Phylogenetic diversity and functional characterization of the Manila clam microbiota: a culture-based approach

According to the hologenome theory, the microbiota contributes to the fitness of the holobiont having an important role in its adaptation, survival, development, health, and evolution. Environmental stress also affects the microbiota and its capability to assist the holobiont in coping with stress factors. Here, we analyzed the diversity of cultivable bacteria associated with Manila clam tissues (mantle, gills, hemolymph) in two non-contaminated sites (Portugal and France) and one metal-contaminated site (Portugal). A total of 240 isolates were obtained. Representative isolates (n = 198) of the overall diversity were identified by 16S rDNA sequencing and subjected to functional characterization. Isolates affiliated with Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes. Proteobacteria (mostly Pseudoalteromonadaceae and Vibrionaceae) were dominant in non-contaminated sites while Actinobacteria (mostly Microbacteriaceae) dominated in the metal-contaminated site. The main factor affecting the microbiota composition was contamination. No significant differences were observed between clam tissues and geographic regions. Several isolates tested positive for antibacterial activity, biofilm formation, protease, and siderophore production. The results show that the Manila clam harbors a diverse microbiota that may contribute to clam protection and overall fitness, as well as to its adaptation to stressful environments. In addition, the Manila clam microbiota is revealed as a promising source of novel probiotics with potential application in aquaculture.     

Method of median semi-variance for the analysis of left-censored data: Comparison with other techniques using environmental data

In environmental monitoring, variables with analytically non-detected values are commonly encountered. For the statistical evaluation of these data, most of the methods that produce a less biased performance require specific computer programs. In this paper, a statistical method based on the median semi-variance (SemiV) is proposed to estimate the position and spread statistics in a dataset with single left-censoring. The performances of the SemiV method and 12 other statistical methods are evaluated using real and complete datasets. The performances of all the methods are influenced by the percentage of censored data. In general, the simple substitution and deletion methods showed biased performance, with exceptions for L/2, Inter and L/√2 methods that can be used with caution under specific conditions. In general, the SemiV method and other parametric methods showed similar performances and were less biased than other methods. The SemiV method is a simple and accurate procedure that can be used in the analysis of datasets with less than 50% of left-censored data.     

Analytical methods and validation for determining trace elements in red wines

The objective of this work is to quantify As, Hg, Cd, Ni and Pb in Portuguese red wines. First, the methods for the quantification of trace elements in red wines were validated. Several pre-treatments were compared, namely a pre-digestion process with HNO₃, a pre-oxidation step with H₂O₂, and a spiking step of wine samples with a known concentration of the trace elements analyzed. Except for As, it was determined that the quantification of the trace elements does not require a pre-digestion process with HNO₃. For all of the trace elements analyzed, a pre-oxidation step with H₂O₂ may enable an accurate quantification. The techniques chosen for the quantification of trace elements were hydride generation atomic absorption spectrometry (HGAAS) for As and Hg, electrothermal atomic absorption spectrometry (ETAAS) for Cd, and flame atomic absorption spectrometry (FAAS) for Ni and Pb. In the second stage of this work, 25 Portuguese red wines spanning all of the red wine-producing regions were analyzed for all of the five trace elements referred to above. Only Cd and Pb have shown concentrations above the limit values defined by the “Organization Internationale de la Vigne et du Vin.” The Target Hazard Quotient (THQ) equation was used to determine in which wine-producing regions that wine consumption can be a problem for public health in terms of the concentrations of the five trace elements analyzed. THQ values have indicated that for the universe of the 25 red wines analyzed no region produces wines that can pose problems for public health, when the Portuguese red wine standard consumption is considered.     

Assessment of metolachlor and diuron leaching in a tropical soil using undisturbed soil columns under laboratory conditions

In the present study, diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] and metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-metoxi-1-methylethyl)acetamide] leaching was studied in undisturbed soil columns collected in a cotton crop area in Mato Grosso State, Brazil. The pesticides were applied to the soil surface in dosages similar to those used in a cotton plantation. To assess the leaching process, soil columns were submitted to simulated rain under laboratory conditions at 25 ± 3°C, in the absence of wind and direct solar radiation. During the rain simulations, leachate solutions were collected and herbicide concentrations were determined. At the end of the experiment, the soil columns were cut into 10 cm sections to determine the remaining herbicide concentrations through the soil profile. Metolachlor was detected in all soil sections, and approximately 4% of the applied mass was leached. Diuron was detected only in the upper two soil sections and was not detected in the leachate. A linear correlation (r > 0.94) between the metolachlor soil concentrations and the organic contents of the soil sections was observed. Mass balance suggests that around 56% of diuron and 40% of metolachlor were degraded during the experiments. Measurements of the water table depth in the area where the samples were collected showed that it varied from 2 to 6 m and is therefore vulnerable to contamination by the studied herbicides, particularly metolachlor, which demonstrated a higher leaching potential.     

