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Hepatitis E virus (HEV) infection is a common cause of acute viral hepatitis in tropical regions. In Brazil, HEV G3 is the only genotype detected to date. Reports on HEV prevalence are heterogeneous. We aimed to compare the prevalence of anti-HEV among three populations living in the Brazilian Amazon basin. Two cross-sectional studies were conducted in urban, rural, and Yanomami indigenous areas. Plasma samples from 428 indigenous and 383 non-indigenous subjects were tested for anti-HEV IgG using enzyme-linked immunosorbent assays. The overall prevalence of anti-HEV was 6.8% (95%CI: 5.25-8.72), with 2.8% (12/428) found in the Yanomami areas, 3% (3/101) in an urban area, and 14.2% (40/282) in a rural area. Multivariate logistic analysis indicated that patients aged 31-45 years or ≥46 years are more likely to present anti-HEV positivity, with a respective aOR of 2.76 (95%CI: 1.09-7.5) and 4.27 (95%CI: 1.58-12.35). Furthermore, residence in a rural area (aOR: 7.67; 95%CI: 2.50-33.67) represents a relevant risk factor for HEV infection. Additional studies detecting HEV RNA in fecal samples from both humans and potential animal reservoirs are necessary to comprehensively identify risk factors associated with HEV exposure.

Hepatitis E virus (HEV) infection is a common cause of acute viral hepatitis in tropical regions. In Brazil, HEV G3 is the only genotype detected to date. Reports on HEV prevalence are heterogeneous. We aimed to compare the prevalence of anti-HEV among three populations living in the Brazilian Amazon basin. Two cross-sectional studies were conducted in urban, rural, and Yanomami indigenous areas. Plasma samples from 428 indigenous and 383 non-indigenous subjects were tested for anti-HEV IgG using enzyme-linked immunosorbent assays. The overall prevalence of anti-HEV was 6.8% (95%CI: 5.25–8.72), with 2.8% (12/428) found in the Yanomami areas, 3% (3/101) in an urban area, and 14.2% (40/282) in a rural area. Multivariate logistic analysis indicated that patients aged 31–45 years or ≥46 years are more likely to present anti-HEV positivity, with a respective aOR of 2.76 (95%CI: 1.09–7.5) and 4.27 (95%CI: 1.58–12.35). Furthermore, residence in a rural area (aOR: 7.67; 95%CI: 2.50–33.67) represents a relevant risk factor for HEV infection. Additional studies detecting HEV RNA in fecal samples from both humans and potential animal reservoirs are necessary to comprehensively identify risk factors associated with HEV exposure.

The Araguari River, one of the most important waterways in the Brazilian state of Amapá, is ecologically relevant and essential for the conservation of Amazonian fish biodiversity. Our previous studies demonstrated contamination of water and fish by metals. In particular, water samples showed genotoxic damage in Danio rerio. Here, we extended our studies of potential genotoxic damage to native fish from sampling sites located in the Araguari River lower section. To accomplish this, we collected samples of fish with different feeding behaviors, but in the same sampling sites, and evaluated the same genotoxicity biomarkers in erythrocytes. All eleven species of fish collected in the Araguari River lower section showed profiles and frequencies of genotoxic damage similar to those previously observed in tests with D. rerio, confirming that genotoxic pollutants present in these waters are also causing damage to native fish species. [ABSTRACT FROM AUTHOR]

Recently, high concentrations of metals have been found in the waters from sampling sites located in the Araguari River lower section. These metals can be bioaccumulated by fish, but also biomagnified, representing a potential risk through human consumption. Here, we aimed to (1) determine the metal concentrations in muscle samples from fish; (2) evaluate bioaccumulation of metals in muscle tissue of fish species with different feeding behaviors; and (3) investigate the risks of single metals, as well as mixture of metals, to human fish consumption. Eleven fish species were sampled (3 carnivorous, 5 omnivorous, and 3 detritivorous). In four fish species, Cd and Pb concentrations exceeded the Brazilian maximum limits (MLs) for human consumption. Individually, Pb and Cd concentrations in the muscle of these fish indicate risks to human health. When considering the mixture of metals, the consumption of all fish species represents risks. The highest risks implicate detritivorous species. For all fish species evaluated, the estimated daily intake (EDI) of Hg shows values above its reference dose (RfD). Our results suggest the need to establish an environmental monitoring program that aims to preserve environmental quality, biodiversity, and human health. It is also necessary to develop actions that aim to educate Amazonian populations on safe fish preparation and consumption. [ABSTRACT FROM AUTHOR]

