Toxic effects of aniline in liver, gills and kidney of freshwater fish Channa punctatus after acute exposure.

Aniline (C6H5NH2) is one of the hazardous aromatic amine where an amino group -NH2) is connected to phenyl ring (C6H5). Based on the evaluation of the 96-hour LC50 of aniline, two sublethal concentrations (4.19 mg/l and 8.39 mg/l) were selected for acute exposure tests in freshwater fish Channa punctatus. The liver, gills and kidney of fish being the principal sites of xenobiotic material accumulation, respiration, biotransformation, and excretion are the focus of the present study. Throughout the exposure time, the comet assay revealed increased tail length and tail DNA percentage indicating maximum damage to liver, gills and kidney of treated group after 96 h. After acute exposure, there was a significant (p ≤ 0.05) increase in the enzymatic activity of glutathione-S-transferase (GST) and acetylcholinesterase (AChE), whereas decline in superoxide dismutase (SOD) and catalase (CAT) activity was observed. Meanwhile, levels of malondialdehyde (MDA) increased over the exposure period for both concentrations. After 96 h of exposure, degree of tissue change (DTC) was evaluated in liver, gill and kidney of aniline exposed fish. Additionally, light microscopy revealed multiple abnormalities in liver, gills and kidney of all the treated groups. Significant changes were observed in the levels of biochemical markers viz., glucose, triglyceride, cholesterol, aspartate transaminase, alanine transaminase and urea following a 96-hour exposure to aniline. Studies using ATR-FTIR and transmission electron microscopy (TEM) revealed changes in biomolecules and structural abnormalities in several tissues of the aniline-exposed groups in comparison to the control group respectively.

Gill arch and raker morphology of common carp (Cyprinus carpio, Linnaeus, 1758) sampled in aquaculture system.

The study aimed to investigate the morphologic aspects of common carp's gill arch and gill rakers (Cyprinus carpio, Linnaeus, 1758), an omnivore and highest-produced aquaculture species. The study used 10 common carp (395.35 ± 45.06 g) grown entirely under aquaculture conditions. The fish tissues were fixed with Glutaraldehyde (2.5%) for scanning electron microscopy and with formalin (10%) for stereomicroscopic examination. In the SEM examination, two types of taste papillae (Type II and Type III) were observed in the pharyngeal mucosa. Microridge-like structures in the epithelial layer were found to have two forms. The study findings indicate a significant decrease in gill arch lengths from cranial to caudal and a significant increase in rakers per unit area, as determined through digital calliper measurements and stereomicroscopic examinations (p < 0.05). However, there was no significant difference in measurements of gill arches and raker numbers between the bilateral symmetry of the gill arches (p > 0.05). In conclusion, it was observed that the epithelial structure on the common carp gill arch contained two types of microridge-like structures: the gill arch length decreased from cranial to caudal, and the rake density on these arches increased caudally.

Gill histology and ultrastructure in Aplysia depilans (Mollusca, Euopisthobranchia).

The gill of Aplysia depilans consists of several wedge-shaped pinnules with a highly folded structure, differing from the typical ctenidial gills of mollusks. Light microscopy and transmission electron microscopy were used to investigate this organ in juveniles and adults. In this species, the gill epithelium comprised ciliated, unciliated, and secretory cells. The ultrastructural analysis suggests other functions for the gill besides respiration. The deep cell membrane invaginations associated with mitochondria in the basal region of epithelium point to a role in ion regulation. Endocytosis and intracellular digestion were other activities detected in epithelial cells. In juveniles, an intranuclear crystalline structure was seen in some ciliated cells. The presence of an intranuclear crystalline structure was frequently associated with chromatin decondensation, swelling of the nuclear envelope and endoplasmic reticulum cisternae, and abundance of Golgi stacks. As these intranuclear inclusions were not found in the gill of the adult specimens, their occurrence in the two juveniles seems likely to be an anomalous condition whose cause cannot be established at the moment. Mucous cells were the most abundant secretory cells in the epithelium, but a few epithelial serous cells were also found. In addition, large protein-secreting subepithelial cells had the main cell body inserted in the connective tissue and a long thin neck crossing the epithelium. Mucous cells can be considered responsible for the production of the mucus layer that protects the epithelium, but the specific functions of the epithelial and subepithelial protein-secreting cells remain elusive. Below the epithelium, a layer of connective tissue with muscle cells lined the narrow hemolymph space. The connective tissue included cells with a large amount of rough endoplasmic reticulum cisternae. Bacteria were found on the surface of the gill, and the most abundant had a thin stalk for attachment to the epithelial cells.

