A field screening test for the assessment of concentrations and mobility of potentially toxic elements in soils: a case study on urban soils from Rome and Novi Sad.

The increasing demand for environmental pollution control results in the development and use of new procedures for the determination of dangerous chemicals. Simple screening methods, which can be used directly in the field for a preliminary assessment of soil contamination, seem to be extremely advantageous. In our laboratory, we developed and optimized a rapid test for a preliminary evaluation of both the concentration and the mobility of some potentially toxic metals in soils. This screening test consists of a single extraction of the soil sample with a buffer solution, followed by the titration of the extracted solution with dithizone to determine the contents of bi-valent heavy metals (such as Pb, Cu, Zn, and Cd). This screening method was then directly applied in the field during the sampling campaign in the framework of an Italian-Serbian collaborative project, finalized in the study of metal availability in soils. The results obtained in the field with the rapid test were compared with those obtained in the laboratory following the conventional procedure commonly used to evaluate metal bioavailability (diethylenetriaminepentaacetic extraction). Moreover, selected samples were analyzed sequentially in the laboratory using the standardized BCR three-step sequential extraction procedure. The screening test gave results conceptually in good agreement with those obtained via the BCR procedure. These preliminary data show that the proposed screening test is a reliable method for the preliminary rapid evaluation of metal total concentrations and of potential metal mobility in soils, supporting sampling activities directly in the field.

The mechanism of iron binding processes in erionite fibres.

Fibrous erionite-Na from Rome (Oregon, USA) was K-exchanged and characterized from the structural point of view. In addition, the modifications experienced after contact with a Fe(II) source were investigated for evaluating if the large potassium ions, blocking off nearly all the erionite cavity openings, might prevent the Fe(II) binding process, which is currently assumed to be one of the reasons of the toxicity of erionite. The K-exchanged sample had a 95% reduction of the BET surface area indicating that it behaves as a mesoporous material. Exchanged K is segregated at K2 and at OW sites commonly occupied by H2O. The latter K cations provide a relevant contribution to the reduction of the surface area. Surprisingly, despite the collapse of its surface area the sample preserves the tendency to bind Fe(II). Therefore, yet in the case of a peculiar and potentially hostile structural environment the Fe(II) ion-exchange process has essentially the same kinetics observed in a typical erionite sample. This is a clear evidence of the very limited effect of the chemical composition of erionite on the Fe(II) binding process and reasonably it does not play a significant role in its toxicity.

Antarctic snow: metals bound to high molecular weight dissolved organic matter.

In this paper we studied some heavy metals (Cu, Zn, Cd, Pb, As, U) probably associated to high molecular weight organic compounds present in the Antarctic snow. Snow-pit samples were collected and analysed for high molecular weight fraction and heavy metals bound to them by means of ultrafiltration treatment. High molecular weight dissolved organic matter (HMW-DOM) recovered by ultrafiltration showed a dissolved organic carbon concentration (HMW-DOC) of about 18-83% of the total dissolved organic carbon measured in Antarctic snow. The characterisation of HMW-DOM fraction evidenced an ageing of organic compounds going from surface layers to the deepest ones with a shift from aliphatic compounds and proteins/amino sugars to more high unsaturated character and less nitrogen content. The heavy metals associated to HMW-DOM fraction follows the order: Zn > Cu > Pb >> Cd âˆ¼ As âˆ¼ U. The percentage fraction of metals bound to HMW-DOM respect to total metal content follows the order: Cu >> Pb > Zn, Cd in agreement with humic substance binding ability (Irwing-William series). Going down to depth of trench, all metals except arsenic, showed a high concentration peak corresponding to 2.0-2.5 m layer. This result was attributed to particular structural characteristic of organic matter able to form different type of complexes (1:1, 1:2, 1:n) with metals.

Biodistribution and acute toxicity of a nanofluid containing manganese iron oxide nanoparticles produced by a mechanochemical process.

