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This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)

O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)

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This short note aims to report in detail a preliminary assessment of the concentrations of Cd, Hg and Pb in tissues of blue crabs Callinectes danae collected from the Cananéia-Iguape-Peruíbe estuarine complex (CIP), in the South of São Paulo State coast, Brazil. In October 2014, blue crabs were collected from CIP. Tissues were removed by dissection and metal determination was performed by GF-AAS and CV-AAS. According to statistical analysis, Pb and Cd concentrations in gills were significantly higher than those found in muscles and hepatopancreas, respectively. There were no significant differences in Hg concentrations between samples. Cd, Hg and Pb concentrations in gills and hepatopancreas were lower than those reported in a previous study performed at CIP. However, Cd concentration in hepatopancreas was higher than the Brazilian limit for consumption and new efforts to monitor Cd concentrations in C. danae tissues must be performed.

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The environmental quality of a Ramsar wetland site located at the Cananéia-Iguape-Peruíbe Protected Area (CIP-PA), in São Paulo, Brazil, was assessed by geochemical analyses and biomarker assessments (GPx, GST, GSH, GST, MT, LPO, DNA damage) performed in swimming crab Callinectes danae Smith, 1869 organs (posterior and anterior gills and hepatopancreas) to estimate sediment contaminant bioavailability. The results indicated that two sampling stations, PT and PM, exhibited the worst environmental conditions, as sediments collected at both points contained metal contamination, while crabs exhibited significant responses for GPx, GST, and LPO (mostly during winter). Sediment contamination tended to be associated to fine sediments (both seasons) and organic matter (winter). During the summer survey, Pb concentrations in sediments of station PT exceeded the Brazilian Sediment Quality Guidelines (SQGs) and the Canadian Interim Marine Sediment Quality Guidelines. Metal concentrations in sediments sampled in winter were higher compared with summer, with Co, Ni, and Pb exceeding SQGs levels at PT, whereas Co, Ni, Hg, Zn, and Pb exceeded SQGs at PM. Biomarker induction during summer appeared to be caused by natural variables (water salinity and temperature, and molting cycle), whereas oxidative stress and tissue damage during winter appeared to be more clearly linked to metal contamination. Anterior gills presented the clearest signs of seasonal variability, being more responsive to sediment contamination. The results suggest that metals originated from the upper Ribeira de Iguape River are transported toward the estuarine system, causing effects on C. danae individuals. Additionally, seasonality is a strong factor concerning CIP-PA toxicity, since the rainfall regime significantly modifies the freshwater flow and, consequently, estuarine water salinity, suspended particle and metal inputs, as well as the location of depositional areas. Thus, efforts to mitigate CIP-PA contamination should be based on the control of upstream pollution sources.

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Shrimp of Farfantepenaeus californiensis (78 groups) and Litopenaeus stylirostris (14 groups) were caught in the northwestern fishing zones in Mexico during the 2014-2015 fishing season (September-February); both shrimp species have high commercial value. Muscle, hepatopancreas and exoskeleton were analyzed to determine their metal contents. For F. californiensis, the highest Cd, Pb and Zn contents were determined in specimens caught off the State of Baja California Sur (BCS) with 22.4 ± 8.9 (hepatopancreas), 2.83 ± 4.63 (muscle), and 748.5 ± 1567 (muscle) µg/g, in the regions off Mulegé, Los Cabos and Los Cabos, respectively. For L. stylirostris, the fishing zone of Comundú (BCS) showed higher Cd (12.3 ± 11.5 µg/g), Cu (569.1 ± 646.5 µg/g) and Zn (549.7 ± 400.7 µg/g) contents; all values were determined in the hepatopancreas. Regarding the hazard quotient and total hazard quotient calculated in this study, the consumption of marine shrimp caught off NW Mexico does not represent a risk to human health (both < 1).

