Contaminant-Activated Visible Light Photocatalysis.

Pristine titanium dioxide (TiO2) absorbs ultraviolet light and reflects the entire visible spectrum. This optical response of TiO2 has found widespread application as white pigments in paper, paints, pharmaceuticals, foods and plastic industries; and as a UV absorber in cosmetics and photocatalysis. However, pristine TiO2 is considered to be inert under visible light for these applications. Here we show for the first time that a bacterial contaminant (Staphylococcus aureus-a MRSA surrogate) in contact with TiO2 activates its own photocatalytic degradation under visible light. The present study delineates the critical role of visible light absorption by contaminants and electronic interactions with anatase in photocatalytic degradation using two azo dyes (Mordant Orange and Procion Red) that are highly stable because of their aromaticity. An auxiliary light harvester, polyhydroxy fullerenes, was successfully used to accelerate photocatalytic degradation of contaminants. We designed a contaminant-activated, transparent, photocatalytic coating for common indoor surfaces and conducted a 12-month study that proved the efficacy of the coating in killing bacteria and holding bacterial concentrations generally below the benign threshold. Data collected in parallel with this study showed a substantial reduction in the incidence of infections.

Influence of Suwannee River humic acid on particle properties and toxicity of silver nanoparticles.

Adsorption of natural organic matter (NOM) on nanoparticles can have dramatic impacts on particle dispersion resulting in altered fate and transport as well as bioavailability and toxicity. In this study, the adsorption of Suwannee River humic acid (SRHA) on silver nanoparticles (nano-Ag) was determined and showed a Langmuir adsorption at pH 7 with an adsorption maximum of 28.6 mg g(-1) nano-Ag. It was also revealed that addition of <10 mg L(-1) total organic carbon (TOC) increased the total Ag content suspended in the aquatic system, likely due to increased dispersion. Total silver content decreased with concentrations of NOM greater than 10mg TOCL(-1) indicating an increase in nanoparticle agglomeration and settling above this concentration. However, SRHA did not have any significant effect on the equilibrium concentration of ionic Ag dissolved in solution. Exposure of Daphnia to nano-Ag particles (50 µg L(-1) and pH 7) produced a linear decrease in toxicity with increasing NOM. These results clearly indicate the importance of water chemistry on the fate and toxicity of nanoparticulates.

Polyhydroxy fullerenes (fullerols or fullerenols): beneficial effects on growth and lifespan in diverse biological models.

Recent toxicological studies on carbon nanomaterials, including fullerenes, have led to concerns about their safety. Functionalized fullerenes, such as polyhydroxy fullerenes (PHF, fullerols, or fullerenols), have attracted particular attention due to their water solubility and toxicity. Here, we report surprisingly beneficial and/or specific effects of PHF on model organisms representing four kingdoms, including the green algae Pseudokirchneriella subcapitata, the plant Arabidopsis thaliana, the fungus Aspergillus niger, and the invertebrate Ceriodaphnia dubia. The results showed that PHF had no acute or chronic negative effects on the freshwater organisms. Conversely, PHF could surprisingly increase the algal culture density over controls at higher concentrations (i.e., 72% increase by 1 and 5 mg/L of PHF) and extend the lifespan and stimulate the reproduction of Daphnia (e.g. about 38% by 20 mg/L of PHF). We also show that at certain PHF concentrations fungal growth can be enhanced and Arabidopsis thaliana seedlings exhibit longer hypocotyls, while other complex physiological processes remain unaffected. These findings may open new research fields in the potential applications of PHF, e.g., in biofuel production and aquaculture. These results will form the basis of further research into the mechanisms of growth stimulation and life extension by PHF.

Optical heating and rapid transformation of functionalized fullerenes.

Irradiating single-walled carbon nanotubes can lead to heat generation or ignition. These processes could be used in medical and industrial applications, but the poor solvent compatibility and high aspect ratios of nanotubes have led to concerns about safety. Here, we show that certain functionalized fullerenes, including polyhydroxy fullerenes (which are known to be environmentally safe and to have therapeutic properties) are heated or ignited by exposure to low-intensity (<10(2 ) W cm(-2)) continuous-wave laser irradiation. We also show that polyhydroxy fullerenes and other functionalized fullerenes can be transformed into single-walled nanotubes, multiwalled nanotubes and carbon onions without the presence of a catalyst by exposure to low-intensity laser irradiation in an oxygen-free environment. To demonstrate the potential usefulness of these processes in applications, we disrupted animal cells dosed with polyhydroxy fullerenes by exposing them to a near-infrared laser for a few seconds, and also ignited an explosive charge in contact with a particle of carboxy fullerenes.

Influence of alternating oxic/anoxic conditions on growth of denitrifying bacteria.