This study aimed to characterize a Klebsiella pneumoniae strain (KP411) recovered from the stool samples of poultry (Gallus gallus) in the Brazilian Amazon Region. The whole-genome sequencing of KP411 revealed the presence of an important arsenal of antimicrobial resistance genes to β-lactams (blaCTX-M-14, blaTEM-1B, blaKPC-2, blaSVH-11), aminoglycosides [aph(3″)- Ib, aph(6)-Id, aph(3′)-Ia], sulfonamides (sul1, sul2), quinolones (oqxAB), fosfomycin (fosAKP), and macrolides [mph(A)]. Furthermore, our analyses revealed that the KP411 strain belongs to the ST258 clonal lineage, which is one of the main epidemic clones responsible for the dissemination of KPC-2 worldwide. Our data suggest that food-producing animals may act as reservoirs of multidrug-resistant K. pneumoniae belonging to the ST258 clone, and, consequently, contribute to their dissemination to humans and the environment. [ABSTRACT FROM AUTHOR]

This study aimed to characterize a Klebsiella pneumoniae strain (KP411) recovered from the stool samples of poultry (Gallus gallus) in the Brazilian Amazon Region. The whole-genome sequencing of KP411 revealed the presence of an important arsenal of antimicrobial resistance genes to β-lactams (blaCTX-M-14, blaTEM-1B, blaKPC-2, blaSVH-11), aminoglycosides [aph(3″)- Ib, aph(6)-Id, aph(3′)-Ia], sulfonamides (sul1, sul2), quinolones (oqxAB), fosfomycin (fosAKP), and macrolides [mph(A)]. Furthermore, our analyses revealed that the KP411 strain belongs to the ST258 clonal lineage, which is one of the main epidemic clones responsible for the dissemination of KPC-2 worldwide. Our data suggest that food-producing animals may act as reservoirs of multidrug-resistant K. pneumoniae belonging to the ST258 clone, and, consequently, contribute to their dissemination to humans and the environment.

The human pressure on nature makes it imperative understanding how environmental issues can influence the world view of society. The aim of the current article is to measure the environmental attitude of the urban population living in the Macapá City, Amapá state, Brazilian Amazon region and to check whether this attitude is influenced by sociodemographic variables. We collected 387 full interviews conducted with individuals. In order to do so, we used the New Ecological Paradigm (NEP) tool composed of 15 items distributed in five facets. Results showed positive correlations between the NEP scale and variables "education level" and "annual income," as well as negative correlation between such scale and variable "age." Urban childhood place recorded higher pro-ecological attitude rate than the rural one. All facets, except for limits, recorded moderate-to-high scores, thus indicating that the population has positive pro-environmental attitude profile, although it did not show much concern about the increased human population on Earth. [ABSTRACT FROM AUTHOR]

We refer to Oswaldo Cruz’s reports dating from 1913 about the necessities of a healthcare system for the Brazilian Amazon Region and about the journey of Carlos Chagas to 27 locations in this region and the measures that would need to be adopted. We discuss the risks of endemicity of Chagas disease in the Amazon Region. We recommend that epidemiological surveillance of Chagas disease in the Brazilian Amazon Region and Pan-Amazon region should be implemented through continuous monitoring of the human population that lives in the area, their housing, the environment and the presence of triatomines. The monitoring should be performed with periodic seroepidemiological surveys, semi-annual visits to homes by health agents and the training of malaria microscopists and healthcare technicians to identify Trypanosoma cruzifrom patients’ samples and T. cruziinfection rates among the triatomines caught. We recommend health promotion and control of Chagas disease through public health policies, especially through sanitary education regarding the risk factors for Chagas disease. Finally, we propose a healthcare system through base hospitals, intermediate-level units in the areas of the Brazilian Amazon Region and air transportation, considering the distances to be covered for medical care.

