Phytostabilization potential of Erica australis L. and Nerium oleander L.: a comparative study in the Riotinto mining area (SW Spain).

Phytostabilization is a green, cost-effective technique for mine rehabilitation and ecological restoration. In this study, the phytostabilization capacity of Erica australis L. and Nerium oleander L. was assessed in the climatic and geochemical context of the Riotinto mining district, southwestern Spain, where both plant species colonize harsh substrates of mine wastes and contaminated river banks. In addition to tolerating extreme acidic conditions (up to pH 3.36 for E. australis), both species were found to grow on substrates very poor in bioavailable nutrients (e.g., N and P) and highly enriched with potentially phytotoxic elements (e.g., Cu, Cd, Pb, S). The selective root absorption of essential elements and the sequestration of potentially toxic elements in the root cortex are the main adaptations that allow the studied species to cope in very limiting edaphic environments. Being capable of a tight elemental homeostatic control and tolerating extreme acidic conditions, E. australis is the best candidate for use in phytostabilization programs, ideally to promote early stages of colonization, improve physical and chemical conditions of substrates and favor the establishing of less tolerant species, such as N. oleander.

Effect of two different composts on soil quality and on the growth of various plant species in a polymetallic acidic mine soil.

The effect of the addition (0-10%) of two types of sewage sludge composts (composted sewage sludge [CS] and sewage sludge co-composted with olive prune wastes [CSO]) on a polymetallic acidic soil from the Riotinto mining area was evaluated by i) a soil incubation experiment and ii) a greenhouse pot experiment using tomato (Solanum lycopersicum Mill.), ryegrass (Lolium perenne L.) and ahipa (Pachyrhizus ahipa (Wedd.) Parodi). Compost addition improved the soil organic carbon content, increased the pH and the electrical conductivity and enhanced enzyme activities and soil respiration, more for CSO than for CS. Plant growth was generally enhanced after compost addition, but not proportionally to the dose. Foliar concentrations of some hazardous elements (As, Cr, Fe) in tomato growing in non-amended soil were above the thresholds, questioning the adequacy of using this plant species. However, leaf concentrations of essential and potentially toxic elements (Fe, As, Cr and Pb) in tomato and/or ryegrass were reduced after the amendment with both composts, generally more for CSO than for CS. Conversely, foliar concentrations in ahipa, a plant species which is able to grow without the need of compost addition, were safe except for As and were only slightly affected by compost addition. This plant species would be a suitable candidate due to its low requirements and due to the limited element translocation to the leaves. Concerning the composts, amelioration of plant and soil properties was better accomplished when using CSO, a compost of sewage sludge and plant remains, than when using CS, which only contained the sludge.

Optimizing the combined application of amendments to allow plant growth in a multielement-contaminated soil.

This study was aimed to 1) properly understand the dynamics of toxic elements (Al, Fe, Mn, Cu, Pb, Zn and As) in a sulphide-mine soil after combined application of compost from urban sewage sludge (SVC) and bottom ashes from biomass combustion (BA) and to 2) optimize the combination of both amendments for vegetation growth. Soil was amended following a D-optimal design and the mixtures (15 in total) were incubated during 30 d. At the end of the incubation, the effects of amendments on the assessed variables as well as the process modelling were evaluated by Response Surface Methodology (RSM). The process modelling confirmed that quadratic models were adequate to explain the behaviour of the assessed variables (R(2) ≥ 0.94 and Q(2) ≥ 0.75). Both amendments significantly increased pH and electrical conductivity, while reduced metal extractability. A different behaviour of As respect to metals was observed and high doses of BA sharply increased its extractability. The optimization process indicated that adequate conditions for vegetation growth would be reached adding the soil with 6.8% of SVC and 3.1% of BA (dry weight). After amendments application the germination and root elongation of three energy crops were significantly increased while lipid peroxidation was decreased. Therefore, the combined application of SVC and BA to a contaminated soil could improve soil conditions and might be expected to have an advantage during plant growth. Moreover, the RSM could be a powerful technique for the assessment of combined amendment effects on soil properties and their effective application in multielement-contaminated soils.

