- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: Selenium (Se) is an essential trace-element that becomes toxic when present at high concentrations. Little is known regarding Se effects on parameters such as oxidative stress biomarkers. The aim of the present study was to investigate the effects of acute selenium exposure on oxidative stress biomarkers in a model organism, zebrafish (Danio rerio). Fish were exposed to selenium selenite at 1mgL(-1). Reduced glutathione (GSH), and metallothionein (MT) concentrations were determined in liver, kidney and brain, with MT also being determined in bile. Essential metals and trace-elements were also determined by inductively coupled mass spectrometry (ICP-MS) in order to verify possible metal homeostasis alterations. GSH concentrations in liver, kidney and brain increased significantly (1.05±0.03μmolg(-1) ww, 1.42±0.03μmolg(-1) ww and 1.64±0.03μmolg(-1) ww, respectively) in the Se-exposed group when compared to the controls (0.88±0.05μmolg(-1) ww, 0.80±0.04μmolg(-1) ww and 0.89±0.03μmolg(-1) ww for liver, kidney and brain, respectively). MT levels in Se-exposed liver (0.52±0.03μmolg(-1) ww) decreased significantly in comparison to the control group (0.64±0.02μmolg(-1) ww), while levels in bile increased, albeit non-significantly. This is in accordance with previous studies that indicate efficient biliary MT action, leading to a rapid metabolism and elimination of contaminants from the body. Levels in the brain increased significantly after Se-exposure (0.57±0.01μmolg(-1) ww) when compared to the control group (0.35±0.03μmolg(-1) ww) since this organ does not present a detoxification route as quick as the liver-gallbladder route. Several metal and trace-elements were altered with Se-exposure, indicating that excess of selenium results in metal dyshomeostasis. This is the first report on metal dyshomeostasis due to Se-exposure, which may be the first step in the mechanism of action of selenium toxicity, as is postulated to occur in certain major human pathophysiologies.
- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: Fish bile metallothioneins (MT) have been recently reported as biomarkers for environmental metal contamination; however, no studies regarding standardizations for their purification are available. Therefore, different procedures (varying centrifugation times and heat-treatment temperatures) and reducing agents (DTT, β-mercaptoethanol and TCEP) were applied to purify MT isolated from fish (Oreochromis niloticus) bile and liver. Liver was also analyzed, since these two organs are intrinsically connected and show the same trend regarding MT expression. Spectrophotometrical analyses were used to quantify the resulting MT samples, and SDS-PAGE gels were used to qualitatively assess the different procedure results. Each procedure was then statistically evaluated and a multivariate statistical analysis was then applied. A response surface methodology was also applied for bile samples, in order to further evaluate the responses for this matrix. Heat treatment effectively removes most undesired proteins from the samples, however results indicate that temperatures above 70°C are not efficient since they also remove MTs from both bile and liver samples. Our results also indicate that the centrifugation times described in the literature can be decreased in order to analyze more samples in the same timeframe, of importance in environmental monitoring contexts where samples are usually numerous. In an environmental context, biliary MT was lower than liver MT, as expected, since liver accumulates MT with slower detoxification rates than bile, which is released from the gallbladder during feeding, and then diluted by water. Therefore, bile MT seems to be more adequate in environmental monitoring scopes regarding recent exposure to xenobiotics that may affect the proteomic and metalloproteomic expression of this biological matrix.
- [Show abstract] [Hide abstract] ABSTRACT: The pacu (Piaractus mesopotamicus) is a hypoxia-tolerant neotropical fish species. There is little or no information in this species regarding biochemical adaptations to waters with different oxygen concentrations, such as the production of reactive oxygen species and antioxidant scavengers, which might be of interest in the study of antioxidant defense mechanisms. Metallothioneins (MT) have been widely applied as biomarkers for metal exposure in fish liver, and, recently, in bile. These metalloproteins, however, have also been reported as free radical scavengers, although studies in this regard are scarce in fish. In this context, normoxic and hypoxic controlled experiments were conducted with pacu specimens and MT levels were quantified in both liver and bile. Reduced glutathione (GSH) indicative of oxidative stress, and thiobarbituric acid reactive substances (TBARS), indicative of lipid peroxidation, were also determined in liver. The results demonstrate that hypoxic fish present significantly lower metallothionein levels in liver and bile and lower reduced glutathione levels in liver, whereas lipid peroxidation was not significantly different between hypoxic and normoxic fish. The results of the present study seem to suggest that metallothioneins may actively participate in redox regulation in hypoxic fish in both bile and liver. MT levels in these organs may be temporarily suppressed, supporting the notion that down-regulation of oxidant scavengers during the oxidative burst is important in defense signaling in these adapted organisms.
