Ultrastructure of intracytoplasmic Rickettsia-like infection of the gills of the teleost Archosargus probatocephalus (Sparidae) in northeastern Brazil / Ultraestrutura de Rickettsia intracitoplasmática infectando as brânquias do teleósteo Archosargus probatocephalus (Sparidae) no Nordeste do Brasil

Abstract A histopathological survey was conducted to investigate the presence of microparasites in fish Archosargus probatocephalus in a river near Maceió, Brazil. Light microscope observations of fragments of gill showed the presence of small cysts containing numerous myxospores that were morphologically identified as Henneguya. Transmission electron microscopy observations further revealed several gill cells containing groups of prokaryotic cells within large cytoplasmic vacuoles. Each infected host cell displayed a single vacuole containing a variable number of Rickettsia-like cells (up to 11), some of which presented the dumbbell shape characteristic of binary fission. The Rickettsia-like cells were pleomorphic, without a nucleus and with chromatin dispersed in the cytoplasm. They had a thin electron-dense wall of Gram-negative type. The morphology of these prokaryotic was similar to those of the order Rickettsiales and was described as a Rickettsia-like organism. Histopathological evaluation showed that several vacuole membranes had a lysed appearance. Some had ruptured, thus allowing direct contact between the Rickettsia-like organism and the cytoplasm of the host cell. The rupturing of the branchial epithelium may have contributed towards reduction of the surface area of the gills, but it is not possible to say that this was the cause of the host's death.

Resumo Um levantamento histopatológico foi realizado para pesquisar a presença de microparasitas, no peixe Archosargus probatocephalus, em um rio próximo a Maceió, Brasil. Observações ao microscópio óptico de fragmentos de brânquias mostraram a presença de pequenos cistos contendo numerosos mixósporos, identificados morfologicamente como Henneguya. Ocasionalmente, na microscopia eletrônica de transmissão, foram observados vários corpos citoplasmáticos de inclusão, grupo aparentemente de células procarióticas que vivem dentro de um grande vacúolo citoplasmático de algumas células branquiais. As células hospedeiras infectadas tinham um único vacúolo contendo um número variável de células do tipo Rickettsia, até 11, algumas das quais em forma do haltere, característica da fissão binária. Essas células eram pleomórficas sem núcleo, tendo a cromatina dispersa no citoplasma e possuíam uma parede densa de elétrons finos do tipo Gram-negativo. A morfologia dessas células procarióticas foi semelhante àquelas da ordem Rickettsiales e foram descritas como organismos tipo Rickettsiae. A histopatologia mostra várias membranas de vacúolos circundantes com aspetos lisados, enquanto outras apresentam rupturas que mostram contato direto do organismos tipo Rickettsiae com o citoplasma da célula hospedeira. A ruptura do epitélio branquial pode ter contribuído para a redução da superfície das brânquias, mas não é possível afirmar que foi a causa da morte do hospedeiro.

An ultrastructural approach for gill responses after pollutants exposure / Un enfoque ultraestructural de las respuestas branquiales después de la exposición a contaminantes

SUMMARY: Morphological and physiological responses of gills exposed to environmental contaminants are widely described in the literature. Among the primary findings are the lamellar fusion, hyperplasia, aneurysms, proliferation of mitochondria-rich cells and mucous cells. This work has as main objectives, to show the main changes caused by a polluted lake in gills, and understand how each gill's cell type responds to pollutants' exposure. To those individuals of the species, Astyanax altiparanae were exposed to water from an urban lake, which receives such contaminants from various sources. The gills were analyzed under a TEM. Our results showed that prior to these changes, commonly found, only the mucous cells did not show structural changes. Among the main results, we observed the emergence of Rodlet cells in the group exposed to contaminants, as well as severe damage to the mitochondria-rich cells. The latter result was extremely relevant for demonstrating that the proliferation of this cell type, widely described as an alternative ionic regulation, may in some cases be completely inefficient because structurally these cells are no longer able to perform their basic metabolic functions.