Epidemiological studies generally use particulate matter measurements with diameter less 2.5 μm (PM 2.5 ) from monitoring networks. Satellite aerosol optical depth (AOD) data has considerable potential in predicting PM 2.5 concentrations, and thus provides an alternative method for producing knowledge regarding the level of pollution and its health impact in areas where no ground PM 2.5 measurements are available. This is the case in the Brazilian Amazon rainforest region where forest fires are frequent sources of high pollution. In this study, we applied a non-linear model for predicting PM 2.5 concentration from AOD retrievals using interaction terms between average temperature, relative humidity, sine, cosine of date in a period of 365,25 days and the square of the lagged relative residual. Regression performance statistics were tested comparing the goodness of fit and R 2 based on results from linear regression and non-linear regression for six different models. The regression results for non-linear prediction showed the best performance, explaining on average 82% of the daily PM 2.5 concentrations when considering the whole period studied. In the context of Amazonia, it was the first study predicting PM 2.5 concentrations using the latest high-resolution AOD products also in combination with the testing of a non-linear model performance. Our results permitted a reliable prediction considering the AOD-PM 2.5 relationship and set the basis for further investigations on air pollution impacts in the complex context of Brazilian Amazon Region. [ABSTRACT FROM AUTHOR]

This study presents an environmental assessment of a bi-fuel Thermal Power Plant (TPP) in the Brazilian Amazon Region. The TPP was originally designed to use Heavy Fuel Oil (HFO), but with the Natural Gas (NG) extraction in the Amazon basin, the TPP was modified to use NG and HFO concomitantly. In order to observe the differences, a comparative study of this power plant was performed in two situations: using only heavy fuel oil and using HFO and NG concomitantly. The life cycle assessment methodology was conducted from “cradle-to-gate”. The CML baseline method was used to assess the midpoint impacts. Primary data were collected by personal visits for environmental emissions, wastewater, fuel used, and technical specifications. Eleven impact categories were considered and Monte Carlo (MC) Analysis was used to assess and to obtain robust results. The study shows that upstream processes have more impacts except global warming potential, eutrophication, acidification, human toxicity and photochemical oxidation where TPP showed the main contribution. The bi-fuel conversion resulted in lower power plant participations in the impacts as, for example, a reduction of 61.1% in Eutrophication Potential. In addition, throughout the life cycle the bi-fuel power plant, when the operation is compared with HFO only, has six potential impacts lower. Furthermore, this study showed that bi-fuel operation has lower local and regional impact, and according to the MC analysis, statistically, bi-fuel operation does not contribute to exacerbate global trends in GHG emissions when compared with monofuel operation. It is an interesting observation that global warming results by MC analysis showed that there are no statistical differences between the two modes of operation. [ABSTRACT FROM AUTHOR]

Recentemente é descrito estado de persistência do vírus da hepatite B denominado hepatite crônica B oculta. Sua prevalência e fisiopatologia são desconhecidas. O objetivo deste estudo foi avaliar a ocorrência dessa entidade clínica em pacientes da Amazônia brasileira. De 51 pacientes anti-HBc total reativos testados pela reação em cadeia da polimerase, 17% foram positivos. Não observamos associação com fatores de risco clássicos de infecção pelo vírus da hepatite B, testes bioquímicos, hematológicos e histopatologia. No entanto, os pacientes ictéricos e reativos para o anti-HIV apresentaram associação com a presença do ADN-vírus da hepatite B. Os resultados demonstram a ocorrência da hepatite crônica B oculta, entre nossos doentes, porém, com taxas de prevalência abaixo do esperado para a região. Acreditamos que, apesar do tamanho da amostra avaliada ser pequeno, sua ocorrência poderia ter sido maior se empregássemos primers para a região S, C e X do genoma do vírus da hepatite B, aumentando a sensibilidade do teste.A persistent form of the hepatitis B virus called occult chronic hepatitis B has recently been described. Its prevalence and physiopathology are unknown. The aim of this study was to evaluate the occurrence of this clinical entity among patients in the Brazilian Amazon region. Out of 51 anti-HBc total-positive patients who were tested using the polymerase chain reaction, 17% were positive. We did not find any associations with classical risk factors for hepatitis B virus infection or with biochemical tests, hematological tests or histological patterns. However, the jaundiced and HIV-positive patients showed a statistical association with the presence of hepatitis B virus-DNA. The results demonstrated that occult hepatitis B occurred among our patients, but at prevalence rates lower than expected for this region. We believe that despite the small sample size, the occurrence might have been found to be greater if we had used primers for the S, C and X regions of the hepatitis B virus genome, thereby increasing the sensitivity of the test.