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BACKGROUND: Cathepsin D is a lysosomal enzyme that is found in all organisms acting in protein turnover, in humans it is present in some types of carcinomas, and it has a high activity in Parkinson's disease and a low activity in Alzheimer disease. In marine organisms, most of the research has been limited to corroborate the presence of this enzyme. It is known that cathepsin D of some marine organisms has a low thermostability and that it has the ability to have activity at very acidic pH. Cathepsin D of the Jumbo squid (Dosidicus gigas) hepatopancreas was purified and partially characterized. The secondary structure of these enzymes is highly conserved so the role of temperature and pH in the secondary structure and in protein denaturation is of great importance in the study of enzymes. The secondary structure of cathepsin D from jumbo squid hepatopancreas was determined by means of circular dichroism spectroscopy. OBJECTIVE: In this article, our purpose was to determine the secondary structure of the enzyme and how it is affected by subjecting it to different temperature and pH conditions. METHODS: Circular dichroism technique was used to measure the modifications of the secondary structure of cathepsin D when subjected to different treatments. The methodology consisted in dissecting the hepatopancreas of squid and freeze drying it. Then a crude extract was prepared by mixing 1: 1 hepatopancreas with assay buffer, the purification was in two steps; the first step consisted of using an ultrafiltration membrane with a molecular cut of 50 kDa, and the second step, a pepstatin agarose resin was used to purification the enzyme. Once the enzyme was purified, the purity was corroborated with SDS PAGE electrophoresis, isoelectric point and zymogram. Circular dichroism is carried out by placing the sample with a concentration of 0.125 mg / mL in a 3 mL quartz cell. The results were obtained in mdeg (millidegrees) and transformed to mean ellipticity per residue, using 111 g/mol molecular weight/residue as average. Secondary-structure estimation from the far-UV CD spectra was calculated using K2D Dichroweb software. RESULTS: It was found that α helix decreases at temperatures above 50 °C and above pH 4. Heating the enzyme above 70°C maintains a low percentage of α helix and increases ß sheet. Far-UV CD measurements of cathepsin D showed irreversible thermal denaturation. The process was strongly dependent on the heating rate, accompanied by a process of oligomerization of the protein that appears when the sample is heated, and maintained a certain time at this temperature. An amount typically between 3 and 4% α helix of their secondary structure remains unchanged. It is consistent with an unfolding process kinetically controlled due to the presence of an irreversible reaction. The secondary structure depends on pH, and a pH above 4 causes α helix structures to be modified. CONCLUSION: In conclusion, cathepsin D from jumbo squid hepatopancreas showed retaining up to 4% α helix at 80°C. The thermal denaturation of cathepsin D at pH 3.5 is under kinetic control and follows an irreversible model.

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This study evaluated lead (Pb) content in mangrove root crabs Goniopsis cruentata and performed a risk assessment based on individual frequency of food consumption of the residents of an estuarine village in Brazil. The Pb content in the raw and two forms of processed crabs were analyzed by graphite furnace atomic absorption spectrometry. Based on consumption by the community, estimates of non-carcinogenic target hazard quotients (THQ) were calculated. The mean (± SD) Pb concentrations in raw tissues were: 0.17 (± 0.36), < 0.012 and 0.05 (± 0.09) mg kg-1 (gills, muscle and hepatopancreas), respectively. In the processed products, the highest levels were found in the traditional stew collected in April (2.84 mg kg-1), surpassing the maximum limit allowed. The THQ for consumption of traditional stew was < 1, classified as negligible risk. The G. cruentata of the Jaguaripe River estuary can be considered a safe food, but cooking should be avoided in lead-glazed pottery.

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The antioxidant defense system (ADS) protects organisms against the potential oxidative stress induced by environmental features, underlying processes of habitat diversification. The anomurans Aegla constitute the most threatened freshwater decapods of South America, occupying pristine habitats with narrow distribution. Using phylogenetic comparative methods, we addressed: Is the variability of habitat physicochemical parameters and tissue ADS phylogenetically structured? How do environmental features correlate with ADS? How do they vary among species? Several physicochemical parameters of water, as well as metals in sediments, were measured in ten aeglid species' habitats. Additionally, metal accumulation and ADS parameters [metallothionein-like proteins (MTLP), antioxidant capacity against peroxyl radicals (ACAP), and glutathione system (GSH-GSSG)] were evaluated in hepatopancreas. Water conductivity and pH showed phylogenetic signal, while all other physicochemical traits demonstrated plastic variability. Metals were present at natural concentrations, which are corroborated by the relative stable GSH/GSSG ratio, and by their absence of correlation with bioaccumulation levels and MTLP, both phylogenetically structured. However, metal variability across species' niches is associated with ACAP, a potential biomarker tool. Thus, the physiological sensitivity of aeglids is environmentally driven but also  phylogenetically constrained, unraveling the importance of systematic framework for cross-species investigations and future monitoring strategies of these conspicuous freshwater animals.