Field approach to mining-dump revegetation by application of sewage sludge co-compost and a commercial biofertilizer.

An approach was devised for revegetation of a mining dump soil, sited in a semiarid region, with basic pH as well as Fe and Mn enrichment. A field experiment was conducted involving the use of co-compost (a mixture of urban sewage sludge and plant remains) along with a commercial biofertilizer (a gel suspension which contains arbuscular mycorrhizal fungus) to reinforce the benefits of the former. Four treatments were studied: unamended soil; application of conditioners separately and in combination. Pistachio, caper, rosemary, thyme and juniper were planted. We evaluated the effects of the treatments using soil quality (physicochemical properties, total content of hazardous elements, nutrient availability, microbial biomass and enzyme activities) and plant establishment indicators (survival, growth, vigor, nutrient content in leaves, nutrient balances and mycorrhizal root colonization). Thyme and juniper did not show a suitable survival rate (<50%) whereas 70-100% of the pistachio, rosemary and caper survived for at least 27 months. In unamended soil, plant growth was severely hampered by P, N, K and Zn deficiencies as well as Fe and Mn excess. Overall, the treatments affected the soil and plant indicators as follows: biofertilizer + co-compost > co-compost > biofertilizer > unamended. The application of co-compost was therefore essential with regard to improving soil fertility; furthermore, it increased leaf N and P content, whereas leaf Fe and Mn concentrations showed a decrease. The combined treatment, however, provided the best results. The positive interaction between the two soil conditioners might be related to the capacity of the biofertilizer to increase nutrient uptake from the composted residue, and to protect plants against Fe and Mn toxicity.

Response of drought and fertilization in Erica andevalensis seed banks: significance for conservation management.

The present study attempts to investigate the size, composition and seedling dynamics of the seed bank of a metalliferous and vulnerable species, Erica andevalensis. Samples were taken during spring and autumn from two different sites. We also studied the effects of nutrient solution, irrigation from the river Tinto and irrigation deficit on seeding establishment and survival. Only E. andevalensis and Poaceae species emerged from the seed banks, although the former was dominant (98%). Germination and seedling establishment was totally inhibited by the waters of the river Tinto. Seed density was high in the soils of both seed banks irrigated with water and nutrient solution. We found no seasonal differences in the seed bank and number of germinated seeds and mortality rate and density were similar. From the standpoint of restoration management, the results indicate that the seed bank is a very important factor for successful species establishment.

Tolerance to high Zn in the metallophyte Erica andevalensis Cabezudo & Rivera.

The tolerance to high Zn was studied in the metallophyte Erica andevalensis Cabezudo & Rivera grown in nutrient solutions at different Zn concentrations (5, 500, 1,000, 1,500 and 2,000 µM Zn). Plant growth and nutrient uptake were determined. Metabolic changes were assessed by the analysis of peroxidase activity, organic metabolites related to metal chelation (amino acids, organic acids (malate, citrate) or protection (polyamines). While plants tolerated up to 1,500 µM Zn, despite presenting of low growth rates, the concentration of 2,000 µM Zn was toxic producing high mortality rates. Roots accumulated high Zn concentration (11,971 mg/kg) at 1,500 µM external Zn) apparently avoiding metal transfer into shoots. After 30 days of treatment with high Zn (1,000 and 1,500 µM Zn), the leaves accumulated high levels of glutamine. Short-term treatment with 500 µM Zn, significantly increased the concentration of asparagine and glutamine in roots. Citrate concentration was also considerably increased when exposing roots to Zn excess. Metal immobilization in the root system, low interference with the uptake of nutrients and an increased production of putative organic ligands (amino acids, citrate) might have provided the Zn tolerance displayed by Erica andevalensis.

Evaluation of lead toxicity in Erica andevalensis as an alternative species for revegetation of contaminated soils.

