RESUMO

In recent decades, the scientific community has widely debated the contamination of fish in the Amazon region by mercury species. As the diet of riverside populations in the Amazon region is based mainly on fish, these populations are exposed to mercurial species that can cause serious and irreversible damage to their health. The risks of consuming fish exposed to mercurial species in the Amazon region have motivated toxicological investigations. However, the effect of mercurial species on protein and enzyme levels is still controversial. In this work, analytical and bioanalytical techniques Two-dimensional polyacrylamide gel electrophoresis [2D-PAGE] Graphite Furnace Atomic Absorption Spectrometry [GFAAS], and Mass Spectrometry in Sequence with Electrospray Ionization [ESI-MS/MS] were used to identify proteins associated with mercury (metal-binding protein) in muscle and liver tissues of the fish species Pinirampus pirinampu from the Madeira River, in the Brazilian Amazon. Enzymatic and lipid peroxidation analyses were also used to assess changes related to oxidative stress. Determinations of total mercury by GFAAS indicated higher concentrations in liver tissue (555 ± 19.0 µg kg-1) when compared to muscle tissue (60 ± 2.0 µg kg-1). The fractionation process of tissue proteomes by 2D-PAGE and subsequent mapping of mercury by GFAAS in the protein spots of the gels identified the presence of mercury in three spots of the liver tissue (concentrations in the range of 0.800 to 1.90 mg kg-1). The characterization of protein spots associated with mercury by ESI-MS/MS identified the enzymes triosephosphate isomerase A, adenylate kinase 2 mitochondrial, and glyceraldehyde-3-phosphate dehydrogenase as possible candidates for mercury exposure biomarkers. The muscle tissue did not show protein spots associated with mercury. Enzymatic activity decreased proportionally to the increase in mercury concentrations in the tissues.

Assuntos

RESUMO

Mercury dynamics in hydroelectric reservoirs have been studied worldwide. In tropical reservoirs, especially in those of the Amazon, the influence of geochemistry on Hg levels along this aquatic system is not well understood. The main objective of the present study was to assess the influence of environmental conditions (physical and chemical water parameters, trace element concentrations of sediment and sediment geochemistry) on mercury levels of sediment along the Balbina Reservoir (Amazon basin, Brazil). Sediment was collected along the reservoir and in the main tributaries in May 2015 (n = 10). These samples were assessed for labile iron (LFe), manganese (LMn), aluminum (LAl) and mercury (LHg) concentrations, total mercury (THg) concentrations, organic matter (OM) content, and granulometry. Concentrations in the sediment were 4-212 (LFe), 2-460 (LAl), 180-613 (LMn), < detection limit-256 (LHg), and 12-307 µg kg-1 dry weight (THg). In general, these concentrations decreased along the reservoir from upstream to downstream, reaching the lowest concentrations in the middle of the lake, and they increased in the sampling points near the dam. The lability and concentrations of mercury were influenced by the concentrations of LFe, LMn, LAl, and the granulometry and OM content of the sediment. Altogether, THg concentrations of sediments of the Balbina Reservoir encompass the range of concentrations of other reservoirs or natural lakes in the Amazon basin (from ten to a few hundred µg kg-1 dry weight).

Assuntos

RESUMO

Fish is an important source of protein, vitamins, and minerals. However, this food is also a major source of human exposure to toxic contaminants such as mercury. Thus, this paper aimed to evaluate mercury-binding proteins for possible application as biomarkers of mercury contamination in hepatic and renal tissues of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish) from the Amazon region using metalloproteomic approach. The proteome of hepatic and renal tissues of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). Finally, the protein spots associated to mercury were characterized by electrospray ionization mass spectrometry (ESI-MS/MS). The activity of antioxidant enzymes (SOD, CAT, GPx, and GST) and lipid peroxidation (LPO) were also determined. The results showed that the highest concentrations of mercury were found in the carnivorous species (P. squamosissimus) and that the accumulation pattern of this metal was higher in hepatic tissues than in renal tissues for both species. A tendency was observed for greater enzymatic activity in the hepatic and renal tissues of P. squamosissimus, the species with the highest concentration of mercury. Only GPx activity in the kidney and GST in the liver were lower for the P. squamosissimus species, and this finding can be explained by the interaction of mercury with these enzymes. The data obtained by ESI-MS/MS allowed for the characterization of the protein spots associated to mercury, revealing proteins involved in energy metabolism, biomolecules transport, protein synthesis and degradation, cell differentiation, gene regulation, and the antioxidant system. The results obtained in the present study can contribute to understanding the physiological processes underlying mercury toxicity and have provided new perspectives on possible candidates for mercury contamination biomarkers in fish.

