Biophysical and monetary quantification of ecosystem services in a mountain region: the case of avoided soil erosion

Environment, Development and Sustainability - Even though ecosystem services have a significant contribution to human well-being, their value is not always recognized. Despite difficulties, the...     

Restoring biochemical activity and bacterial diversity in a trichloroethylene-contaminated soil: the reclamation effect of vermicomposted olive wastes

Background, aim, and scope  In this work, the potential for using olive-mill solid waste as an organic amendment for biochemical and biological restoration of a trichloroethylene-contaminated soil, which has previously been stabilized through vermicomposting processes, has been explored. Materials and methods  Trichloroethylene-contaminated water was pumped into soil columns with a layer of vermicompost at 10-cm depth (biobarrier system). The impacts of the trichloroethylene on the microbial community were evaluated by determining: (1) the overall microbial activity (estimated as dehydrogenase activity) and enzyme activities related to the main nutrient cycles (β-glucosidase, o-diphenoloxidase, phosphatase, urease, and arylsulphatase activities). In addition, isoelectric focusing of the soil extracellular humic-β-glucosidase complexes was performed to study the enzymatically active humic matter related to the soil carbon cycle. (2) The soil bacterial diversity and the molecular mechanisms for the bacterial resistance to organic solvents were also determined. For this, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to detect changes in bacterial community structure and PCR-single-strand conformational polymorphism (SSCP) was developed and optimised for detection and discrimination of the resistance-nodulation-division (RND) genes amplified from the contaminated soils. Results  Vermicompost reduced, with respect to the unamended soil, about 30% of the trichloroethylene leaching during the first month of the experiment. Trichloroethylene had a marked negative effect on soil dehydrogenase, β-glucosidase, urease, phosphatase, and arylsulphatase activities. Nevertheless, the vermicompost tended to avoid this toxic effect. Vermicompost also displays stable humic-β-glucosidase complexes that increased the extracellular activity related to C-cycle in the contaminated soils. The isoelectric focusing technique showed a more biochemically active humic matter in the soil sampled under the vermicompost. The behaviour of the three main phyla of bacteria isolated from the DGGE bands was quite different. Bands corresponding to Actinobacteria disappeared, whereas those affiliated with Proteobacteria remained after the trichloroethylene contamination. The disappeared Actinobacteria became visible in the soil amended with the vermicompost. Bands corresponding to Bacteriodetes appeared only in columns of contaminated soils. In this study, six types of RND proteins were detected by PCR-SSCP in the natural soil, three in the trichloroethylene-contaminated soil and 7/5 in trichloroethylene-contaminated soil above/below the vermicompost in the biobarrier columns. Trichloroethylene tended to reduce or eliminate all the clones detected in the uncontaminated soil, whereas new efflux pumps appeared in the biobarrier columns. Discussion  Although enzymes incorporated into the humic substances of vermicomposted olive wastes are quite stable, trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity in the amended soils. The decrease was less severe in the biobarrier system, but in any case, no relation was found between the levels of trichloroethylene in soil and extracellular β-glucosidase activity, or between the latter and the quantity of humic carbon in soils. The isoelectric focusing technique was carried out in the humic fraction to determine whether the loss of activity occurred in overall extracellular β-glucosidase or in that linked to stable humic substances (humic–enzyme complexes). The contaminated soils showed the lower enzyme activities, whereas contaminated and amended soils presented greater quantity of focalised (and therefore stable) humic carbon and spectra heterogeneity: very different bands with higher enzyme activities. No clear relationship between trichloroethylene concentration in soil and diversity of the bacterial population was noted. Similar patterns could be found when the community structures of bacteria and microbial activity were considered. Since the use of the dehydrogenase assay has been recognised as a useful indicator of the overall measure of the intensity of microbial metabolism, these results could be attributed to PCR-DGGE methodology, since the method reveals the presence of dominant populations regardless of their metabolic state. Trichloroethylene maintained or even increased the number of clones with the DNA encoding for RND proteins, except for the contaminated soil located above the vermicompost. However, the main effect of trichloroethylene was to modify the structure of the community in contaminated soils, considering the type of efflux pumps encoded by the DNA extracted from soil bacteria. Conclusions  Trichloroethylene inhibited specific functions in soil and had a clear influence on the structure of the autochthonous bacterial community. The organic matter released by the vermicomposted olive waste tended to avoid the toxic effect of the contaminant. Trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity, even when vermicompost was present. In this case, the effect of the vermicompost was to provide and/or to stimulate the humic-β-glucosidase complexes located in the soil humic fraction >10⁴, increasing the resistance of the enzyme to the inhibition. The bacterial community from the soil presented significantly different mechanisms to resistance to solvents (RND proteins) under trichloroethylene conditions. The effect of the vermicompost was to induce these mechanisms in the autochthonous bacterial community and/or incorporated new bacterial species, able to grow in a trichloroethylene-contaminated ambient. Coupled biochemical and molecular methodologies are therefore helpful approaches in assessing the effect of an organic amendment on the biochemical and biological restoration of a trichloroethylene-contaminated soil. Recommendations and perspectives  Since the main biochemical and biological effects of the organic amendment on the contaminated soil seem to be the incorporation of biochemically active humic matter, as well as new bacterial species able to grow in a trichloroethylene-contaminated ambient, isoelectric focusing and PCR-SSCP methodologies should be considered as parts of an integrated approach to determine the success of a restoration scheme.     