Although revegetation using native flora is a low cost way to stabilize soil and restore the landscape contaminated with metals, little is known regarding the Pb-tolerance of many of these species. For this purpose, we evaluated the tolerance of Erica andevalensis to Pb by growing plants in nutrient solutions with increasing concentrations of Pb (up to 100 microM). Plant growth and different physiological parameters were determined to ascertain tolerance to metal stress. Additionally, an electron microscopy study coupled with EDX-analysis was performed to get clues on the Pb uptake and translocation from roots into stem and leaves. The LOEC (the lowest observed effect concentration) of Pb was 40 microM while the IC50 (inhibition concentration) was 80 microM Pb. Chemical analysis revealed a root > stem > leaf accumulation pattern. There was a severe reduction in fresh biomass and chlorophyll concentration at the highest Pb dose. The SEM-EDX study indicated that Pb was mostly located in root epidermal tissues. The blockage of Pb on the root probably avoided its toxic effects by limiting Pb transport to other tissues.

Assessment of phenylurea herbicides sorption on various Mediterranean soils affected by irrigation with wastewater.

The retention values of two herbicides, chlorotoluron and isoproturon, in five Mediterranean soils were assessed by two different approaches, a dynamic method, using a batch technique (BT) and a static method, using a soil saturated paste (SP). The SP method led in all cases to lower herbicide sorption when compared with BT, although pesticide distribution constants from both methods were linearly related for the set of used soils (R(2)⩾0.99) showing that both methods similarly reflected the behaviour of the different soils. Low-quality water, evaluated by employing recycled urban wastewater, did not modify herbicide sorption when compared with high quality water, in any soil and with any method.

Desorption of two organophosphorous pesticides from soil with wastewater and surfactant solutions.

A batch test was used to evaluate the extent of desorption of diazinon and dimethoate, preadsorbed on a calcareous agricultural soil, representative of the Mediterranean area. Urban wastewater from a secondary treatment and seven surfactant solutions, at concentrations ranging from 0.75 mg L(-1) to 10 gL(-1), were used. The surfactants assayed were cationic (hexadecyl trimethyl ammonium bromide (HD)), anionic (sodium dodecyl sulfate (SDS), Aerosol 22 (A22) and Biopower (BP)), and nonionic (Tween 80 (TW), Triton X 100 (TX) and Glucopon 600 (G600)). Desorption of dimethoate was either not affected or only slightly by the nonionic and anionic surfactants tested, while desorption of diazinon from the soil was only enhanced by A22, BP and TW. This desorption increase correlated significantly with the surfactant concentration of the solution used for desorption and with the concurrent increase in the supernatant of the dissolved organic carbon, in particular that originating from the surfactant. This parameter did not vary with the use of SDS, G600 and TX. The cationic surfactant HD was retained on the soil surface, as confirmed by an increase in soil organic carbon, resulting in a fall in desorption rate for both pesticides. Comparing treatment by wastewater with control water, there was no difference in desorption rate for either pesticide. Mixed TW/anionic surfactant solutions either did not modify or slightly increased desorption of both pesticides in comparison with individual surfactant solutions.

Persistence of two neonicotinoid insecticides in wastewater, and in aqueous solutions of surfactants and dissolved organic matter.

Wastewater treatment plants receive organic contaminants, such as pesticides, which reach the sewage system from domestic, industrial or agricultural activities. In wastewater, which is a complex mixture of organic and inorganic compounds, biotic or abiotic degradation of contaminants can be affected by the presence of co-solutes. The photodecomposition in natural sunlight of two neonicotinoid insecticides, thiamethoxam and thiacloprid, was investigated in wastewater, aqueous extracts of sewage sludge and in aqueous surfactant solutions, which are abundant in wastewater. Dissipation in the dark was also studied in wastewater, due to reduction of transmitted sunlight in wastewater ponds. With regard to photolysis, thiamethoxam degraded rapidly in all the aqueous solutions. Among them sewage sludge extracts slightly modified (average half-life 17.6h), wastewater increased (13.7h) and non-ionic surfactants led, as a family, to the highest dissipation rates (average 6.2h), with respect to control water (18.7h). Additionally this pesticide also underwent a slower biodegradation process in wastewater in the dark under anaerobic conditions (around 25d). A metabolite of thiamethoxam from the biological decomposition in wastewater was identified by HPLC/MS. On the other hand thiacloprid was found to be resistant to photo- and biodecomposition and remained almost unchanged during the experimental periods in all the tested media.