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Total mercury (THg) concentrations measured in two freshwater shrimp species (Macrobrachium depressimanum and Macrobrachium jelskii) showed a relationship with the location of artisanal and small-scale gold mining (ASGM) from the Madeira River Basin, Western Amazon. Between August 2009 and May 2010, 212 shrimp samples were collected in the confluence of the Madeira River with three of its tributaries (Western Amazon). THg concentration was quantified in the exoskeleton, hepatopancreas and muscle tissue of the shrimps by cold vapor atomic absorption spectrophotometry. There were no significant differences between the two shrimp species when samples came from the Madeira River, but Hg concentrations were significantly lower in a tributary outside the influence of the gold mining area. Average THg concentrations were higher in the hepatopancreas (up to 160.0 ng g-1) and lower in the exoskeleton and muscle tissue (10.0-35.0 ng g-1 and < 0.9-42.0 ng g-1, respectively). Freshwater shrimps from the Madeira River respond to local environmental levels of Hg and can be considered as biomonitors for environmental Hg at this spatial scale. These organisms are important for moving Hg up food webs including those that harbor economic significant fish species and thus enhancing human exposure.

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We determined total Hg and Se contents of hepatopancreas, exoskeleton, and muscle, and the Se:Hg molar ratios in the muscle of shrimps Farfantepenaeus californiensis and Litopenaeus stylirostris caught in NE Pacific Mexican waters. Total Hg mean values in muscle, hepatopancreas, and exoskeleton were 0.31 ± 0.26, 0.28 ± 0.29, and 0.24 ± 0.06 µg g-1, and 0.46 ± 0.46, 0.41 ± .034, and 0.24 ± 0.06 µg g-1 for F. californiensis and L. stylirostris, respectively. In all tissues, the mean concentrations of Se tended to be close to one order of magnitude higher than the respective Hg values. In F. californiensis, the hepatopancreas of the larger commercial size had significantly (p < 0.05) higher Hg content than smaller sizes, but correlations size-Hg concentration calculated for each tissue of either species were not significant. The Hg content of the muscle of all commercial sizes of both species was lower than the permissible limit and their Se:Hg ratios in all sizes were higher than 1, indicating low risk for human consumption.

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The mangrove crab Ucides cordatus is widespread in the Brazilian coast, which has an important role in nutrient cycling. This species reproduces in summer and females carry eggs about a month, when they maintain contact with water and sediments. It remains unclear if trace metals can be absorbed or adsorbed by the eggs during development. The present study aims to investigate, for the first time, trace metal concentrations in ovigerous female tissues and eggs of U. cordatus in two areas with different metal pollution levels in the Southeastern Brazil. Samples were collected in two different mangroves, Guanabara Bay (GB) highly polluted environment and Paraíba do Sul River (PSR). In both populations, we observed significant increase of V, Cr, and Mn concentrations along eggs maturation. The higher metals averages were found in PSR population. This trend was reported since the 1990s and lower concentrations in GB marine organisms were attributed to reducing conditions, high organic load, and the presence of sulfide ions. These conditions restrict the bioavailability of metals in the bay, with exception of Mn. No significant differences were observed in gills and muscles. In both populations of the present study, V, Zn, As, and Pb were higher in eggs of initial stage, whereas Mn, Ni, Cu, and Cd were higher in hepatopancreas. Beside this, V, Cr, Mn, As, and Pb showed an increase concerning egg development. Thus, V, As, and Pb in eggs come from two sources previous discussed: females and environment. Zinc came mainly from females due to essential function. Those new information should be considered as one of the mechanisms of trace metal transfer to the trophic chain, between benthonic and pelagic environment.

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A method based on matrix solid-phase dispersion (MSPD) and gas chromatography-mass spectrometry to determine pesticides in fish liver and crab hepatopancreas was optimized. Ethyl acetate and acetonitrile were evaluated as elution solvents and their volumes were also checked. The best results were obtained with 1.0 g reused C18 as sorbent, using 5 mL acetonitrile as the elution solvent. Analytical recoveries ranged between 57 and 107% with RSD lower than 26% in fish liver and between 56 and 122% with RSD lower than 21% in crab hepatopancreas. The LOQ values for these compounds ranged from 0.05 to 0.5 mg kg(-1) for crab hepatopancreas and from 0.125 to 1.25 mg kg(-1) for fish liver. MPSD was shown to be easy and fast to use, with a clear advantage regarding costs because it does not need any expensive instrument. The proposed method was successfully applied to determine dimethoate, atrazine, clomazone, fenitrothion, malathion, fipronil and tebuconazole in fish liver and crab hepatopancreas samples.