Assuntos

RESUMO

The high concentrations of mercury found in Amazon have been intensively studied by the scientific community in the last decades. These mercurial species bind preferentially to proteins. Therefore, this work proposal sought to obtain the fractionation, identification and study of mercury - bound proteins present in samples of muscular and hepatic tissue from fish collected in the reservoir of the Jirau Hydroelectric Power Plant - on the Madeira River. Two-dimensional electrophoresis (2D-PAGE) for protein fractionation, graphite furnace atomic absorption spectrometry (GFAAS) for the quantification of mercury and Mass Spectrometry (ESI-MS/MS) were used for the identification of proteins. Concluding the work with analysis of graphics from the Blast2go program. Two mercury - bound proteins were identified as triosephosphate isomerase A and Protein FAM45A. The data generated by the bioinformatics programs confirm the tendency of these proteins to be linked to mercury and elucity the possibles existing physiological and cellular interactions.

Assuntos

RESUMO

High concentrations of mercury found in soils, sediments, fish, and humans of the Amazon region have gained prominence in scientific studies during the last decade. However, studies related to the elucidation of mercury toxicity mechanisms in ichthyofauna at the molecular and metallomic levels that seek to elucidate physiological and functional aspects, as well as the search for biomarkers of mercury exposure, are still sparse. In the search for these answers, the present study analyzed the hepatic tissue proteome of the Arapaima gigas (pirarucu) fish species collected in the Jirau hydroelectric power plant reservoir in the state of Rondônia state, Brazil, in order to identify mercury-related metal-binding proteins and to elucidate their physiological and functional aspects. The proteomic profile of the hepatic tissue of Arapaima gigas was obtained by two-dimensional electrophoresis (2D-PAGE) and the presence of mercury was mapped in the protein SPOTS by graphite furnace atomic absorption spectrometry(GFAAS). Mercury was detected in 18 protein SPOTS with concentrations ranging from 0.13 ±â€¯0.003 to 131.00 ±â€¯3 mg kg-1. The characterization of the protein SPOTS associated with mercury was performed by electrospray ionisation tandem mass spectrometry (ESI-MS/MS), and 10 proteins were identified. Bioinformatics analyses showed that most of the proteins found linked to mercury were involved in cellular component processes and biological processes. For the most part, protein sequences have cellular functions comprising catalytic, binding, sense of localization, and metabolic processes.

Assuntos

RESUMO

In the affiliation section, Luiz Fabricio Zara's affiliation "Pontifical Catholic University of Goiás (PUC), Goiânia, GO, Brazil" was incorrect. The correct affiliation is College of Planaltina, UnB - University of Brasília, Distrito Federal, Brazil.

RESUMO

This study presents data on the extraction and characterization of proteins associated with mercury in the muscle and liver tissues of jaraqui (Semaprochilodus spp.) from the Madeira River in the Brazilian Amazon. Protein fractionation was carried out by two-dimensional electrophoresis (2D-PAGE). Mercury determination in tissues, pellets, and protein spots was performed by graphite furnace atomic absorption spectrometry (GFAAS). Proteins in the spots that showed mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The highest mercury concentrations were found in liver tissues and pellets (426 ± 6 and 277 ± 4 µg kg-1), followed by muscle tissues and pellets (132 ± 4 and 86 ± 1 µg kg-1, respectively). Mercury quantification in the protein spots allowed us to propose stoichiometric ratios in the range of 1-4 mercury atoms per molecule of protein in the protein spots. The proteins characterized in the analysis by ESI-MS/MS were keratin, type II cytoskeletal 8, parvalbumin beta, parvalbumin-2, ubiquitin-40S ribosomal S27a, 39S ribosomal protein L36 mitochondrial, hemoglobin subunit beta, and hemoglobin subunit beta-A/B. The results suggest that proteins such as ubiquitin-40S ribosomal protein S27a, which have specific domains, possibly zinc finger, can be used as biomarkers of mercury, whereas mercury and zinc present characteristics of soft acids.

Assuntos

RESUMO

The aim was to compare the trace element concentrations found in samples of water (cadmium, copper, manganese, lead and zinc) from ten water catchments and muscle tissues (aluminum, iron, manganese, lead and zinc) from six fish trophic guilds, both samples collected in watercourses of the Upper Tocantins River, Goiás, Central Brazil. Trace element concentrations did not differ among the water catchments, except for Cu and Mn that exceeded the concentrations allowed by the environmental Brazilian legislation. The highest concentrations (Al, Fe, Mn, Pb e Zn) were found in fish of the guild detritivores, omnivores and terrestrial insectivores, while the aquatic invertivores showed the lowest ones; no correlation was observed between the concentrations of Mn, Pb and Zn of the water samples and fish muscle tissues. It suggests that the hydrological and chemical characteristics of watercourses sampled influence on a similar way the availability of trace elements in the water column. The differences of trace element concentrations observed among the trophic guilds do not display a trend based on fish feed strategy, but on the availability of the trace element in the environment, finding reinforced by the absence of relationship between trace elements concentrations of water and fish muscle tissue.

