António O.S.S. Rangel

Remediation of heavy metal contaminated soils: phytoremediation as a potentially promising clean-up technology.

Increased soil pollution with heavy metals due to various human and natural activities has led to a growing need to address environmental contamination. Some remediation technologies have been developed to treat contaminated soil, but a biology-based technology, phytoremediation, is emerging. Phytoremediation includes phytovolatilization, phytostabilization, and phytoextraction using hyperaccumulator species or a chelate-enhancement strategy. To enhance phytoremediation as a viable strategy, microbiota from the rhizosphere can play an important role, but the use of genetic engineering can also increase the success of the technique. Here we review the key information on phytoremediation, addressing both potential and limitations, resulting from the research established on this topic.

Cell membrane damage induced by phenolic acids on wine lactic acid bacteria.

The aim of this work was to investigate the effect of phenolic acids on cell membrane permeability of lactic acid bacteria from wine. Several phenolic acids were tested for their effects on the cell membrane of Oenococcus oeni and Lactobacillus hilgardii by measuring potassium and phosphate efflux, proton influx and by assessing culture viability employing a fluorescence technique based on membrane integrity. The experimental results indicate that hydroxycinnamic acids (p-coumaric, caffeic and ferulic acids) induce greater ion leakages and higher proton influx than hydroxybenzoic acids (p-hydroxibenzoic, protocatechuic, gallic, vanillic, and syringic acids). Among the hydroxycinnamic acids, p-coumaric acid showed the strongest effect. Moreover, the exposure of cells to phenolic acids caused a significant decrease in cell culture viability, as measured by the fluorescence assay, in both tested strains. The results agree with previous results obtained in growth experiments with the same strains. Generally, phenolic acids increased the cell membrane permeability in lactic acid bacteria from wine. The different effects of phenolic acids on membrane permeability could be related to differences in their structure and lipophilic character.

Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis.

Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD(5)) (from an inlet of 420 to 1000 mg L(-1)) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L(-1)), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P<0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha(-1)d(-1) and 529 kg BOD(5)ha(-1)d(-1) were achieved for a loading ranging from 242 to 1925 kg COD ha(-1)d(-1) and from 126 to 900 kg BOD(5)ha(-1)d(-1). Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation.

Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment

This study focused on the diversity of bacterial communities from two series of two-stage constructed wetlands (CWs) treating tannery wastewater, under different hydraulic conditions. Series were separately planted with Typha latifolia and Phragmites australis in expanded clay aggregates and operated for 31 months. The effect of plant species, hydraulic loading and unit stage on bacterial communities was addressed through bacterial enumeration and denaturating gradient gel electrophoresis (DGGE). Diverse and distinct bacterial communities were found in each system unit, which was related in part to the type of plant and stage position (first or second unit in the series). Numerical analysis of DGGE profiles showed high diversity in each unit with an even distribution of species. No clear relation was established between the sample collection time, hydraulic loading applied and the bacterial diversity. Isolates retrieved from plant roots and substrates of CWs were affiliated with gamma-Proteobacteria, Firmicutes, alpha-Proteobacteria, Sphingobacteria, Actinobacteria and Bacteroidetes. Both series were effective in removing organic matter from the inlet wastewater, however, based on batch degradation experiments it seems that biodegradation was limited by the recalcitrant properties of the wastewater.

Inoculating Helianthus annuus (sunflower) grown in zinc and cadmium contaminated soils with plant growth promoting bacteria – Effects on phytoremediation strategies

Plant growth promoting bacteria (PGPR) may help reducing the toxicity of heavy metals to plants in polluted environments. In this work the effects of inoculating metal resistant and plant growth promoting bacterial strains on the growth of Helianthus annuus grown in Zn and Cd spiked soils were assessed. The PGPR strains Ralstonia eutropha (B1) and Chrysiobacterium humi (B2) reduced losses of weight in metal exposed plants and induced changes in metal bioaccumulation and bioconcentration - with strain B2 decreasing up to 67% Zn accumulation and by 20% Zn bioconcentration factor (BCF) in the shoots, up to 64% Zn uptake and 38% Zn BCF in the roots, and up to 27% Cd uptake and 27% Cd BCF in plant roots. The impact of inoculation on the bacterial communities in the rhizosphere of the plant was also assessed. Bacterial community diversity decreased with increasing levels of metal contamination in the soil, but in rhizosphere soil of plants inoculated with the PGPR strains, a higher bacterial diversity was kept throughout the experimental period. Inoculation of sunflower, particularly with C. humi (B2), appears to be an effective way of enhancing the short term stabilization potential of the plant in metal contaminated land, lowering losses in plant biomass and decreasing aboveground tissue contamination.

A review on sequential injection methods for water analysis

The development of fast, automatic and less expensive methods of analysis has always been the main aim of flow methodologies. The search for new procedures that still maintain the reliability and accuracy of the reference procedures is an ever growing challenge. New requirements are continually added to analytical methodologies, such as lower consumption of samples and reagents, miniaturisation and portability of the equipment, computer interfaces for full decision systems and so on. Therefore, the development of flow methodologies meeting the extra requirements of water analysis is a challenging work. Sequential injection analysis (SIA) presents a set of characteristics that make it highly suitable for water analysis. With sequential injection analysis, most routine determinations in waters can be performed more quickly with much lower reagent consumption when compared to reference procedures. Additionally, SIA can be a valuable tool for analyte speciation and multiparametric analysis. This paper critically reviews the overall work in this area.