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The freshwater crayfish Cherax quadricarinatus is a tropical species of great interest for aquaculture. Vitellogenin (Vg), a lipoprotein precursor of the vitellum accumulated in spawned eggs, can be synthesized in the ovary and/or hepatopancreas of most crustaceans, being the hemolymph the way for transporting Vg throughout the reproductive cycle. Concentration of Vg in hemolymph, ovary and hepatopancreas of Cherax quadricarinatus adult females was measured by means of ELISA, specifically developed after purifying the native Vg. Measurements were made at four periods of the reproductive cycle: pre-reproductive, mid-reproductive, late reproductive and post-reproductive. Besides, both hepatosomatic (HSI) and gonadosomatic (GSI) indexes were determined in each period. Significant variations in Vg levels were detected in both hemolymph and hepatopancreas, being the highest values observed during the mid-reproductive period. Besides, such variations were positively correlated to the HSI. A positive correlation between Vg levels in hepatopancreas and ovary was also seen. These results support previous evidences about the central role of the hepatopancreas as a site of Vg synthesis in the studied species, together with the relevancy of hemolymph for transporting Vg from the hepatopancreas to the ovary. For aquaculture purposes, Vg monitoring in hemolymph could be used as a non-injurious method, to check the reproductive activity of C. quadricarinatus females.

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The freshwater crayfish Cherax quadricarinatus is a tropical species of great interest for aquaculture. Vitellogenin (Vg), a lipoprotein precursor of the vitellum accumulated in spawned eggs, can be synthesized in the ovary and/or hepatopancreas of most crustaceans, being the hemolymph the way for transporting Vg throughout the reproductive cycle. Concentration of Vg in hemolymph, ovary and hepatopancreas of Cherax quadricarinatus adult females was measured by means of ELISA, specifically developed after purifying the native Vg. Measurements were made at four periods of the reproductive cycle: pre-reproductive, mid-reproductive, late reproductive and post-reproductive. Besides, both hepatosomatic (HSI) and gonadosomatic (GSI) indexes were determined in each period. Significant variations in Vg levels were detected in both hemolymph and hepatopancreas, being the highest values observed during the mid-reproductive period. Besides, such variations were positively correlated to the HSI. A positive correlation between Vg levels in hepatopancreas and ovary was also seen. These results support previous evidences about the central role of the hepatopancreas as a site of Vg synthesis in the studied species, together with the relevancy of hemolymph for transporting Vg from the hepatopancreas to the ovary. For aquaculture purposes, Vg monitoring in hemolymph could be used as a non-injurious method, to check the reproductive activity of C. quadricarinatus females.

La langosta de agua dulce Cherax quadricarinatus es una especie tropical de gran interés para la acuicultura. Se midió la concentración de vitelogenina (Vg) en hemolinfa, ovario y hepatopáncreas de hembras adultas de esta especie, por medio de ELISA. Las mediciones fueron hechas en los cuatro períodos del ciclo reproductivo: pre-reproductivo, reproductivo medio, reproductivo tardío y post-reproductivo. Se detectaron variaciones significativas en los niveles de Vg tanto en hemolinfa como en hepatopáncreas, se observó el mayor valor durante el período reproductivo medio. Además, tales variaciones se correlacionaron positivamente con el índice hepatosomático. Se observó además una correlación positiva de los niveles de Vg entre hepatopáncreas y ovario. Estos resultados apoyan evidencias previas sobre el papel central del hepatopáncreas como sitio de síntesis de Vg, en esta especie, y también enfatizan la importancia de la hemolinfa para el transporte de la Vg del hepatopáncreas al ovario. Para propósitos de acuicultura, la medición de Vg en hemolinfa podría ser utilizada como un método no lesivo, con el fin de constatar la actividad reproductiva de hembras de C. quadricarinatus.