Objetivou-se comparar as concentrações dos elementos-traço encontrados em amostras de água (cádmio, cobre, manganês, chumbo e zinco) de dez sub-bacias e tecido muscular (alumínio, ferro, manganês, chumbo e zinco) de peixes pertencentes a seis guildas tróficas, ambas as amostras coletadas em cursos de água do alto rio Tocantins, Goiás, Brasil Central. As concentrações dos metais traços da água não diferiram entre sub-bacias, exceto para Cu e Mn que ultrapassaram estas indicadas pela legislação ambiental brasileira. As maiores concentrações (Al, Fe, Mn, Pb e Zn) foram encontradas nos peixes detritívoros, onívoros e insetívoros terrestres, enquanto que os invertívoros aquáticos apresentaram as menores concentrações, não se observou correlação entre as concentrações de Mn, Pb e Zn na água e nos peixes. Isto sugere que as características hidrológicas e químicas dos cursos de água amostrados influenciam de maneira semelhante a disponibilidade de elementos-traço na coluna de água. As diferenças de concentrações dos elementos-traço observadas entre as guildas tróficas não apresentam uma tendência com base na estratégia de alimentação dos peixes, mas relacionada à disponibilidade do elemento-traço no ambiente, resultado reforçado pela ausência de relação entre a concentração dos elementos-traço da água e do tecido muscular dos peixes.

Assuntos

RESUMO

The aim was to compare the trace element concentrations found in samples of water (cadmium, copper, manganese, lead and zinc) from ten water catchments and muscle tissues (aluminum, iron, manganese, lead and zinc) from six fish trophic guilds, both samples collected in watercourses of the Upper Tocantins River, Goiás, Central Brazil. Trace element concentrations did not differ among the water catchments, except for Cu and Mn that exceeded the concentrations allowed by the environmental Brazilian legislation. The highest concentrations (Al, Fe, Mn, Pb e Zn) were found in fish of the guild detritivores, omnivores and terrestrial insectivores, while the aquatic invertivores showed the lowest ones; no correlation was observed between the concentrations of Mn, Pb and Zn of the water samples and fish muscle tissues. It suggests that the hydrological and chemical characteristics of watercourses sampled influence on a similar way the availability of trace elements in the water column. The differences of trace element concentrations observed among the trophic guilds do not display a trend based on fish feed strategy, but on the availability of the trace element in the environment, finding reinforced by the absence of relationship between trace elements concentrations of water and fish muscle tissue.(AU)

Objetivou-se comparar as concentrações dos elementos-traço encontrados em amostras de água (cádmio, cobre, manganês, chumbo e zinco) de dez sub-bacias e tecido muscular (alumínio, ferro, manganês, chumbo e zinco) de peixes pertencentes a seis guildas tróficas, ambas as amostras coletadas em cursos de água do alto rio Tocantins, Goiás, Brasil Central. As concentrações dos metais traços da água não diferiram entre sub-bacias, exceto para Cu e Mn que ultrapassaram estas indicadas pela legislação ambiental brasileira. As maiores concentrações (Al, Fe, Mn, Pb e Zn) foram encontradas nos peixes detritívoros, onívoros e insetívoros terrestres, enquanto que os invertívoros aquáticos apresentaram as menores concentrações, não se observou correlação entre as concentrações de Mn, Pb e Zn na água e nos peixes. Isto sugere que as características hidrológicas e químicas dos cursos de água amostrados influenciam de maneira semelhante a disponibilidade de elementos-traço na coluna de água. As diferenças de concentrações dos elementos-traço observadas entre as guildas tróficas não apresentam uma tendência com base na estratégia de alimentação dos peixes, mas relacionada à disponibilidade do elemento-traço no ambiente, resultado reforçado pela ausência de relação entre a concentração dos elementos-traço da água e do tecido muscular dos peixes.(AU)

Assuntos

RESUMO

This study aimed to identify metalloproteins that lose their metal ions in the presence of mercury (Hg) and bind to Hg in breast milk samples collected from the riverine population of the Madeira River, a tributary of the Amazon River. Initially, total Hg was determined from the hair of lactating women to identify individuals who were contaminated followed by a proteomic analysis of breast milk samples through two-dimensional polyacrylamide gel electrophoresis after acetone precipitation. The presence of Hg in the obtained protein spots was determined by cold vapor atomic absorption spectrometry and cold vapor atomic fluorescence spectrometry. These determinations indicated the presence of Hg in one protein spot, which was then characterized through electrospray ionization tandem mass spectrometry. Based on searches in the UniProt database, this protein spot was identified as lysozyme C.

Assuntos