Enhanced Natural Attenuation of Groundwater Cr(VI) Pollution Using Electron Donors: Yeast Extract vs. Polyhydroxybutyrate.

Remediation interventions based on the native bacteria's capability to reduce Cr(VI) represent a valid strategy in terms of economic and environmental sustainability. In this study, a bioremediation test was carried out using viable microcosms set with groundwater and deep soil (4:1), collected from the saturated zone of an industrial site in Southern Italy that was polluted by ~130 µg L-1 of Cr(VI). Conditions simulating the potential natural attenuation were compared to the enhanced natural attenuation induced by supplying yeast extract or polyhydroxybutyrate. Sterile controls were set up to study the possible Cr(VI) abiotic reduction. No pollution attenuation was detected in the unamended viable reactors, whereas yeast extract provided the complete Cr(VI) removal in 7 days, and polyhydroxybutyrate allowed ~70% pollutant removal after 21 days. The incomplete abiotic removal of Cr(VI) was observed in sterile reactors amended with yeast extract, thus suggesting the essential role of native bacteria in Cr(VI) remediation. This was in accordance with the results of Pearson's coefficient test, which revealed that Cr(VI) removal was positively correlated with microbial proliferation (n = 0.724), and also negatively correlated with pH (n = -0.646), dissolved oxygen (n = -0.828) and nitrate (n = -0.940). The relationships between the Cr(VI) removal and other monitored parameters were investigated by principal component analysis, which explained 76.71% of the total variance.

Camouflaged Fluorescent Silica Nanoparticles Target Aggregates and Condensates of the Amyloidogenic Protein Tau.

Intrinsically disordered proteins (IDPs) are increasingly found to be associated with irreversible neurodegenerative disorders. The protein tau is a prototypical IDP whose abnormal aggregation into insoluble filaments is a major hallmark of Alzheimer's disease. The view has emerged that aggregation may proceed via alternative pathways involving oligomeric intermediates or phase-separated liquid droplets. Nanoparticles (NPs) offer significant potential for probing the mechanisms of protein fibrillation and may be capable of redirecting conformational transitions. Here, we camouflaged dye-doped silica NPs through functionalization with tau molecules to impart them the ability to associate with protein assemblies such as aggregates or condensates. The prepared NP-tau conjugates showed little influence on the aggregation kinetics and morphology of filamentous aggregates of tau but were found to associate with the filaments. Moreover, NP-tau conjugates were recruited and concentrated into polyanion-induced condensates of tau, driven by multivalent electrostatic interactions, thereby illuminating liquid droplets and their time-dependent transformation, as observed by fluorescence microscopy. NP-tau conjugates were capable of entering human neuroglioma cells and were not cytotoxic. Hence, we propose that NP-tau conjugates could serve as nanotracers for in vitro and in-cell studies to target and visualize tau assemblies and condensates, contributing to an explanation for the molecular mechanisms of abnormal protein aggregation.

Uptake and Intracellular Trafficking Studies of Multiple Dye-Doped Core-Shell Silica Nanoparticles in Lymphoid and Myeloid Cells.

INTRODUCTION: Since most biologically active macromolecules are natural nanostructures, operating in the same scale of biomolecules gives the great advantage to enhance the interaction with cellular components. Noteworthy efforts in nanotechnology, particularly in biomedical and pharmaceutical fields, have propelled a high number of studies on the biological effects of nanomaterials. Moreover, the determination of specific physicochemical properties of nanomaterials is crucial for the evaluation and design of novel safe and efficient therapeutics and diagnostic tools. In this in vitro study, we report a physicochemical characterisation of fluorescent silica nanoparticles (NPs), interacting with biological models (U937 and PBMC cells), describing the specific triggered biologic response. METHODS: Flow Cytometric and Confocal analyses are the main method platforms. However TEM, NTA, DLS, and chemical procedures to synthesize NPs were employed. RESULTS: NTB700 NPs, employed in this study, are fluorescent core-shell silica nanoparticles, synthesized through a micelle-assisted method, where the fluorescence energy transfer process, known as FRET, occurs at a high efficiency rate. Using flow cytometry and confocal microscopy, we observed that NTB700 NP uptake seemed to be a rapid, concentration-, energy- and cell type-dependent process, which did not induce significant cytotoxic effects. We did not observe a preferred route of internalization, although their size and the possible aggregated state could influence their extrusion. At this level of analysis, our investigation focuses on lysosome and mitochondria pathways, highlighting that both are involved in NP co-localization. Despite the main mitochondria localization, NPs did not induce a significant increase of intracellular ROS, known inductors of apoptosis, during the time course of analyses. Finally, both lymphoid and myeloid cells are able to release NPs, essential to their biosafety. DISCUSSION: These data allow to consider NTB700 NPs a promising platform for future development of a multifunctional system, by combining imaging and localized therapeutic applications in a unique tool.

