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Fish bile has been applied as a biomarker for environmental contamination for several decades, and several pollutants are known to be excreted in this matrix. With the advent of the proteomic field, however, the discovery of protein biomarkers of response to pollutants has become the highlight, and fish bile shows very high potential in this regard. A proteomic case study carried out in Southeastern Brazil with mullet bile indicates the importance of assessing bile colour, as different feeding statuses lead to differential proteomic profiles as observed by 2D SDS-PAGE analyses. In addition, several heat-stable proteins displaying a differential gel profile were also observed in tilapia bile when compared a contaminated and reference site. Therefore, the bile proteome displays the potential to offer a more sensitive and informative method to analyse the presence and effects of contaminants in aquatic ecosystems.

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One of the metal detoxifying mechanisms that occurs in fish is metallothionein (MT) induction and metal binding. Hepatic MT induction has been well described, but biliary MT metal detoxification has only recently been described in fish. In this scenario, metal-metal interactions have been increasingly evaluated to further understand the behavior of these contaminants regarding homeostasis and biological functions, as well as their toxic effects. Studies, however, have been mainly conducted concerning the elemental pair Se-Hg, and scarce reports are available concerning other metal pairs. Therefore, this study aimed to evaluate biliary and hepatic MT metal detoxification mechanisms in a territorial neotropical cichlid, Geophagus brasiliensis. Fish were sampled from the anthropogenically impacted estuarine Rodrigo de Freitas Lagoon, located in Southern Rio de Janeiro, and trace elements and MT were determined by inductively coupled plasma mass spectrometry (ICP-MS) and UV-Vis spectrophotometry, respectively, in fish liver and bile. MT in bile were significantly lower than in liver. Significant differences between bile and liver were observed for many trace elements, and, although most were higher in liver, Cd and Ni were significantly higher in bile, indicating efficient excretion from the body via the biliary route. A significant correlation was observed between MT and Fe in bile, and between MT in liver and Cu and Zn in bile. Molar ratio calculations demonstrated protective elements effects against Al, As, Cd, Hg, Pb and V in both bile and liver, as well as some novel interrelationships, indicating the importance of these investigations regarding the elucidation of element detoxifying mechanisms. Furthermore, investigation of other elemental associations may aid in decision-making processes regarding environmental contamination scenarios linked to public health.

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UV filters (UV-Fs) constitute a heterogeneous group of chemicals used as protection against the effects of UV radiation, widely used in all sort of goods and ubiquitous in the environment. The presence of these chemicals in fish is a matter of concern, because many UV-Fs display hormonal activity. In this study, muscle, gills, and liver from 11 Mugil liza individuals from the highly urbanized Guanabara Bay (Rio de Janeiro, Brazil) were analysed in order to detect eight UV-Fs and metabolites (4-dihydroxybenzophenone [BP1] (2-hydroxy-4-methoxybenzophenone [BP3], 4-methylbenzylidiene camphor [4MBC], ethylhexyl methoxycinnamate [EHMC], ethylhexyl dimethyl p-aminobenzoic acid [ODPABA], octocrylene [OC], 4-hydroxybenzophenone [4HB], and 4,4'-dihydroxybenzophenone [4DHB]) using liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Results showed that both target UV-Fs and metabolites were ubiquitous in the analysed tissues. Lower concentrations were observed in muscle and gills (3.07-31.6ngg-1 dry weight (dw)), whereas in liver significant amounts of metabolites (5.47-451ngg-1 dw) were present. With the concentrations determined in the fish, an estimation of the daily intake revealed that consumption of muscle in the diet represent from 0.3 to 15.2ng UV-Fs (kg body weight-1) d-1, higher than those reported in fish for selected persistent organic pollutants (POPs).

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The present study aimed to investigate metal bioaccumulation in mullet (M. liza) from a tropical bay located in Southeastern Brazil, comparing a previously considered reference site to a known contaminated area of the bay, as well as to conduct human health risk assessments with regard to the consumption of this species. The metal concentrations were compared to the maximum residue level (MRL) in foods established by the different national and international regulatory agencies, and the Provisional Tolerable Daily Intake (PTDI) was determined and compared to reference values. Chromium (Cr), Zinc (Zn), Copper (Cu), Manganese (Mn), Nickel (Ni), Cadmium (Cd) and Lead (Pb) concentrations were determined in the gills, muscle and liver of 28 mullet by ICP-MS after acid digestion. Certain metals exceeded MRL guidelines established by different regulatory agencies, indicating human health risks associated to these metals. PTDI values, however, did not exceed corresponding metal values proposed by the World Health Organization. The metal concentrations found in the mullet samples indicate that the previously considered reference site is now showing signs of anthropogenic contamination.