Chlorpyrifos caused necroptosis via MAPK/NF-κB/TNF-α pathway in common carp (Cyprinus carpio L.) gills.

Chlorpyrifos (CPF) is an organophosphate insecticide and can cause cell death of animals. In the study, the common carp were exposed to CPF at 0 µg/L (the control group), 1.16 µg/L (the low dose group), 11.6 µg/L (the medium dose group), and 116 µg/L (the high dose group), respectively. The carp were euthanized at the 30th day and gills were collected immediately. The ultrastructural and histopathological observations showed obvious necrosis characteristics and inflammatory injury in the CPF-treated groups. CPF exposure activated the MAPK pathway, in which the mRNA and protein expressions of extracellular signal-regulated (ERK), p38 MAP kinase (p38), and c-Jun N-terminal kinase (JNK) were increased; the mRNAs and proteins of NF-κB and TNF-α were activated; and the mRNAs and proteins of necroptosis related genes were changed (the mRNA and protein expression of RIPK1, RIPK3, MLKL, and FADD were increased and caspase-8 was decreased) with concentration dependency. Taken together, we concluded that CPF exposure activated the MAPK/NF-κB/TNF-α pathway, promoted inflammatory injure and evoked necroptosis in common carp gills. In addition, CPF-induced inflammation and necroptosis was concentration dependency. The toxic effects of CPF on gills provided data for both aquaculture and toxicological studies.

Dietary Phytogenics and Galactomannan Oligosaccharides in Low Fish Meal and Fish Oil-Based Diets for European Sea Bass (Dicentrarchus labrax) Juveniles: Effects on Gill Structure and Health and Implications on Oxidative Stress Status.

An effective replacement for fish meal (FM) and fish oil (FO) based on plant-based raw materials in the feed of marine fish species is necessary for the sustainability of the aquaculture sector. However, the use of plant-based raw materials to replace FM and FO has been associated with several negative health effects, some of which are related to oxidative stress processes that can induce functional and morphological alterations in mucosal tissues. This study aimed to evaluate the effects of dietary oligosaccharides of plant origin (5,000 ppm; galactomannan oligosaccharides, GMOS) and a phytogenic feed additive (200 ppm; garlic oil and labiatae plant extract mixture, PHYTO) on the oxidative stress status and mucosal health of the gills of juvenile European sea bass (Dicentrarchus labrax). The experimental diets, low FM and FO diets (10%FM/6%FO) were supplemented with GMOS from plant origin and PHYTO for 63 days. GMOS and PHYTO did not significantly affect feed utilization, fish growth, and survival. GMOS and PHYTO downregulated the expression of ß-act, sod, gpx, cat, and gr in the gills of the fish compared with that in fish fed the control diet. The expression of hsp70 and ocln was upregulated and downregulated, respectively, in the GMOS group compared with that in the control group, whereas the expression of zo-1 was downregulated in the PHYTO group compared with that in the GMOS group. The morphological, histopathological, immunohistochemical, and biochemical parameters of the fish gills were mostly unaffected by GMOS and PHYTO. However, the PHYTO group had lower incidence of lamellar fusion than did the control group after 63 days. Although the tissular distribution of goblet cells was unaffected by GMOS and PHYTO, goblet cell size showed a decreasing trend (-11%) in the GMOS group. GMOS and PHYTO significantly reduced the concentration of PCNA+ in the epithelium of the gills. The above findings indicated that GMOS and PHYTO in low FM/FO-based diets protected the gill epithelia of D. labrax from oxidative stress by modulating the expression of oxidative enzyme-related genes and reducing the density of PCNA+ cells in the gills of the fish.