RESUMEN: Las respuestas morfológicas y fisiológicas de las branquias expuestas a contaminantes ambientales se describen ampliamente en la literatura. Entre los hallazgos principales se encuentran la fusión laminar, la hiperplasia, los aneurismas, la proliferación de células ricas en mitocondrias y las células mucosas. Este trabajo tiene como objetivo principal, mostrar los principales cambios en las branquias, causados por un lago contaminado y comprender cómo responde el tipo de célula de las branquias a la exposición de los contaminantes. Individuos de la especie Astyanax altiparanae fueron expuestos al agua de un lago urbano, receptor de contaminantes de diversas fuentes. Las branquias se analizaron bajo un TEM. Nuestros resultados mostraron que antes de estos cambios, comúnmente encontrados, solo las células mucosas no mostraban cambios estructurales. Entre los resultados principales, observamos la aparición de células Rodlet en el grupo expuesto a contaminantes, así como el daño severo a las células ricas en mitocondrias. Este último resultado fue extremadamente relevante para demostrar que la proliferación de este tipo de células, ampliamente descrito como una regulación iónica alternativa, en algunos casos puede ser completamente ineficiente porque estructuralmente estas células ya no son capaces de realizar sus funciones metabólicas básicas.

Gill morphology in two bottom feeder Mediterranean Sea fishes: Grey Gurnard Fish (Eutrigla gurnardus, Linnaeus, 1758) and Striped Red Mullet Fish (Mullus barbatus surmuletus, Linnaeus, 1758) by scanning electron microscopy / Morfología de las branquias en dos peces que se alimentan en el fondo del Mar Mediterráneo: Gurnard Grises (Eutrigla gurnardus, Linnaeus, 1758) y salmonete rojo rayado (Mullus barbatus surmuletus,Linnaeus, 1758) por microscopía electrónica de barrido

The purpose of the current investigation was to describe the gill morphology of two bottom feeders Mediterranean Sea fishes: Striped red mullet fish (M. surmuletus) and grey gurnard fish (E. gurnardus) that showed the same feeding habits. The gill system of the grey gurnard consisted of four pairs of gill arches while consisted of three pairs of gill arches in the striped red mullet. Our study focused on the scanning electron microscopical (SEM) features of the gills, where some differences in the gill arches and gill rakers surface in both species were observed. Our results marked that there was an interbranchial septum carrying a four transverse elevated crest in grey gurnard while in striped red mullet carrying a median longitudinal elevated crest. There are some similar structure on the gill arch of two fishes as; no angle between a ceratobranchial part and epibranchial part and also the gill arch has a region of many longitudinal microridges demarcated the region between gill rakers and gill filaments. By SEM, in striped red mullet, the smooth surface of gill arch and gill rakers was characterized by the presence of high number of taste buds. By SEM in grey gurnard, gill raker appeared as a round short projected body with high number of curved apex spines. Furthermore, in striped red mullet, gill raker appeared as short projected body with high number of rod-like spines. Gill filaments were long at middle and short at extremities of gill arch in both species.

El propósito de este trabajo fue describir la morfología de las branquias de dos peces que se alimentan en el fondo del mar Mediterráneo: Gurnard grises (E. gurnardus) y salmonete rojo rayado (M. surmuletus) los cuales mostraron los mismos hábitos alimenticios. El sistema de branquias del E. gurnardus gris consiste en cuatro pares de arcos branquiales, mientras que en el salmonete rayado son tres pares. El estudio se centró en la observación de las características de las branquias por microscopía electrónica de barrido (MEB), donde se visualizaron algunas diferencias en ambas especies tanto en los arcos branquiales, como en la superficie de las ramas branquiales. Nuestros resultados marcaron la presencia de un septum interbranchial que presentaba cuatro crestas elevadas a nivel transversal en el E. gurnardus gris, mientras que el salmonete rayado presentaba una cresta elevada mediana longitudinal. Hay una estructura similar en el arco branquial de los dos tipos de peces, pero no hay ángulo entre una parte ceratobranquial y la parte epibranquial. Además, el arco branquial tiene una región de muchas microrredes longitudinales que delimitan la región entre las ramas branquiales y los filamentos branquiales. Por MEB, en el salmonete rayado, la superficie lisa del arco branquial y el de las ramas branquiales se caracterizaron por la presencia de un alto número de papilas gustativas. Por MEB en E. gurnardus gris, las branquias aparecieron como un cuerpo corto proyectado con un alto número de espinas de vértice curvo. Además, en el salmonete rojo rayado, el rastrillo de las branquias apareció como un cuerpo corto proyectado con un alto número de espinas tipo bastón. En ambas especies los filamentos branquiales eran largos en el centro y cortos en los extremos del arco branquial.

