BackgroundA survey on endoparasitic helminths from freshwater fishes in the Pantanal Region (Mato Grosso do Sul, Brazil) revealed the occurrence of third-larval stage of the nematode Brevimulticaecum sp. (Heterocheilidae) in most organs of Gymnotus inaequilabiatus (Gymnotidae) also known by the local name tuvira. The aim of the present study was to examine Brevimulticaecum sp.-infected tuvira liver at the ultrastructural level and clarify the nature of granulomas and the cellular elements involved in the immune response to nematode larvae.MethodsThirty-eight adult specimens of tuvira from Porto Morrinho, were acquired in January and March 2016. Infected and uninfected liver tissues were fixed and prepared for histological and ultrastructure investigations.ResultsThe prevalence of infection of tuvira liver by the nematode larvae was 95 %, with an intensity of infection ranging from 4 to 343 larvae (mean ± SD: 55.31 ± 73.94 larvae per liver). In livers with high numbers of nematode larvae, almost entire hepatic tissue was occupied by the parasites. Hepatocytes showed slight to mild degenerative changes and accumulation of pigments. Parasite larvae were surrounded by round to oval granulomas, the result of focal host tissue response to the infection. Each granuloma was typically formed by three concentric layers: an outer layer of fibrous connective tissue with thin elongated fibroblasts; a middle layer of mast cells entrapped in a thin fibroblast-connective mesh; and an inner layer of densely packed epithelioid cells, displaying numerous desmosomes between each other. Numerous macrophage aggregates occurred in the granulomas and in the parenchyma.ConclusionsOur results in tuvira showed that the larvae were efficiently sequestered within the granulomas, most of the inflammatory components were confined within the thickness of the granuloma, and the parenchyma was relatively free of immune cells and without fibrosis. Presumably this focal encapsulation of the parasites permits uninfected portions of liver to maintain its functions and allows the survival of the host.

The aim of this study was to determine the morphometric measures and morphological aspects of nematode-cysts in Gymnotus inaequilabiatus, and the presence of melanomacrophage centers (MMCs) associated with the periphery of cysts and in the liver parenchyma. Adult specimens, 34 female (123.1 ± 43.9g) and 45 male (135.5 ± 43.4g), from Paraguay River, Corumbá, Brazil, were used. The number of nematode-cysts was determined in 79 livers and 25 of them randomly selected for histopathological analysis and morphometric measures of nematode-cysts (mean diameter, thickness of collagen layer, and cyst-wall layer). The percentage of cysts with MMCs on the periphery and density in the liver parenchyma was estimated. The average number of macroscopic cysts was of 48.7 ± 2.78. Granulomatous reaction was observed surrounding the cysts. Diameter, collagen layer and cyst-wall measurements were 293.0 ± 75.18 (µm), 17.72 ± 6.01 (µm) and 12.21 ± 9.51 (µm), respectively. The number of nematode-cysts was correlated with hepatosomatic index, (r=0.26, P<0.05). Collagen layer was correlated with cyst diameter (r=0.62, P<0.01). Pericystic and parenchymatous MMCs were moderately (r=0.48) and highly (r=0.90) correlated with nematode-cysts number. Morphological characteristics of hepatic tissue and cysts-nematodes measures suggest that G. inaequilabiatus acts as a paratenic host to nematodes in the larval stage.Keywords: Brevimulticaecum sp., Gymnotiformes, nematoda, melanomacrophages. ResumoO objetivo deste estudo foi determinar as características morfológicas e morfométricas de cistos de nematodas em fígado de Gymnotus inaequilabiatus, e a presença de centros de melanomacrófagos (CMMs) pericísticos e no parênquima hepático. Espécimes adultos, 34 fêmeas (123,1 ± 43,9 g) e 45 machos (135,5 ± 43,4g), provenientes do Rio Paraguai, Corumbá, Brasil foram utilizados. Os cistos foram quantificados em 79 fígados. Em 25 fígados foi realizada a análise histopatológica e morfometria dos cistos (diâmetro, espessura da camada de colágeno e da parede). Estimou-se a porcentagem de cistos com CMMs periférico e a densidade de CMMs no parênquima. A média de cistos macroscópicos foi de 48,7 ± 2,78. Observou-se reação granulomatosa pericística. O diâmetro, a espessura da camada de colágeno e da parede do cisto foi de 293,0 ± 75,18 (µm), 17,72 ± 6,01 (µm) e 12,21 ± 9,51 (µm), respectivamente. O número de cistos foi associado ao índice hepatossomático, (r=0,26, P <0,05). A camada de colágeno correlacionou-se com o diâmetro do cisto (r=0,62, P <0,01). CMMs pericísticos e do parênquima apresentaram correlação moderada (r=0,48) e alta (r=0,90) com o número de cistos, respectivamente. As características morfológicas do tecido hepático e a morfometria dos cistos de nematodas sugerem que G. inaequilabiatus atua como hospedeiro paratênico para larvas de nematodas.Palavras-chave: Brevimulticaecum sp., Gymnotiformes, nematoda, melanomacrófagos.

The Gymnotus inaequilabiatus is a Neotropical fish widely distributed in marginal areas of bays. The aim of this study was to describe the main histological and histopathological traits in the head and exocrine kidney. Here, histometric and structural density techniques were associated with renosomatic index (RSI). The kidney was processed for light microscopy. Lipofuscin and hemosiderin content were visually estimated in the melonomacrophages centers (MMCs). All the biometric body variables were correlated with RSI, especially the kidney weight and gross lesions count. The general architecture of head and exocrine kidney was similar to that described for other teleost species. MMCs were prevalent in both portions and correlated with RSI in the head and exocrine kidney. Granulomatous structures were often observed in both portions; however, they were associated only in the exocrine kidney with RSI. Of all the structures hystometrically estimated, only proximal tubular diameter and thickness, and distal tubular thickness were correlated to renosomatic index. The RSI is an useful biometric variable that represent some physiological and morphological characteristics of kidney in G. inaequilabiatus. These findings may be used in future studies to evaluate the effects of environmental stressors on the renal adaptative physiological process.

This study describes the hematological and biometric characteristics of male and female Gymnotus species from the Pantanal, Mato Grosso do Sul state, Brazil. Fifty adult specimens of Gymnotus inaequilabiatus were weighed, measured, and then euthanized. Blood was collected by puncturing the celiac mesenteric vein to determine the hematocrit, hemoglobin content, number of erythrocytes, mean corpuscular volume, mean corpuscular hemoglobin concentration, glucose level, absolute value of leukocytes, and relative value of leukocytes and thrombocytes. Body weight and relative condition factor did not differ (P > 0.05) between the sexes, as well as erythrogram and the blood glucose values. Hematocrit ranged from 18.0% to 54.0%; hemoglobin from 1.1 to 14.7 g dL -1 ; number of erythrocytes from 0.2 × 10 6 to 3.8 ×10 6 µL -1 ;MCV from 24.2 to 321.7 fL; and MCHC from 4.2 to 44.5 g dL -1 . In the differential count were identified thrombocytes, lymphocytes, neutrophils, monocytes, basophils, immature leukocytes, and PAS-positive granular leukocyte (PAS-GL). Females had a higher percentage of immature leukocytes (P < 0.05) than males. Glucose levels, erythrogram, leukogram, and the morphology of defense cells are comparable to other fish species of the Pantanal. Thrombocytes were the most frequent defense cells, followed by lymphocytes and neutrophils.