Superparamagnetic iron oxide nanoparticles are candidate contrast agents for magnetic resonance imaging and targeted drug delivery. Biodistribution and toxicity assessment are critical for the development of nanoparticle-based drugs, because of nanoparticle-enhanced biological reactivity. Here, we investigated the uptake, in vivo biodistribution, and in vitro and in vivo potential toxicity of manganese ferrite (MnFe2O4) nanoparticles, synthesized by an original high-yield, low-cost mechanochemical process. Cultures of murine Balb/3T3 fibroblasts were exposed for 24, 48, or 72 hours to increasing ferrofluid concentrations. Nanoparticle cellular uptake was assessed by flow-cytometry scatter-light measurements and microscopy imaging after Prussian blue staining; cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony-forming assays. After a single intravenous injection, in vivo nanoparticle biodistribution and clearance were evaluated in mice by Mn spectrophotometric determination and Prussian blue staining in the liver, kidneys, spleen, and brain at different posttreatment times up to 21 days. The same organs were analyzed for any possible histopathological change. The in vitro study demonstrated dose-dependent nanoparticle uptake and statistically significant cytotoxic effects from a concentration of 50 µg/mL for the MTT assay and 20 µg/mL for the colony-forming assay. Significant increases in Mn concentrations were detected in all analyzed organs, peaking at 6 hours after injection and then gradually declining. Clearance appeared complete at 7 days in the kidneys, spleen, and brain, whereas in the liver Mn levels remained statistically higher than in vehicle-treated mice up to 3 weeks postinjection. No evidence of irreversible histopathological damage to any of the tested organs was observed. A comparison of the lowest in vitro toxic concentration with the intravenously injected dose and the administered dose of other ferrofluid drugs currently in clinical practice suggests that there might be sufficient safety margins for further development of our formulation.

Assessment of the applicability of a "toolbox" designed for microbially assisted phytoremediation: the case study at Ingurtosu mining site (Italy).

The paper describes the fieldwork at the Italian test site of the abandoned mine of sphalerite and galena in Ingurtosu (Sardinia), with the aim to assess the applicability of a "toolbox" to establish the optimized techniques for remediation of soils contaminated by mining activities. A preliminary characterization-including (hydro)geochemistry, heavy metal concentration and their mobility in soil, bioprospecting for microbiology and botany-provided a data set for the development of a toolbox to deliver a microbially assisted phytoremediation process. Euphorbia pithyusa was selected as an endemic pioneer plant to be associated with a bacterial consortium, established with ten selected native strains, including metal-tolerant bacteria and producers of plant growth factors. The toolbox was firstly assessed in a greenhouse pot experiment. A positive effect of bacterial inoculum on E. pithyusa germination and total plant survival was observed. E. pithyusa showed to be a well-performing metallophyte species, and only inoculated soil retained a microbial activity with a high functional diversity, expanding metabolic affinity also towards root exudates. These results supported the decision to proceed with a field trial, investigating different treatments used singly or in combination: bioaugmentation with bacterial consortia, mycorrhizal fungi and a commercial mineral amendment. Microbial activity in soil, plant physiological parameters and heavy metal content in plants and in soil were monitored. Five months after the beginning, an early assessment of the toolbox under field conditions was carried out. Despite the cold season (October-March), results suggested the following: (1) the field setup as well as the experimental design proved to be effective; (2) plant survival was satisfactory; (3) soil quality was increased and bioaugmentation improved microbial activity, expanding the metabolic competences towards plant interaction (root exudates); and (4) multivariate analysis supported the data provided that the proposed toolbox can be established and the field trial can be carried forward.

Beryllium natural background concentration and mobility: a reappraisal examining the case of high Be-bearing pyroclastic rocks.

