Gill structure and respiratory ability of Euchiloglanis kishinouyei (Osteichthyes: Siluriformes: Sisoridae).

Glyptosternoid fishes are distributed in the torrent environment of alpine canyons, where they often leave the water to climb rocky cliffs. As one of the most primitive species of glyptosternoid fishes, Euchiloglanis kishinouyei was examined in the current study to analyse its gill microstructure and respiratory ability. We first found that the oxygen consumption rate was relatively high and negatively correlated with body mass and that the average oxygen consumption at night was higher than during the day. The asphyxiation point of E. kishinouyei (5.05 ± 0.22 g) was c. 1.93 mg/L. Subsequently, the surface morphology, gross gill tissue structure, and ultra-microstructure of gill lamellae were investigated using optical microscopy and SEM. The gills showed an overall trend of regression, with five pairs of gill arches in each gill cavity. The adjacent gill filaments had large gaps, and the gill lamellae were thick. The gill filaments were closely arranged on the gill arches, their folded respiratory surface was highly vascularized with no tiny crest, and there were obvious tiny crests, grooves, pits, and pores on the nonrespiratory surface. The gill lamellae were closely embedded on both sides of gill filaments, which were composed of flat epithelial cells, basement membrane, pillar cells, and mucous cells. The gill total respiratory area correlated positively with body mass and length, whereas the gill relative respiratory area correlated negatively with body mass. We comprehensively analysed the gill microstructure and respiratory capacity of E. kishinouyei to provide fundamental data for the adaptive evolution of the gill structures of bimodally respiring fishes and offer insights into further study on the accessory air-breathing function of skin.

Localization and Bioreactivity of Cysteine-Rich Secretions in the Marine Gastropod Nucella lapillus.

Marine biodiversity has been yielding promising novel bioproducts from venomous animals. Despite the auspices of conotoxins, which originated the paradigmatic painkiller Prialt, the biotechnological potential of gastropod venoms remains to be explored. Marine bioprospecting is expanding towards temperate species like the dogwhelk Nucella lapillus, which is suspected to secrete immobilizing agents through its salivary glands with a relaxing effect on the musculature of its preferential prey, Mytilus sp. This work focused on detecting, localizing, and testing the bioreactivity of cysteine-rich proteins and peptides, whose presence is a signature of animal venoms and poisons. The highest content of thiols was found in crude protein extracts from the digestive gland, which is associated with digestion, followed by the peribuccal mass, where the salivary glands are located. Conversely, the foot and siphon (which the gastropod uses for feeding) are not the main organs involved in toxin secretion. Ex vivo bioassays with Mytilus gill tissue disclosed the differential bioreactivity of crude protein extracts. Secretions from the digestive gland and peribuccal mass caused the most significant molecular damage, with evidence for the induction of apoptosis. These early findings indicate that salivary glands are a promising target for the extraction and characterization of bioactive cysteine-rich proteinaceous toxins from the species.

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.

Biomarcadores enzimáticos e histológicos em brânquias de Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) indicativos de impactos ambientais em uma região portuária do nordeste do Brasil / [Enzymatic and histological biomarkers in Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) gills indicative of environmental impacts in a port region of northeastern Brazil]

Objetivou-se neste estudo analisar biomarcadores histológicos e bioquímicos em brânquias de U. cordatus indicativos de impactos na Baía de São Marcos. Caranguejos foram coletados em quatro áreas na Baía de São Marcos: A1= Ilha dos Caranguejos (com baixo impacto); A2= Coqueiro, A3= Porto Grande, A4= Cajueiro (áreas potencialmente impactadas). Mediram-se os dados biométricos de cada exemplar de caranguejo. Amostras de brânquias foram submetidas à técnica histológica padrão e homogeneizadas em tampão fosfato, e o sobrenadante foi utilizado para análise das enzimas glutationa-S-transferase (GST) e catalase (CAT). A biometria indicou que os caranguejos de A1 são significativamente (P<0,05) maiores e mais pesados do que os caranguejos das áreas A2, A3 e A4. As alterações branquiais (rompimento das células pilastras, deformação do canal marginal, deslocamento da cutícula e necrose) foram significativamente (P˂0,05) mais frequentes em caranguejos de A2, A3 e A4 do que nos caranguejos de A1. As atividades enzimáticas da GST e CAT nos caranguejos apresentaram diferença significativa (P<0,05) entre as áreas de coletas, com padrão similar ao observado para as alterações branquiais. Os biomarcadores analisados mostraram que os caranguejos estão sob diferentes níveis de impactos (A4>A3>A2>A1) ao longo da Baía de São Marcos.(AU)