The transmission of malaria in Brazil is heterogeneous throughout endemic areas and the presence of asymptomatic Plasmodium sp. carriers (APCs) in the Brazilian Amazon has already been demonstrated. Malaria screening in blood banks is based on the selection of donors in respect to possible risks associated with travel or residence, clinical evidence and/or inaccurate diagnostic methods thereby increasing the probability of transfusion-transmitted infection. We evaluated the frequency of APCs in four blood services in distinct areas of the Brazilian Amazon region. DNA was obtained from 400 human blood samples for testing using the phenol-chloroform method followed by a nested-PCR protocol with species-specific primers. The positivity rate varied from 1 to 3% of blood donors from the four areas with an average of 2.3%. All positive individuals had mixed infections for Plasmodium vivax and Plasmodium falciparum. No significant differences in the results were detected among these areas; the majority of cases originated from the transfusion centres of Porto Velho, Rondônia State and Macapá, Amapá State. Although it is still unclear whether APC individuals may act as reservoirs of the parasite, efficient screening of APCs and malaria patients in Brazilian blood services from endemic areas needs to be improved.A transmissão da malária no Brasil é heterogênea em todas as áreas endêmicas e a presença de portadores assintomáticos de Plasmodium sp. (PAPs) na Amazônia brasileira já foi demonstrada. A triagem de pacientes maláricos em bancos de sangue é baseada na seleção dos doadores com relação aos riscos possíveis associados com residência, evidência clínica e/ou os métodos diagnósticos não acurados que aumentam a probabilidade da infecção transmitida por transfusão. Avaliamos a freqüência de PAPs em quatro bancos de sangue em áreas distintas da região Amazônica brasileira. O DNA foi obtido a partir de 400 amostras de sangue humano usando o método do fenol-clorofórmio, seguido por um protocolo de nested-PCR com oligonucleotídeos espécie-específicos. A taxa de positividade variou de 1 a 3% de doadores do sangue das quatro áreas, com uma média de 2,3%. Todos os indivíduos positivos tinham infecções mistas entre o Plasmodium vivax e o Plasmodium falciparum. Nenhuma diferença significativa nos resultados foi detectada entre estas áreas; a maioria dos casos originou dos Hemocentros de Porto Velho, do Estado de Rondônia e de Macapá, Estado do Amapá. Embora ainda não esteja claro se os indivíduos PAPs possam agir como reservatórios do parasito, a triagem eficiente de PAPs e de pacientes com malária em bancos de sangue no Brasil das áreas endêmicas necessita ser implementada.

The characterization of behavioral patterns allows a better understanding of the transmission dynamics and the design of more effective malaria vector control strategies. This study analyzed the behavioral patterns of the Anopheles species of the Coração district situated in the northeast of the Brazilian Amazon region. The behavioral patterns of the anopheline species were measured based on the 36 collection sites of this district from December 2010 to November 2011. Collections of four hours for three consecutive nights each month and four 12-h collections, comprising two in the rainy season and two in the dry season, were performed. Furthermore, to infer the anthropophily and zoophily indexes, four additional four-hour collections were performed. The samples were also evaluated for parity rate and natural infectivity for Plasmodium spp. A total of 1689 anophelines were captured, comprising of nine species and two subgenera ( Nyssorhynchus – six species, and Anopheles – three species). Anopheles darlingi was the most abundant and widely distributed species in the area, followed by A. braziliensis and A.marajoara . Anopheles darlingi and A. marajoara were the only species present in the four collections of 12-h, but only A. darlingi showed activity throughout night. Anopheles darlingi was the most anthropophilic species (AI = 0.40), but the zoophily index was higher (ZI = 0.60), revealing an eclectic and opportunistic behavior. Of the six most frequent species, A. nuneztovari s.l. was the most zoophilic species (ZI = 1.00). All captured species showed predominance towards biting in outdoor environments. Anopheles darlingi and A. braziliensis showed multimodal biting peaks, whereas A. marajoara revealed a stable pattern, with the biting peak after sunset. Using the PCR technique, no anopheline was found infected with the malaria parasite. Since A. darlingi and A. marajoara are recognized as important vectors in this region, the district of Coração may be considered as a highly potent area for transmission of malaria, therefore, the prevention and surveillance measures should be taken constantly to prevent the same. The role of A. braziliensis as malaria vector needs to be urgently investigated. [ABSTRACT FROM AUTHOR]