Beryllium is widely distributed in soils at low levels, but it can also occur naturally in higher concentrations in a variety of materials exploited for many industrial applications. Beryllium is also one of the most toxic natural elements and is known to be a human carcinogen. A concise account of the literature data on baseline concentrations of Be in soils illustrates the possibility of worldwide presence of areas with a high natural background concentration of Be (up to 300 mg/kg), the crustal abundance of which is generally estimated to be in the range 2-6 mg/kg. Nevertheless, the number of available data is rather limited in comparison with those about other toxic elements such as Pb, Cd and Cr. This has probably caused the choice of low values of concentration level as the reference for the definition of soil contamination: these values are not always realistic and are not applicable to large areas. As a case study, we report and analyse a diffuse, unusually high (up to 80 mg/kg, average approximately 20 mg/kg), natural occurrence of beryllium in loose and poorly consolidated pyroclastic layers related to the Pleistocene activity of the Vico volcano. Additionally, the analysis of Be leachability has been carried out, providing evidence of a not negligible mobility in contrast with the scarce data presented in the literature that usually indicate beryllium as an element with low mobility in oxidising surface environmental conditions. This research marks the beginning of a possible reappraisal of beryllium geochemical behaviour and background levels, providing more realistic reference values for risk assessment and land management.

Fluorine in the rocks and sediments of volcanic areas in central Italy: total content, enrichment and leaching processes and a hypothesis on the vulnerability of the related aquifers.

Rock, sediment and water samples from areas characterised by hydrothermal alterations in the Sabatini and Vico Volcanic Districts, near Rome and the large city of Viterbo, respectively, were collected and analysed to determine the total fluorine (F) content and to understand the F geochemical background level in the volcanic districts of central Italy. Leaching and alteration processes controlling the high concentration of F in water were also investigated. Fluorine concentrations were directly determined (potentiometrically) by an F selective electrode in water samples, while the procedure for rock samples included preliminary F dissolution through alkaline fusion. F concentrations higher than 800 mg kg(-1) were commonly found in the analysed rocks and sediments; the concentration depended on the lithology and on the distance from the alteration areas. A specific successive sampling campaign was conducted in three areas where the F content in sediments was particularly high; in the same areas, measurements of CO(2) flux were also performed to investigate the possible deep origin of F. To verify the relationships among the high F contents in rocks and sediments, the leaching processes involved and the presence of F in the aquifer, we also collected water samples in the western sector of the Sabatini Volcanic District, where hydrothermal manifestations and mineral springs are common. The data were processed using a GIS system in which the F distribution was combined with morphological and geological observations. The main results of our study are that (1) F concentrations are higher in volcanic and recently formed travertine (especially in hydrothermally altered sediments) than in sedimentary rocks and decrease with distance from hydrothermal alteration areas, (2) F is more easily leached from hydrothermally altered rocks and from travertine and (3) sediments enriched with F may indicate the presence of deep regional fractures that represent direct pathways of hydrothermal fluids from the crust to the surface.

Cadmium, lead and their mixtures with copper: Paracentrotus lividus embryotoxicity assessment, prediction, and offspring quality evaluation.

The aim of this research was to assess the combined effects of three heavy metals (copper, lead, cadmium) on the fertilization and offspring quality of the sea urchin Paracentrotus lividus at EC50, NOEL, and EC1 concentrations. The observed data were compared with the predictions derived from approaches of Concentration Addition (CA) and Independent Action (IA) in order to evaluate the proper prediction of the observed mixture toxic effect. The P. lividus embryotoxicity of trace metals decreases as follows: Cu > Pb > Cd at all toxicity concentration tested. EC50 mixture revealed less toxic only than Cu; EC50 was 0.80 (± 0.07) mg/l, the offspring malformations were mainly P1 type (skeletal alterations) up to 20% mixture concentration, and P2 type from 70% concentration. The NOEL and EC1 mixtures evidenced that all compounds contribute to the overall toxicity, even if present at low concentrations: the former EC50 was 0.532 (± 0.058) mg/l and the latter was 1.081 (± 0.240) mg/l. The developmental defects observed were mainly P1 type in both mixtures. Both CA and IA models did not accurately predict mixture toxicity for EC50 and NOEL mixtures. Instead, EC1 mixture effects seemed well represented by the IA model. The protective action of the CA model, although quite accurate when applied to simple biological systems like algae and bacteria, but failed to represent the worst-case in this study with more complex organisms. It would be useful to introduce in the models one or more factors that take into account the complexity of these biological systems.