Enhancement of Chromium (VI) Reduction in Microcosms Amended with Lactate or Yeast Extract: A Laboratory-Scale Study.

A laboratory-scale study was carried out to evaluate the groundwater bioremediation potential of hexavalent chromium (Cr(VI)), taking into account the chromate pollution of an industrial site located in Southern Italy (Apulia Region). The reduction of Cr(VI) was studied on laboratory microcosms, set up in different experimental conditions, namely: ABIO (soil and water sterilized), BIO (soil and water not sterilized), LATT (with the addition of lactate), and YE (with the addition of yeast extract). Control test lines, set up by using sterilized matrices and amendments, were employed to assess the occurrence of the pollutant reduction via chemical processes. By combining molecular (microbial abundance, specific chromate reductase genes (ChR) and the Shewanella oinedensis bacterial strain) with chemical analyses of chromium (VI and III) in the matrices (water and soil) of each microcosm, it was possible to investigate the response of microbial populations to different experimental conditions, and therefore, to assess their bioremediation capability in promoting Cr(VI) reduction. The overall results achieved within this work evidenced the key role of amendments (lactate and yeast extract) in enhancing the biological reduction of hexavalent chromium in the contaminated aqueous phase of laboratory microcosms. The highest value of Cr(VI) removal (99.47%) was obtained in the YE amended microcosms at seven days.

Oxidation of azo and anthraquinonic dyes by peroxymonosulphate activated by UV light.

The photochemical degradation of two azo and two anthraquinonic dyes was performed using potassium peroxymonosulphate (Oxone®) activated by UV radiation. The fast decolourization of all dyes was observed within 6 min of UV irradiation, with corresponding dye decays higher than 80%. The kinetic rate constants of the dyes' decay were determined, along with the energetic efficiency of the photochemical treatment, taking into account the influence of a few anions commonly present in real wastewaters (i.e., chloride, nitrate, carbonate/bicarbonate and phosphate ions). Chloride and carbonate/bicarbonate ions enhanced dye degradation, whereas phosphate ions exerted an inhibitory effect, and nitrates did not have a predictable influence. The dye decolourization was not associated with efficient mineralization, as suggested by the lack of a significant total organic carbon (TOC) decrease, as well as by the low concentrations of a few detected low molecular weight by-products, including nitrate ions, formaldehyde and organic acids. High molecular weight by-products were also detected by mass spectrometry analysis. The investigated process may be proposed as a convenient pre-treatment to help dye degradation in wastewater during combined treatment methods.

Biodegradation of UV-filters in marine sediments.

The degradation of two of the most frequently used UV-filters was investigated through microcosm studies. Marine sediments sampled from two sites in Italy (La Spezia harbour and Sarno river estuary, S1 and S2 respectively) were used to set up aerobic and anaerobic sets of reactors. The sediments were spiked with a methanol solution of 3-(4-methylbenzylidene)camphor (4-MBC) and 2-ethylhexyl 4-(dimethylamino)benzoate (EH-DPAB), at concentrations of either 25 or 50mgkg-1 each. Methanol (6.3g/L) also served as an organic amendment and growth substrate for improving microbial activity. Monitoring of the biotic and abiotic degradation of the selected contaminants over 16months revealed that 4-MBC biodegradation was very slow and incomplete, whereas over 90% of EH-DPAB was degraded both in the aerobic and the anaerobic reactors by the natural microbial communities of both sediments. Repeated spikes of EH-DPAB were followed by complete decay, characterised by first-order kinetics. The calculated kinetic rate constants under aerobic and anaerobic conditions were similar. In reactors inoculated with the S1 sediment the degradation rate constants progressively increased after each spike, up to the value of 0.039d-1. For the S2 sediment the rate constant was around 0.020d-1 throughout the duration of the experiment. Mass spectrometry analysis of sediment extracts allowed detection of potential transformation products of EH-DPAB and 4-MBC. Moreover, the natural microbial community of the sediments was studied using the CAtalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) both in the initial sediments and after degradation under aerobic and anaerobic conditions.

Amorphous boron-doped sodium titanates hydrates: Efficient and reusable adsorbents for the removal of Pb2+ from water.

