Acclimated green microalgae consortium to treat sewage in an alternative urban WWTP in a coastal area of Central Italy.

This study exposed a microalgal consortium formed by Auxenochlorella protothecoides, Tetradesmus obliquus, and Chlamydomonas reinhardtii to six mixed wastewater media containing different proportions of primary (P) or secondary (S) effluents diluted in centrate (C). Algae could grow at centrate concentrations up to 50 %, showing no significant differences between effluents. After acclimation, microalgae cultivated in 50%P-50%C and 50%S-50%C grew at a rate similar to that of control cultures (0.59-0.66 d-1). These results suggest that the consortium acclimated to both sewage streams by modulating the proportion of the species and their metabolism. Acclimation also altered the photosynthetic activity of wastewater-grown samples compared to the control, probably due to partial photoinhibition, changes in consortium composition, and changes in metabolic activity. No major differences were observed between the two streams with respect to biochemical composition, biomass yield, or bioremediation capacity of the cultivated algae but algae grown in the secondary effluent showed qualitatively higher exopolysaccharides (EPS) production than algae grown in primary. Regarding wastewater remediation, microalgae grown in both WW media showed proficient nutrient removal efficiencies (close to 100 %); however, the final pH value (close to 11) would be controversial if the system were upscaled as it is over the legal limit and would cause phosphorus precipitation, so that CO2 addition would be required. The theoretical scale-up of the microalgae system could achieve water treatment costs of 0.109 €·m-3, which was significantly lower than the costs of typical activated sludge systems.

Study of the stability of a paramagnetic label linked to mesoporous silica surface in contact with rat mesothelial cells in culture.

Stable radicals detectable by electron paramagnetic resonance (EPR) may be use in the investigation of early events in cell-particle toxicity. Piperidine-N-oxyl derivatives (nitroxides), covalently linked to the surface of a high surface area silica (used as model solid for the technique), served as probes in the investigation of the effects of incubation of silica particles with mesothelial cells. A mesoporous silica (MCM-41), prepared by precipitation from a micellar solution, was the most appropriate silica-based particle for this purpose, as its channels allow direct contact with small molecules but not with macromolecules. The cytotoxicity of this amorphous silica is very low, allowing relatively high particle loading in the cell cultures. Both the high surface area of the sample and the large amount of inorganic material extracted from the cell culture provide enough material to run reasonably intense EPR spectra. Computer-aided analysis of the EPR spectra of silica-bound nitroxides provided information on the sensitivity of the labeled silica monitoring different environments, e.g., to follow the path of particles in a mammalian cell culture. Upon contact of the particles with mesothelial cells, the mean distance among the labels at the silica surface decreased as a consequence of the release of oxidizing and/or radical moieties from the cells.

Use of Nitroxides as Topological Monitors of the Interaction of Silica-Based Particles with Components of the Biological Environment

The interaction of solid particles, such as silica and vitreous fibers, with different surrounding media which well mimic the various environments in a biological medium, such as inhaled in vivo or in a cell culture, has been studied by means of the electron paramagnetic resonance (EPR) spectra of spin labels attached to the solid surface or spin probes inserted in the surrounding medium. Among the solid particles, a MCM-41 type mesoporous silica was found to be very suitable for investigating the binding between the labels and different molecules, due to the high surface area and the availability of interacting sites in the internal channels of the structure. The computer-aided analysis of the spectral lineshape allowed the evaluation of structural and dynamic parameters. A model has been proposed which describes the interactions of the solid surface with: (a) pure solvents at different polarities; (b) molecules present in biological fluids, which mimic the effect of physiological solutions; (c) the components of the cell membrane (phospholypid or proteins in water solution); and (d) a model phospholypid membrane, to mimic the interaction between the solid particles and the cell membrane. The hydration of the surface lets the labels interact preferentially with the water molecules with respect to the surface itself, or the other labels. Apolar molecules decreased the mobility of the labels attached to the surface. Phospholipid bilayers were formed at the solid surface, whose internal structure was more fluid with respect to noninteracting bilayers, whereas the external polar groups trapped probe and label molecules in restricted space at the surface. The labels were partially extracted from the wet surface of the vitreous fibers by the interaction with a protein (albumin) and distributed in two different environments (at different polarities).