Bioremediation of diesel oil in a co-contaminated soil by bioaugmentation with a microbial formula tailored with native strains selected for heavy metals resistance.

The aim of the work is to assess the feasibility of bioremediation of a soil, containing heavy metals and spiked with diesel oil (DO), through a bioaugmentation strategy based on the use of a microbial formula tailored with selected native strains. The soil originated from the metallurgic area of Bagnoli (Naples, Italy). The formula, named ENEA-LAM, combines ten bacterial strains selected for multiple resistance to heavy metals among the native microbial community. The biodegradation process of diesel oil was assessed in biometer flasks by monitoring the following parameters: DO composition by GC-MS, CO2 evolution rate, microbial load and composition of the community by T-RFLP, physiological profile in Biolog ECOplates and ecotoxicity of the system. The application of this microbial formula allowed to obtain, in the presence of heavy metals, the complete degradation of n-C(12-20), the total disappearance of phenantrene, a 60% reduction of isoprenoids and an overall reduction of about 75% of the total diesel hydrocarbons in 42 days. Concurrently with the increase of metabolic activity at community level and the microbial load, the gradual abatement of the ecotoxicity was observed. The T-RFLP analysis highlighted that most of the ENEA-LAM strains survived and some minor native strains, undetectable in the soil at the beginning of the experiment, developed. Such a bioaugmentation approach allows the newly established microbial community to strike a balance between the introduced and the naturally present microorganisms. The results indicate that the use of a tailored microbial formula may efficiently facilitate and speed up the bioremediation of matrices co-contaminated with hydrocarbons and heavy metals. The study represents the first step for the scale up of the system and should be verified at a larger scale. In this view, this bioaugmentation strategy may contribute to overcome a critical bottleneck of the bioremediation technology.

Investigating heavy metal resistance, bioaccumulation and metabolic profile of a metallophile microbial consortium native to an abandoned mine.

Contaminated sites represent new ecological niches where historical pollution has originated an unusual microbial biodiversity. The knowledge of these microorganisms contributes to the discovery of new pathways and metabolic networks and may offer potential solutions for damaged areas. In the present work seven microbial consortia have been isolated from an abandoned mine of blend and galena (Ingurtosu, Italy) through a selection for resistance to zinc (tested up to 40 mM in solution). All the consortia were able to accumulate zinc and the best accumulator, named Ing5, has been studied for the following characteristics: resistance and accumulation of Zn, Cd, Hg, bioaccumulation mechanisms of Zn, and influence of Zn and Cd on the metabolic profile. The results indicate that the consortium Ing5 bears resistance systems for Cd and Hg as well as Zn and that, for some of the 5 isolates belonging to Ing5, the resistance thresholds are higher in consortium than in pure culture. The prevalent mechanism for zinc accumulation can be reasonably considered to be metabolism-dependent, inducible and regulated by metal concentrations. The study on the metabolic profile, carried out by the Biolog system, shows that Zn exerts a very low influence on the metabolic profile and that this influence can also be positive; Cd has a stronger negative influence but that, despite this, the consortium is able to maintain a wide metabolic potential in the presence of heavy metals. These features of Ing5 make it a good candidate for biotechnological applications and for further investigation of the degradation of organic pollutants in the presence of metals.

Effect of urea on degradation of terbuthylazine in soil.

Pesticide and nitrate contamination of soil and groundwater from agriculture is an environmental and public health concern worldwide. The herbicide terbuthylazine (CBET) has replaced atrazine in Italy and in many other countries because the use of the latter has been banned because of its adverse environmental impacts. Unlike atrazine, knowledge about the fate of CBET in soil is still not extensive, especially regarding its transformation products, but recent monitoring data show its occurrence and that of its main metabolite, desethyl-terbuthylazine (CBAT), in groundwater above the limit of 0.1 microg/L established by European Union Directive and Italian legislation. The objective of this work was to investigate if the presence of the fertilizer urea affects CBET degradation in the soil. Laboratory CBET degradation experiments in the presence/absence of urea were performed with microbiologically active soil and sterilized soil. Terbuthylazine degradation rates under the different experimental conditions were assessed, and the formation, degradation, and transformation of the metabolite CBAT were also studied. Terbuthylazine degradation was affected by the presence of urea, in terms both of a higher disappearance time of 50% of the initial concentration and of a lower amount of CBAT formed. These findings have practical implications for the real-life assessment of the environmental fate of triazine herbicides in agricultural areas since these herbicides are frequently applied to soils receiving ureic fertilizers.