The objective of this study was to analyze histological and biochemical biomarkers in U. cordatus gills indicative of impacts in São Marcos Bay. Crabs were collected from four areas in São Marcos Bay: A1=Ilha dos Caranguejos (with low impact); A2=Coqueiro, A3=Porto Grande, A4=Cajueiro (potentially impacted areas). The biometric data of each specimen was measured. Gill samples were submitted to standard histological technique and homogenized in phosphate buffer, and the supernatant was used for analysis of glutathione S-transferase (GST) and catalase (CAT) enzyme activity. Biometric data indicated that crabs found in A1 are significantly (P<0.05) larger and heavier than crabs found in A2, A3 and A4 areas. Gill alterations (rupture of pilaster cells, Dilation of the marginal channel, Cuticle Rupture and necrosis) were significantly (P˂0.05) more frequent in the crabs in A2, A3 and A4 than crabs in A1. The enzymatic activities of GST and CAT showed significant difference (P<0.05) between the sampling areas, similar to that observed for gill alterations. The biomarkers analyzed showed that the crabs are under different impact levels (A4> A3> A2> A1) along the São Marcos Bay.(AU)

Biomarcadores histológicos em Sciades herzbergii (Pisces, Ariidae) para avaliação de impactos em ambientes estuarinos da Baía de São Marcos, Maranhão / [Biomarkers histological in Sciades herzbergii (Pisces, Ariidae) for impact assessment in estuarine environments of São Marcos Bay, Maranhão]

Neste estudo, objetivou-se avaliar o impacto ambiental em áreas estuarinas da Baía de São Marcos, Maranhão, utilizando-se alterações histológicas nas brânquias e no fígado de Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período chuvoso (março de 2018) e de estiagem (julho de 2018), em que foram capturados 40 exemplares de S. herzbergii em duas regiões distintas da Baía de São Marcos: A1) Ilha dos Caranguejos (área referência); A2) Terminal de Pesca Industrial de Porto Grande (área potencialmente contaminada). As estações foram georreferenciadas por GPS e, em cada local de captura dos peixes, registraram-se as seguintes variáveis abióticas: temperatura, salinidade, pH e oxigênio dissolvido. Em campo, os organismos foram pesados e medidos, as brânquias e os fígados foram retirados e fixados. Posteriormente, foram mantidos em álcool a 70% até o procedimento histológico padrão. Resultados indicam que as duas áreas são semelhantes em termos de características físico-químicas da água e do sedimento. Todavia, a análise semiquantitativa das alterações histológicas observadas revelou que as brânquias foram o órgão mais afetado nos peixes de A2, indicando que os peixes desse local apresentam danos teciduais maiores resultantes de estresse acentuado por impactos antrópicos.(AU)

This study aimed to evaluate the environmental impact in estuarine areas of São Marcos Bay, Maranhão, using histological changes in the gills and liver of Sciades herzbergii as biomarkers of aquatic contamination. Samples were collected during the rainy (March 2018) and dry (July 2018) seasons, in which 40 specimens of S. herzbergii were captured in two distinct regions of São Marcos Bay: A1) Crab Island (reference area); A2) Porto Grande Industrial Fishing Terminal (potentially contaminated area). The stations were georeferenced by GPS and at each fish capture site the following abiotic variables were recorded: temperature, salinity, pH and dissolved oxygen. In the field, the organisms were weighed and measured and the gills and livers removed and fixed. Afterwards, they were kept in 70% alcohol until the standard histological procedure. Results indicate that the two areas are similar in terms of physicochemical characteristics of water and sediment. However, the semi-quantitative analysis of the observed histological alterations revealed that the gills were the most affected organ in A2 fish, indicating that the fish in this site have greater tissue damage resulting from stress accentuated by anthropic impacts.(AU)

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.