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Microcystin (MC) accumulation and depuration in environmentally exposed tilapia (Oreochromis niloticus) at a chronically contaminated eutrophic lagoon was studied. This is one of the scarce reports on microcystin accumulation in bile of environmentally exposed fish, and gonad MC accumulation in the field, in contrast to laboratory exposure experiments. Results show that preferential MC accumulation in the environment occurred in tilapia fish muscle, followed by gonads, liver and, finally, bile. Biliary MC excretion in in situ conditions indicates elimination from the body to a certain degree. High gonad MC bioaccumulation is of concern, since this could indicate potential reproductive problems in this species. This study also demonstrated that tilapia shows similar oxidative stress responses (in the form of reduced glutathione, GSH) in the environment as those observed in laboratory exposed fish. MC dietary intake values for tilapia muscle and liver were above the limits imposed by international legislations, indicating that the local human population should exercise care when ingesting this species as a part of their diet and that human ingestion of MC-contaminated samples should be carefully monitored.

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Objetivo: determinar la influencia de contaminantes atmosféricos sobre el agua de lluvia. Métodos: utilización de la deposición húmeda en un estudio de largo plazo, realizado en el Estado de Acre, Amazonia Occidental, 2005-2010. Se monitorearon 185 eventos de lluvia, se determinó en ellos el pH, la conductividad eléctrica y la concentración de carbono orgánico total. Se monitoreó la carga de aerosoles atmosféricos mediante fotometría solar y las condiciones meteorológicas a través de mediciones del vapor de agua, temperatura, vientos y lluvias en su comportamiento estacional. Resultados: la concentración de contaminantes según la profundidad óptica de aerosoles varió desde valores muy altos, entre 3 y 4 durante la seca, hasta valores muy bajos, aproximadamente de 0,08, durante la estación lluviosa. El 25 % de las lluvias fue ácida con pH entre 3 y 4,7 y un promedio de 5,4. Los valores de conductividad eléctrica generalmente fueron inferiores a 10 µS cm-1. El carbono orgánico total alcanzó valores relativamente altos, entre 20 y 30 mg L-1. Conclusiones: durante la seca los incendios forestales responden por la alta contaminación de la atmósfera, que queda limpia durante la estación lluviosa por efecto de la deposición húmeda. Durante la seca la carga de aerosoles atmosféricos aumenta en más de 40 veces, en comparación con el estado de la atmósfera en la estación lluviosa. Son necesarias medidas más eficientes de control de la degradación ambiental en la Amazonia y de monitoreo sistemático de la contaminación de su atmósfera, en beneficio de la salud humana y de otras formas de vida.

Objective: to determine the influence of air pollutants on the rainwater. Methods: use of wet deposition in a long-term study conducted in the state of Acre, in Western Amazon, from 2005 to 2010. One hundred and eightyfive rainy events were monitored to determine the pH value, the electrical conductivity and the concentration of total organic carbon. The atmospheric aerosol load was measured by solar photometry whereas the water vapors, temperature, winds and rains were monitored to observe weather conditions. Results: the pollutant concentration, depending on the aerosol optical depth, varied from very high values of 3 and 4 during the dry season, to very low values of about 0.08 during the rainy season. Twenty five percent of rainfalls were acidic with pH ranging 3 to 4.7, being 5.4 the average value. The electrical conductivity values were generally lower than 10 µS cm-1. The total organic carbon reached relatively high values of 20 to 30 mg L-1. Conclusions: in the dry season, the forest fires are responsible for high degree of air pollution, but the situation changes in the rainy season when the environment is clean because of the wet deposition. During the dry season, the air aerosol load increases 40 times if compared to the environmental conditions in the dry season. Therefore, more efficient control measures are needed to stop environmental degradation in the Amazon as well as a systematic monitoring of the air pollution for the benefit of human health and other forms of life.

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Fish accumulate several trace elements in muscle, gills and liver, however studies also indicate that metals can be excreted through bile. Since metal contamination leads to modifications in bile composition, biliary excretion offers an alternative way to evaluate the presence of trace-elements. Bile is easier to obtain than other organs and presents a simpler matrix, making it easier for chemical pre-treatment. To verify if bile can be useful as a biomonitoring tool for metal contamination, liver and bile trace element concentrations were determined and correlated. The Artificial Neural Networks statistical technique was used to verify if liver trace-element quantification could be substituted by bile analysis. Results show that significant correlations were obtained between trace elements in bile and liver and the ANN validated the hypothesis that certain trace-elements in bile could be utilized instead of liver trace-elements. Further studies in this field are of interest to further validate this biomarker.