Mutagenic properties of PM2.5 urban pollution in the Northern Italy: The nitro-compounds contribution

PM2.5 is the breathable fraction of the particulate matter and some adverse health effects, such as respiratory functionality, cardiological diseases and cancer, can be in some measure attributable to this risk factor exposure. Some of the most carcinogen compounds transported by PM2.5 are nitro-compounds. In this study, a strengthened in vitro bioassay — able to predict the mutagenic/carcinogenic activity of the environmental mixtures — was conducted on PM2.5 organic extracts to define the nitro-compounds burden. PM2.5 air pollution was daily monitored, during 2006, in three cities located in the Northern part of Italy (Torino, Pavia and Verona) and the mutagenic properties of the PM2.5 organic extracts were assessed with the Ames test. The bacterial used in this study were three Salmonella typhimurium strains: TA98, nitroreductase-less mutant TA98NR and YG1021 carrying a nitroreductase-producing plasmid. The annual PM2.5 mean level measured in Torino was 46.5 (± 31.6) μg/m³, in Pavia 34.8 (± 25.1) μg/m³, and in Verona 37.3 (± 27.8) μg/m³, while the mutagenicity expressed as TA98 net reverants/m³ was 28.0 (± 22.1), 28.3 (± 24.9), and 34.2 (± 30.9) respectively. Monthly pool bioassays, conducted with the three different strains, showed a greater mutagenic response of the YG1021 in each city. The relationship among the mutagenic answers for YG1021:TA98:TA98NR was about 6:3:1 (p < 0.001). Over nitroreductase activity enhanced the response of 2.2, 2.0 and 1.7 times for Torino, Pavia, and Verona (ANOVA Torino p < 0.05) respectively. Without nitroreductase activity the genotoxicity was limited. These biological findings are able to describe a relevant role played by the nitro compounds in the mutagenic properties of the urban PM2.5 in the Padana plain; moreover the bacterial nitroreductase plays a predominant role in DNA interaction primarily for Torino PM2.5 extracts.     

Monitoring pollution in River Mureş, Romania, part II: metal accumulation and histopathology in fish

As a part of an exposure and effect monitoring conducted along the river Mureş, Western Romania in 2004, the health status of two indigenous fish species, sneep (Chondrostoma nasus) and European chub (Leuciscus cephalus) was investigated upstream and downstream the city of Arad. In fish, histopathology was assessed in liver and gills, and heavy metals (cadmium, copper, lead and zinc) were analyzed in liver samples. In both fish species, histopathological reactions in the gills (epithelial lifting, focal proliferation of epithelial cells of primary and secondary lamellae and resulting fusion of secondary lamellae, hyperplasia and hypertrophy of mucous cells, focal inflammation and necrosis of epithelial cells) were most severe at the two sampling sites upstream Arad city, which were shown to be polluted by copper, cadmium, faecal coliforms and streptococci in a parallel study. At these two sites, also histopathology in the liver of L. cephalus was more prominent than at the two downstream sites. In C. nasus, symptoms in the liver (focal inflammation with lymphocytic infiltrations, macrophage aggregates and single cell necrosis) were also highly pronounced at the sampling site located directly downstream the municipal sewage treatment plant of Arad. With the exception of copper accumulation in L. cephalus caught at the most upstream sampling site, in both fish species cadmium and copper accumulation were exceptionally high and did not differ significantly between the four sampling sites.     