Amorphous titanium hydroxide and boron-doped (B-doped) sodium titanates hydrates were synthetized and used as adsorbents for the removal of Pb2+ from water. The use of sodium borohydride (NaBH4) and titanium(IV) isopropoxide (TTIP) as precursors permits a very easy synthesis of B-doped adsorbents at 298K. The new adsorbent materials were first chemically characterized (XRD, XPS, SEM, DRIFT and elemental analysis) and then tested in Pb2+ adsorption batch experiments, in order to define kinetics and equilibrium studies. The nature of interaction between such sorbent materials and Pb2+ was also well defined: besides a pure adsorption due to hydroxyl interaction functionalities, there is also an ionic exchange between Pb2+ and sodium ions even working at pH 4.4. Langmuir model presented the best fitting with a maximum adsorption capacity up to 385mg/g. The effect of solution pH and common ions (i.e. Na+, Ca2+ and Mg2+) onto Pb2+ sorption were also investigated. Finally, recovery was positively conducted using EDTA. Very efficient adsorption (>99.9%) was verified even using tap water spiked with traces of Pb2+ (50ppb).

Sorption properties of an amorphous hydroxo titanate towards Pb(2+), Ni(2+), and Cu(2+) ions in aqueous solution.

Titanates may be selectively used as inorganic adsorbents for heavy metal ions owing to their stability and fast adsorption kinetics. Nevertheless, the synthesis of such materials usually requires extreme reaction conditions. In this work, a new titanium-based material was rapidly synthesized under mild laboratory conditions. The obtained amorphous hydroxo titanate was tested for heavy metal sorption through kinetic and equilibrium batch tests, which indicated that the new material had high adsorption rates and adsorption capacities towards Cu(2+), Ni(2+) and Pb(2) ions. Adsorption kinetics were pseudo-second order, and equilibrium data fitted the Langmuir isotherm model. The calculated maximum adsorption capacities of Cu(2+), Ni(2+) and Pb(2+) in deionized water were around 1 mmol g(-1), and they decreased for Cu(2+) and Ni(2+) in the presence of Na(+), Ca(2+) and Mg(2+) ions, whereas the alkali metal ions did not influence Pb(2+) uptake. The efficiency of adsorption and recovery of lead ions were evaluated through column dynamic tests, by feeding the column with groundwater and tap water spiked with Pb(2+). The high performance of the hydroxo titanate over several cycles of retention and elution suggested that the product is potentially useful for the solid phase extraction of lead at trace levels in natural water samples, with potential use in metal pre-concentration for analytical applications.

Soil amendment with olive mill wastes: impact on groundwater.

Two sets of soil lysimeters were amended with solid and liquid olive mill wastes and the composition of leachate was analysed. Five treatments were carried out using: olive mill wastewater (OMW) at two different rates (80 and 320 m(3)/ha); OMW pre-treated by catalytical digestion with MnO2; compost obtained by exhausted olive pomace; freshwater as the control. Electric conductivity, pH, potassium, total polyphenols and nitrates were monitored in the leachate as indexes of potential groundwater contamination. The study demonstrated that the impact of all the selected amendments on groundwater was the minimum. OMW was safely applied to soil even at four times the rate allowed by the Italian law, and pre-treatment by catalytical digestion was not necessary to further reduce the impact on groundwater. The application of olive pomace compost was equally safe.

Simultaneous Cr(VI) reduction and non-ionic surfactant oxidation by peroxymonosulphate and iron powder.

Some industrial wastewaters contain both hexavalent chromium and surfactants. In this work, their removal from aqueous solution by zero-valent iron (ZVI) and peroxymonosulphate (PMS) was studied using Brij 35 as a representative non-ionic surfactant. The performance of the ZVI/PMS system in the simultaneous removal of both pollutants was compared to that achieved with control solutions containing either Cr(VI) or Brij 35 separately. Reactions were carried out over 24h at initial pH=2.3 with variable initial amounts of Cr(VI) and Brij 35. The results showed that surfactant removal was enhanced in the system also containing Cr(VI). Surfactant degradation followed zero-order kinetics and produced formic acid as the main by-product, together with hydroxylated aldehydes, formates and alcohols that were identified by LC/MS. The presence of surfactant similarly enhanced Cr(VI) reduction, which also followed zero-order kinetics. Chromium removal was quantitative only when the initial chromium concentration was lower than 140 mg L(-1). Reduced chromium was mainly in the solution phase together with dissolved iron. Precipitation with NaOH was therefore required to definitively remove dissolved metals from the investigated system.

Peroxymonosulfate-Co(II) oxidation system for the removal of the non-ionic surfactant Brij 35 from aqueous solution.