Histopathologic alterations of gill tissue in Siluriformes and Characiformes from the Middle Tocantins River in the Brazilian Amazon / Alterações histopatológicas do tecido branquial em Siluriformes e Characiformes do médio rio Tocantins, na Amazônia brasileira

As águas do rio Tocantins têm sofrido descargas de dejetos tóxicos que ameaçam a biota aquática. Para tanto, a investigação das brânquias de peixes atua como biomonitoramento, visto que esses órgãos respondem por meio de adaptações a xenobióticos. Este trabalho teve como objetivo avaliar a ocorrência de alterações morfológicas das espécies de Psectrogaster amazonica, Pimelodina flavipinnis e Pimelodus blochii. As coletas ocorreram nas estações chuvosa e seca. As brânquias de todos os espécimes coletados foram processadas de acordo com metodologia específica para protocolo de hematoxilina e eosina. As principais alterações histológicas observadas no período chuvoso foram: hiperplasia, destacamento do epitélio filamentar, fusão parcial e total das lamelas secundárias, congestão, aneurisma e encurtamento das lamelas respiratórias. Em relação ao período seco, as principais lesões observadas foram: ruptura do epitélio, aneurisma, hiperplasia e necrose. Considerando a diversidade ictiológica do rio Tocantins, o presente estudo propiciará o conhecimento da condição das brânquias, órgão imprescindível para a saúde do peixe, e consequentemente a compreensão das implicações sobre a qualidade das águas do rio Tocantins.(AU)

Morphological and histological characterization of an ectopically mineralized structure in a gilthead sea bream Sparus aurata with opercular deformation.

In this study, we describe an abnormal ectopically mineralized structure (EMS) that was found inside the skull of a juvenile Sparus aurata that also showed a bilateral opercular deformation. The overall phenotype and tissue composition were studied using micro-CT scanning and histological analyses. The ectopic structure occupies a large volume of the brain cavity, partially extruding into the gill cavity. It shows a dense mineralization and an extracellular matrix-rich phenotype, with variation in both the morphology and size of the cell lacunae, combined with an irregular fibre organization inside the matrix. This study is the first to report such an EMS in a juvenile teleost fish, where the tissue does not resemble any other connective tissue type described in bony fish so far. The tissue phenotype seems to rule out that the EMS corresponds to a tumorous cartilage. Yet, it is rather reminiscent of a highly mineralized structure found in cartilaginous fish, where it is suggested to be associated with damage repair.

Gill remodelling during terrestrial acclimation in the amphibious fish Polypterus senegalus.

Fishes are effectively weightless in water due to the buoyant support of the environment, but amphibious fishes must cope with increased effective weight when on land. Delicate structures such as gills are especially vulnerable to collapse and loss of surface area out of water. We tested the 'structural support' hypothesis that amphibious Polypterus senegalus solve this problem using phenotypically plastic changes that provide mechanical support and increase stiffness at the level of the gill lamellae, the filaments, and the whole arches. After 7 d in terrestrial conditions, enlargement of an inter-lamellar cell mass filled the water channels between gill lamellae, possibly to provide structural support and/or reduce evaporative water loss. Similar gill remodelling has been described in several other actinopterygian fishes, suggesting this may be an ancestral trait. There was no change in the mechanical properties or collagen composition of filaments or arches after 7 days out of water, but 8 months of terrestrial acclimation caused a reduction in gill arch length and mineralized bone volume. Thus, rather than increasing the size and stiffness of the gill skeleton, P. senegalus may instead reduce investment in supportive gill tissue while on land. These results are strikingly similar to the evolutionary trend of gill loss that occurred during the tetrapod invasion of land, raising the possibility that genetic assimilation of gill plasticity was an underlying mechanism.