Having a river basin as the study site, it is of fundamental importance to comprehend the sediment dynamics of river systems, in order to diagnose and understand them, aiming to carry out predictions. The observance of the sediment supply carried by the river systems is related to the erosion, transport and deposition, through rainfall and dynamic land use and cover in hillside environments. The Purus River basin is part of this context, the study area of this paper and it is located in the Amazon biome. This study aims to start the first steps to understand the suspended sediment dynamics of that basin, because of its expressive environmental variations. For this purpose was considered the sediment present in the river systems as an indicator which can be understood and used in order to support soil and water conservation measures in the Purus River basin, distinguishing the natural dynamics and anthropogenic dynamics. Thus, this study has the objective of evaluating the suspended sediments discharge in the Purus River basin, in order to collaborate with action planning and watershed management, with the objective of generating knowledge about the suspended sediment dynamics of that spring. [ABSTRACT FROM PUBLISHER]

Knowledge of the embryonic development of species such as quail is important for our understanding of its production and reproduction. Quail provides a practical solution to the animal protein shortage problem in developing countries and is an excellent alternative to chicken (Shanaway, 1994). This study evaluated quail embryonic viability by comparing the main morphological changes that occur over the development in a homemade incubator with previous data; the incubator was built using easily accessible and low-cost materials, which allows small producers and communities to develop quail farms as an activity to increase income or even produce a source of animal protein for the community in the Brazilian Amazon region. The homemade incubator measured 40 cm × 42 cm × 32 cm and was constructed with wooden boards, a digital thermostat, and an incandescent lamp. A total of 24 fertile eggs were incubated at a temperature of 37.5 °C and 60% relative humidity; age "0" was set at the start of incubation. Two eggs were opened on each day of development, starting from day 5 to day 16 of incubation. After gently opening the eggs, the embryos were removed, separated from the placenta and amniotic fluid, washed with running water, and weighed on a digital scale (SHIMADZU AUY 220 brand and UNI BLOC model). Crown-rump length and other measurements were performed with a digital caliper and the morphological structures were observed using a stereomicroscope (NOVA brand and NOVA ZTX-E model). All eggs opened contained live embryos, indicating that the homemade incubator temperature, relative humidity, and manual turning were adequate to maintain the quail's viability. The morphometric data of the embryos at different developmental time points were similar to those described in the literature. Embryos weighed 0.0069 g at day 5 and 4.7863 g at day 16, and the crown-rump length (CRL) means were 0.368 cm and 3.657 cm, respectively. Weight and CRL increased by 0.434 g and 2.593 cm per day of incubation, respectively. In conclusion, after 16 days of incubation in a homemade incubator, the embryos presented all phases of the development cycle, with appropriate development of weight and height based on the days of incubation without anomaly or external interference. These findings demonstrate that the homemade incubator does not alter the embryonic development of quail embryos in the Brazilian Amazon region., Competing Interests: Conflict of interests: ACM, GCS, RTO, TSF, AALB and ASLS – No conflict of interest.

Blood sampling on filter paper is a current practice seroepidemiological studies by indirect fluorescent antibody test (IFAT). There is, however, scant comparative information about the use of bloodspot eluates for detection of malarial IgG antibodies simultaneously by IFAT and enzyme immunoassay (ELISA). Here we report data obtained by both serological methods done on 219 bloodspot eluate samples collected in a rural community in Brazilian Amazon Basin (Alto Paraíso, Ariquemes municipality) where malaria is endemic. Plasmodium falciparum and P. vivax thick smear antigens were used in the IFAT; a detergent-soluble P. falciparum antigen was prepared for ELISA. Substantial agreement of results (Kappa coefficient k = 0.686) was observed when P. falciparum antigen was used in both tests, and IFAT titers were found to be strongly correlated ELISA antibody units (Spearman correlation coeficient rs = 0.818, p < 0.0001). Only moderate agreement (k = 0.467) between IFAT with P. vivax antigen and ELISA with P. falciparum antigen was observed. Spearman correlation coefficient value between quantitative results (IFAT titers and ELISA antibody units) in this case was numerically lowe (rs = 0.540, p < 0.0001). Our results suggest that, with P. falciparum antigen, both IFAT and ELISA performed on bloodspot eluates are equivalent for seropidemiological purposes.