Simazine biodegradation in soil: analysis of bacterial community structure by in situ hybridization.

Pesticide and nitrate contamination of soil and groundwater from agriculture is an environmental and public health concern worldwide. Simazine, 6-chloro-N2,N4-diethyl-1,3,5-triazine-2,4-diamine, is a triazine herbicide used in agriculture for selective weed control with several types of crops and it is frequently applied to soils receiving N-fertilizers. Degradation experiments were performed in the laboratory to assess whether the biodegradation of simazine in soil may be influenced by the presence of urea. Simazine degradation rates under different experimental conditions (presence/absence of urea, microbiologically active/sterilized soil) were assessed together with the formation, degradation and transformation of its main metabolites in soil. Simazine degradation was affected by the presence of urea, in terms both of a smaller half-life (t(1/2)) and of a higher amount of desethyl-simazine formed. The soil bacterial community was also studied. Microbial abundances were determined by epifluorescence direct counting. Moreover in situ hybridization with rRNA-targeted fluorescent oligonucleotide probes was used to analyze the bacterial community structure. Fluorescent in situ hybridization (FISH) was used to detect specific groups of bacteria such as the alpha,beta,gamma-subdivisions of Proteobacteria, Gram-positive bacteria with a high G + C DNA content, Planctomycetes, Betaproteobacterial ammonia-oxidizing bacteria and nitrifying bacteria. The presence of the herbicide and/or urea affected the bacterial community structure, showing that FISH is a valuable tool for determining the response of bacterial populations to different environmental conditions.

Reuse of a treated red mud bauxite waste: studies on environmental compatibility.

Red mud is the major solid waste produced in the process of alumina extraction from bauxite (Bayer process). Environmental "compatibility" of a treated red mud was studied in order to evaluate its possible recycling in environmental compartments. The leaching test requested by the Italian law on treated solid waste to be "re-introduced in the environment" was performed on this material. Moreover, in order to better evaluate the environmental compatibility, three different types of eco-toxicological tests were applied (Microtox test, ASTM microalgae toxicity test and sea urchin embryo toxicity test). These "chemical" and eco-toxicological tests gave encouraging results. The possibility to use this material for treating contaminated waters and soils was evaluated, again with particular attention to the Italian regulatory system, through experiments on the treated red mud metal trapping ability and on the subsequent release of trapped metals, at low pH conditions. The treated red mud showed a general high metal trapping capacity and the release at low pH was generally low.

Comparison of three sequential extraction procedures (original and modified 3 steps BCR procedure) applied to sediments of different origin.

The 3 steps sequential extraction procedure proposed by the Standards Measurements and Testing program (SM&T--formerly BCR) of the European Union has been applied for the speciation of metals in sediments. Results obtained by the application of the BCR standardized procedure were compared to those of two four step sequential extraction procedures, which are different from the BCR procedure only for the introduction of an additional step with NaOCl, as 2nd and 3rd step respectively. Five different metals have been taken into consideration: Cd, Cu, Ni, Pb and Zn. The analytical performances of the laboratory have been evaluated using three certified reference materials: the BCR 601 lake sediment for the BCR sequential extraction procedure, PACS-1 and MESS-1 for total metal concentration. Results showed that the efficiency of NaOCl treatment is higher or at least equal to that of H2O2 treatment and that its selectivity is quite satisfying. Moreover the NaOCl treatment doesn't significantly influence the extraction of the easily reducible fraction.