Morphological Studies on the Gills of Puffer Fish (Lagocephalus sceleratus, Gmelin, 1789) / Estudios Morfológicos en Branquias del Pez Globo (Lagocephalus sceleratus, Gmelin, 1789)

In the present study, we focused on the morphology of L. sceleratus gills using gross anatomy, scanning electron microscopy as well as light microscopy. Results of this study revealed that the gill openings appeared as simple slits anterior to the pectoral fin without distinct opercular cover. The gill system consisted of three pairs of gill arches carrying two rows of gill rakers on its concave border and gill filaments on its convex border. SEM showed that all surfaces of the gill arch were characterized by the presence of the longitudinal ridges with many taste buds in addition to many spines around the rakers. Histologically, the gill arch was composed of curved bar of hyaline cartilage with slightly elevated area corresponding to the sites of gill rakers. Each filament was formed of a thin central cartilaginous core surrounded by peripheral cartilaginous matrix and covered by primary epithelial layer with abundant mucous cells. The chloride cells appeared mainly near to the base of secondary lamellae. Each gill filament gave rise to a very large number of secondary lamellae on both sides. The epithelial lining of the secondary lamellae comprised epithelial pavement cells, few mucous cells and pillar cells. The latter interposed the enriched blood capillaries. These findings suggest that L. sceleratus gills have characteristic morphological features that are related to adaptive functions for feeding habits, osmoregulation and respiratory mechanism with in their living environment.

El objetivo fue estudiar la morfología de las branquias de Sceleratus L. desde la anatomía macroscópica, microscopía electrónica de barrido, así como la microscopía de luz. Los resultados revelaron que las aberturas branquiales aparecían como simples rendijas por delante de la aleta pectoral sin una cubierta opercular distinta. El sistema branquial consistió en tres pares de arcos branquiales con dos filas de branquiespinas en sus filamentos branquiales frontales y cóncavos en el margen. La microscopía de barrido mostró que todas las superficies del arco branquial se caracterizaron por la presencia de crestas longitudinales con muchas papilas gustativas, además de una cantidad importante de espinas alrededor de los rastrillos. Histológicamente, el arco branquial se compone de una barra curva de cartílago hialino con una zona ligeramente elevada, correspondiente a los sitios de branquiespinas. Cada filamento se formó por un delgado núcleo central cartilaginoso rodeado de matriz cartilaginosa periférica y cubierto por una capa epitelial primaria con abundantes células mucosas. Las células de cloruro aparecieron principalmente cerca a la base de laminillas secundaria. Cada filamento de las branquias en ambos lados dio origen a un gran número de laminillas secundarias. El revestimiento epitelial de laminillas secundarias estaba formado de células epiteliales, algunas células mucosas y células sostenedoras. Estos hallazgos sugieren que las branquias de L. sceleratus tienen características morfológicas que están relacionadas con las funciones de adaptación de los hábitos de alimentación, la osmorregulación y el mecanismo respiratorio de acuerdo a su entorno.

Surface ultrastructure of the gills of Odontesthes bonariensis (Valenciennes, 1835)(Teleostei: Atheriniformes) from a temperate shallow lake / Estudio ultraestructural de las branquias de Odontesthes bonariensis (Valenciennes, 1835) (Teleostei: Atheriniformes) de un lago somero templado

We studied the gill epithelium of juveniles of Odontesthes bonariensis using scanning electron microscopy with a particular interest in the relationship between cell morphology and the different functional aspects of gills. The specimens used were collected in Laguna de los Padres, Buenos Aires Province, Argentina. The gill filaments or primary lamellae are arranged in two rows along the convex outer face of the gill arch forming the anterior and posterior hemibranchiae. On its concave inner face, each arch supports two rows of gill rakers forming the pharyngeal region wall. The primary lamellae are lined by a stratified epithelium made up of polygonal cells with well-defined outlines and cell membrane microfolds. Several mucous and chloride cells were observed in the primary lamellae. The epithelial cells of the secondary lamellae have ill-defined outlines and few microfolds. Both the arches and gill rakers are covered by a stratified epithelium of polygonal cells, among which mucous cells and taste corpuscles are abundant. The ultrastructural analysis of gills of O. bonariensis contributes to the advancement of knowledge of the organ histophysiology and of future histopathological studies.