Abstract: The aim of this study was to assess the prevalence pattern of mixed-Plasmodium falciparum malaria infections in Brazil by molecular diagnosis and to address its clinically important features. DNA was extracted from 115 thick blood film P. falciparum human blood positive samples using the phenol–chloroform method, followed by a semi-nested PCR protocol with species-specific primers. Seventy-three percent of P. falciparum single infections and 26.95% of mixed infections were found. Amongst mixed infections, the majority was double infection (96.77%). Our results suggest that the prevalence of one species over the other can be important on weakening P. falciparum malaria clinical symptoms. We confirm that P. falciparum co-infections frequently occur in Brazilian malaria endemic areas, with underestimated diagnosis. The results point to the need of improving microscopy or changing for another accurate diagnosis technique to differentiate among human malaria species, as this is essential to choose the best treatment and control measure for malaria. More investigations are necessary in order to clarify the role of mixed-infections in the severity of P. falciparum disease. [Copyright &y& Elsevier]

We compared the serological phenotypic frequencies of ABO, MNSs, and Duffy in 417 blood donors and 309 malaria patients from four Brazilian Amazon areas. Our results suggest no correlation between ABO phenotype and malaria infection in all areas studied. We observed significant correlation between the S+s+, S+s -, and S-s+ phenotypes and malaria infection in three areas. Some of the Duffy phenotypes showed significant correlation between donors and malaria patients in different areas. These data are an additional contribution to the establishment of differential host susceptibility to malaria. [ABSTRACT FROM AUTHOR]

Carbon isotope data on soil organic matter (SOM) were collected along an ecosystem transect 90 m in length that includes a tropical forest on the plateau, a transitional forest-savanna and savanna in a depression. Total organic carbon data show a significant increase in carbon content from sites representing forest ecosystem to sites representing savanna ecosystem. It was hypothesized that carbon accumulation in the depression is controlled by flooding conditions that slow down carbon decomposition and in part by carbon transport from the upper part of the transect (the savanna and the transition forest-savanna areas) into the depressions by water during the rainy season. The origin of the carbon was confirmed by using soil 13 C analysis. The savanna sites located in the depression showed d 13 C values between –19.5‰ and –22.5‰ indicating a mixture of C3 and C4 plants. The vegetation cover in the depression is predominantly C3 grasses with d 13 C values of about –27‰ and –26‰. In the site under savanna located at an elevation slightly higher, the d 13 C value was more enriched (–16‰) showing the predominance of C4 plants ( d 13 C of –13.6‰). At the forest-savanna transition and in the forest ecosystem the d 13 C values were characteristic of C3 plants (–25‰ and –28.1‰). 14 C and 13 C data indicate that the organic matter of mixed origin has been deposited for at least the last 7000 years in the savanna depressions. The 13 C pattern observed in the soil organic matter profiles indicate a predominance of C3 plants in the early part of the Holocene. About 7000 to 4000 years ago, the data show the influence of C4 plants, indicating forest regression associated with a drier climate than at present. The more recent 13 C records suggest forest expansion, and the return to a climate similar to the present. [ABSTRACT FROM AUTHOR]

JC virus (JCV) is a member of the Polyomaviridae family and is associated to a severe disease known as progressive multifocal leukoencephalopathy, PML, which is gradually increasing in incidence as an opportunistic infection among AIDS patients. The present study aimed to investigate the occurrence of JCV among HIV-1 carriers including their types and molecular subtypes and the possible association with disease. Urine samples from 66 HIV-1 infected subjects were investigated for the presence of the virus by amplifying VP1 (215bp) and IG (610bp) regions using the polymerase chain reaction. JCV was detected in 32% of the samples. The results confirmed the occurrence of type B (subtype Af2); in addition, another polyomavirus, BKV, was also detected in 1.5% of samples of the HIV-1 infected subjects. Apparently, there was no significant difference between mono- (HIV-1 only) and co-infected (HIV-1/JCV) subjects regarding their TCD4(+)/TCD8(+) lymphocyte counts or HIV-1 plasma viral load. Self admitted seizures, hearing and visual loses were not significantly different between the two groups., (Copyright © 2016 Elsevier Editora Ltda. All rights reserved.)