Role of iron in the reactivity of mineral fibers.

Any foreign body containing iron may be (or become) highly toxic in vivo. If its solubility in water is poor, surface chemistry governs the reactivity at the solid-liquid interface. Iron toxicity thus increases with the extent of exposed surface. Iron of endogenous origin may also be deposited on the particle surface and be activated under particular circumstances. The chemical processes that implicate surface iron as a primary cause of toxicity are: free radical release, mobilization by chelators, iron-catalyzed reactions. Three kinds of solids are compared: (i) well-known toxic materials, for example asbestos; (ii) non-toxic iron oxides; and (iii) model solids with surface exposed iron prepared for investigations on the reactivity of iron in biological media. The iron content of the solid is not directly related to the biological response: only a small fraction of ions, in a well-defined coordination and redox state, appears involved in the toxicity of the mineral dust.

Free radical generation at the solid/liquid interface in iron containing minerals.

The potential for free radical release has been measured by means of the spin trapping technique on three kinds of iron containing particulate: two asbestos fibers (chrysotile and crocidolite); an iron-exchanged zeolite and two iron oxides (magnetite and haematite). DMPO (5,5'-dimethyl-1-pirroline-N-oxide), used as spin trap in aqueous suspensions of the solids, reveals the presence of the hydroxyl and carboxylate radicals giving rise respectively to the two adducts [DMPO-OH] and [DMPO-CO2], each characterized by a well-defined EPR spectrum. Two target molecules have been considered: the formate ion to evidence potential for hydrogen abstraction in any biological compartment and hydrogen peroxide, always present in the phagosome during phagocytosis. The kinetics of decomposition of hydrogen peroxide has also been measured on all solids. Ferrozine and desferrioxamine, specific chelators of Fe(II) and Fe(III) respectively, have been used to remove selectively iron ions. Iron is implicated in free radical release but the amount of iron at the surface is unrelated to the amount of radicals formed. Only few surface ions in a particular redox and coordination state are active. Three different kinds of sites have been evidenced: one acting as H abstracter, the other as a heterogeneous catalyst for hydroxyl radical release, the third one related to catalysis of hydrogen peroxide disproportionation. In both mechanisms of free radical release, the Fe-exchanged zeolite mimics the behaviour of asbestos whereas the two oxides are mostly inert. Conversely magnetite turns out to be an excellent catalyst for hydrogen peroxide disproportionation while haematite is inactive also in this reaction. The results agree with the implication of a radicalic mechanism in the in vitro DNA damage and in the in vivo toxicity of asbestos.

Physicochemical mechanism of the interaction between cobalt metal and carbide particles to generate toxic activated oxygen species.

Hard metal alloys (or cemented carbides) are made of a mixture of tungsten carbide particles (WC, more than 80%) cemented in cobalt metal powder (Co, 5-10%). The inhalation of hard metal particles may cause an interstitial pulmonary disease, the mechanism of which involves an interaction between Co and WC particles. Some epidemiological data also suggest that hard metal dust can induce lung cancer in workers. In a macrophage culture model, butylated hydroxytoluene (1 mM) protected from the cytotoxicity of hard metal particles, suggesting a possible involvement of lipid peroxidation in the toxicity of these powders. In a biochemical system, a mixture of Co and WC particles, but not Co or WC alone, stimulated the production of thiobarbituric acid-reactive substances from arachidonic acid. Using a spin trapping system applied to aqueous particulate suspensions and electrochemical techniques, we present experimental evidence that the association of Co and carbide particles represents a specific toxic entity producing large amounts of activated oxygen species. The mechanism of this interaction proceeds through the oxidation of cobalt metal catalyzed at the surface of carbide particles and resulting in the reduction of dissolved oxygen. This physicochemical property of hard metal particles provides a new basis for interpreting their inflammatory action and their possible carcinogenic effect on the lung.