Se estudió el epitelio branquial de juveniles de Odontesthes bonariensis utilizando microscopía electrónica de barrido con un interés particular en la relación entre la morfología celular y los distintos aspectos funcionales de las branquias. Los ejemplares utilizados fueron colectados en la Laguna de los Padres, Provincia de Buenos Aires, Argentina. Los filamentos branquiales o laminillas primarias están dispuestos en dos filas a lo largo de la cara exterior convexa del arco branquial formando las hemibranquias anterior y posterior. En su cara interior cóncava, cada arco sostiene dos hileras de branquiespinas que forman la pared de la región faríngea. Las laminillas primarias están revestidas por un epitelio estratificado constituido por células poligonales con contornos bien definidos y micropliegues de la membrana celular. Se observaron numerosas células clorhídricas y mucosas en las laminillas primarias. Las células epiteliales de las laminillas secundarias tienen contornos poco definidos y escasos micropliegues. Tanto los arcos como las branquiespinas están cubiertos por un epitelio estratificado de células poligonales, entre las cuales hay abundantes células mucosas y corpúsculos gustativos. El análisis ultraestructural de las branquias de O. bonariensis proporciona información para avanzar en el conocimiento de la histofisiología del órgano y para realizar futuros estudios histopatológicos.

The effect of salinity on osmoregulation and development of the juvenile fat snook, Centropomus parallelus (POEY) / O efeito da salinidade sobre a osmoregulacao e o desenvolvimento do juvenil de robalo-peva, Centropomus parallelus (POEY)

Eurihaline fish support waters with different salt concentration. However, numerous studies have shown that salinity can affect fish development. Thus, the effect of salinity change from 20 to 5 and 35 on survival, weight, length, gill chloride cell ultrastructure and gill Na+, K+ ATPase activity was evaluated in Centropomus parallelus following short-term (6, 24 and 96 hours) and long-term exposure (30 and 60 days). Salinity did not affect C. parallelus survival, final weight and length. The quantity of chloride cells increased visibly at salinities of 5 and 35, with the cells exhibiting the typical features of uptake and secretory cells, respectively. Na+, K+ ATPase activity in the gill of the C. parallelus was significantly greater at a salinity of 5 than at a salinity of 20 or 35 after 96 hours, but not after 30 or 60 days. These results indicate that salinity change from high to low salt water induces gill chloride cell and Na+, K+ ATPase activity adaptations after short-term exposure. However, after long-term exposure at salinity 5, gill Na+, K+ ATPase activity is no more necessary at high levels. The increase in salinity to 35 does not induce significant change in gills. Juveniles of C. parallelus may thus be capable of acclimating to salinities of 5 to 35 for 60 days without significant effects on development.

Peixes eurihalinos suportam águas com diferentes concentrações de sal. Contudo, muitos estudos têm mostrado que a salinidade pode afetar o desenvolvimento do peixe. Portanto, o efeito da mudança de salinidade de 20 para 5 e 35 na taxa de sobrevivência, peso, comprimento, morfologia das células de cloreto branquiais e atividade da Na+, K+ ATPase foram avaliadas no Centropomus parallelus após curto (6, 24 e 96 horas) e longo tempo de exposição (30 e 60 dias). A salinidade não afetou a sobrevivência, o peso e comprimento final do robalo-peva. A quantidade de células de cloreto aumentou visivelmente nas salinidades 5 e 35, exibindo morfologias típicas de células que absorvem e secretam sal, respectivamente. A atividade da Na+, K+ ATPase nas brânquias do C. parallelus foi significativamente maior na salinidade 5 do que nas salinidades 20 ou 35 após 96 horas, mas não após 30 e 60 dias. Esses resultados indicam que a mudança de alta para baixa salinidade provoca adaptações nas células de cloreto e na atividade da Na+, K+ ATPase branquial em curto prazo. Contudo, após longa exposição na salinidade 5, a alta atividade da Na+, K+ ATPase branquial não é mais necessária. O aumento de salinidade para 35 não induz mudanças significativas nas brânquias. Portanto, juvenis de C. parallelus possuem a capacidade de aclimatação nas salinidades de 5 a 35 semefeitos significativos no desenvolvimento após 60 dias.