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Fish are well-recognized bioindicators of environmental contamination. Several recent proteomic studies have demonstrated the validity and value of using fish in the search and discovery of new biomarkers. Certain analytical tools, such as comparative protein expression analyses, both in field and lab exposure studies, have been used to improve the understanding of the potential for chemical pollutants to cause harmful effects. The metallomic approach is in its early stages of development, but has already shown great potential for use in ecological and environmental monitoring contexts. Besides discovering new metalloproteins that may be used as biomarkers for environmental contamination, metallomics can be used to more comprehensively elucidate existing biomarkers, which may enhance their effectiveness. Unfortunately, metallomic profiling for fish has not been explored, because only a few fish metalloproteins have thus far been discovered and studied. Of those that have, some have shown ecological importance, and are now successfully used as biomarkers of environmental contamination. These biomarkers have been shown to respond to several types of environmental contamination, such as cyanotoxins, metals, and sewage effluents, although many do not yet possess any known function. Examples of successes include MMPs, superoxide dismutases, selenoproteins, and iron-bound proteins. Unfortunately, none of these have, as yet, been extensively studied. As data are developed for them, valuable new information on their roles in fish physiology and in inducing environmental effects should become available.

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Metallothioneins (MT) are a validated biomarker for metal exposure. In fish they are usually determined in the liver, while some studies have observed MT in kidney, muscle and gills. There are, however, no reports on MT in fish bile. This study aimed to verify if these proteins are present in fish bile, if they can be used as biomarkers for metal exposure and to characterize the metal speciation present in the different species. Three independent methods to verify MT presence in fish bile were conducted: 1D/2D gel electrophoresis, spectrophotometric analyses and SEC-HPLC-ICP-MS. Results show that all methods verified the presence of these proteins in fish bile, while SEC-HPLC-ICP-MS also characterized also characterized metals responsible for the contamination of the studied sites. Quantification was successfully conducted by two methods, densitometry (1D electrophoresis) and spectrophotometry, and the data indicates that bile MTs follow the same trend as in liver. Therefore, Biliary MT show the potential to be used the same way as MT from other matrices in metal exposure studies, with the advantage of being a simpler matrix and showing the possibility of not sacrificing the animals, as chronic duct cannulation is available.

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On April 26th, 2005, an accident caused a leak of 60,000L of Diesel Oil Type "B", freighted by train wagons upstream on a mangrove area within Guanabara Bay, Southeast Brazil. After the accident, samples from animals with different biological requirements were collected in order to monitor polycyclic aromatic hydrocarbons concentrations for the following 12months. Sessile, mobile, carnivorous, omnivorous, organic detritus feeders, planktivorous and suspension feeders were some of the attributes compared. Concentrations of PAHs did not vary in relation to different dietary habits and the best response was from the sessile suspensivorous barnacles. A background level of <50microgkg(-1) was suggested based on the reference site and on values observed in the following months after the accident. The highest values of PAH concentrations were observed in barnacles in the first month immediately after the spill, decreasing to background levels after few months. Barnacles are suggested as a sentinel species.

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Owing to the importance of dissolution and weathering processes following oil spills, this work focused on the operational (quantitative) aspects related to the dissolution of petroleum-derived products, as well as the influence of solar light on both dissolution and the photoproduction of hydrogen peroxide. Four Brazilian crude oil samples were used to study the transfer process of organic compounds from the crude oil film to the aqueous phase (natural seawater) over a period of up to 45 days. Dissolved organic carbon (DOC), measured by non-dispersive infrared spectroscopy followed by high temperature catalytic combustion, was used to follow the partitioning between the two phases. Aqueous DOC values increased as a function of time (up to 15 days) until equilibrium was reached at concentrations ranging from 5 to 45 mg C L(-1). The final DOC concentration as well as the rate of dissolution depends on the nature of the crude oil. When exposed to sunlight, the dissolution was enhanced by up to 67.3%, and inorganic peroxides were generated in the concentration range from 4.5 up to 8.0 micromol L(-1) after 7.3 h irradiation. These results indicate that there is a need for a standard procedure for the production of the WSF in order to generate a more reliable tool to assess the impact of oil spills on the marine environment.

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