Metabolic and bacterial diversity in soils historically contaminated by heavy metals and hydrocarbons

The aim of this study was to characterize soils contaminated by different levels of heavy metals and hydrocarbons (Madonna Dell'Acqua, Pisa, Italy). The soils were chemically and biochemically analysed by measuring the standard chemical properties and some enzyme activities related to microbial activity (dehydrogenase activity) and the soil carbon cycle (total and extracellular beta-glucosidase activities). The metabolic capacities of soil microorganisms to degrade hydrocarbons through catechol 2,3-dioxygenase were also described. The microbial diversity of contaminated and uncontaminated soils was estimated by denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences. The PCR/single-strand conformation polymorphism (PCR/SSCP) method was used to estimate the genetic diversity of PAH-degrading genes in both contaminated and uncontaminated soils. A greater bacterial diversity and lower catechol 2,3-dioxygenase activity was detected in unpolluted soils. The complexity of the microbial community (Shannon and Simpson indices) as well as the dehydrogenase soil activity negatively correlated with contamination levels. The greatest PAH-degrading gene diversity and the most intense catechol 2,3-dioxygenase activity were found in the soils with the highest levels of hydrocarbons. Heavy metals and hydrocarbon pollution has caused a genetic and metabolic alteration in microbial communities, corresponding to a reduction in microbial activity. A multi-technique approach combining traditional biochemical methods with molecular-based techniques, along with some methodological improvements, may represent an important tool to expand our knowledge of the role of microbial diversity in contaminated soil.     

Metallothionein levels in Algerian mice (Mus spretus) exposed to elemental pollution: an ecophysiological approach

The potential use of metallothioneins (MTs) as biomarkers of trace metal contamination was evaluated for the first time in the Algerian mouse (Mus spretus). Mice were collected seasonally in an abandoned mining area (Aljustrel) and in a reference area, both located in southern Portugal. MT levels were quantified in liver and kidney by differential pulse polarography and hepatic elemental concentrations (Mn, Fe, Cu, Zn, Se) were determined by particle-induced X-ray emission. Hepatic iron and selenium concentrations were elevated in mice from Aljustrel mine when compared to reference animals. MTs levels were averagely higher in mice from Aljustrel than those originated from the reference area. A season-dependent significant effect was found on the hepatic and renal MT concentrations, characterized by higher levels in winter and lower in autumn. In contaminated mice positive relationship between liver elemental contents (Cu in autumn and Fe in winter) and MTs were found. The seasonal variation of MT suggests that probably physiological and environmental factors could influence hepatic and renal MT induction. Results seem to imply that some environmental disturbance occur in the vicinity of the Aljustrel mine. Therefore, for the management purposes MT levels should be followed in liver of M. spretus, especially in winter. Furthermore, other physiological factors that could influence MT expression and turnover in Algerian mouse should also be monitored.     

Comparative study of hotplate wet digestion methods for the determination of mercury in biosolids