The non-ionic surfactant Brij 35 was effectively removed from concentrated aqueous solution by the peroxymonosulfate/Co(II) system, using oxone (2KHSO(5)·KHSO(4)·K(2)SO(4)) as a source of peroxymonosulfate. At pH=2.3 and initial Brij 35 concentration in the range 680-2410 mg L(-1), 86-94% removal was achieved after 24 h, using Co(II)=15 µM and oxone=5.9 mM. The effectiveness of removal did not change when initial pH was in the range 2.3-8.2. After five subsequent additions of Co(II) and oxone to the solution, COD and TOC removals increased up to 64% and 33%, respectively. Radical quenching tests confirmed that sulfate radical was the dominant radical species in the system. The main identified by-products from surfactant degradation were: (a) low molecular weight organic acids; (b) aldehydes and formates with shorter ethoxy chain than Brij 35; (c) alcohol ethoxylates carrying hydroxyl groups bonded to ethoxy chain. By-products identification allowed to hypothesize the pathways of Brij 35 degradation.

Degradation of chlorobenzene by Fenton-like processes using zero-valent iron in the presence of Fe3+ and Cu2+.

Batch and column tests were conducted to compare the efficiencies of three Fenton-like systems in the degradation of chlorobenzene. In the investigated systems, iron powder was the source of Fe2+ ions, and either Fe3+ or Cu2+ were added in order to enhance the degradation process. Optimum pH and concentrations of Fe3+, Cu2+ and hydrogen peroxide were assessed by treating synthetic chlorobenzene solutions both in batch and in column mode, thus achieving the minimum residual concentration of chlorobenzene. Optimum conditions assessed by means of column tests were then applied to column treatment of real chlorobenzene-contaminated groundwater. Results showed that, when the TOC content of water was above 12 mg L(-1), the performance of the Fe0/H2O2 system was enhanced by the addition of Fe3+, leading to 10-20 microg L(-1) of residual chlorobenzene.

A combined PHREEQC-2/parallel fracture model for the simulation of laminar/non-laminar flow and contaminant transport with reactions.

A combination of a parallel fracture model with the PHREEQC-2 geochemical model was developed to simulate sequential flow and chemical transport with reactions in fractured media where both laminar and turbulent flows occur. The integration of non-laminar flow resistances in one model produced relevant effects on water flow velocities, thus improving model prediction capabilities on contaminant transport. The proposed conceptual model consists of 3D rock-blocks, separated by horizontal bedding plane fractures with variable apertures. Particle tracking solved the transport equations for conservative compounds and provided input for PHREEQC-2. For each cluster of contaminant pathways, PHREEQC-2 determined the concentration for mass-transfer, sorption/desorption, ion exchange, mineral dissolution/precipitation and biodegradation, under kinetically controlled reactive processes of equilibrated chemical species. Field tests have been performed for the code verification. As an example, the combined model has been applied to a contaminated fractured aquifer of southern Italy in order to simulate the phenol transport. The code correctly fitted the field available data and also predicted a possible rapid depletion of phenols as a result of an increased biodegradation rate induced by a simulated artificial injection of nitrates, upgradient to the sources.

Enhanced bioremediation of methyl tert-butyl ether (MTBE) by microbial consortia obtained from contaminated aquifer material.

A microcosm study was carried out to evaluate the potential for biodegradation of methyl tert-butyl ether (MTBE) impacting groundwater at a former oil refinery site located in Naples (SW Italy). A screening of aerobic, anaerobic and co-metabolic aerobic conditions was carried out by triplicate batch reactors, using contaminated soil and groundwater from the study site. All microcosms were amended with ammonium and phosphate salts and, if aerobic, they were supplied with excess oxygen throughout the static incubation period of 6 months. Propane, pentane and n-hexane were selected as the primary substrates for co-metabolic treatments. After the initial lag phase (40-60d), quantitative MTBE decay was repeatedly observed in the aerobic set amended only with nitrogen and phosphorus and further fed with MTBE, thus suggesting that the indigenous soil bacteria have the ability to degrade MTBE. All other treatments, i.e., anaerobic and co-metabolic aerobic, resulted unsuccessful after incubation extending up to 190d. Bacterial consortia in the active microcosms were later enriched and further studied through second and third generation batch reactors with no soil, operated under continuous mixing for 5-7 months. MTBE degradation rate progressively increased with reactor operating time, following a zero order kinetics in the concentration range 1-10mgL(-1) and leading to a residual concentration of less than 10microgL(-1). The calculated maximum biodegradation rate was 20mg(MTBE)g(VSS)(-1)h(-1). An accumulation of nitrite ions also occurred after long operating times, thus inhibiting MTBE degradation. This effect was minimized by replacing ammonium with nitrate. Identified degradation intermediates were tert-butyl alcohol and tert-butyl formate. Fluorescent in situ hybridization was applied for a preliminary microbiological screening of the consortia, suggesting that the detected cocci (about 0.5 and 1.5microm diameter, respectively) and long bacilli with a narrow diameter might be as yet undescribed species.