Copper impair autophagy on zebrafish (Danio rerio) gill epithelium.

Copper (Cu) is an essential element for organism's metabolism, being controversially listed as a priority pollutant. Importantly, the toxicity of Cu has been linked to several cell death pathways. Thus, this study aimed to assess if macroautophagic pathways are triggered by Cu in zebrafish gill, the main target of waterborne pollutants. The electron microscopy findings indicated that Cu induced profound impacts on zebrafish gill structure and functions, being this tissue a biomarker sensitive enough to indicate early toxic effects. The findings also support a clear impairment of autophagy, througth the absence of phagossomes and the significant down-regulation mRNA transcript levels of microtubule-associated protein light chain 3 (LC3). The reduction of LC3 levels was often associated to an increase of apoptotic activation, indicating that the inhibition of macroautophagy triggers apoptosis in zebrafish gills. This study highlighted that the autophagic down-regulation might be affected through the activation of other cell death signaling pathway.

A practical guide to unbiased quantitative morphological analyses of the gills of rainbow trout (Oncorhynchus mykiss) in ecotoxicological studies.

Rainbow trout (Oncorhynchus mykiss) are frequently used as experimental animals in ecotoxicological studies, in which they are experimentally exposed to defined concentrations of test substances, such as heavy metals, pesticides, or pharmaceuticals. Following exposure to a broad variety of aquatic pollutants, early morphologically detectable toxic effects often manifest in alterations of the gills. Suitable methods for an accurate and unbiased quantitative characterization of the type and the extent of morphological gill alterations are therefore essential prerequisites for recognition, objective evaluation and comparison of the severity of gill lesions. The aim of the present guidelines is to provide practicable, standardized and detailed protocols for the application of unbiased quantitative stereological analyses of relevant morphological parameters of the gills of rainbow trout. These gill parameters inter alia include the total volume of the primary and secondary gill lamellae, the surface area of the secondary gill lamellae epithelium (i.e., the respiratory surface) and the thickness of the diffusion barrier. The featured protocols are adapted to fish of frequently used body size classes (300-2000 g). They include well-established, conventional sampling methods, probes and test systems for unbiased quantitative stereological analyses of light- and electron microscopic 2-D gill sections, as well as the application of modern 3-D light sheet fluorescence microscopy (LSFM) of optically cleared gill samples as an innovative, fast and efficient quantitative morphological analysis approach. The methods shown here provide a basis for standardized and representative state-of-the-art quantitative morphological analyses of trout gills, ensuring the unbiasedness and reproducibility, as well as the intra- and inter-study comparability of analyses results. Their broad implementation will therefore significantly contribute to the reliable identification of no observed effect concentration (NOEC) limits in ecotoxicological studies and, moreover, to limit the number of experimental animals by reduction of unnecessary repetition of experiments.

Fish telocytes and their relation to rodlet cells in ruby-red-fin shark (rainbow shark) Epalzeorhynchos frenatum (Teleostei: Cyprinidae).

Telocytes comprise the major constituents of the supportive interstitial framework within the various organs. They form a 3D network between different types of stromal and non-stromal cells, which makes them distinctively vital. We have previously explored the origin of the peculiar rodlet cells, especially on their differential stages in aquatic species. The current study aimed at highlighting the relation of telocytes with different rodlet stages. Samples of fish, olfactory organs, and gills were processed for semi thin sections, transmission electron microscopy, and immunohistochemistry. It was evident in the study that telocytes formed a 3D interstitial network, entrapping stem cells and differentiating rodlet cells, to establish direct contact with stem cells. Differentiated stem cells and rodlet progenitor cells, practically in the granular and transitional stages, also formed ultrastructure junctional modifications, by which nanostructures are formed to establish cell contact with telocytes. Telocytes in turn also connected with macrophage progenitor cells. Telocytes (TCs) expressed CD34, CD117, VEGF, and MMP-9. In conclusion, telocytes established direct contact with the stem and rodlet cells in various differential stages. Telocytes may vitally influence stem/progenitor cell differentiation, regulate rodlet cell function, and express MPP-9 that may regulate immune cells functions especially, including movement and migration ability.