Stress responses of Erica andevalensis Cabezudo & Rivera plants induced by polluted water from Tinto River (SW Spain).

The effects of Tinto River water on Erica andevalensis growth, biochemical indicators and elemental concentration and distribution were investigated under laboratory conditions. High levels of toxic elements such as B, Fe and S and acidic pH characterized the river water. Plant analysis revealed that the concentration of Al, B, S and Fe increased in all plant organs reaching in some cases values in the toxicity range. Plants transferred into river water stopped growing and stress was manifested by plant water loss, increase in peroxidase activity and decrease of chlorophyll a concentration. Significant decreases of free amino acid concentration were found in shoots and roots of plants grown in diluted river water. The results indicated that Tinto River water acidity and its excess in soluble elements produced altogether severe alterations in roots affecting plant water and nutrient uptake and leading to the massive entry of some metals (e.g. Fe, Al) with toxic effects. Scanning-electron microscopy (cryoSEM and ESEM) observations showed that E. andevalensis had not exclusion mechanisms of Cu, Fe and S therefore it was not able to reduce translocation to aerial parts.

Design of experiments in environmental chemistry studies: example of the extraction of triazines from soil after olive cake amendment.

The disposal of natural, composted and vermicomposted olive cake for modifying the fate of triazine herbicides with different physicochemical properties (terbuthylazine, cyanazine, simazine and prometryn) has been tested. Experimental design (surface response methodology plus desirability function) to multicriteria optimization was carried out to evaluate both dose and type of amendment to retain the cited analytes and to develop two simple and low cost analytical methods for extracting triazines from soil. From a methodological point of view, classical and D-optimal designs were employed depending on the problem. Thus, the best combination of soil amount and solvent ratio, the most important parameters affecting triazine extraction from soil, was looked for by means of Central Composite Designs. Under the optimized conditions, the range of triazines recovery was 75-85% for shaking extraction and 87-107% for ultrasonic extraction. Regarding the amendment assay, D-optimal design was selected to keep the reliability of the estimations. Natural olive cake added to the soil at a high dose (8%) reduced herbicide recovery to ca. 50-60% for terbuthylazine, prometryn and simazine, while cyanazine recovery was negligible. Design of experiment provides an efficient working strategy to explore those conditions which ensure the optimum or target value of several responses evaluated simultaneously.

Strategies of heavy metal uptake by plants growing under industrial emissions.

Total concentrations of Al, Ba, Ca, Cr, Cu, Fe, K, Mg, Mn, Pb and Zn have been estimated in soil (A-horizon) and in leaves and stem samples of two Mediterranean species (Nerium oleander L. and Pinus pinea L.) growing in an industrial area in Spain (Huelva). Both species showed a different behaviour for the elements studied. Bark and leaves of both species acted as excluders of Al, Ba, Cr, Fe and Pb, N. oleander acted as indicator of Cu and Zn and, needles and bark of P. pinea behaved as accumulators of Cu. The enrichment ratio data indicated that Cu in soil and plant was enhanced with anthropogenic activities, with industrial activities being the primary contributor for Cu. All the other elements studied were controlled by natural source variations, but Pb could also be anthropogenically enhanced. Wood did not accumulate pollutants, with the translocation from bark being rather reduced. Uptake patterns of metals into foliage and bark tissues were more or less the same in both species for almost all the studied elements, which indicates that both plant parts could be indifferently used as biomonitors.

A simple methodology for the evaluation of groundwater pollution risks.

