Paula M. L. Castro

Phytoremediation of polyaromatic hydrocarbons, anilines and phenols.

Phytoremediation technologies based on the combined action of plants and the microbial communities that they support within the rhizosphere hold promise in the remediation of land and waterways contaminated with hydrocarbons but they have not yet been adopted in large-scale remediation strategies. In this review plant and microbial degradative capacities, viewed as a continuum, have been dissected in order to identify where bottlenecks and limitations exist. Phenols, anilines and polyaromatic hydrocarbons (PAHs) were selected as the target classes of molecule for consideration, in part because of their common patterns of distribution, but also because of the urgent need to develop techniques to overcome their toxicity to human health.Depending on the chemical and physical properties of the pollutant, the emerging picture suggests that plants will draw pollutants including PAHs into the plant rhizosphere to varying extents via the transpiration stream. Mycorrhizosphere-bacteria and -fungi may play a crucial role in establishing plants in degraded ecosystems. Within the rhizosphere, microbial degradative activities prevail in order to extract energy and carbon skeletons from the pollutants for microbial cell growth. There has been little systematic analysis of the changing dynamics of pollutant degradation within the rhizosphere; however, the importance of plants in supplying oxygen and nutrients to the rhizosphere via fine roots, and of the beneficial effect of microorganisms on plant root growth is stressed.In addition to their role in supporting rhizospheric degradative activities, plants may possess a limited capacity to transport some of the more mobile pollutants into roots and shoots via fine roots. In those situations where uptake does occur (i.e. only limited microbial activity in the rhizosphere) there is good evidence that the pollutant may be metabolised. However, plant uptake is frequently associated with the inhibition of plant growth and an increasing tendency to oxidant stress. Pollutant tolerance seems to correlate with the ability to deposit large quantities of pollutant metabolites in the ‘bound’ residue fraction of plant cell walls compared to the vacuole. In this regard, particular attention is paid to the activities of peroxidases, laccases, cytochromes P450, glucosyltransferases and ABC transporters. However, despite the seemingly large diversity of these proteins, direct proof of their participation in the metabolism of industrial aromatic pollutants is surprisingly scarce and little is known about their control in the overall metabolic scheme. Little is known about the bioavailability of bound metabolites; however, there may be a need to prevent their movement into wildlife food chains. In this regard, the application to harvested plants of composting techniques based on the degradative capacity of white-rot fungi merits attention.

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.

4-Chlorophenol degradation by a bacterial consortium: development of a granular activated carbon biofilm reactor

Abstract A bacterial consortium that can degrade chloro- and nitrophenols has been isolated from the rhizosphere of Phragmitis communis. Degradation of 4-chlorophenol (4-CP) by a consortium attached to granular activated carbon (GAC) in a biofilm reactor was evaluated during both open and closed modes of operation. During the operation of the biofilm reactor, 4-CP was not detected in the column effluent, being either adsorbed to the GAC or biodegraded by the consortium. When 4-CP at 100 mg l−1 was fed to the column in open mode operation (20 mg g−1 GAC total supply), up to 27% was immediately available for biodegradation, the rest being adsorbed to the GAC. Biodegradation continued after the system was returned to closed mode operation, indicating that GAC bound 4-CP became available to the consortium. Biofilm batch cultures supplied with 10–216 mg 4-CP g−1 GAC suggested that a residual fraction of GAC-bound 4-CP was biologically unavailable. The consortium was able to metabolise 4-CP after perturbations by the addition of chromium (Cr VI) at 1–5 mg l−1 and nitrate at concentrations up to 400 mg l−1. The development of the biofilm structure was analysed by scanning electron microscopy and confocal laser scanning microscopy (CLSM) techniques. CLSM revealed a heterogeneous structure with a network of channels throughout the biofilm, partially occupied by microbial exopolymer structures.

Application of a statistical design to the optimization of culture medium for recombinant interferon-gamma production by Chinese hamster ovary cells.

SummaryThe importance of serum-free medium components on the growth of Chinese hamster ovary (CHO) cells and production of recombinant human interferon(IFN)-gamma was investigated. The complexity of the medium led to the adoption of a statistical optimization approach based on a Plackett-Burman design. From this analysis a set of nutritional components was identified as important for cell growth and recombinant protein production. Glycine was identified as an important determinant of specific growth rate, whereas for cell production bovine serum albumin (BSA), phenylalanine and tyrosine were also identified as important. BSA, sodium pyruvate, glutamate, methionine, proline, histidine, hydroxyproline, tyrosine and phenylalanine were shown to be important for IFN-gamma production. Other medium components, such as insulin, arginine, aspartate and serine produced an inhibitory effect on both cell growth and IFN-gamma production. The effect of the stimulatory nutrients as a whole group was tested by increasing their concentration in the medium. A significant improvement in specific cell growth rate, cell production and IFN-gamma production (up to 45%) was achieved on both shake-flask and fermentor cultures. An increase in the medium concentration of the negative variables had only a small inhibitory effect (approximately 10%) on the same parameters. Analysis of the effects of the group of stimulatory amino acids and BSA on CHO cell growth showed that the effect of the former was independent of BSA.