Mechanosensory system of the lateral line in the subantarctic Patagonian blenny Eleginops maclovinus.

This study describes the cephalic and trunk lateral line systems in Patagonian blenny Eleginops maclovinus juveniles, providing morphological details for pores, canals and neuromasts. Eleginops maclovinus juveniles possess a complete laterodorsal lateral line that extends from the upper apex of the gill opening along the trunk as far as the caudal fin. The lateral line was ramified through pores and canals. The following pores were recorded: four supraorbital pores, with two along the eye border and two on the snout; seven infraorbital pores, with three on the lacrimal bone and four being infraorbital; five postorbital pores, with three along the preopercular border (upper preoperculum branch) and two on the bone curvature (inferior preoperculum branch); and four mandibular pores aligned along the jaw. Furthermore, five narrow-simple and interconnected canals were found (i.e. preopercular, mandibular, supraorbital and infraorbital canals). Histologically, the dorsal lateral line presented thin neuromasts (350 µm) with short hair cells. By contrast, the cranial region presented long, thick neuromasts. Infraorbital and mandibular neuromasts had a major axis length of 260 µm and respective average diameters of 200 and 185 µm. Sensory system variations would be due to a greater concentration of neuromasts in the cranial region, allowing for a greater perception of changes in water pressure. Scarce morphological information is available for the lateral sensory system in Eleginopsidae, particularly compared to Channichthyidae, Bovichthydae, Artedidraconidae and Bathydraconidae. Therefore, the presented results form a fundamental foundation of knowledge for the lateral-line system in juvenile E. maclovinus and provide a basis for future related research in this taxon as well as within the Notothenioidei suborder.

Gill Morphology and Na+/K+-ATPase Activity of Gobionellus oceanicus (Teleostei: Gobiidae) in an Estuarine System.

Gobionellus oceanicus is a species widely distributed on the Atlantic coast and may be susceptible to anthropic effects. This study evaluated the morphology and the Na+/K+-ATPase activity of G. oceanicus gills considering the concentration of metals in the fish and Subaé River estuary. Although the metal concentrations detected in the water and sediment did not exceed certain limits, CONAMA (Brazilian Environment Council), TEL (Threshold Effect Level), and PEL (Probable Effect Level), the metals levels in gills plus muscle and skin of G. oceanicus were above the permitted setting of the Ministry of Health, Brazil. The pavement epithelial cells (PVC) of the gill filament was observed that there were long microridges either in the apical surface or in the lamella; especially, microridges degeneration was shown in some PVC from filament epithelium. The number of ionocyte and the volume density were (0.02 ± 0.001)/µm and (0.38 ± 0.27) %, respectively. The activity of Na+/K+-ATPase was 1.13 ± 0.76 µM Pi mg protein-1 h-1. We describe the volume density and number of ionocytes and Na+/K+-ATPase enzymatic activity in G. oceanicus for the first time, which is useful for basic and comparative future studies to support aquatic biomonitoring.

Interactive effects of salinity and temperature acclimation on gill morphology and gene expression in threespine stickleback.

Colonization of freshwater habitats from marine environments exposes organisms to novel combinations of temperature and salinity, but little is known about physiological responses to the interactive effects of these stressors. Here, we examined the effects of temperature (14 versus 4 °C) and salinity (11 versus 0.3 ppt) on gill gene expression in marine, anadromous, and freshwater populations of threespine stickleback (Gasterosteus aculeatus). Expression of the epithelial calcium channel was not affected by temperature or salinity, but had significantly higher expression in the freshwater ecotype. The combination of low temperature and low salinity had non-additive effects on the expression of the Na+/H+ exchanger. Fish exposed to the combination of low temperature and low salinity had expression levels similar to fish exposed to either factor in isolation. Expression of Na+,K+-ATPase α-subunit was greater in fish exposed to low temperature and low salinity than in fish exposed to the factors separately, and this effect was the most pronounced in the marine ecotype. We also examined the interactive effects of salinity and temperature on gill morphology in the marine ecotype, and observed non-additive effects. Low temperature increased the size of the interlamellar cell mass in fish held at 11 ppt, but not at 0.3 ppt, and the effect of low salinity was in the opposite direction in fish at high and low temperatures. These data demonstrate interactive effects of temperature and salinity and highlight that overwintering in cold freshwater was likely a physiological challenge for marine stickleback as they colonized freshwater following the last glaciation.