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While invertebrates store neutral lipids as their major energy source, little is known about triacylglycerol (TAG) class composition and their differential catabolism in aquatic arthropods. This study focuses on the composition of the main energy source and its catabolism by lipase from the midgut gland (hepatopancreas) of the crustacean Macrobrachium borellii. Silver-ion thin-layer chromatography of prawn large TAG deposit (80% of total lipids) and its subsequent fatty acid analysis by gas chromatography allowed the identification of 4 major fractions. These are composed of fatty acids of decreasing unsaturation and carbon chain length, the predominant being 18:1n-9. Fraction I, the most unsaturated one, contained mainly 20:5n-3; fraction II 18:2n-6; fraction III 18:1n-9 while the most saturated fraction contained mostly 16:0. Hepatopancreas main lipase (Mr 72 kDa) cross-reacted with polyclonal antibodies against insect lipase, was not dependent on the presence of Ca(2+) and had an optimum activity at 40°C and pH 8.0. Kinetic analysis showed a Michaelis-Menten behavior. A substrate competition assay evidenced lipase specificity following the order: 18:1n-9-TAG>PUFA-enriched-TAG>16:0-TAG different from that in vertebrates. These data indicate there is a reasonable correspondence between the fatty acid composition of TAG and the substrate specificity of lipase, which may be an important factor in determining which fatty acids are mobilized during lipolysis for oxidation in crustaceans.

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The ghost crab, Ocypode quadrata, is found on sandy beaches from the United States to Southern Brazil. Because there is still little information about the metabolism and reproduction of O. quadrata on the southern coast of Brazil, the objectives of this study were to (i) evaluate the effect of seasonal variations on the carbohydrate, lipid, and protein metabolism of O. quadrata at Rondinha Beach, a beach with high anthropogenic activity, and to compare it with data from Siriú Beach, which has less human activity; and (ii) describe the effect of seasonal variations on the histological characteristics of male and female gonads, in order to assess the reproductive capacity of the crabs. The gonads of male crabs showed no significant variations in the gonadosomatic index (GSI) and glycogen levels; however, histological analysis of the testes revealed that they are mature in the summer. In females, the GSI and glycogen values are higher in summer, concomitantly with the presence of mature oocytes. These results suggest that the reproductive peak of O. quadrata occurs in summer. The seasonal analysis of the biochemical parameters, as well as comparison with Siriú Beach, demonstrated that the ghost crabs of Rondinha Beach have a different pattern of metabolism than those of Siriú. This difference may be a consequence of differences in the environmental conditions as well as in the anthropogenic pressures, such as vehicle traffic and the increase in human population at the beach in summer.

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We analyzed the dietary copper effects in the estuarine crab Neohelice (Chasmagnathus) granulata and its interaction with water salinity. Crabs were maintained at 2 per thousand and 30 per thousand salinity for 5 weeks and they were fed with commercial food supplemented with the green alga Scenedesmus vacuolatus previously exposed to copper. No mortalities were observed, but crabs maintained at 2 per thousand salinity accumulated on average 40% more copper compared to animals maintained at 30 per thousand salinity. At 2 per thousand salinity, superoxide dismutase (SOD) activity and reduced glutathione (GSH) levels were increased at the first and second weeks, respectively, while lipid peroxidation and protein oxidation were evident after 4 weeks of copper exposure. At 30 per thousand salinity, all measured variables increased progressively but were significantly higher only at the end of the assay (5th week), except for protein oxidation that remained unchanged throughout the experiment. The hepatosomatic index (HSI) was significantly decreased in response to copper exposure, but only in crabs acclimated to 2 per thousand. These findings have suggested that dietary copper exposure induces greater metal accumulation and larger oxidative stress responses in crabs maintained at 2 per thousand salinity.

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Phosphate-rich amorphous mineral granules (AMG) have been studied in a number of organisms, and show different physical and chemical properties according to their organic and mineral composition. We studied AMG isolated from the hepatopancreas of the land crab Ucides cordatus, which were subjected to different pHs in order to mimic the possible effects of H(+) on these structures. We used scanning and transmission electron microscopy (SEM, TEM), energy-filtering transmission electron microscopy (EFTEM), energy-dispersive X-ray analysis (EDXA) and nuclear magnetic resonance (NMR). TEM showed that granules were structurally disrupted when subjected to pH 5. The granules contain a soluble fraction that is rich in orthophosphate, which was the most abundant form of phosphate, although pyrophosphate and glucose-6-phosphate were also detected by (31)P NMR analysis. The redistribution of elements in the structure and pH conditions is discussed, focusing on their possible implications for AMG structure, function and dynamics.