- [Show abstract] [Hide abstract] ABSTRACT: The present study analyzed metallothionein (MT) excretion from liver to bile in Nile Tilapia (Oreochromis niloticus) exposed to sub-lethal copper concentrations (2mgL(-1)) in a laboratory setting. MTs in liver and bile were quantified by spectrophotometry after thermal incubation and MT metal-binding profiles were characterized by size exclusion high performance liquid chromatography coupled to ICP-MS (SEC-HPLC-ICP-MS). Results show that liver MT is present in approximately 250-fold higher concentrations than bile MT in non-exposed fish. Differences between the MT profiles from the control and exposed group were observed for both matrices, indicating differential metal-binding behavior when comparing liver and bile MT. This is novel data regarding intra-organ MT comparisons, since differences between organs are usually present only with regard to quantification, not metal-binding behavior. Bile MT showed statistically significant differences between the control and exposed group, while the same did not occur with liver MT. This indicates that MTs synthesized in the liver accumulate more slowly than MTs excreted from liver to bile, since the same fish presented significantly higher MT levels in liver when compared to bile. We postulate that bile, although excreted in the intestine and partially reabsorbed by the same returning to the liver, may also release MT-bound metals more rapidly and efficiently, which may indicate an efficient detoxification route. Thus, we propose that the analysis of bile MTs to observe recent metal exposure may be more adequate than the analysis of liver MTs, since organism responses to metals are more quickly observed in bile, although further studies are necessary.
- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: This study presents a classification method combining logistic regression and fuzzy logic in the determination of sampling sites for feral fish, Nile Tilapia (Tilapia rendalli). This method statistically analyzes the variable domains involved in the problem, by using a logistic regression model. This in turn generates the knowledge necessary to construct the rule base and fuzzy clusters of the fuzzy inference system (FIS) variables. The proposed hybrid method was validated using three fish stress indices; the Fulton Condition Factor (FCF) and the gonadossomatic and hepatossomatic indices (GSI and HSI, respectively), from fish sampled at 3 different locations in the Rio de Janeiro State. A multinomial logistic regression allowed for the FIS construction of the proposed method and both statistical approaches, when combined, complemented each other satisfactorily, allowing for the construction of an efficient classification method regarding feral fish sampling sites that, in turn, has great value regarding fish captures and fishery resource management.
- [Show abstract] [Hide abstract] ABSTRACT: A remediação de uma amostra de Latossolo Vermelho-Amarelo contaminado artificialmente com petróleo (5.000 mg kg-1) foi estudada comparando diferentes oxidantes químicos: KMnO4 0,10 mol L-1; K2S2O8 0,10 mol L-1; H2O2 0,10 mol L-1; H2O2/Fe2+ 0,10/0,080 mol L-1 (reação de Fenton); e H2O2/Fe2+/UV 0,10/0,080 mol L-1 (reação de foto-Fenton). A remediação foi praticamente completa aos 30 min para o processo de foto-Fenton (99 %). Todos os outros oxidantes não atingiram essa taxa de dissipação até 180 min. O processo em que se usou H2O2 0,10 mol L-1 (70,51 %) foi o de menor eficiência. A adição de íons Fe aumentou a taxa de dissipação para 86,98 % (H2O2/Fe2+ 0,10/0,080 mol L-1). Os métodos convencionais apresentaram taxas de dissipação de 76,58 % (KMnO4 0,10 mol L-1) e 93,85 % (K2S2O8 0,10 mol L-1).
- [Show abstract] [Hide abstract] ABSTRACT: Mullet (Mugil liza) were sampled in five different areas along the Guanabara Bay, southeastern Brazil, classified as non-contaminated, moderately contaminated and contaminated. Morphometric (Fulton condition factor, relative condition factor and weight to length scaling coefficient) and organosomatic (hepatosomatic index) indices of environmental stress were analysed. Fish from the differentially contaminated areas show statistically different Fulton and relative condition factors and hepatosomatic indices, but not the weight to length scaling coefficient. The Kn and the FCF followed the same trend, with fish from São Gonçalo (1.07 ± 0.04 and 0.89 ± 0.03), Itaipu (0.84 ± 0.01 and 0.86 ± 0.01) and the Rodrigo de Freitas Lagoon (1.03 ± 0.01 and 0.87 ± 0.20) showing higher FCFs than fish from Magé (0.96 ± 0.01 and 0.81 ± 0.01). Fish from Itaipu showed significantly higher HSI values than the other sampling sites (1.68 ± 0.07), with fish from Olaria and Ipiranga showing the lowest (1.56 ± 0.12 and 1.60 ± 0.07, respectively).
