis of Henneguya multiplasmodialis n. sp infecting Pseudoplatystoma spp. in Brazilian Pantanal wetland

A new species of the genus Henneguya (Henneguya multiplasmodialis n. sp.) was found infecting the gills of three of 89 specimens (3.3%) of Pseudoplatystoma corruscans and two of 79 specimens (2.6%) of Pseudoplatystoma reticulatum from rivers in the Pantanal wetland, Brazil. Partial sequencing of the 18S rDNA gene of the spores obtained from one plasmodium from the gills of P. corruscans and other one from the gills of P. reticulatum, respectively, resulted in a total of 1560 and 1147 base pairs. As the spores of H. multiplasmodialis n. sp. resemble those of Henneguya corruscans, which is also a parasite of P. corruscans, sequencing of the 18S rDNA gene of the spores of H. corruscans found on P. corruscans caught in the Brazilian Pantanal wetland was also provided to avoid any taxonomic pendency between these two species, resulting in 1913 base pairs. The sequences of H. multiplasmodialis n. sp. parasite of P. corruscans and P. reticulatum and H. corruscans did not match any of the Myxozoa available in the GenBank. The similarity of H. multiplasmodialis n. sp. obtained from P. corruscans to that from P. reticulatum was of 99.7%. Phylogeny revealed a strong tendency among Henneguya species to form clades based on the order and/or family of the host fish. H. multiplasmodialis n. sp. clustered in a clade with Henneguya eirasi and H. corruscans, which are also parasites of siluriforms of the family Pimelodidae and, together with the clade composed of Henneguya spp. parasites of siluriforms of the family Ictaluridae, formed a monophyletic clade of parasites of siluriform hosts. The histological study revealed that the wall of the plasmodia of H. multiplasmodialis n. sp. were covered with a stratified epithelium rich in club cells and supported by a layer of connective tissue. The interior of the plasmodia had a network of septa that divided the plasmodia into numerous compartments. The septa were composed of connective tissue also covered on both sides with a stratified epithelium rich in club cells. Inflammatory infiltrate was found in the tissue surrounding the plasmodia as well as in the septa.

Host–parasite–environment relationship, morphology and molecular analyses of Henneguya eirasi n. sp. parasite of two wild Pseudoplatystoma spp. in Pantanal Wetland, Brazil☆

A new myxosporean species, Henneguya eirasi n. sp., is described parasitizing the gill filaments of Pseudoplatystoma corruscans and Pseudoplatystoma fasciatum (Siluriformes: Pimelodidae) caught in the Patanal Wetland of the state of Mato Grosso, Brazil. The parasite formed white, elongated plasmodia measuring up to 3mm. Mature spores were ellipsoidal in the frontal view, measuring 37.1 ± 1.8 μm in total length, 12.9 ± 0.8 μm in body length, 3.4 ± 0.3 μm in width, 3.1 ± 0.1 μm in thickness and 24.6 ± 2.2 μm in the caudal process. Polar capsules were elongated and equal in size, measuring 5.4 ± 0.5 μm in length and 0.7 ± 0.1 μm in width. Polar filaments had 12-13 coils. Histopathological analysis revealed that the parasite developed in the sub-epithelial connective tissue of the gill filaments and the plasmodia were surrounded by a capsule of host connective tissue. The plasmodia caused slight compression of the adjacent tissues, but no inflammatory response was observed in the infection site. Ultrastructure analysis revealed a single plasmodial wall connected to the ectoplasmic zone through numerous pinocytotic canals. The plasmodial wall exhibited numerous projections and slightly electron-dense material was found in the ectoplasm next to the plasmodial wall, forming a line just below the wall. Partial sequencing of the 18S rDNA gene of H. eirasi n. sp. obtained from P. fasciatum resulted in a total of 1066 bp and this sequence did not match any of the Myxozoa available in the GenBank. Phylogenetic analysis revealed the Henneguya species clustering into clades following the order and family of the host fishes. H. eirasi n. sp. clustered alone in one clade, which was the basal unit for the clade composed of Henneguya species parasites of siluriform ictalurids. The prevalence of the parasite was 17.1% in both fish species examined. Parasite prevalence was not influenced by season, host sex or host size.

Phylogeny, ultrastructure, histopathology and prevalence of Myxobolus oliveirai sp. nov., a parasite of Brycon hilarii (Characidae) in the Pantanal wetland, Brazil

This paper presents the morphological, histological and ultrastructural characteristics of Myxobolus oliveirai sp. nov., a parasite of the gill filaments in Brycon hilarii from the Brazilian Pantanal. Out of 216 B. hilariispecimens examined (126 wild and 90 cultivated), 38.1% of wild specimens (n = 48) were infected. The parasites form elongated plasmodia primarily in the tip of gill filaments, reaching about 3 mm in length. A thorough comparison with all the Myxobolus species described from South American hosts, as well as nearly all the Myxobolus species described so far is provided. Partial sequencing of the 18S rDNA gene revealed a total of 1,527 bp. The Myxobolus species parasite of B. hilarii did not match any of the Myxozoa available in GenBank. In the phylogenetic analysis, M. oliveirai sp. nov. composed a monophyletic group with eight other species: five species of Myxobolus parasites of mugilid fishes, two parasites of pangasiid and one of centrarchid. Infection prevalence values of the parasite revealed no significant differences between wet and dry seasons or between males and females. The importance of the infection to the farming of the host species is emphasized.