Groundwater represents a very significant source of fresh water for irrigation and drinking purposes and therefore preserving the availability and quality of this resource is extremely important. A first assessment of the amount of pollutants that can be exported to groundwater via soil drainage can be made by a) measuring the amount of contaminants present in the soil solution at the bottom of the soil after a prolonged simulated rainfall event, and b) estimating the amount of drainage water passing the soil bottom during a period of time long enough to include sufficient instances of both, wet and dry episodes inherent to the local climate. Drainage water was estimated by means of a simple infiltration model ("bucket model") which computes on a daily basis the inputs and outputs of soil water through rainfall and evapotranspiration generated by a stochastic model of the local climate along a period of 50-100 years. The methodology was applied in the Guadiamar valley after the toxic spill of a pyrite mine in Aznalcóllar (Spain). The results show that Zn is the dominant contaminant at the site with a 1.2 g m(-2)year(-1) contribution to groundwater. The presence of a gravel rich horizon below 50 cm depth reveals an increase in drainage and the threat to groundwater.

Guidelines for improving organic carbon recovery by the wet oxidation method.

The study of soil degradation processes as well as the monitoring of environmental quality often yields large sample batches in which organic carbon is the key property to be determined. A description is given for a rapid method to analyse organic carbon by wet oxidation; the reaction is carried out with 3ml K(2)Cr(2)O(7) and 6ml H(2)SO(4) in small vials, and Cr(3+) resulting from organic C oxidation is determined by spectrophotometry. No special equipment is needed and handling is reduced to a minimum, enabling over 100 samples per day to be analysed. There is one important detail in the operation protocol: the vials are placed inside an insulator block during oxidation. Heat insulation retards the cooling of vials, which results in a high organic C recovery (over 0.90). The improved method is feasible, simple, and easy to perform. Reproducibility was lower than 10% and recoveries from certified reference materials were about 90%. The simplified method is recommended for widespread organic C content sampling.

Heavy metals content in N. oleander leaves as urban pollution assessment.

Nerium oleander L. (Oleander) leaves grown in Palermo city (Sicily, Italy) were collected from six sampling sites representing either areas of high traffic and urbanisation density or areas far away from traffic (e.g. city gardens). Concentration of Al, Ba, Cr, Cu, Fe, Pb, Mg, Mn, and Zn were determined in leaf samples during two years. Multivariate analysis classified the sampling sites in four groups based on the metal content in vegetal leaves in agreement with traffic and human activity site. Many elements studied (Al, Ba, Fe, Mn Mg) arise from the soil composition and others such as Cr, Cu, Pb and Zn as pollutant of the soil. On the other hand, about 30% of Al, Fe Cr, Cu and Pb originate from aerial deposition on leaves. Although the results presented should be handled with caution N. oleander can be considered as a means of assessing dust contamination in the urban environment.

Ultrasound-assisted extraction of pesticides from olive branches: a multifactorial approach to method development.

A rapid and simple method has been developed for the analysis in olive branches of two insecticides currently used in olive pest control, dimethoate and alpha-cypermethrin. The effects of analytical conditions on pesticide recovery and the optimal extraction conditions were evaluated by means of a factorial design. The use of this chemometric tool in analytical method development allows the identification of the principal and interaction effects of the extraction conditions on the recovery of pesticides. It also gives information about the location of pesticide maximum recovery with minimal experimental investment. Extraction was carried out with an ultrasonic bath and the experimental conditions studied included the volume of extractant, the time of extraction, the number of extraction steps and the sample weight. The sample was further cleaned up using a Florisil solid-phase extraction cartridge. For the overall extraction procedure, recoveries of 99 % for alpha-cypermethrin and 90 % for dimethoate from the spiked samples were found for 1 g of sample extracted three times with 35 mL hexane, sonicating for 2 min in each step. The complete process including ultrasonic extraction and filtration will not require more than 15-20 min, in contrast with several hours for conventional liquid-solid extraction techniques. The proposed method allows a high sample throughput, as commonly required in monitoring studies.