The re-use of biosolids is becoming increasingly popular for land applications. However, biosolids may contain elevated levels of metals and metalloids (including mercury) relative to background environmental concentrations. Consequently, reliable mercury analysis is important to allow classification of biosolids and to determine appropriate options for beneficial uses. This paper reports on a comparative study of 12 hotplate wet digestion methods for their suitability for the determination of mercury in biosolids. The methods were applied to mercury biosolids samples from four localities of two different sewage treatment plants in the State of Victoria, Australia. Samples were also spiked with methylmercury chloride and mercury sulphide to evaluate the Hg recovery in each hotplate digestion method. Aqua regia (HCl:HNO(3)=3:1), reverse aqua regia (HCl:HNO(3)=1:3), nitric, hydrochloric, sulphuric acid and their combinations with or without hydrogen peroxide were studied as wet digestion solutions. The method providing the best mercury recoveries was optimized. Under optimal conditions the corresponding analytical procedure consisted of 1h pre-digestion of 0.4 g biosolids sample with 10 ml reverse aqua regia with temperature increasing to 110 degrees C and 3h digestion at this temperature. In the last 10 min of the digestion step, 2 ml hydrogen peroxide were added to ensure complete decomposition of all mercury containing compounds. After filtering and dilution with deionised water (1:10), the concentration of mercury was determined by cold vapour atomic absorption spectrometry. It is expected, that the wet acid digestion method developed in this study will be also applicable to biosolids from other sewage treatment plants and to other types of solid mercury samples with elevated levels of organic matter.     

Soil vapor extraction in sandy soils: influence of airflow rate

Airflow rate is one of the most important parameters for the soil vapor extraction of contaminated sites, due to its direct influence on the mass transfer occurring during the remediation process. This work reports the study of airflow rate influence on soil vapor extractions, performed in sandy soils contaminated with benzene, toluene, ethylbenzene, xylene, trichloroethylene and perchloroethylene. The objectives were: (i) to analyze the influence of airflow rate on the process; (ii) to develop a methodology to predict the remediation time and the remediation efficiency; and (iii) to select the most efficient airflow rate. For dry sandy soils with negligible contents of clay and natural organic matter, containing the contaminants previously cited, it was concluded that: (i) if equilibrium between the pollutants and the different phases present in the soil matrix was reached and if slow diffusion effects did not occur, higher airflow rates exhibited the fastest remediations, (ii) it was possible to predict the remediation time and the efficiency of remediation with errors below 14%; and (iii) the most efficient remediation were reached with airflow rates below 1.2 cm(3)s(-1) standard temperature and pressure conditions.     

Hydrolytic degradation of azimsulfuron, a sulfonylurea herbicide

The chemical degradation of the herbicide azimsulfuron was investigated in aqueous solutions at different pH values. The hydrolysis rate, determined by HPLC analyses, was pH dependent and was much faster in acidic than in neutral or weakly basic conditions. The metabolites formed at different pH values were compared with standards when possible or isolated and identified using ESI-LC-MS/MS, (1)H NMR and (13)C NMR. The two main products of hydrolysis in mild acidic solution were identified as 2-amino-4,6-dimethoxy-pyrimidine and 2-methyl-4-(2-methyl-2H-tetrazol-5-yl)-2H-pyrazole-3-sulfonamide, both produced as a result of the sulfonylurea bridge cleavage. Under basic conditions, a new product, a substituted 2-pyrimidinamine, deriving from the contraction of the sulfonylurea bridge, was isolated and completely characterized for the first time.     

The environmental behaviour of polychlorinated phenols and its relevance to cork forest ecosystems: a review

Pentachlorophenol (PCP) has been used as a herbicide, biocide and preservative worldwide since the 1930s and as a result, extensive and prolonged contamination exists. The environmental impact increases when its many degradation products are taken into consideration. A number of chloroanisols and their related chlorophenols have been found in cork slabs collected from Portuguese oak tree forests before stopper manufacturing, and contamination by PCP and polychlorinated anisole (PCA) has been detected in Canadian forests. It is suggested that the use of polychlorinated phenols, in particular PCP, is thought to be a cause of the cork taint problem in wine, a major socio-economic impact not only for industry but on sensitive and highly biodiverse ecosystems. It also highlights particular issues relating to the regional regulation of potentially toxic chemicals and global economics world wide. To fully understand the impact of contamination sources, the mechanisms responsible for the fate and transport of PCP and its degradation products and assessment of their environmental behaviour is required. This review looks at the current state of knowledge of soil sorption, fate and bioavailability and identifies the challenges of degradation product identification and the contradictory evidence from field and laboratory observations. The need for a systematic evaluation of PCP contamination in relation to cork forest ecosystems and transfer of PCP between trophic levels is emphasised by discrepancies in bioaccumulation and toxicity. This is essential to enable long term management of not only transboundary contaminants, but also the sustainable management of socially and economically important forest ecosystems.