Light, electron microscopy and histopathology of Myxobolus salminus n. sp., a parasite of Salminus brasiliensis from the Brazilian Pantanal.

In this report, we describe the morphology and histopathology of Myxobolus salminus n. sp., a parasite of the gill filaments of wild Salminus brasiliensis (dourado) from the Brazilian Pantanal. The small polysporic plasmodia were approximately 100 microm in diameter and the development was asynchronous. The mature spores were oval to pear shaped and had a smooth wall. The spore measurements were (mean+/-S.D., with range in parentheses): length 10.1+/-0.4 microm (9.6-10.5), width 6.1+/-0.4 microm (5.8-6.6) and thickness 5.0+/-0.6 microm (4.7-5.3). The polar capsules were elongated and of equal size: length 4.6+/-0.2 microm (4.3-4.8) and width 1.7+/-0.1 microm (1.5-1.9). The histological analysis revealed numerous plasmodia in the blood vessels of the gill filaments. The site of parasite development was the wall of the large-caliber blood vessel of the gill filament, with progressive growth towards the lumen, resulting in the obstruction of blood flow, congestion and perivascular edema. The ultrastructural study revealed that the plasmodial wall was composed of two membranes, had numerous pinocytic canals and was in direct contact with the basement membrane of the vessel. The development of the parasite was asynchronous, with mature spores, immature spores and young developmental stages randomly distributed throughout the plasmodium. The prevalence of the parasite was 4.4%, with male and female fish being infected.

Light and scanning electron microscopy of Henneguya arapaima n. sp. (Myxozoa: Myxobolidae) and histology of infected sites in pirarucu (Arapaima gigas: Pisces: Arapaimidae) from the Araguaia River, Brazil

In this report, we describe Henneguya arapaima n. sp., a parasite of the gill arch and gall bladder of Arapaima gigas (pirarucu) collected in the Araguaia River, in the municipality of Nova Crixás, Goiás State, central Brazil. The plasmodia were white, round or ellipsoidal and measured 200-600 microm. Parasite development was asynchronous and the mature spores were fusiform and had smooth wall. The spores measurements were (range, with means+/-S.D. in parentheses): total length-48.4-53.1 microm (51.6+/-3.4 microm), body length-13.5-15.2 microm (14.2+/-0.8 microm), body width-5.1-6.1 microm (5.7+/-0.5 microm), body thickness-4.7-5.3 microm (4.9+/-0.2 microm) and caudal process length-38.0-41.2 microm (38.3+/-2.9 microm). The polar capsules were elongated and of unequal size, with lengths of 6.3-6.8 microm (6.5+/-0.2) and 6.2-6.6 microm (6.3+/-0.1) for the longest and shortest axes, respectively. Capsule width was 1.4-1.6 microm (1.5+/-0.1). Histological analysis showed that the plasmodia occurred in the tunica adventitia of the gall bladder and were delimited by a thin capsule of connective tissue. In the gill arch, the plasmodia were also surrounded by connective tissue similar to the endomesium of striated skeletal muscle cells. Sixty-five juvenile specimens of A. gigas weighing 1.0-25.0 kg were examined, 17 (26.1%) of which were infected. Of these, 14 (82.3%) had cysts in the gall bladder, two (11.7%) had cysts in the gill arch and only one (5.9%) had cysts in both organs. When the fish were grouped by weight, the prevalence of infection in fish weighing up to 10.0 kg (20.7%) was significantly lower than in fish weighing 10.1-25.0 kg (50%) (G=3.93; d.f.=1; p<0.05).

Differential effect of chronic aflatoxin B1 intoxication on the growth performance and incidence of hepatic lesions in triploid and diploid rainbow trout (Oncorhynchus mykiss)

El proposito del presente estudio fue comparar el crecimiento y la incidencia de lesiones hepaticas en truchas triploides y diploides tratadas con aflatoxina B1(AFB1). 240 truchas fueron divididas en 4 grupos: DC: truchas diploides alimentadas con racion sin AFB1; TC: truchas triploides alimentadas con racion sin AFB1; DT: truchas tratadas con racion con 80 ppb de AFB1 y TT: truchas triploides alimentadas con racion con 80 ppb de AFB1. Durante doce meses, mensualmente, cinco ejemplares de cada grupo fueron anestesiados y sacrificados. Con posteiroridad a la obtencion del peso y medicion del tamano de los pesces, muestras hepaticas fueron fijadas en solucion de formalina salina 10% y procesadas para analisis histopatologico. El analisis comparativo del rendimiento en crecimiento indicaron diferencias significativas entre truchas diploides del grupo control y del tratado, sugiriendo que AFB1 afecta el crecimiento de las truchas diploides. En truchas triploides no se observaron diferencias entre pesces del grupo control y los pesces tratados. El analisis histopatologico senalo que truchas triploides son mas resistentes a AFB1, ya que ambos grupos tratados presentaron lesiones preneoplasicas, sin embargo, el grupo TT demostro menor incidencia de lesiones e igualmente un desarrollo mas lento de las mismas. En cuanto a la ocurrencia de neoplasia, en el grupo DT 4 pesces desarrolaron carcinoma hepatocelular en el ultimo trimestre del experimento, mientras que ningun animal triploide desarrollo lesion neoplasica