Mitochondria-rich cells changes induced by nitrite exposure in tambaqui (Colossoma macropomum Cuvier, 1818)

ABSTRACT The gill mitochondria-rich cells of the juvenile Amazonian fish Colossoma macropomum were analyzed using light and scanning and transmission electron microscopy after 96 h exposure to 0.04 and 0.2 mM nitrite. Although the number of mitochondria-rich cells decreased significantly in the lamellar epithelium, no decrease was found in the interlamellar region of the gill filament. Nitrite exposure caused significant reduction on the apical surface area of individual mitochondria-rich cells (p < 0.05), with a resulting reduction of the fractional area of these cells in both the lamellar and filament epithelium. Swelling of endoplasmic reticulum cisternae, nuclear envelope and mitochondria were the main changes found in the mitochondria-rich cells. Cristae lysis and matrix vacuolization characterized the mitochondrial changes. The overall ultrastructural changes indicated cellular functional disruption caused by exposure to nitrite. The changes observed in the gill indicate that the cellular structures involved in the process of energy production become severely damaged by exposure to nitrite indicating irreversible damage conducting to cell death.

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.

Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo / The influence of the aquatic environment in tanks sequetially interconnected with PVC pipes on the gill epithelium and lamellas of tilapia (Oreochromis niloticus)

Objetivou-se analisar o comportamento do epitélio branquial de tilápias, cultivadas em tanques posicionados em diferentes altitudes e interconectados por tubos de PVC. Filamentos branquiais de quatro espécimes de quatro tanques (T1, T2, T3 e T4) interconectados seqüencialmente foram submetidos à rotina histológica, para obtenção de cortes de 5µm de espessura, os quais foram corados com Hematoxilina-Eosina, ou submetidos à técnica histoquímica para glicoconjugados: PAS + solução de diástase ou Alcian Blue pH 2,5 ou Alcian Blue pH 1,0. Considerando as regiões basal, intermediária e apical dos filamentos, mensurou-se a área lamelar e contou-se o número de células mucosas em cada uma dessas regiões, o que correspondia a 0,56 mm². Verificou-se que a concentração de oxigênio, pH e a temperatura se reduziam progressivamente com a passagem da água de um tanque para outro. Em função disso, constatou-se um aumento abrupto do número de células mucosas e da área lamelar no T2, e uma redução progressiva destas medidas nos tanques que recebiam água do T2. Além disso, observou-se nos animais do T2, T3 e T4, descolamento do epitélio branquial, hiperplasia celular no espaço interlamelar e telangectasias. Conclui-se que o ambiente aquático de tanques interconectados seqüencialmente por tubos de PVC se altera ao passar de um tanque para o outro, e que estas flutuações físico-químicas se refletem no comportamento do epitélio branquial através de variações da área lamelar e do número de células mucosas.

The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

Thyroid status alters gill ionic metabolism and chloride cell morphology as evidenced by scanning electron microscopy in a teleost Anabas testudineus (Bloch): short and long term in vivo study.

Gill is the main organ of osmotic regulation in teleosts and chloride cells are the sites of ion transport across gill epithelium. Thyroid hormones are implicated in the regulation of osmotic balance in teleosts also. Treatment with 6-propyl thiouracil (6-PTU) inhibited the membrane bound enzyme Na+K+ ATPase in the gill while triiodothyronine (T3) injection stimulated it in a short-term in vivo study in the teleost Anabas testudineus. Na+, K+ and Ca2+ ions were also decreased in the 6-PTU treated fish and the T3 treatment increased their concentrations in the gill lamellae. The gill morphology also changed according to the thyroid status in the long term study. 6-PTU treatment altered the typical serrated morphology of the gill lamellae, while the T3 treatment reversed it. T3 injection increased the density of pavement and chloride cells as evidenced by scanning electron microscopy. The results demonstrate that physiological status of the thyroid influences gill Na+ pump activity and chloride cell morphological changes. Further, the study suggests a regulatory role of T3 on gill ions (Na+, K+ and Ca2+), Na+K+ and Ca2+ ATPase activity and the different gill cell types in A. testudineus.

