Antibacterial activity of silver nanoparticles grafted on stone surface

Microbial colonization has a relevant impact on the deterioration of stone materials with consequences ranging from esthetic to physical and chemical changes. Avoiding microbial growth on cultural stones therefore represents a crucial aspect for their long-term conservation. The antimicrobial properties of silver nanoparticles (AgNPs) have been extensively investigated in recent years, showing that they could be successfully applied as bactericidal coatings on surfaces of different materials. In this work, we investigated the ability of AgNPs grafted to Serena stone surfaces to inhibit bacterial viability. A silane derivative, which is commonly used for stone consolidation, and Bacillus subtilis were chosen as the grafting agent and the target bacterium, respectively. Results show that functionalized AgNPs bind to stone surface exhibiting a cluster disposition that is not affected by washing treatments. The antibacterial tests on stone samples revealed a 50 to 80 % reduction in cell viability, with the most effective AgNP concentration of 6.7 μg/cm². To our knowledge, this is the first report on antimicrobial activity of AgNPs applied to a stone surface. The results suggest that AgNPs could be successfully used in the inhibition of microbial colonization of stone artworks.     

Capability of the natural microbial community in a river water ecosystem to degrade the drug naproxen

The present work aims at evaluating the ability of the River Tiber natural microbial community to degrade naproxen in water samples collected downstream from a wastewater treatment plant. For this purpose, different water microcosms were set up (microbiologically active vs sterile ones) and treated with naproxen (100 μg/L) alone or in the co-presence of gemfibrozil in order to evaluate if the co-presence of the latter had an influence on naproxen degradation. The experiment was performed in the autumn and was compared with the same experimental set performed in spring of the same year to highlight if seasonal differences in the river water influenced the naproxen degradation. Pharmaceutical concentrations and microbial analysis (total cell number, viability, and microbial community composition) were performed at different times in the degradation experiments. The overall results show that the natural microbial community in the river water had a key role in the naproxen degradation. In fact, although there was a transient negative effect on the natural microbial community in all the experiments (3 h after adding the pharmaceutical), the latter was able to degrade naproxen within about 40 days. On the contrary, no decrease in the pharmaceutical concentration was observed in the sterile river water. Moreover, the co-presence of the two drugs lengthened the naproxen lag phase. As regards the natural microbial community composition detected by Fluorescence in situ Hybridization, Alpha and Gamma-Proteobacteria increased when the pharmaceutical halved, suggesting their role in the degradation. This study shows that with the concentration studied, naproxen was degraded by the natural microbial populations collected from a river chronically contaminated by this pharmaceutical.     

Illuminating reactive microbial transport in saturated porous media: demonstration of a visualization method and conceptual transport model

We demonstrate a method to study reactive microbial transport in saturated translucent porous media using the bacteria Pseudomonas fluorescens 5RL genetically engineered to carry a plasmid with bioluminescence genes inducible by salicylate. Induced bacteria were injected into a cryolite grain filled chamber saturated with a sterile non-growth-promoting (phosphorus limited) chemical mixture containing salicylate as an aromatic hydrocarbon analogue. The amount of light produced by the bacteria serves as an estimator of the relative efficiency of aerobic biodegradation since bioluminescence is dependent on both salicylate and oxygen but only consumes oxygen. Bioluminescence was captured with a digital camera and analyzed to study the evolving spatial pattern of the bulk oxygen consuming reactions. As fluid flow transported the bacteria through the chamber, bioluminescence was observed to initially increase until an oxygen depletion zone developed behind the advective front. Bacterial transport was modeled with the advection dispersion equation and oxygen concentration was modeled assuming bacterial consumption via Monod kinetics with consideration of additional effects of rate-limited mass transfer from residual gas bubbles. Consistent with previous measurements, bioluminescence was considered proportional to oxygen consumed. Using the observed bioluminescence, model parameters were fit that were consistent with literature values and produced results in good agreement with the experimental data. These findings demonstrate potential for using this method to investigate the complex spatial and temporal dynamics of reactive microbial transport in saturated porous media.     