Biomarcadores histológicos em brânquias de Sciades herzbergii (Siluriformes, Ariidae) capturados no Complexo Estuarino de São Marcos, Maranhão / Histological biomarkers in gills of Sciades herzbergii (Siluriformes, Ariidae) caught in the Estuarine Complex of São Marcos, Maranhão

Neste estudo, objetivou-se avaliar a qualidade ambiental de duas áreas do complexo estuarino de São Marcos, utilizando-se as alterações branquiais em Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período de estiagem (setembro/dezembro de 2015) e no período chuvoso (março/junho de 2016). Foram capturados 40 exemplares de S. herzbergii em dois locais distintos do complexo estuarino: A1) Porto do Itaqui Bacanga (área potencialmente impactada); A2) Igarapé do Puca, Alcântara (área controle). Foram registradas as variáveis abióticas: salinidade, temperatura, pH e oxigênio dissolvido. No laboratório, as brânquias de cada espécime foram fixadas em formol a 10% e mantidas em álcool a 70% até a execução da técnica histológica usual. As alterações branquiais identificadas nos peixes da área potencialmente contaminada foram: deslocamento do epitélio (DE), congestão (CO), aneurisma (AN), fusão lamelar (FL), hiperplasia (HI), dilatação capilar (DC), necrose (NE) e proliferação de células epiteliais (PCE). As alterações na região controle foram: deslocamento do epitélio (DE), fusão lamelar (FL), necrose (NE) e proliferação de células epiteliais (PCE). As alterações branquiais foram mais frequentes na região portuária (84%) do que na região controle (16%). Esses resultados indicam que os peixes da área potencialmente contaminada estão apresentando respostas biológicas ao estresse local.(AU)

This study aimed to compare an environmental quality of two areas of the estuarine complex of São Marcos using gills of changes in Sciades herzbergii as biomarkers of aquatic contamination. Samples were made during the dry season (September/ December 2015) and rainy season (March/ June 2016). They were captured 40 specimens of S. herzbergii at two different points of the estuarine complex: A1) Itaqui port (potentially contaminated area); A2) Igarapé do Puca, Alcântara (control area). Abiotic variables were recorded: salinity, temperature, pH and dissolved oxygen. In the laboratory, the gills of each species were fixed in 10% formaldehyde and kept in 70% alcohol until the usual histological technique was performed. The gill alterations identified in the fish of the potentially contaminated area were: epithelial displacement (DE), congestion (CO), aneurysm (AN), lamellar fusion (FL), hyperplasia (HI), capillary dilation (DC), necrosis and epithelial cell proliferation (PCE). The changes in the control region were: epithelial displacement (DE), lamellar fusion (FL), necrosis (NE) and epithelial cell proliferation (PCE). Gill alterations were more frequent in the port region (84%) than in the control region (16%). These results indicate that fish from the potentially contaminated area are presenting biological responses to local stress.(AU)

Histological characteristics of gills and dorsal skin in ambystoma leorae and ambystoma rivulare: morphological changes for living at high altitude / Características histológicas de la piel del dorso y las branquias en ambystoma leorae y ambystoma rivulare: cambios morfológicos para vivir a gran altitud

SUMMARY: Vertebrates exhibit structural changes in their cardiovascular and gas exchange systems in response to hypoxic conditions in high altitude environments. In highland neotenic mole salamanders, as other amphibians, the majority of gases exchange is carried out for skin and gills. But, in high altitude environments, the available oxygen is lower than it is in the air thus, the scarcity of oxygen limits the survival of organisms. Many studies on this subject have focused on understanding the hematological mechanisms that amphibians exhibit in response to hypoxia. However, little is known about possible morphological changes in respiratory structures that may permit increased gas exchange during respiration in high altitude amphibians like Ambystoma leorae and A. rivulare, two threatened Mexican salamander species. The aim of the present study was to describe and compare the histological characteristics of the gills and dorsal skin of A. leorae and A. rivulare from populations at low and high altitudes. We found that, in comparison to lowland organisms, highland ones exhibited more pronounced skin folds, greater numbers of secondary branches in the gills, thinner dorsal and gill epidermises, and greater quantity of melanin surrounding the gill blood vessels. These differences permit a greater capacity for gas exchange and also increase thermoregulatory capacity in high altitude environments.