Karyotype description of Pomacea patula catemacensis (Caenogastropoda, Ampullariidae), with an assessment of the taxonomic status of Pomacea patula

Mitotic chromosomes of the freshwater snail Pomacea patula catemacensis (Baker 1922) were analyzed on gill tissue of specimens from the type locality (Lake Catemaco, Mexico). The diploid number of chromosomes is 2n = 26, including nine metacentric and four submetacentric pairs, therefore, the fundamental number is FN = 52. No sex chromosomes could be identified. The same chromosome number and morphology were already reported for P. flagellata, i.e., the other species of the genus living in Mexico. The basic haploid number for family Ampullariidae was reported to be n = 14 in the literaure; so, its reduction to n = 13 is probably an apomorphy of the Mexican Pomacea snails. Lanistes bolteni, from Egypt, also shows n = 13, but its karyotype is much more asymmetrical, and seems to have evolved independently from P. flagellata and P. patula catemacencis. The nominotypical subspecies, P. patula patula (Reeve 1856), is a poorly known taxon, whose original locality is unknown. A taxonomical account is presented here, and a Mexican origin postulated as the most parsimonious hypothesis.

Primary cell culture from fish gills and kidney using fish serum.

A novel, cost effective and time saving technique for primary cell culture from gills and kidney using fish serum has been developed. Single cell suspension of gills and kidney of Clarias gariepinus was obtained by trypsinization. Minimum essential medium supplemented with 10% fish serum in place of 10% foetal calf serum and 20% fish muscle extract, yielded confluent monolayer on 6th and 8th day in gill and kidney tissue respectively at 28 degrees C. Fish serum can be successfully used as media supplement for cultivation and maintenance of primary cell culture from fishes.

Epithelial gill cells in the armored catfish, Hypostomus cf. plecostomus (Loricariidae)

Epithelial gill cell morphology and distribution were investigated in the armored catfish, Hypostomus cf. plecostomus, which lives in soft ion-poor Brazilian freshwaters. Pavement cells are the most abundant type of cell on both filament and lamellar epithelia and there are a great number of mucous and chloride cells between them. Mucous cells are almost covered by adjacent pavement cells and have large packed granules showing electrondense differences. No mucous cells were found on the lamellar epithelium. Chloride cell were distributed throughout both epithelia and usually have large apical surface facing the external medium and may exhibit short and sparsely distributed microvilli. The presence of chloride cells on the lamellar epithelium may be an adaptation to low ion concentrations in the water, allowing for improved ion-transport capacity of the gill. The large size of these cells increases the water-blood barrier and may affect the transference of respiratory gases. However, the negative effect on the respiratory process may be minimized by this species' ability to resort to atmospheric air to fulfill its oxygen requirements

Structural and morphological features of Piaractus mesopotamicus (Holmberg, 1887) gills

The pacu Piaractus mesopotamicus (Holmberg, 1887), is a commercially important serrasalmid fish endemic to the Paraná-Paraguay River basin, and one of the most widely cultivated native fish species in the Neotropics. As a migratory species, also inhabiting temporary pools subjected to hypoxic conditions, the species presents a high plasticity concerning respiratory adaptations. In order to supplement basic knowledge on the respiratory physiology and morphology of this species, some structural features of pacu gills, such as filament and lamellae structure, and circulatory pattern are described in this paper

K+ balance in rainbow trout gill and liver tissue, cell suspensions and cultured cells

Both intact gill and liver tissue from rainbow trout accumulated K+, as determined by 86Rb+ uptake, a process largely inhibited by ouabain, indicating the presence of functional NaKATPase. Cell suspensions, produced by disaggregation of gill or liver tissues, accumulated very little K+ compared to tissues (Less than 10 percent). Disaggregation resulted in depolarisation of cells with loss of intracellular K+ and although NaKATPase, as measured by 86Rb+ uptake rate, remained functional and inhabitable by ouabain, the activity was insufficient to replace the rapid K+ loss. While attached, cultured gill and liver cells showed normal K+/Na+ ratios and NaKATPase activity, but release from the substratum resulted in depolarisation and rapid K+ loss as seen in cell suspensions. These results suggest that care is required in interpreting ionic regulatory and other results from cell suspensions and that further research should be directed towards systems where cells can maintain normal ionic balance.