Evolution of clog formation with time in columns permeated with synthetic landfill leachate

Laboratory column tests conducted to gain insight regarding the biological and chemical clogging mechanisms in a porous medium are presented. To seed the porous medium with landfill bacteria, a mixture of Keele Valley Landfill and synthetic leachate permeated through the column under anaerobic conditions for the first 9 days of operation. After this, 100% synthetic leachate was used. The synthetic leachate approximated Keele Valley Landfill leachate in chemical composition but contained negligible suspended solids and bacteria compared with real leachate. The removal of volatile fatty acids (VFAs), primarily acetate, in leachate as it passed through the medium was highly correlated with the precipitation of calcium carbonate (CaCO(3(s))) from solution. The columns experienced a decrease in drainable porosity from an initial value of about 0.38 to less than 0.1 after steady state chemical oxygen demand (COD) removal, resulting in a five-order magnitude decrease in hydraulic conductivity. The decrease in drainable porosity prior to steady state COD removal was primarily due to the growth of a biofilm on the medium surface. After steady state COD removal, calcium precipitation was at least equally responsible for the decrease in drainable porosity as biofilm growth. Clog composition analyses showed that CaCO(3(s)) was the dominant clog constituent and that 99% of the carbonate in the clog material was bound to calcium.     

Study of the biodegradation process of polychlorinated biphenyls in liquid medium and soil by a new isolated aerobic bacterium (Janibacter sp.)

We have isolated and characterised a novel aerobic bacterial strain, designated MS3-02, belonging to the genus Janibacter sp. The capability of this new strain to degrade polychlorinated biphenyls (PCBs) in a commercial mixture (Aroclor 1242) in liquid medium and in soil (sterile and non sterile soil), under laboratory scale, has been evaluated. MS3-02 was isolated from the soil around of an incinerator, located in the east of Madrid (Spain). Gas-chromatographic analysis showed that MS3-02 was able to reduce most peaks observed in the chromatogram between 70% and 100% after seven days of incubation in a culture mineral medium containing yeast extract, but without the addition of biphenyl. The presence of biphenyl in the culture medium decreased the rate of PCB degradation by this bacterium. Comparing the performance of the MS3-02 in liquid culture medium and in soil, degradation was less efficient in sterile soil and still less efficient in non sterile soil. Under the best conditions (sterile soil and 20 weeks of incubation) MS3-02 was able to reduce, between 50% and 100%, nine of the main gas-chromatographic peaks in Aroclor 1242.     

The influence of substrate and electron acceptor availability on bioactive zone dynamics in porous media

Two approaches were used to investigate the influence of dissolved oxygen (DO) and substrate availability on the formation and dynamics of "bioactive zones" in a water-saturated porous medium. A bioactive zone is defined as a region where a microbial community is sufficiently active to metabolize bioavailable substrates. In the first approach, microbial activity was characterized by monitoring the spatial and temporal variability of DO and aqueous substrate (salicylate and naphthalene) concentrations during miscible-displacement experiments. In the second approach, microbial activity was monitored using multiple fiber optics emplaced in the porous medium to detect luminescence produced by Pseudomonas putida RB1353, a bioluminescent reporter organism that produces light when salicylate (an intermediate of naphthalene degradation) is present. The results of both approaches show that the location and size of the bioactive zones were influenced by in situ DO and substrate availability. When DO was not a limiting factor (i.e., lower substrate input concentrations), the bioactive zone encompassed the entire column, with the majority of the microbial activity occurring between the inlet and midpoint. However, as the availability of DO became limiting for the higher substrate input experiments, the size of the bioactive zone shrank and was ultimately limited to the proximity of the column inlet.     