Speciation and quantification of Hg in sediments contaminated by artisanal gold mining in the Gualaxo do Norte River,Minas Gerais,SE, Brazil

The Iron Quadrangle in SE Brazil was, in the eighteenth century, one of the most important Au producing regions of Brazil. In this region, gold is produced, even today, by artisanal methods that use Hg to increase the extraction efficiency with no control of Hg release to water systems and the atmosphere. In this context, the Gualaxo do Norte River is of particular interest; its springs are located in the Doce River basin, an important Brazilian basin that supplies water for 3.5 million people. The main goal of this work was to quantify and speciate the Hg in the sediments of the Gualaxo do Norte River using a direct mercury analyzer and gas chromatography-pyrolysis-atomic fluorescence detection system. Statistical analyses consisted of principal component analysis, aiming to assess interactions among elements and species and to group the variables in factors affecting the properties of sediment. The results show that total Hg (THg) and methylmercury (CH₃Hg⁺) concentrations in samples ranged from 209 to 1207 μg kg^?1 and from 0.07 to 1.00 μg kg^?1, respectively (methylation percentages from 0.01 to 0.27%). Thermal desorption analysis showed that mercury is mainly present in the oxidized form, and correlation analyses pointed to a relationship between THg and MnO, indicating that manganese can oxidize and/or adsorb Hg. Together, MO and CH₃Hg⁺ are important parameters in the third principal component, indicating the influence of OM on the methylation process. This first investigation on Hg methylation in this small-scale gold mining area points to the possibility of Hg bioaccumulation and to the need of better understanding the biogeochemical cycle of Hg in this area. Samples were collected in 2012, prior to the 2015 Fundão Dam disaster. The results are also a record of the characteristics of the sediment prior to that event.     

Influence of composition on the biomethanation potential of restaurant waste at mesophilic temperatures

A synthetic waste was used to study the effect of waste composition on anaerobic degradation of restaurant waste. It was made by blending melted pork lard, white cabbage, chicken breast, and potato flakes, to simulate lipids, cellulose, protein, and carbohydrates, respectively. Four blends of the four constituents with an excess of each component were assayed and compared with a fifth blend containing an equal amount of chemical oxygen demand (COD) of each of the four components. The methane production and the time course of soluble COD and volatile fatty acids were assessed in batch assays. A high reduction of volatile solids (between 94% and 99.6%) was obtained in all the assays. The methane yield was between 0.40 m(3) CH(4)/kg VS(initial) (excess of carbohydrates) and 0.49 m(3) CH(4)/kg VS(initial) (excess of lipids). The degradation of the lipid-rich assays differed from the others. Fifty percent of the biochemical methane potential was obtained after 3-6 days for all of the assays, except for the one with excess of lipids which achieved 50% methanation only after 14.7 days of incubation. In the assay with excess of lipids, a considerable fraction of COD remained in the liquid phase, suggesting an inhibition of the methanogenic process that was likely due to the accumulation of long chain fatty acids. The hydrolysis rate constants, assuming first order kinetics, over the first 6 days were between 0.12d(-1) (excess of lipids) and 0.32 d(-1) (excess of carbohydrates). The results indicate that anaerobic digestion facilities with large variations in lipid input could have significant changes in process performance that merit further examination.     