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.

The mycorrhizal status of Phragmites australis in several polluted soils and sediments of an industrialised region of Northern Portugal

Abstract Roots of Phragmites australis from three polluted soils and sediments (a periodically flooded stream bank containing organic pollutants, a high-pH drying sedimentation pond and an acidic, periodically flooded sand polluted by industrial effluents) were sampled over a 1-year cycle of plant growth to assess the degree of colonisation by arbuscular mycorrhizal fungi (AMF). At the dry sedimentation pond, root samples of Juncus effusus and Salix atrocinerea were also taken to assess the presence of AMF throughout the year. Root colonisation was low (<5% root length colonised) but arbuscule presence peaked in P. australis during the spring and autumn prior to flowering. These changes in arbuscule abundance were also seen in a parallel greenhouse trial using seed taken from one of the sites. Roots of J. effusus contained mainly vesicular colonisation but arbuscule activity peaked during the winter months (December–March). S. atrocinerea roots were found to be ectomycorrhizal throughout the year but the fine feeder roots were colonised by AMF. The results confirm that semi-aquatics, like P. australis, can become arbuscular mycorrhizal but that this status changes during the year depending on soil moisture content and plant phenology. The influence of AMF in these polluted soils is uncertain but the potential exists to establish a more diverse plant ecosystem during the landscaping of these areas (phytostabilisation) by management of adapted plant and AMF ecotypes.

Metal Uptake by Microalgae: Underlying Mechanisms and Practical Applications

Metal contamination of a few aquatic, atmospheric, and soil ecosystems has increased ever since the industrial revolution, owing to discharge of such elements via the effluents of some industrial facilities. Their presence to excessive levels in the environment will eventually lead to serious health problems in higher animals owing to accumulation throughout the food web. Current physicochemical methods available for recovery of metal pollutants (e.g., chemical precipitation, oxidation/reduction, or physical ion exchange) are either expensive or inefficient when they are present at very low concentrations. Consequently, removal of toxic metals by microorganisms has emerged as a potentially more economical alternative. Microalgae (in terms of both living and nonliving biomass) are an example of microorganisms suitable to recover metals and able to attain noteworthy percent removals. Their relatively high metal‐binding capacities arise from the intrinsic composition of their cell walls, which contain negatively charged functional groups. Consequently, microalgal cells are particularly efficient in uptake of those contaminants when at low levels. Self‐defense mechanisms developed by microalgal cells to survive in metal‐containing media and environmental factors that affect their removal (e.g., pH, temperature, and biomass concentration) are reviewed here in a comprehensive way and further discussed in attempts to rationalize this form of remediation vis‐a‐vis with conventional nonbiological alternatives.

Bacterial community dynamics in horizontal flow constructed wetlands with different plants for high salinity industrial wastewater polishing

This study is focused on the diversity of bacterial communities from two series of horizontal subsurface flow constructed wetlands (CW) polishing high salinity tannery wastewater. Each series was planted with Arundo donax or Sarcocornia sp. in a substrate composed by expanded clay and sand. Chemical and biochemical oxygen demand removal efficiencies were similar in each series, varying between 58 and 67% (inlet COD 218 ± 28 mg L(-1)) and 60 and 77% (inlet BOD(5) 37 ± 6 mg L(-1)), respectively. High numbers of culturable bacteria were obtained from substrate and root samples - 5.75 × 10(6)-3.95 × 10(8) CFU g(-1) recovered on marine agar and 1.72 × 10(7)-8.46 × 10(8) CFU g(-1) on nutrient agar. Fifty bacterial isolates were retrieved from the CW, related phylogenetically to Firmicutes, Actinobacteria, Bacteroidetes, α-, β-, and γ-Proteobacteria. Changes in the bacterial communities, from roots and substrate of each series, related to the plant species, hydraulic loading rates and along CW operation were examined using denaturating gradient gel electrophoresis (DGGE). The clustering analysis suggested that a diverse and distinct bacterial community inhabits each series, which was related to the type of plant present in each CW.