Microbial community dynamics in uranium contaminated subsurface sediments under biostimulated conditions with high nitrate and nickel pressure

BACKGROUND, AIM, AND SCOPE: The subsurface at the Oak Ridge Field Research Center represents an extreme and diverse geochemical environment that places different stresses on the endogenous microbial communities, including low pH, elevated nitrate concentrations, and the occurrence of heavy metals and radionuclides, including hexavalent uranium [U(VI)]. The in situ immobilization of U(VI) in the aquifer can be achieved through microbial reduction to relatively insoluble U(IV). However, a high redox potential due to the presence of nitrate and the toxicity of heavy metals will impede this process. Our aim is to test biostimulation of the endogenous microbial communities to improve nitrate reduction and subsequent U(VI) reduction under conditions of elevated heavy metals. MATERIALS AND METHODS: Column experiments were used to test the possibility of using biostimulation via the addition of ethanol as a carbon source to improve nitrate reduction in the presence of elevated aqueous nickel. We subsequently analyzed the composition of the microbial communities that became established and their potential for U(VI) reduction and its in situ immobilization. RESULTS: Phylogenetic analysis revealed that the microbial population changed from heavy metal sensitive members of the actinobacteria, alpha- and gamma-proteobacteria to a community dominated by heavy metal resistant (nickel, cadmium, zinc, and cobalt resistant), nitrate reducing beta- and gamma-proteobacteria, and sulfate reducing Clostridiaceae. Coincidentally, synchrotron X-ray absorption spectroscopy analyses indicated that the resulting redox conditions favored U(VI) reduction transformation to insoluble U(IV) species associated with soil minerals and biomass. DISCUSSION: This study shows that the necessary genetic information to adapt to the implemented nickel stress resides in the endogenous microbial population present at the Oak Ridge FRC site, which changed from a community generally found under oligotrophic conditions to a community able to withstand the stress imposed by heavy metals, while efficiently reducing nitrate as electron donor. Once nitrate was reduced efficient reduction and in situ immobilization of uranium was observed. CONCLUSIONS: This study provides evidence that stimulating the metabolism of the endogenous bacterial population at the Oak Ridge FRC site by adding ethanol, a suitable carbon source, results in efficient nitrate reduction under conditions of elevated nickel, and a decrease of the redox potential such that sulfate and iron reducing bacteria are able to thrive and create conditions favorable for the reduction and in situ immobilization of uranium. Since we have found that the remediation potential resides within the endogenous microbial community, we believe it will be feasible to conduct field tests. RECOMMENDATIONS AND PERSPECTIVES: Biostimulation of endogenous bacteria provides an efficient tool for the successful in situ remediation of mixed-waste sites, particularly those co-contaminated with heavy metals, nitrate and radionuclides, as found in the United States and other countries as environmental legacies of the nuclear age.     

Limestone Dissolution in Flue Gas Scrubbing: Effect of Sulfite

Batch limestone dissolution experiments were carried out in a pH stat apparatus at 55°C with CO2 sparging and dissolved sulfite. Particle size distribution, utilization, sulfite in solution, limestone type, and the approach to calcite equilibrium were all found to contribute to the limestone reactivity. In the absence of sulfite, limestone dissolution was controlled solely by mass transfer. For a given stone under mass transfer control, film thickness was found to be independent of pH. The dissolution rate in the presence of sulfite was controlled by a combined surface kinetics/mass transfer regime. SEM micrographs supported this conclusion. A surface rate correlation was developed which accounted for observed inhibition by an inverse dependence on calcium sulfite concentration at the limestone surface. While the form of the rate expression was applicable to all stones, the surface rate constant was stone dependent. A computer code which accounted for mass transfer with surface kinetics was tested against experimental observations of four limestone types. Changes in pH and the concentrations of calcium, carbonate, sulfite, sulfate, and adipic acid were accurately modeled.     

Effect of acid rain on building material of the El Tajín archaeological zone in Veracruz, Mexico

Three limestone slabs (approx. 20 cm2 each) were extracted from the El Tajin archaeological zone in Veracruz, Mexico. X-ray diffraction analysis revealed three components: calcite (81.2%), quartz (17.9%) and feldspar (0.9%). Calcite content by x-ray diffraction analysis was slightly higher than that determined by chemical reaction between the limestone sample and nitric acid. The latter analysis, carried out in triplicate, yielded a calcite content of 77.1%. Mean water absorption, density and porosity of the limestone samples were also determined. Dissolution of limestone samples was investigated using an experimental rainfall simulation chamber, in which the stone samples were irrigated with artificial rain matching the pH and the ionic composition with 40 rainfall samples collected at El Tajin from August 18, 2002, to April 9, 2003. According to calcium and bicarbonate net concentrations found in the effluent of the chamber, a chemical mechanism by which limestone at El Tajin is dissolved by acid rain is proposed. A model used to investigate the air transport pathways corresponding to precipitation events at El Tajin shows that air parcels come mainly from the Gulf of Mexico, although no directional preference is evident for acidic vs. non-acidic events.     