Denitrifying bacteria that are switched from oxic to anoxic conditions can experience diauxic lag, which is the time required for re-synthesis of nitrate reductase and other denitrifying enzymes. Pseudomonas denitrificans were exposed to alternating oxic/anoxic phases in a continuous flow reactor with either 4-h or 8-h anoxic phase lengths, in comparison to a measured diauxic lag of 9.5h following steady-state oxic conditions. The P. denitrificans were unable to sustain anoxic growth at either of the anoxic phase lengths tested. Diauxic lag observed after several cycles of alternating oxic/anoxic phases was significantly longer than the diauxic lag measured after steady-state oxic conditions. This may be attributed to increase of cell maintenance energy requirements due to substrate accumulation during anoxic phases and concomitant high specific growth rates during oxic phases.

Effect of periplasmic nitrate reductase on diauxic lag of Paracoccus pantotrophus.

Paracoccus pantotrophus expresses two nitrate reductases-membrane bound nitrate reductase (Nar) and periplasmic nitrate reductase (Nap). In growth experiments with two denitrifying species (Paracoccus pantotrophus and Alcaligenes eutrophus) that have both Nap and Nar and two species (Pseudomonas denitrificans and Pseudomonas fluorescens) with Nar only, it was found that diauxic lag is shorter for bacteria that express Nap. In P. pantotrophus, napEDABC encodes the periplasmic nitrate reductase. To analyze the effect of Nap on diauxic lag, the nap operon was deleted from P. pantotrophus. The growth experiments with nap(-) mutant resulted in increased diauxic lag when switched from aerobic to anoxic respiration, suggesting Nap is responsible for shorter lags and helps in adaptation to anoxic metabolism after transition from aerobic conditions.

Effect of UVA irradiance on photocatalytic and UVA inactivation of Bacillus cereus spores.

The current study is the first to delineate the contribution of photocatalysis to inactivation of Bacillus cereus endospores on dry surfaces over a broad range (0-153W/m(2)) of UVA irradiance. Inactivation of spores at low UVA irradiance (30W/m(2)) was primarily due to photocatalysis, whereas at higher UVA irradiance inactivation was primarily due to UV alone. A linear relationship between UVA irradiance and the rate of spore inactivation was observed in the absence of photocatalyst. The rate of photocatalytic inactivation was non-linear with respect to UVA irradiance, exhibiting a maximum at 30W/m(2).

Determination of diauxic lag in continuous culture.

A procedure was developed to characterize diauxic lag of bacteria switching between electron acceptors in continuous culture. In this procedure, a virtual batch growth curve is developed by integrating the time-dependent net specific growth rates of bacteria observed under continuous flow conditions. The length of diauxic lag and the highest net specific growth rate following lag are conveniently estimated from the virtual batch curve. The procedure was found to give reproducible diauxic lag lengths and highest net specific growth rates when applied to experimental data from replicate continuous culture trials.

Estimation of nitrate reductase enzyme parameters in activated sludge using an extended Kalman filter algorithm.

This paper demonstrates an application of the extended Kalman filter to a wastewater plant using real process data to estimate parameters in a semi-mechanistic model. This technique for parameter identification allows a semi-mechanistic model developed for pure cultures to be applied to a mixed culture population where isolation of enzyme kinetic parameters is not practical. This paper demonstrates an application in which a wastewater treatment facility with three physical unit operations and an unusual operating profile is modeled as 11 reactor units with recirculation streams. On-line nutrient probe data from two locations in the aerobic reactor are combined with operational data for flows and feed composition. This paper demonstrates the successful application of a biochemical model developed at the bench scale to this facility through the use of an extended Kalman filter. The estimated biological model parameters and the covariance matrix diagonal converge to stable values, indicating a successful implementation.

Dominance of endospore-forming bacteria on a Rotating Activated Bacillus Contactor biofilm for advanced wastewater treatment.

The bacterial diversity inherent to the biofilm community structure of a modified rotating biological contactor wastewater treatment process, referred to as the Rotating Activated Bacillus Contactor (RABC) process, was characterized in this study, via both culture-dependent and culture-independent methods. On the basis of culture-dependent methods, Bacillus sp. were found to exist in large numbers on the biofilm (6.5% of the heterotrophic bacteria) and the microbial composition of the biofilms was quite simple. Only three phyla were identified-namely, the Proteobacteria, the Actinobacteria (High G+C Gram-positive bacteria), and the Firmicutes (Low G+C Gram-positive bacteria). The culture-independent partial 16S rDNA sequence analysis revealed a considerably more diverse microbial composition within the biofilms. A total of eight phyla were recovered in this case, three of which were major groups: the Firmicutes (43.9%), the Proteobacteria (28.6%), and the Bacteroidetes (17.6%). The remaining five phyla were minor groups: the Planctomycetes (4.4%), the Chlorobi (2.2%), the Actinobacteria (1.1%), the Nitrospirae (1.1%), and the Verrucomicrobia (1.1%). The two most abundant genera detected were the endospore-forming bacteria (31.8%), Clostridium and Bacillus, both of which are members of the Firmicutes phylum. This finding indicates that these endospore-forming bacteria successfully colonized and dominated the RABC process biofilms. Many of the colonies or clones recovered from the biofilms evidenced significantly high homology in the 16S rDNA sequences of bacteria stored in databases associated with advanced wastewater treatment capabilities, including nitrification and denitrification, phosphorus accumulation, the removal of volatile odors, and the removal of chlorohydrocarbons or heavy metals. The microbial community structures observed in the biofilms were found to correlate nicely with the enhanced performance of advanced wastewater treatment protocols.