RESUMEN: Los anfibios que viven en ambientes de altitud se enfrentan a factores abióticos que limitan la vida, tales como la disminución de la presión barométrica con la consecuente disminución de la presión parcial de oxígeno (O2). Conocer los mecanismos que optimizan la obtención del O2 en estos animales es de gran importancia para entender las diferencias en la sensibilidad a la hipoxia de las diferentes especies. Ambystoma rivulare y A. leorae son anfibios endémicos del Estado de México que viven en ambientes de alta altitud por lo que se cree presentan estrategias fenotípicas para asimilar eficazmente el O2 y poder subsistir en los ambientes de altitud. El objetivo de este trabajo fue analizar las características histológicas de branquias y piel cefálica (que son las principales estructuras que se encargan del intercambio gaseoso) provenientes de tres poblaciones con diferente altitud. Nuestros resultados muestran que los organismos que habitan a mayor altitud tienden a aumentar la superficie de intercambio gaseoso, como es el caso de pliegues epidérmicos y ramas branquiales secundarias. Las diferencias histológicas de branquias y piel cefálica tanto interespecíficas como intraespecíficas respecto a la altitud parecen apoyar la idea de que los organismos modifican sus estructuras para contrarrestar las limitantes de la vida en ambientes de altitud.

Protective role of phenylalanine on the ROS-induced oxidative damage, apoptosis and tight junction damage via Nrf2, TOR and NF-κB signalling molecules in the gill of fish.

This study explored the possible preventive effects of dietary phenylalanine (Phe) on antioxidant responses, apoptosis and tight junction protein transcription in the gills of young grass carp (Ctenopharyngodon idella). Fish were fed six different experimental diets containing graded levels of Phe (3.4-16.8 g kg-1) for 8 weeks. The results showed that Phe deficiency induced protein oxidation and lipid peroxidation by decreasing the glutathione content and the activities and mRNA levels of Cu/Zn superoxide dismutase (SOD1), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST) in fish gill (P < 0.05). These results may be ascribed to the downregulation of NF-E2-related factor 2 (Nrf2), target of rapamycin (TOR) and ribosomal protein S6 kinase 1 (S6K1), and the upregulation of Kelch-like-ECH-associated protein 1 a (Keap1a) expression in grass carp gills (P < 0.05). Additionally, Phe deficiency induced DNA fragmentation via the up-regulation of Caspase 3, Caspase 8 and Caspase 9 mRNA expression (P < 0.05). These results may be ascribed to the improvement in reactive oxygen species (ROS) levels in the fish gills (P < 0.05). Furthermore, the results indicated that Phe deficiency decreased Claudin b, Claudin 3, Occludin and ZO-1 transcription and increased Claudin 15 expression in the fish gills (P < 0.05). These effects were partly due to the downregulation of interleukin 10 (IL-10), transforming growth factor ß (TGF-ß) and inhibitor factor κBα (iκBα) and the upregulation of relative mRNA expression of interleukin 1ß (IL-1ß), interleukin 8 (IL-8), tumour necrosis factor-α (TNF-α) and nuclear transcription factor-κB p65 (NF-κB p65) (P < 0.05). Taken together, the results showed that Phe deficiency impaired the structural integrity of fish gills by regulating the expression of tight junction proteins, cytokines, antioxidant enzymes, NF-κB p65, iκBα, TOR, Nrf2, Keap1 and apoptosis-related genes in the fish gills.

Air breathing and aquatic gas exchange during hypoxia in armoured catfish.