Straining phenomena in bacteria transport through natural porous media

Background, aim, and scope  

Transport of bacteria through natural porous media is an issue of increasing concern arising in several very important environmental processes. These include the percolation of bacteria from fecal waste to drinking water reservoirs, thus leading to a risk for human health, or the bioremediation of contaminated soils in which the bacteria are expected to travel long distances underground in order to reach contaminated areas and degrade chemicals originating from accidental spills. An understanding of bacterial retention and transport mechanisms in porous media would be of great help in the development of models able to predict the distance covered by bacterial suspensions in these situations.     

Mineralization of PAHs in coal-tar impacted aquifer sediments and associated microbial community structure investigated with FISH

The microbial community structure and mineralization of polycyclic aromatic hydrocarbons (PAHs) in a coal-tar contaminated aquifer were investigated spatially using fluorescence in situ hybridization (FISH) and in laboratory-scale incubations of the aquifer sediments. DAPI-detected microbial populations in the contaminated sediments were three orders of magnitude greater than nearby uncontaminated sediments, suggesting growth on coal-tar constituents in situ. Actinobacteria, beta- and gamma-Proteobacteria, and Flavobacteria dominated the in situ aerobic (>1 mg l(-1) dissolved oxygen) microbial community, whereas sulfate-reducing bacteria comprised 37% of the microbial community in the sulfidogenic region of the aquifer. Rapid mineralization of naphthalene and phenanthrene were observed in aerobic laboratory microcosms and resulted in significant enrichment of beta- and gamma-Proteobacteria potentially explaining their elevated presence in situ. Firmicutes, Flavobacteria, alpha-Proteobacteria, and Actinobacteria were also enriched in the mineralization assays, but to a lesser degree. Nitrate- and sulfate-limited mineralization of naphthalene in laboratory microcosms occurred to a small degree in aquifer sediments from locations where groundwater chemistry indicated nitrate- and sulfate-reduction, respectively. Some iron-limited mineralization of naphthalene and phenanthrene was also observed in sediments originating near groundwater measurements of elevated ferrous iron. The results of this study suggest that FISH may be a useful tool for providing a much needed link between laboratory microcosms and groundwater measurements made in situ necessary to better demonstrate the potential for natural attenuation at complex PAH contaminated sites.     

Coupling permanganate oxidation with microbial dechlorination of tetrachloroethene.

For sites contaminated with chloroethene non-aqueous-phase liquids, designing a remediation system that couples in situ chemical oxidation (ISCO) with potassium permanganate (KMnO4) and microbial dechlorination may be complicated because of the potentially adverse effects of ISCO on anaerobic bioremediation processes. Therefore, one-dimensional column studies were conducted to understand the effect of permanganate oxidation on tetrachloroethene (PCE) dechlorination by the anaerobic mixed culture KB-1. Following the confirmation of PCE dechlorination, KMnO4 was applied to all columns at a range of concentrations and application velocities to simulate varied distances from oxidant injection. Immediately following oxidation, reductive dechlorination was inhibited; however, after passing several pore volumes of sterile growth medium through the columns after oxidation, a rebound of PCE dechlorination activity was observed in every inoculated column without the need to reinoculate. The volume of medium required for a rebound of dechlorination activity differed from 1.1 to 8.1 pore volumes (at a groundwater velocity of 4 cm/d), depending on the specific condition of oxidant application.     