Ultrastructure of intracytoplasmic Rickettsia-like infection of the gills of the teleost Archosargus probatocephalus (Sparidae) in northeastern Brazil.

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.

Modifications in the gills of hill stream Moth catfish, Hara hara (Erethistidae, Siluriformes): A light and scanning electron microscope investigation.

Present study reports significant modifications in surface ultrastructure, histological organization, and histochemical localization of glycoproteins (GPs) in the gills of a hill stream catfish, Hara hara. Punctate microridges on free surface of epithelial cells covering gill arches, gill rakers, gill filaments and secondary lamellae are considered to provide adaptive plasticity to gills in relation to the environment inhabited by fish. Short and stout gill rakers are considered to prevent food particles to pass in opercular chamber along with respiratory current that could damage delicate gill filaments. Mucous goblet cells show presence of different classes of glycoproteins. GPs with oxidizable vicinal diols are considered to control acidity of acidic GPs. GPs with carboxyl groups have been implicated with defensive mechanism against microorganisms. GPs with O-sulphate esters are associated to trap and to lubricate food particles for easy swallowing. Taste buds on gill arches and gill rakers function to select palatable food particles. Occurrence of taste buds on the gill filaments is regarded significant adaptation to analyse the chemical nature of water. This study could play a significant role to understand adjustment of gills in the hill stream fish.

Morphological studies on the gills of the European hake (Merluccius merluccius, Linnaeus, 1758).

The current work gives concern to study the morphology of the Merluccius merluccius gills by using gross morphology, scanning electron microscopy (SEM), and light microscopy. The findings of the present study revealed that the gill system consisted of four pairs of gill arches which carry the gill filaments on the convex border and gill rakers on the concave border of them. SEM results revealed that the rakers and the spines distribution on the first gill arch differed from that of the other three gill arches on the lateral and medial surfaces. On the surface the gill filaments, there were longitudinal ridges that carried pores of chloride cells and mucous cells. The histological examination revealed that, the gill arch composed of hyaline cartilage that presented in the form of cups. Each cup consisted of central cartilagenous core and peripheral cartilagenous matrix. The gill filaments composed of cartilaginous bar of peripheral cartilaginous matrix and central cartilaginous core extended from the gill arches and covered by an epithelial layers with a few mucous cells permeate it, and chloride cells were straggly in the interlamellar epithelium. Each gill filament carried several leaves like secondary lamellae on both sides of it. The epithelium, which lined the secondary lamellae, composed of epithelial pavement cells, some mucous cells, and pillar cells.

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.

Changes in distribution, morphology and ultrastructure of chloride cell in Atlantic salmon during an AGD infection.

Amoebic gill disease (AGD) is emerging as one of the most significant health challenges affecting farmed Atlantic salmon in the marine environment. It is caused by the amphizoic amoeba Neoparamoeba perurans, with infestation of gills causing severe hyperplastic lesions, compromising overall gill integrity and function. This study used histology, transmission electron microscopy (TEM), immunohistochemistry and transcript expression to relate AGD-associated pathological changes to changes in the morphology and distribution of chloride cells (CCs) in the gills of Atlantic salmon (Salmo salar L.) showing the progression of an AGD infection. A marked reduction in numbers of immunolabelled CCs was detected, and a changing pattern in distribution and morphology was closely linked with the level of basal epithelial hyperplasia in the gill. In addition, acute degenerative ultrastructural changes to CCs at the lesion site were observed with TEM. These findings were supported by the early-onset downregulation of Na+ /K+ -ATPase transcript expression. This study provides supportive evidence that histological AGD lesion assessment was a good qualitative tool for AGD scoring and corresponded well with qPCR genomic Paramoeba perurans quantification. Ultrastructural changes induced in salmon CCs as a result of AGD are reported here for the first time.

The histopathological, cytopathological and ultrastructural effects of carbaryl on gills of Oreochromis niloticus (Linnaeus, 1758).