Air breathing in fish is commonly believed to have arisen as an adaptation to aquatic hypoxia. The effectiveness of air breathing for tissue O2 supply depends on the ability to avoid O2 loss as oxygenated blood from the air-breathing organ passes through the gills. Here, we evaluated whether the armoured catfish (Hypostomus aff. pyreneusi)-a facultative air breather-can avoid branchial O2 loss while air breathing in aquatic hypoxia, and we measured various other respiratory and metabolic traits important for O2 supply and utilization. Fish were instrumented with opercular catheters to measure the O2 tension (PO2) of expired water, and air breathing and aquatic respiration were measured during progressive stepwise hypoxia in the water. Armoured catfish exhibited relatively low rates of O2 consumption and gill ventilation, and gill ventilation increased in hypoxia due primarily to increases in ventilatory stroke volume. Armoured catfish began air breathing at a water PO2 of 2.5 kPa, and both air-breathing frequency and hypoxia tolerance (as reflected by PO2 at loss of equilibrium, LOE) was greater in individuals with a larger body mass. Branchial O2 loss, as reflected by higher PO2 in expired than in inspired water, was observed in a minority (4/11) of individuals as water PO2 approached that at LOE. Armoured catfish also exhibited a gill morphology characterized by short filaments bearing short fused lamellae, large interlamellar cell masses, low surface area, and a thick epithelium that increased water-to-blood diffusion distance. Armoured catfish had a relatively low blood-O2 binding affinity when sampled in normoxia (P50 of 3.1 kPa at pH 7.4), but were able to rapidly increase binding affinity during progressive hypoxia exposure (to a P50 of 1.8 kPa). Armoured catfish also had low activities of several metabolic enzymes in white muscle, liver, and brain. Therefore, low rates of metabolism and gill ventilation, and a reduction in branchial gas-exchange capacity, may help minimize branchial O2 loss in armoured catfish while air breathing in aquatic hypoxia.

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.

Salinity effects on osmoregulation and gill morphology in juvenile Persian sturgeon (Acipenser persicus).

The effect of abrupt and 5-day gradual salinity transfers from freshwater (FW) to 11 ‰ Caspian Sea brackish water (BW) was investigated in juvenile Persian sturgeon Acipenser persicus with three different weight groups: 1-2 g (1.62 ± 0.27 g), 2-3 g (2.55 ± 0.41 g) and 3-5 g (4.28 ± 0.76 g). Mortality rates, blood osmotic pressure, gill morphology and branchial Na+, K+-ATPase (NKA) activity were measured 4 and 10 days after abrupt transfer and 9 and 15 days after the initial gradual transfer (i.e. 4 and 10 days after reaching Caspian Sea salinity). Fish under 3 g could not survive increased salinity, and the blood osmotic pressure of the remaining surviving fish increased and remained elevated. However, heavier fish were able to survive and successfully acclimate, even to rapid salinity change with osmotic pressure reduced to Caspian Sea osmolality levels. At the gill level, the developmental increase in chloride cell volume and a higher NKA content most probably allow juveniles weighing more than 2 g to sharply increase NKA activity if the fish are transferred to BW. The rapid chloride cell proliferation occurring with increased salinity should strengthen this acclimation response. Therefore, a drastic physiological change occurs when fish weigh more than 2 g that allows migration to higher salinities. The triggering signal on chloride cells must be further investigated in order to optimize this functional step.

Gill morphology in the red swamp freshwater crayfish procambarus clarkii (Crustacea: Decapoda: Cambarids) (Girard 1852) from the River Nile and its Branches in Egypt / Morfología de las branquias del cangrejo rojo de pantano de agua dulce procambarus clarkii (Crustacea: Decapoda: Cambarids) (Girard 1852) del Río Nilo y sus Ramas en Egipto

The aim of the present study was focused on and describes the gross morphological and scanning electron microscopical features of the gill of the red swamp freshwater crayfish. Our results noted that: all gills have the same general structure and appearance. The gill consists of axis with numerous finger-like filaments, having three morphological types; round, pointed and somewhat hooked shaped. There is a variation in the direction of filaments according to their position, in middle part were nearly perpendicular to gill axis while in the apex were nearly parallel to axis. There were characteristic system of gill spines on central axis, basal plate, setobranch and on the bilobed epipodal plate. There are four shape of spinated-like parts of setobranch seta, two pointed processes and two broad processes. The bilobed epipodal plate is devoid from any filaments and under SEM, its apical part has serrated free border and corrugated surface while the middle part has no serrated free border.