Assessment of airborne heavy metal pollution by aboveground plant parts.

Italian stone pine (Pinus pinea L.) and oleander (Nerium oleander L.) leaves, bark and wood samples were collected at different sites around an industrial area (Huelva, SW Spain) and compared with samples of the same species from a background site. Samples were analysed with respect to the following pollutants: Al, Ba, Cr, Cu, Fe and Pb by ICP-AES. The suitability of different plant parts as biomonitors of pollution was investigated. In pine samples from the polluted sites the ratio of concentrations between bark and wood was high for Al, Ba, Cu and Fe, whereas no differences were found in samples from the unpolluted area. No differences were detected in oleander for the same ratio. In the oleander species, the ratio between leaves and wood concentration allowed to distinguish between control and polluted sites. The ratio of the concentration between leaves and wood was elevated for Al, Ba and Fe in pine samples from the polluted sites. The ratio of the concentration in bark or leaves to their concentration in wood might be useful to detect inorganic atmospheric pollutants.

Chemical and physical factors affecting the extractability of methidathion from soil samples.

Determination of methidathion in soil samples using Soxhlet extraction has been studied. Several factors were investigated for their effect on methidathion recovery. Some were related to the extraction procedure, for example solvent type used for the extraction (acetone or hexane/toluene), extraction time, soil humidity, and type of sterilisation system employed. Other factors tested included the addition of organic matter to the soil, for example urban sewage sludge and the cationic surfactant TDTMA. Experimental designs were used to determine the effects of the different factors. Acetone resulted in higher recoveries and was less affected by the presence of water. Autoclaving was the most appropriate sterilisation method. Thimerosal resulted in a decrease in insecticide recovery. Methidathion recovery increased as the amount of cationic surfactant was increased, but decreased as the amount of sewage sludge added to the soil was increased. In general, because the factors studied were not always independent of each other, a clear description of the methodology used is needed when analysing pollutants in environmental samples.

Serum selenium and risk of large size colorectal adenomas in a geographical area with a low selenium status.

OBJECTIVE: Selenium is a fundamental nutrient to human health that might have anticarcinogenic effects. Previous studies have assessed the possible relationship of selenium status to colorectal adenomas with controversial results. We primarily aimed to assess the relationship of serum selenium status with the presence of large size colorectal adenomas in subjects living in a poor selenium region. The serum selenium status in colorectal cancer was also evaluated. METHODS: Serum selenium levels were measured in 28 patients with large size sporadic adenomatous polyps, 24 patients with colorectal adenocarcinomas, and 35 age-matched healthy individuals. A logistic regression analysis was performed to assess the relationship of serum selenium to colorectal adenomatous polyps after adjusting for confounding variables (age, sex, smoking habit, and alcohol drinking). RESULTS: Among subjects aged < or = 60 yr, mean serum selenium levels were significantly lower in both patient groups (adenoma, 57.9 +/- 4.3 microg/L; cancer, 43.7 +/- 6.6 microg/L) than in healthy controls (88.9 +/- 8 microg/L) (p = 0.0001). There were no difference among subjects > 60 yr old. A significant inverse association between selenium status and the diagnosis of large size adenomatous polyps after adjusting for confounding variables was found (adjusted p = 0.029). Subjects with higher selenium status (> or = 75th percentile value of 82.11 microg/L) had a lower probability (OR = 0.17, 95% CI = 0.03-0.84) to be in the adenoma group than subjects with lower selenium status (< 82.11 microg/L). This association was more marked in subjects aged < or = 60 yr (adjusted p value = 0.04, OR = 0.08, 95% CI = 0.007-0.91), and was not significant in older subjects. CONCLUSIONS: Results suggest that high selenium status may decrease the risk of large size adenomas in a low selenium region, and that this preventive effect seems to be exclusive to subjects < or = 60 yr. These results will need to be confirmed in additional epidemiological studies before recommending selenium supplementation in patients with colon adenomas.