Chlorinated herbicide (triallate) dehalogenation by iron powder.

The reductive degradation of a chlorinated herbicide by iron powder was investigated at lab scale. The studied substrate was triallate (S-2,3,3-trichloroallyl di-isopropyl thiocarbamate) which contains a trichloroethylene moiety potentially reducible by zero-valent iron. Degradation reactions were carried out in batch, at 25 degrees C, in the absence of oxygen, by contacting electrolytic iron powder (size range: 20-50 microm) with a triallate aqueous solution (2.5 mgl(-1)). Herbicide decay, corresponding evolutions of TOC, TOX and chloride ion release were regularly monitored throughout the reactions. Furthermore, the main degradation by-products were identified by HPLC/MS. The results showed that, after 5 days, herbicide degradation extent was about 97% and that the reaction proceeded through the formation of a dechlorinated alkyne by-product (S-2-propinyl di-isopropyl thiocarbamate) resulting from the complete dechlorination of triallate. The subsequent reduction of such an alkyne intermediate gave S-allyl di-isopropyl thiocarbamate as main end by-product. The identified by-products suggested that dechlorination took place mainly via reductive beta-elimination. However, as traces of dichloroallyl di-isopropyl thiocarbamate were also detected, a role, although minor, was assigned even to hydrogenolysis in the overall dechlorination process.

Fenton's pre-treatment of mature landfill leachate.

The aim of this study was to check the effectiveness of the Fenton's reagent (Fe2+ + H2O2 + H+) for the pre-treatment of a municipal landfill leachate with the objective of improving its overall biodegradability, evaluated in terms of BOD5/COD ratio, up to a value compatible with biological treatment. The leachate came from a municipal sanitary landfill located in southern Italy and the average values of its main parameters were: pH=8.2; COD=10,540 mgl(-1); BOD5=2,300 mgl(-1); TOC=3,900 mgl(-1); NH4-N=5210 mgl(-1); conductivity=45,350 microScm(-1); alkalinity=21,470 mgl(-1) CaCO3. The effect of initial pH value on the pre-treatment effectiveness was evaluated by titrating the amount of acidic by-products formed. The extent of leachate oxidation was monitored and controlled by both pH and redox potential measurements. The best operational conditions for achieving the desired goal (i.e., BOD5/COD> or =0.5) resulted: Fe2+=275 mgl(-1); H2O2=3,300 mgl(-1); initial pH=3; reaction time=2 h. At the end of the Fenton's pre-treatment, in order to permit a subsequent biological treatment, residual ferric ions were removed increasing the pH up to 8.5 by adding 3 gl(-1) of Ca(OH)2 and 3 mgl(-1) of a cationic polyelectrolyte, the latter as an aid to coagulation. This final step also resulted in a further modest removal of residual COD due to co-precipitation phenomena.

Olive husk: an alternative sorbent for removing heavy metals from aqueous streams.

Sorption properties of olive husk were investigated under equilibrium (batch tests) and dynamic (column tests) conditions in order to assess the possibility of using such a waste material for removing heavy metals from aqueous streams. Husk samples were contacted, at 25 degrees C, with aqueous solutions of nitric salts of Pb, Cd, Cu, and Zn. Sorption isotherms obtained from equilibrium data were fitted and interpreted by the Freundlich model. Metals-saturated husk samples resulting from column tests were air-dried and incinerated to simulate combustion in order to assess the fate of sorbed metals. The results demonstrated that, under both equilibrium and dynamic conditions, metal sorption capacity of the husk was in the sequence Pb>Cd>Cu>Zn. For all the metals, calculated Freundlich constants decreased by increasing initial metal concentration or decreasing solution pH. In dynamic tests, a significant reduction of sorption capacity was recorded (except for copper) when a metal was fed simultaneously to the others: Pb (77%); Cd (93%); Zn (68%). Combustion tests carried out on metals-saturated husk samples showed that the average losses of lead and cadmium, as volatile species, were always three to four times greater than the losses of copper and zinc, in both single-metal- and multimetal-saturated samples.