Interaction between water flow and spatial distribution of microbial growth in a two-dimensional flow field in saturated porous media

Bacterial growth and its interaction with water flow was investigated in a two-dimensional flow field in a saturated porous medium. A flow cell (56 x 44 x 1 cm) was filled with glass beads and operated under a continuous flow of a mineral medium containing nitrate as electron acceptor. A glucose solution was injected through an injection port, simulating a point source contamination. Visible light transmission was used to observe the distribution of the growing biomass and water flow during the experiment. At the end of the experiment (on day 31), porous medium samples were destructively collected and analyzed for abundance of total and active bacterial cells, bacterial cell volume and concentration of polysaccharides and proteins. Microbial growth was observed in two stripes along the length of the flow cell, starting at the glucose injection port, where highest biomass concentrations were obtained. The spatial distribution of biomass indicated that microbial activity was limited by transverse mixing between glucose and nitrate media, as only in the mixing zone between the media high biological activities were achieved. The ability of the biomass to change the flow pattern in the flow cell was observed, indicating that the biomass was locally reducing the hydraulic conductivity of the porous medium. This bioclogging effect became evident when the injection of the glucose solution was turned off and water flow still bypassed the area around the glucose injection port, preserving the flow pattern as it was during the injection of the glucose solution. As flow bypass was possible in this system, the average hydraulic properties of the flow cell were not affected by the produced biomass. Even in the vicinity of the injection port, the total volume of the bacterial cells remained below 0.01% of the pore space and was unlikely to be responsible for the bioclogging. However, the bacteria produced large amounts of extracellular polymeric substances (EPS), which likely caused the observed bioclogging effects.     

Methanotrophic activity in a diffusive methane/oxygen counter-gradient in an unsaturated porous medium

Microbial methane (CH4) oxidation is a main control on emissions of this important greenhouse gas from ecosystems such as contaminated aquifers or wetlands under aerobic onditions. Due to a lack of suitable model systems, we designed a laboratory column to study this process in diffusional CH4/O2 counter-gradients in unsaturated porous media. Analysis and simulations of the steady-state CH4, CO2 and O2 gas profiles showed that in a 15-cm-deep active zone, CH4 oxidation followed first-order kinetics with respect to CH4 with a high apparent first-order rate constant of approximately 30 h(-1). Total cell counts obtained using DAPI-staining suggested growth of methanotrophic bacteria, resulting in a high capacity for CH4 oxidation. This together with apparent tolerance to anoxic conditions enabled a rapid response of the methanotrophic community to changing substrate availability, which was induced by changes in O2 concentrations at the top of the column. Microbial oxidation was confirmed by a approximately 7 per thousand enrichment in CH4 stable carbon isotope ratios along profiles. Using a fractionation factor of 1.025+/-0.0005 for microbial oxidation estimated from this shift and the fractionation factor for diffusion, simulations of isotope profiles agreed well with measured data confirming large fractionation associated with microbial oxidation. The designed column should be valuable for investigating response of methanotrophic bacteria to environmental parameters in future studies.     

Volatile metabolites from mold growth on building materials and synthetic media

Mold species which were isolated from damp buildings were grown on sterile building materials and some synthetic media in order to study the microbial volatile organic compounds produced. Patterns of the microbial volatile organic compounds (MVOC) were very media dependent but media which favor terpene biosynthesis may give patterns unique enough for identification of dominant indoor molds.     

Investigations on mobility of carbon colloid supported nanoscale zero-valent iron (nZVI) in a column experiment and a laboratory 2D-aquifer test system

Nanoscale zero-valent iron (nZVI) has recently gained great interest in the scientific community as in situ reagent for installation of permeable reactive barriers in aquifer systems, since nZVI is highly reactive with chlorinated compounds and may render them to harmless substances. However, nZVI has a high tendency to agglomerate and sediment; therefore it shows very limited transport ranges. One new approach to overcome the limited transport of nZVI in porous media is using a suited carrier colloid. In this study we tested mobility of a carbon colloid supported nZVI particle “Carbo-Iron Colloids” (CIC) with a mean size of 0.63 μm in a column experiment of 40 cm length and an experiment in a two-dimensional (2D) aquifer test system with dimensions of 110?×?40?×?5 cm. Results show a breakthrough maximum of 82 % of the input concentration in the column experiment and 58 % in the 2D-aquifer test system. Detected residuals in porous media suggest a strong particle deposition in the first centimeters and few depositions in the porous media in the further travel path. Overall, this suggests a high mobility in porous media which might be a significant enhancement compared to bare or polyanionic stabilized nZVI.     