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The effect of water salinity and ions on metallothionein-like proteins (MTLP) concentration was evaluated in the blue crab Callinectes sapidus. MTLP concentration was measured in tissues (hepatopancreas and gills) of crabs acclimated to salinity 30 ppt and abruptly subjected to a hypo-osmotic shock (salinity 2 ppt). It was also measured in isolated gills (anterior and posterior) of crabs acclimated to salinity 30 ppt. Gills were perfused with and incubated in an isosmotic saline solution (ISS) or perfused with ISS and incubated in a hypo-osmotic saline solution (HSS). The effect of each single water ion on gill MTLP concentration was also analyzed in isolated and perfused gills through experiments of ion substitution in the incubation medium. In vivo, MTLP concentration was higher in hepatopancreas than in gills, being not affected by the hypo-osmotic shock. However, MTLP concentration in posterior and anterior gills significantly increased after 2 and 24 h of hypo-osmotic shock, respectively. In vitro, it was also increased when anterior and posterior gills were perfused with ISS and incubated in HSS. In isolated and perfused posterior gills, MTLP concentration was inversely correlated with the calcium concentration in the ISS used to incubate gills. Together, these findings indicate that an increased gill MTLP concentration in low salinity is an adaptive response of the blue crab C. sapidus to the hypo-osmotic stress. This response is mediated, at least in part, by the calcium concentration in the gill bath medium. The data also suggest that the trigger for this increase is purely branchial and not systemic.

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The effect of the water-soluble fraction (WSF) of crude oil on lipid contents, lipid classes, FA, and PC molecular species was studied in high-phospholipid (hepatopancreas) and low-phospholipid (egg) tissues of a freshwater crustacean. After a 21-d exposure to a sublethal concentration of WSF, a significant decrease in shrimp total lipids was observed, although no alterations could be detected in the hepatopancreas or egg lipid contents. TAG/phospholipid ratios increased in the hepatopancreas and decreased in the eggs, suggesting alterations either in the mobilization of TAG to phospholipid pools or in the energy balance. The FA composition of phosphoglycerides in the hepatopancreas and eggs was dominated by PUFA, whereas the n-3/n-6 ratio was not affected by WSF exposure, although there was a significant increase in hepatopancreas 18:1n-9. Analysis of the PC molecular species by HPLC-ELSD showed the presence of 15 species, with 16:0/18:1, 18:1/18:2, 16:0/20:5, and 16:1/20:5 being the major species in the hepatopancreas. The PC molecular species in the eggs showed a different pattern, dominated by 16:0/18:1 and 18:1/18:2. Of the PC molecular species, 10 contained 22:6n-3, 20:5n-3, and 20:4n-6. Small amounts of di-PUFA species were also found. Exposure to WSF altered the PC molecular species in both tissues. The four major hepatopancreas molecular species and most of the ones containing PUFA decreased. This was compensated for by an increase in 16:1/18:1 (152%) and 18:1/18:1 (50%). The two major egg PC molecular species decreased, whereas the PUFA-containing ones increased. The contrasting responses of both tissues to WSF contamination suggests the presence of different homeostatic mechanisms.

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We investigated the transport of (14)C-methylaminoisobutyric acid ((14)C-MeAIB) and (14)C-alanine oxidation in hepatopancreas and jaw muscle of Chasmagnathus granulata submitted to 24, 72, and 144 h of hypo- or hyperosmotic stress. While (14)C-MeAIB uptake increased in jaw muscle and hepatopancreas from crabs submitted to hyperosmotic stress, it did not change in tissues from animals submitted to hypo-osmotic stress. Incubation of jaw muscle and hepatopancreas from control groups with 1 mM ouabain did not decrease (14)C-MeAIB uptake. However, ouabain prevented (14)C-MeAIB uptake in hepatopancreas at 24 h of hyperosmotic stress. In contrast, in jaw muscle from crabs submitted to the same conditions, (14)C-MeAIB uptake was not prevented by ouabain in the incubation medium. Jaw muscle from the control group produced four times more (14)CO(2) from (14)C-alanine than the hepatopancreas. During hypo-osmotic stress, amino acid oxidation does not seem to be one of the pathways implicated in the decrease of the amino acid pools in hepatopancreas and jaw muscle. In contrast, during hyperosmotic stress the reduction in (14)C-alanine oxidation appears to be one of the mechanisms involved in the increase of the amino acid pool in the hepatopancreas.

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