Water quality of the Ribeirão Preto Stream, a watercourse under anthropogenic influence in the southeast of Brazil

It is known that Brazil still has a privileged position of water quantity and quality, but water use has not proceeded in a responsible manner and often results in impairment of quality. This study aims to evaluate limnological parameters, parasites and bacteria, and concentrations of heavy metals (Cd, Pb, Cu, Cr, Mn, Hg, and Zn) in surface water of Ribeirão Preto Stream. The Ribeirão Preto Stream is located in urban areas under anthropogenic influence. The results showed that the levels of dissolved oxygen values were lower than those established by the National Environmental Council (CONAMA Resolution No 357/2005). The reading of electrical conductivity showed values typical of impacted environments. The parasitological analysis revealed the presence of nematode larvae. The bacteriological analysis showed higher values for total coliform and Escherichia coli than those set by the Brazilian National Environment Council (CONAMA). The heavy metals Cd, Pb, Cu, Cr, Mn, Hg, and Zn showed concentrations in accordance with the guidelines established by CONAMA. The results provide data on the quality of these waters and showed the necessity to protect the watercourse from point sources of contamination, recommending their continued monitoring.     

Distribution and sources of PAHs using three pine species along the Ebro River

Needles of three pine species (Pinus halepensis, Pinus pinea and Pinus nigra) were analysed to assess the occurrence of polycyclic aromatic hydrocarbons (PAHs) in 34 sites located throughout the Ebro River, in Northeast Spain. Overall, the concentration varied between 55 and 808 ng g^− 1 (dry weight). The three- and four-ring PAHs were the most representative, with phenanthrene having 43% of the total PAH load and naphthalene showing a high incidence in rural areas. Despite matrix apparent similarities, P. halepensis needles revealed higher entrapment levels than P. nigra and P. pinea, the latter showing the lowest levels. The assessment of possible sources using PAH ratios (phenanthrene/anthracene and fluoranthene/pyrene) did not reveal a clear tendency regarding the distinction of petrogenic and pyrogenic sources in general, reflecting heterogeneous sources of PAHs in the Ebro area.     

Mobility and ecological risk assessment of trace metals in polluted estuarine sediments using a sequential extraction scheme

The three-stage sequential extraction procedure for the fractionation of Cd, Cr, Cu, Ni, Pb, and Zn, proposed by the Commission of the European Communities Bureau of Reference, was applied to sediment five samples collected from the Sal estuary, Sergipe State, northeast Brazil, in September 2009. The method showed satisfactory recoveries, detection limits, and standard deviations for determinations of trace metals in the sediments. Cd and Pb were the metals most prevalent in the bioavailable fractions (carbonates, Fe and Mn oxides, organic matter, and sulfides), while Ni, Zn, Cu, and Cr showed higher percentages in the inert fraction. The order of mobility of the metals was Cd (66 %) > Pb (65 %) > Zn (59 %) > Ni (57 %) = Cr (57 %) > Cu (56 %). Possible toxicity related to these metals was examined using the risk assessment code, and by comparing the chemical data with sediment quality guideline ERL-ERM values. Results obtained using the two methods were in agreement, and showed low to medium risk for all metals, indicating that adverse effects on aquatic biota should rarely occur.     

Utilization of UV‐A and UV‐C radiation in advanced oxidation processes with the intention of removing color and organic matter from the gross leachate of a stabilized sanitary landfill

The present work investigated color and biochemical oxygen demand (COD) removal from treated landfill leachate via advanced oxidative processes (AOPs) artificially emitted. The AOP H₂O₂/UV, TiO₂/UV, and photolysis were tested in two bench‐scale photoreactors: The first one with UV‐C lamp and the other with UV‐A lamp associated or not with coagulation / flocculation pretreatment. Crude leachate samples with pH ranging from 8 to 3 were used, and time varied in 30, 45, and 60 minutes. Experiments were performed in two stages: step 1 with a 20 mL volume from each sample and step 2 with repetition of the best results from the previous step, adopting the 150 mL volume. In step 1, the AOP applied in the crude leachate sample showed the best results, standing out H₂O₂/UV‐C with 30 minutes and TiO₂/UV‐A with 60 minutes. In step 2, H₂O₂/UV‐C had a 60% color removal and 25% COD reduction, while TiO₂/UV‐A had a 10% color removal and 20% COD reduction. Therefore, the H₂O₂/UV‐C process was the most efficient, because the complex characteristics of the effluent interfered in the TiO₂/UV‐A efficiency, but it is necessary to eliminate the process interferences. The use of artificial radiation is a viable alternative; however, it can be costly, being relevant in the associations between processes with artificial UV and solar UV, as the natural radiation becomes more attractive and allows the process operating costs reduction.