Deformación de branquias en salmónidos: análisis macroscópico, histológico, ultraestructural y de elementos / Gill deformation in salmonids: macroscopic, histological, ultrastructural and elements analysis

Las branquias son los órganos destinados al proceso de respiración en los peces, estas están consideradas, entre las estructuras más delicadas del cuerpo de los teleósteos ya que están expuestas a sufrir daños de cualquier agente irritante, los cuales constituyen la causa más frecuente y significativa de sus cambios patológicos. Los objetivos del presente estudio son caracterizar las alteraciones branquiales desde los puntos de vista anatómico, histológico, además, determinar niveles de Ca, P y Zn del tejido óseo de la columna vertebral y cráneo de salmones afectados. Se utilizaron 47 salmones, divididos en dos grupos: control y con deformaciones, a los cuales se les realizaron análisis macroscópico, histológico, ultraestructural y de elementos. Se observaron alteraciones tanto en el contexto anatómico como histológico, así como gran presencia de bacterias y hongos que afectaban principalmente al primer arco, pudiendo concluir que los peces tendrían una susceptibilidad génica a la deformación branquial.

Gills act as the respiratory organ in the respiration process in fish. They are considered among the most delicate body structures of teleost fish as they are subject to damage from any irritant, which are the most frequent and significant causes of their pathological changes. The objective of this study is to characterize gill alterations from the anatomical and histological standpoint, and to determine levels of Ca, P and Zn in bone tissue of the spine and skull of affected salmon. For purposes of this study 47 salmon were used in two groups: control and deformation, subjected to complete histological, ultrastructural and element inspection. Abnormalities were observed in both the anatomical and histological context and extensive presence of bacteria and fungi affecting mainly the first arc were found. Therefore, we can conclude that these fish have a genetic susceptibility to gill deformation.

Descripción anatómica, histológica y ultraestructural de la branquia e hígado de tilapia (Oreochromis niloticus) / Anatomical, histological and ultraestructural description of the gills and liver of the tilapia (Oreochromis niloticus)

La tilapia (Oreochromis niloticus) es un pez presente en muchos ecosistemas de agua dulce, en diversas regiones de Colombia. Esta especie es una fuente de alimentación de alta demanda de la población en general, por lo cual es de gran interés que su hábitat acuícola esté libre de contaminantes, con el fin que el especimen se desarrolle y crezca normalmente y así pueda ser consumido por el ser humano sin ningún perjuicio. Las branquias y el hígado de peces son considerados como los órganos primarios donde detectar efectos de polución. Este estudio muestra la morfología normal de las branquias e hígado de tilapia, a través de microscopías: de luz, óptica de alta resolución (MOAR), electrónica de transmisión (TEM) y de barrido (MEB); como un estudio básico para futuras investigaciones, en las cuales se puedan comparar tejidos sanos y tejidos expuestos a contaminantes ambientales; por lo cual, este análisis podría ser utilizado como referente para la aplicación como biomarcador histopatológico de estrés ambiental. Para esta investigación se recolectaron especímenes en un afluente del Río Patía (Departamento del Cauca). Sacrificado el animal, se disecó el arco branquial izquierdo y el hígado. Las biopsias se fijaron con las técnicas estándares para microscopías de luz y electrónica. En los cortes obtenidos se observa la organización en forma detallada de los diferentes tejidos que componen a las branquias y al hígado de especímenes sanos.

The tilapia (Oreochromis niloticus) is a fish that can be found in many of the sweet water ecosystems throughout various regions of Colombia. This species is a high demand food source for the population in general and therefore it is important that its aquaculture habitat remains free of contaminating pollutants, in order that the species develop and grow normally for adequate human consumption. The branchia and the liver of fish are considered the primary organs wherein the effects of pollution can be detected. This study shows the normal morphology of the branquia and the liver of the tilapia through light microscopy, high resolution optical microscopy , electronic transmission microscopy, and electronic scanning microscopy, as basic research for future investigation in which healthy tissue and tissue exposed to environmental pollutants can be compared. This analysis is thus used as reference for the application as a histopathological marker of environmental stress. For purposes of this research specimens of a tributary of the Patia river (Departamento de Cauca) were collected. The animal was sacrificed, the left branquial arch and the liver were dissected; the biopsies were set for standard light microscopy as well as electronic microscopy. The organization is shown in detail in the slices obtained in the different tissue which compose the branquia and the liver of healthy specimen.