Ultrastructural and histopathological reponses in the organs of living organisms are important and useful tools to determine the health condition and the effects of pollutants, such as pesticides, on the organisms. The aim of this study is to determine possible histopathological, cytopathological and ultrastructural alterations in gills of Oreochromis niloticus individuals exposed to 850 µg/L carbaryl standart at 7th, 14th and 21st days with light and electron microscopes. The fish were exposed to carbaryl for 21 days and the histopatological, ultrastructural and cytopathological alterations occuring in the gill tissues of organisms were determined by light, Scanning and Transmission Electron Microscopes (SEM and TEM). At the end of the study, it was observed that carbaryl caused both histopathological and cytopathological changes in the gills of O. niloticus. It has been determined that the most of the pathological changes in the exposed organisms are the metabolic defence reactions.

Functionalized silver nanoparticles depress aerobic metabolism in the absence of overt toxicity in brackish water killifish, Fundulus heteroclitus.

Engineered nanomaterials (ENMs) tend to precipitate in saline waters so the majority of aquatic toxicity studies have focused on freshwaters, where bioavailability is presumed to be higher. Recent studies have illustrated that some ENM formulations are bioavailable and bioactive in salt water and that their effects are more pronounced at the physiological than biochemical level. These findings raise concerns regarding the effects of ENMs on marine organisms. Therefore, our goal was to characterize the effects of polyvinylpyrolidone-functionalized silver ENMs (nAg) on aerobic performance in the killifish (Fundulus heteroclitus), a common euryhaline teleost. Fish were exposed to 80 µg L-1 of 5 nm nAg for 48 h in brackish water (12 ppt) and routine (MO2min) and maximum (MO2max) rates of oxygen consumption were quantified. Silver dissolution was minimal and nAg remained well dispersed in brackish water, with a hydrodynamic diameter of 21.0 nm, compared to 19.3 in freshwater. Both MO2min and MO2max were significantly lower (by 53 and 30%, respectively) in killifish exposed to nAg and a reduction in MO2 variability suggested spontaneous activity was suppressed. Neither gill Na+/K+-ATPase activity, nor various other biochemical markers were affected by nAg exposure. The results illustrate that a common ENM formulation is bioactive in salt water and, as in previous studies on functionalized copper ENMs, that effects are more pronounced at the whole animal than the biochemical level.

Histopathological, biochemical and molecular studies on the toxic effect of used engine oil on the health status of Oreochromis niloticus.

The accidental spilling of petroleum oils into natural water resources expose fishes in the effluent area to serious problems.. Oreochromis niloticus were used in the current study as a model to investigate the toxicity of used engine oil and to evaluate the protective role of vitamin C against this toxicity. The oil concentration used in this study was previously determined to be 0.25 ml/l by 96 h-LC50. After 21 days of engine oil exposure, haematological and biochemical analyses revealed significant reduction in RBCs counts, haemoglobin concentrations and total proteins. However, ALT, AST and glucose levels were significantly increased by the end of the experiment indicating the damaging effects of the oil on fish tissues. Oxidative stress biomarkers were also measured; liver CAT activity was significantly decreased in the oil exposed group compared to control group, while MDA levels were significantly elevated. Histopathological examination showed the presence of several alterations in hepatic and branchial tissues in exposed group compared to the control group. Significant elevations in CYP1 A1 mRNA expression levels in hepatic tissue were also detected in the group exposed to used engine oil compared to the control group. However, supplementation of fishexposed to used engine oil with vitamin Csignificantly enhance the biochemical, oxidative and histological parameters.

Lead toxicity in seawater teleosts: A morphofunctional and ultrastructural study on the gills of the Ornate wrasse (Thalassoma pavo L.).