- [Show abstract] [Hide abstract] ABSTRACT: Glutathione S-transferase (GST) assays in non-mammalian organisms are usually conducted inappropriately, since no previous standardization of the optimal concentrations of proteins and substrates and adequate pH is conducted. Standardization is a key task to adjust enzyme assays at their kinetically correct maximal initial velocities, if one wants these velocities to indicate the amount of enzyme in a sample. In this paper GST assays were standardized in liver cytosol to compare seasonal GST levels in liver of mullet from two contaminated lagoons in the Rio de Janeiro to those from a reference bay. GST potential as a biomarker of sublethal intoxication in this species was also evaluated. Mullet liver GST levels assayed with substrates that corresponded to three different GST isoenzymes varied throughout the year. The differences indicated that mullets are suffering from sublethal intoxication from contaminants in these lagoons. Seasonal variations of activity were relevant, since these could indicate differences in xenobiotic input into the areas. An analysis of overall mullet health condition using a morphological index (the Fulton Condition Factor) and macroscopic abnormalities corroborated the differences in GST levels, with fish from one of the sites in worse overall health condition showing lower and significantly different FCF when compared to the reference site. Therefore, GST standardized activity levels are useful biomarkers of environmental contamination for mullet.
- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: The objective of this work was to obtain organic compounds similar to the ones found in the organic matter of anthropogenic dark earth of Amazonia (ADE) using a chemical functionalization procedure on activated charcoal, as well as to determine their ecotoxicity. Based on the study of the organic matter from ADE, an organic model was proposed and an attempt to reproduce it was described. Activated charcoal was oxidized with the use of sodium hypochlorite at different concentrations. Nuclear magnetic resonance was performed to verify if the spectra of the obtained products were similar to the ones of humic acids from ADE. The similarity between spectra indicated that the obtained products were polycondensed aromatic structures with carboxyl groups: a soil amendment that can contribute to soil fertility and to its sustainable use. An ecotoxicological test with Daphnia similis was performed on the more soluble fraction (fulvic acids) of the produced soil amendment. Aryl chloride was formed during the synthesis of the organic compounds from activated charcoal functionalization and partially removed through a purification process. However, it is probable that some aryl chloride remained in the final product, since the ecotoxicological test indicated that the chemical functionalized soil amendment is moderately toxic.
- [Show abstract] [Hide abstract] ABSTRACT: Gallbladder bile from 2 fish species, mullet (Mugil liza) and tilapias (Tilapia rendalli), contain substantial matrix metalloproteinases (MMPs). Extensive purification studies were conducted in order to obtain workable samples for SDS-PAGE and zymography analysis. Proteinase activities were assayed by gelatin substrate zymography. Several protein bands were observed, corresponding to molecular weights of 200, 136, 43, 36, 34, 29, 23 and 14 kDa in mullet bile and 179, 97, 79, 61, 54, 45, 36, 33 and 21 kDa in tilapia bile. Specific inhibitor studies were conducted, in which MMPS were inhibited by EDTA and 1,10 phenanthroline, but not by serine and cysteine protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF) and transepoxysuccinyl-l-leucylamido-l-guanidino butane (E-64), confirming the proteinase identities as MMPs. Differences in proteinase expression were observed in fish from a contaminated and reference site. Some studies regarding MMPs in different fish tissues exist, however this is the first study conducted in fish bile, and their involvement in detoxification processes and organism protection against the effects of aquatic contaminants may be a possibility.
- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: 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.
- [Show abstract] [Hide abstract] ABSTRACT: This study demonstrates a methodology for evaluating the theoretical antimalarial potential of 18 phenazine derivatives, some of which have no known biological activities. Semi-empirical, density functional and statistical methods for analytical data treatment were used to establish a quantitative structure-activity relationship (QSAR). A set of 16 descriptors were obtained. Theoretical tools regarding inferential statistics were used to analyze and interpret the information necessary for the application of a multivariate analysis. We classified four types of phenazines into distinct clusters taking into account including planarity, ring-attached substituents, geometric arrangement and conjugation effects, energy maps as well as frontier orbitals energies. The experimental data and crystallochemical analysis obtained in the present study were able to substantiate the chemo-theoretical analysis. Our classifications agree well with experimental evidence regarding compounds that have already been studied. it would also be desirable to conduct new experimental studies in order to further evaluate the predicting capability of our calculations.
- [Show abstract] [Hide abstract] ABSTRACT: This study investigated pyrene adsorption on two contrasting Brazilian soils: a Kandiudult and a Vertisol. It was found that the time taken to reach thermodynamic equilibrium depended on the soil type. The curves for different pyrene-to-soil mass ratios for Vertisol soil showed significant differences. This is probably related to the presence of 2:1 clays that may increase the adsorption of pyrene due to the resulting interlamellar space. The adsorption of pyrene on the Kandiudult showed, in general, good agreement with the Langmuir isotherm. In the case of the Vertisol, there was good agreement with the linear isotherm. The kinetic model that best explains the adsorption in Kandiudult was the pseudo second-order model. For the Vertisol, the Morris Weber model best explains the behavior of pyrene. KeywordsKinetic–Adsorption–Hydrocarbons
Pontifícia Universidade Católica do Rio de Janeiro
Rio de Janeiro, Rio de Janeiro, Brazil
- Department of Chemistry (QUI)
University of Campinas
Conceição de Campinas, São Paulo, Brazil
- Institute of Chemistry