Effect of carbon substrate on electron acceptor diauxic lag and anoxic maximum specific growth rate in species with and without periplasmic enzyme.

The effect of oxidation state of carbon substrate on the diauxic lag of facultative anaerobic denitrifying bacteria growing aerobically upon switching to anoxic growth was studied. Also studied was the effect on the anoxic maximum specific growth rate. Two pure bacteria cultures were used, Paracoccus pantotrophus, denitrifying bacteria containing a periplasmic nitrate reductase (Nap), and Pseudomonas denitrificans, denitrifying bacteria lacking the periplasmic nitrate reductase. The anoxic maximum specific growth rate of both cultures following a period of aerobic growth with identical dilution up to steady-state was indeed affected by the oxidation state of the carbon, with the most oxidized substrate yielding the highest anoxic maximum specific growth rate. The diauxic lags for Paracoccus pantotrophus were considerably shorter than those for Pseudomonas denitrificans, something expected due to the presence of Nap, an enzyme not affected by aerobiosis. Since the activity of Nap in Paracoccus pantotrophus under aerobic conditions has been shown to increase with the extent of reduction of the carbon substrate, it was also expected that the diauxic lag length for these bacteria would decrease as the reduction state of the carbon substrate increased. This could not be demonstrated, as no significant lags were observed for this species. Pseudomonas denitrificans exhibited a shorter diauxic lag with the more oxidized carbon source.

Enhancement of titanium dioxide photocatalysis by water-soluble fullerenes.

Fullerenes are known for their unique electronic properties including high electron affinity. Although use of fullerenes for scavenging photo-generated electrons from titanium dioxide particles has been demonstrated, no attempts have been made to utilize the unique properties of fullerenes to increase the efficacy of photocatalysis. The present study has demonstrated that a mixture of water-soluble polyhydroxy fullerenes (PHF) and titanium dioxide (anatase polymorph) enhances photocatalytic degradation of organic dye. The PHF molecules adsorbed to the surface of titanium dioxide due to electrostatic forces, with adsorption density being higher at lower pH values. The surface coverage of titanium dioxide nanoparticles by PHF molecules determined the extent of enhancement, with an optimum dosed weight ratio of PHF to titanium dioxide at 0.001. Hydroxylation and concomitant solubilization of fullerenes allow their unique electronic properties to be harnessed for photocatalysis.

A simple model for diauxic growth of denitrifying bacteria.

A simple model has been formulated to simulate diauxic growth of denitrifying bacteria. It is capable of fitting the experimental results of batch growth experiments with Pseudomonas denitrificans under various conditions. It successfully predicts the observed lags when a pure culture of this bacterium switches from oxygen to nitrate as terminal electron acceptor. The model includes the effect of carbon substrate limitation and length of aerobic phase and does not run into problems when switching from anoxic to aerobic conditions, unlike prior models of diauxic growth.

Structured model for denitrifier diauxic growth.

We present a model for diauxic growth of denitrifying bacteria in which nitrate reductase synthesis kinetics dominate the overall growth kinetics. The model is based on the assumption of the existence of a nitrate respiration operon, thereby linking the rate of nitrate uptake to the activity of nitrate reductase. We show that this approach can model diauxic growth of Pseudomonas denitrificans by conducting experiments in which nitrate reductase activity was measured during both lag and ensuing exponential growth phases. We consistently observed the pattern of low nitrate reductase enzyme activity during the lag phase, increasing before the onset of growth. By fitting model parameters we were able to successfully match experimental data for growth, nitrate uptake, and enzyme activity level.

Nitrite inhibition of aerobic growth of Acinetobacter sp.

Nitrite inhibition of Acinetobacter sp. growing under aerobic conditions was studied. Specific growth rates under non-limiting concentrations of acetate and dissolved oxygen averaged 0.62h(-1). Growth and phosphate uptake by Acinetobacter sp. were both inhibited by increasing nitrite concentrations. The median inhibitory concentration (IC50) of free nitrous acid (FNA) was 0.10 mg/L and the IC10 of FNA was 0.05 mg/L. Removing nitrite from cultures reversed the inhibitory effect. Comparison of the IC10 of FNA for Acinetobacter sp. to inhibitory concentrations for other wastewater heterotrophic bacteria suggests that Acinetobacter sp. are relatively sensitive to this compound.