Lead is one of the most alarming toxic pollutants for the environment due to its acute toxicity and high bioaccumulation tendency. Despite legislative efforts, world lead production has more than doubled since the early 1970s to 2016. Due to extensive exploitation and human activity, the coastal and estuarine regions belong to marine environments that are mostly and more rapidly deteriorated by pollutants including lead. A limited number of studies examined the effects of lead in fishes, compared to other aquatic models and even fewer studies have been dedicated to seawater fishes especially regarding Pb adsorption and accumulation in specific organs. Fish gills, key compartments involved in several crucial functions such as gas exchange, osmoregulation, and excretion, are also the organs in which Pb is mainly accumulated. Herein, we investigated the morphofunctional and ultrastructural modifications in the gills of a marine teleost (Thalassoma pavo) after acute exposure (48, 96, 192 h) to three lead concentrations. We showed that pathological alterations can be detected in all the examined samples. The most common modifications observed were: the curling of the lamellae and the dilation of their apical tips, the lamellar edema, the proliferation and the hypertrophy of CCs, the progressive epithelial disorganization with detachment of the epithelium from connective tissue. This study also demonstrates that there is a weak influence on the expression pattern of Na+/K+-ATPase and AQP3 biomarker enzymes while high metallothioneins expression has been observed. The described alterations may adversely affect gas exchange and ionic balance, with a long chain of cascading effects. This is the first evidence of the effects exerted by lead on gills of seawater fishes that highlights the harmful properties of this metal, even at low concentration.

Ultrastructure features of the surface of the gills and the lower pharyngeal jaw of the Tilapia Zilli, redbelly tilapia (Coptodon Zillii, Gervais, 1848).

The present investigation was designed to describe the surface ultrastructure of the gill system of tilapia Zilli. The gill system is formed from four gill arches and each gill arch carries a row of gill filaments on its convex border and two rows of the gill rakers on its concave border. The quadrilateral interbranchial septum has elevated part at the level of the third gill arch. By SEM observations, the gill arch was divided into three regions: rostral, middle and caudal region. The caudal region contained two characteristic structures: oval leaf-like structure and rounded-shaped structure. Each oval leaf-like structure carried two lateral rows of the triangular pointed spines separated by a median groove. All surfaces of gill arches, rakers and filaments were covered with a mosaic of the polygonal pavement cells, in addition to the opening of chloride cells and mucous cells. The gill arch and gill raker had only one appearance of taste buds named type I. Meanwhile, the filaments contained two types of different appearance of the taste buds named: type I and type II. Type I was the main common and similar to that present in gill arch and raker and characterized by its blunt end, while type II had hair-like structures that projected from the volcano-shaped depression. The gill rakers were formed from central axis surrounded by two lateral lobulated regions which carry pointed spines, taste buds and the opening of chloride cells. The surface of triangular lower pharyngeal jaw carries numerous teeth-like papillae which originated from the socket-like depression.

Ontogenetic changes in cutaneous and branchial ionocytes and morphology in yellowfin tuna (Thunnus albacares) larvae.

The development of osmoregulatory and gas exchange organs was studied in larval yellowfin tuna (Thunnus albacares) from 2 to 25 days post-hatching (2.9-24.5 mm standard length, SL). Cutaneous and branchial ionocytes were identified using Na+/K+-ATPase immunostaining and scanning electron microscopy. Cutaneous ionocyte abundance significantly increased with SL, but a reduction in ionocyte size and density resulted in a significant decrease in relative ionocyte area. Cutaneous ionocytes in preflexion larvae had a wide apical opening with extended microvilli; however, microvilli retracted into an apical pit from flexion onward. Lamellae in the gill and pseudobranch were first detected ~ 3.3 mm SL. Ionocytes were always present on the gill arch, first appeared in the filaments and lamellae of the pseudobranch at 3.4 mm SL, and later in gill filaments at 4.2 mm SL, but were never observed in the gill lamellae. Unlike the cutaneous ionocytes, gill and pseudobranch ionocytes had a wide apical opening with extended microvilli throughout larval development. The interlamellar fusion, a specialized gill structure binding the lamellae of ram-ventilating fish, began forming by ~ 24.5 mm SL and contained ionocytes, a localization never before reported. Ionocytes were retained on the lamellar fusions and also found on the filament fusions of larger sub-adult yellowfin tuna; however, sub-adult gill ionocytes had apical pits. These results indicate a shift in gas exchange and NaCl secretion from the skin to branchial organs around the flexion stage, and reveal novel aspects of ionocyte localization and morphology in ram-ventilating fishes.

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.