Rodlet cells from the gills and kidneys of two brazilian freshwater fishes: an ultrastructural study

Rodlet cells (RCs) are fish cells considered to be regulatory elements, ion transportation cells, secretory cells, parasitic cells, transport units of genetic material, non-specific immune cells and endogenous in nature cells. In this report, we describe the ultrastructure of RCs collected from the gills and kidneys of two species of freshwater teleosts (family Curimatidae) in Brazil: Curimata macrops Eigenmann & Eigenmann, 1889 from the Poty river, near the city of Teresina in the State of Piauí, and Curimata inornata Vari, 1989 from the Amazon river near the city of Belém in the State of Pará. A variable number of RCs was observed in these tissues, with a higher frequency in gills compared to the kidneys. No other organs were investigated. RCs were observed in healthy fish and in fish parasitized by a myxosporean of the genus Henneguya. The RCs consisted of a thick-layered capsule enclosing a variable number of small, dense rodlets surrounded by several vacuoles and a nucleus. The capsule was a cytoplasmic structure composed of thick fibrillar elements surrounded externally by the plasmalemma. The capsule and surrounding plasmalemma had a smooth, undulating surface with several microvilli projecting towards the surrounding cells. Some of the microvilli located in the apical zone of the RCs were in contact with the disorganized microfibrils of the capsules. The nucleus was located laterally or basally and showed condensed chromatin at the periphery. The ultrastructural organization of the apical zone of the RCs suggested that these cells may be involved in secretory functions. This is the first report of RCs in these two species of Brazilian fish.

Histological and electron microscopic studies of gills of Nile tilapia oreochromis niloticus from two wetlands differing in water quality

Histological and electron microscopic studies of gill tissue were carried out in specimens from two wild populations of Nile Tilapia Oreochromis niloticus, the first from River Nile and the other from a pond located in south Egypt [Assiut Governorate]. These two wetland ecosystems differ in. water quality. Scanning electron microscopy revealed that the arrangement of microridges on the pavement cell surfaces of gill epithelia was variable among two populations. From River Nile, pavement cells of gill filament and secondary lamnellae were similar for their surface architecture. Also, these cells of both filament and secondary lamella exhibit numerous long microridges with a fingerprint-like pattern. From the pond, Pavement cells of filament may vary from cells on which microridges form a complex pattern of concentric whorls to cells with a smooth central part without microridges, while pavement cells from the secondary lamellae are smooth surfaced with clearly defined cell boundaries by a long microridge. On the other hand, the transmission electron microscopy revealed that the pavement cells of secondary lamellae from the pond are thinner than those from River Nile. Differences in morphology of the pavement cells from two populations may be related to structural rather than functional differences

Microhabitat and surface ultrastructure of neothoracocotyle commersoni and pricea multae [monogenea: microcotyloidea] from gills of scombrid fish, scomberomorus commerson

The microhabitat and surface topography of microcotylid monogenean parasites Neothoracocotyle commersoni and Pricea multae from gills of Spanish mackerels, Scomberomorus commerson, was investigated. The results showed that N. commersoni was aggregated at the first gill sector, while P. Multae was aggregated at both first and second sectors in single infestation. On contrary, in mixed infestation, N. commersoni occupied the first sector of gills and P. Multae was restricted only at the fourth sector with a high intensity of infestation compared with N. commersoni. The aggregation of single species of monogenea in restricted habitat may aid in the cross- fertilization, hence produce hybrids strains of parasites which might be more potent and vicious causing severe damage to the host. The surface ultrastructure of the representative monogenea showed that the tegument was provided with microvillus-like projections that suggested metabolic exchange and absorption of micromolecular nutrients from the surrounding environment. The architecture of clamps of N. Commersoni differed from that of P. Multae in possessing microvillous-like projections, which may act as an organ of nutrition, beside its principle attachment function. The ultrastructure of the clamps of both species provided an aid in understanding of the parasite habitat and survival