El objetivo del presente estudio fue describir las características morfológicas macroscópicas y mediante microscopio electrónico de barrido las branquias del cangrejo rojo de pantano de agua dulce. Nuestros resultados señalan que todas las branquias tienen la misma estructura y apariencia general. Las branquias se componen de ejes con numerosos filamentos similares a dedos, que tiene tres tipos morfológicos; redondo, punteado y con forma de gancho. Hay una variación en la dirección de los filamentos de acuerdo con su posición, en la parte media eran casi perpendicular al eje branquial, mientras que en el ápice fueron casi paralelas al eje. Hubo un sistema característico de espinas branquiales sobre el eje central, placa basal, espinas dorsales y sobre las placas epipodales bilobuladas. Se observaron cuatro formas de las ramas similares a espinas, dos procesos apuntados y dos procesos amplios. La placa epipodal bilobulada estaba desprovista de filamentos bajo microscopía electrónicas, su parte apical tiene una margen libre aserrado, con una superficie ondulada, mientras que la parte media no tiene margenes aserrados.

External morphological features of the gills of Ichthyophis bannanicus embryo (Amphibia, Gymnophiona) / Características morfológicas externas de las branquias del embrión de Ichthyophis bannanicus (Amphibia, Gymnophiona)

The gills of Ichthyophis bannanicus have yet to be investigated. This paper describes the external morphological features of the gills of mature I. bannanicus embryos exhibiting three pairs of gills on their neck region. Each gill is composed of an axis and filaments. In newly released embryos, the filaments and the axis form at approximately 90° relative to each other; eventually, this angle decreases and the color of the gill fades. The filaments on the axis are arranged alternately, and their spacing varies. The mid-pair of gills is significantly longer by nearly twofold than the front and rear pairs. Likewise, the lengths of the front and rear pairs of gills are not significantly different (P >0.05); for the same pair of gills, the lengths of the left and right parts are not significantly different (P >0.05). The number of filaments is greater in the mid-pair of gills than in the front and rear pairs (P <0.05); the number of filaments in the front pair is not significantly different from that of the rear pair (P >0.05); the number of filaments in the left part does not significantly differ from that of the right parts (P >0.05). Results showed that the gills of I. bannanicus embryo are more similar to those of other species in Ichthyophiidae than to those of species in other families.

Las branquias del Ichthyophis bannanicus aún no se han investigado. En este trabajo se describen las características morfológicas externas de las branquias de embriones maduros de I. bannanicus, que exhiben tres pares de branquias en la región del cuello. Cada branquia está compuesta de un eje y filamentos. En embriones recién liberados los filamentos y la forma del eje es de aproximadamente 90° respecto a la otra; finalmente, este ángulo disminuye y el color de las branquias se desvanece. Los filamentos en el eje están dispuestos en forma alternada y su separación varía. La media de par de branquias es significativamente más larga, por casi el doble que los pares anterior y posterior. Del mismo modo, las longitudes de la parte delantera y posterior de pares de branquias no son significativamente diferentes (p >0,05); para el mismo par de branquias, las longitudes de las partes izquierda y derecha no son significativamente diferentes (p >0,05). El número de filamentos es mayor en el par medio de branquias, que en la parte delantera y posterior de pares (p <0,05); el número de filamentos en el par frontal no es significativamente diferente de la del par trasero (p >0,05); el número de filamentos en la parte izquierda no varía significativamente del de las partes derechas (p >0,05). Los resultados mostraron que las branquias del embrión de I. bannanicus son más similares a los de otras especies en Ichthyophiidae que de otras especies.