Arsenic, lead and nickel accumulation in Rubus ulmifolius growing in contaminated soil in Portugal.

This work investigates the potential of Rubus ulmifolius, indigenous to a metal contaminated site--Esteiro de Estarreja--for phytoremediation purposes. The site has a long history of metal contamination. The accumulation of lead (Pb), arsenic (As) and nickel (Ni) in different sections--roots, stems and leaves - of the plant was assessed and compared to the levels of those metals in the soil and in the available fraction. The distribution of metals throughout the area was quite heterogeneous, presenting levels of As, Pb and Ni of up to 3078, 1400 and 135 mg kg(-1), respectively, and the metal content in the sections of R. ulmifolius collected in the banks of the stream varied among sites of collection. Levels of metals were higher in the plant roots: As levels (mgkg(-1)) ranged from 277 to 1721 in the roots, 30 to 110 in the stems, and 60 to 265 in the leaves; Pb concentrations (mgkg(-1)) ranged from 248 to 1178 in the roots, 35 to 133 in the stems, and 25 to 149 in the leaves; and Ni (mgkg(-1)) ranged from 48 to 151 in the roots. Significant correlations were found between the total levels of Pb and As in the soil and the levels in the roots of the plant; further correlations between total and available levels in the soil and metals in other plant tissues were generally found as non-significant. According to the metal accumulation patterns of R. ulmifolius, this species seems to be valuable for application in phytostabilisation strategies.

Evaluation of different substrates to support the growth of Typha latifolia in constructed wetlands treating tannery wastewater over long-term operation.

The aim of this study was to investigate the performance of horizontal subsurface flow constructed wetlands planted with Typha latifolia treating tannery wastewater under long-term operation. Two expanded clay aggregates (Filtralite MR3-8-FMR and Filtralite NR3-8-FNR) and a fine gravel-FG were used as substrate for the constructed wetland units plus one unit with FMR was left as an unvegetated control. The systems were subject to three hydraulic loadings, 18, 8 and 6cmd(-1), and to periods of interruption in the feed. The relationship between the substrate, plant development and removal efficiency, especially of organic matter, was investigated. Organic loadings up to 1800kg BOD(5)ha(-1)d(-1) and 3849kg COD ha(-1)d(-1) were applied leading to mass removals of up to 652kg BOD(5)ha(-1)d(-1) and 1869kg COD ha(-1)d(-1), respectively. The three different substrates were adequate for the establishment of T. latifolia, although the clay aggregates allowed for higher plant propagation levels. The units with FNR and FMR achieved significantly higher COD and BOD5 removal when compared to the FG and to the unplanted units. The systems proved to be tolerant to high organic loadings and to interruptions in feed suggesting this technology as a viable option for the biological treatment of tannery wastewater.

Spectrophotometric determination of iron and boron in soil extracts using a multi-syringe flow injection system.

In the last decade, significant advances in flow analysis have been reported, namely the extensive use of computer-controlled devices to enhance the autonomy and performance of analysers. In the present work, computer-controlled multi-syringe flow injection systems are proposed to perform the spectrophotometric determination of available iron and boron in soil extracts. The methodologies were based on the formation of ferroin complex (determination of iron) and azomethine-H reaction (determination of boron). Both determinations were performed in manifolds with similar configurations by changing the reagents present in the different syringes. In the determination of iron, elimination of Schlieren effect in the detection system was achieved through the binary sampling approach, where a three-way valve was actuated to intercalate small slugs of sample and reagent, promoting better mixing conditions for solutions with different values of refractive index. In the determination of boron, in-line sample blank measurement was attained by omitting the introduction of reagent through software control, without manifold reconfiguration. Linear calibration curves were established between 0.50 and 10.0mgFel(-1) and between 0.20 and 4.0mgBl(-1). No systematic difference was found when soil extracts were analysed by the proposed methodologies and compared to the respective reference procedures.

Evaluation of green coffee beans quality using near infrared spectroscopy: A quantitative approach

Characterisation of coffee quality based on bean quality assessment is associated with the relative amount of defective beans among non-defective beans. It is therefore important to develop a methodology capable of identifying the presence of defective beans that enables a fast assessment of coffee grade and that can become an analytical tool to standardise coffee quality. In this work, a methodology for quality assessment of green coffee based on near infrared spectroscopy (NIRS) is proposed. NIRS is a green chemistry, low cost, fast response technique without the need of sample processing. The applicability of NIRS was evaluated for Arabica and Robusta varieties from different geographical locations. Partial least squares regression was used to relate the NIR spectrum to the mass fraction of defective and non-defective beans. Relative errors around 5% show that NIRS can be a valuable analytical tool to be used by coffee roasters, enabling a simple and quantitative evaluation of green coffee quality in a fast way.