Zn biomineralization processes and microbial biofilm in a metal-rich stream (Naracauli,Sardinia)

Several decades after the closure of the Ingurtosu mine (SW Sardinia), a variety of seasonal Zn biomineralizations occurs. In this work, waters, microbial consortia, and seasonal precipitates from the Naracauli stream were sampled to investigate chemical composition of stream waters and biominerals, and microbial strain identity. Molecular and morphological analysis revealed that activity of dominant cyanobacterium Leptolyngbya frigida results in precipitation of Zn silicate. The activity of the cyanobacterium was associated to other bacteria and many kind of diatoms, such as Halamphora subsalina and Encyonopsis microcephala, which are trapped in the process of biomineral growth. In this work, the precipitation process is shown to be the result of many different parameters such as hydrologic regime, microbial community adaptation, and biological mediation. It results in a decrease of dissolved Zn in the stream water, and is a potential tool for Zn pollution abatement.     

Soil microbial parameters and luminescent bacteria assays as indicators for in situ bioremediation of TNT-contaminated soils

In situ bioremediation is increasingly being discussed as a useful strategy for cleaning up contaminated soils. Compared to established ex situ procedures, meaningful and reliable approaches for monitoring the remediation processes and their efficiency are of special importance. The subject of this study was the significance of two bioassays for monitoring purposes. The work was performed within the scope of a research project on the in situ bioremediation of topsoil contaminated with 2,4,6-trinitrotoluene (TNT). To evaluate changes within different experimental fields during a 17-month remediation period, the results of soil microbial assays and luminescent bacteria assays were compared with chemical monitoring data. The luminescent bacteria assays showed a significant reduction of the water-soluble soil toxicants in the treated fields. This bioassay proved to be a sensitive screening indicator of toxicity and may effectively aid the ecotoxicological interpretation of chemical monitoring data. Microbial biomass (C(mic)), the metabolic quotient (qCO2), and the ratio of microbial to organic carbon (C(mic)/C(org)) showed a highly significant correlation with total concentrations of TNT in the soil. But, in contrast to luminescent bacteria assays, this approach did not reveal any recovery of the soil at the end of the remediation period. There is clear evidence for persistent adverse effects of chronic TNT contamination on the site-specific microbial community and the local carbon cycle in the soil. The study clearly exhibits the differences between, as well as the complementary value of both bioassay approaches for monitoring short-term and long-term effects of soil contamination and the efficiency of remediation.     

Conditions d'acceptation ou de rejet des valeurs obtenues apres denombrement de virus ou bacteries sur milieux de culture solides

Enumeration of viruses or bacteria on solid culture media does not ever give statistically acceptable results. When counting is impossible, we must do it again. For accepting or not the counting, the authors established some probability Tables (95% and 99%) according to microbial densities.     

Water transformation in the media of biofilters controlled by Rhodococcus fascians in treating an ethyl acetate-contaminated airstream

Biofilters do not provide much water for bacteria to grow. To use them efficiently and properly, it is essential to understand the kinetics of water transformation and to control moisture levels. This study aims to clarify whether the metabolism of microorganisms will improve the water-holding capacity of media or will intensify drying. This experiment was conducted in duplicate, that is, both with and without bacterial inoculation. Both the constant water content mode and the declining water content mode show that microbial growth in a log phase will enhance drying. In contrast, the bacteria growing in a logarithmic decline phase will improve water-holding capacity. Basically, water evaporation can result from the latent heat obtained from microbial respiration or from the physical temperature difference between the unsaturated air and the wet media. Two ways that biofilters can gain water are from water incorporated into bacteria cells and from water obtained from the oxidation of volatile organic compounds (VOCs).