The purpose of this study was to evaluate the performance of scanning mobility particle sizers in the characterization of nanoaerosols. A sampling chamber was constructed from aluminum and tempered glass, had a volume of 4.6 cubic feet, and was designed for the introduction of aerosols and dilution air, maintenance of aerosol concentration, and continuous exhaust of chamber air. Penetration and aerosol distribution tests were conducted within the chamber. An aerosol generation and measurement system comprised of nitrogen gas, BGI 3 jet Collison Nebulizer, diffusion dryer, aerosol charge neutralizer, mixing chamber, critical orifice, hygrometer, condensation particle counter, scanning mobility particle sizer, air sampling pump, air sampling cassettes, and a vacuum pump was assembled. A BGI 3 jet Collison Nebulizer was used to generate the nanoparticle aerosols. The two types of nanoparticle aerosols utilized in the experiment were salt (NaCl) and polystyrene latex (PSL) spheres. Relative humidity and temperature measurements were obtained within the chamber. Real-time, direct-reading particle measurement instruments including a condensation particle counter (CPC) (TSI, Model 3007), and three scanning mobility particle sizer (SMPS) instruments (Particle Measuring Systems, Nano-ID NPS500; TSI, NanoScan SMPS Nanoparticle Sizer Model 3910) were used for particle measurements. For each test run, two air samples were collected on membrane filters for electron microscopy (EM) analysis. Eight trials were conducted using NaCl nanoaerosols, and twelve trials were conducted using PSL spheres. The selected particle sizes for the experiments were 57 nm, 92 nm, 147 nm, and 220 nm. For the NaCl nanoaerosol suspensions, the SMPS lines of fit were log-normally distributed and predominantly parallel. The geometric standard deviation (GSD) of these distributions was approximately 1.7, which confirms that the distributions were approximately the same. In these experiments, instrument 3 identified a higher percentage of NaCl particles within the size range intervals of the selected NaCl size parameter, and the count median diameters (CMDs) for the instrument 3 measurements were closer to the selected NaCl size parameter more often than the other instruments. This suggests that instrument 3 was more responsive than the other instruments to the selected size range and the selected NaCl size parameters. The electron microscopy (EM) lines of fit for the NaCl experiments were predominantly parallel with the SMPS lines of fit, suggesting that the log-normally distributions are similar. The GSD of EM distributions was approximately 1.8, which confirms that the distributions were approximately the same as the SMPS distributions. Results from the regression plots demonstrated that the main effects and interaction were statistically significant with a p<0.0001. The coefficient of determination, R2, for the regression lines was 0.87. The post-hoc Tukey HSD results identified a significant difference between the instrument 3 dataset, and the datasets for instruments 1 and 2. For the PSL nanoaerosol suspensions, the SMPS lines of fit were log-normally distributed and predominantly parallel. The GSD of these distributions was approximately 1.3, which confirms that the distributions were approximately the same. In these experiments, instrument 2 identified a higher percentage of PSL particles within the size range intervals of the selected PSL size parameter, and instrument 2 CMDs were closer to the selected PSL size parameter more often than the other instruments. This suggests that instrument 2 was more responsive than the other instruments to the selected size range and the selected PSL size parameters. Results from the regression plots demonstrated that the main effects and interaction were statistically significant with a p<0.01. The coefficient of determination, R2, for the regression lines was 0.44. The post-hoc Tukey HSD test identified a significant difference between the instrument 3 dataset and the instrument 1 dataset. Potential sources of variability include solution water background contamination, surfactants in the PSL solution, and agglomeration. The performance of all the scanning mobility particle sizers compared in these experiments was acceptable for research and field applications, but caution should be taken when comparing the measurements of SMPS, especially SMPS from different manufacturers.

Engineered nanomaterials (ENMs) have been increasingly used in various applications. Often, the ENMs are functionalised with a surface coating to enhance their properties. Decades of research has provided information on mostly pristine and unmodified ENMs, while ecotoxicity of coated ENMs and how their hazard changes with age in soils is still uncertain. The thesis aimed to determine the toxic effects and bioaccumulation potential of CuO ENMs and CdTe quantum dots (QDs) with different chemical coatings (carboxylate, COOH; polyethylene glycol, PEG; ammonium, NH4+) on the earthworm (Eisenia fetida), and compare the effects to their metal salt (CuSO4) or micron-sized counterpart. Then, to determine if any observed toxicity was altered after ageing the soils for up to one year. Incidental plant growth was studied in the exposure soils to maximise the scientific value of the earthworm tests. Toxic effects of CuO ENMs were also assessed in Caenorhabditis elegans exposed in liquid and soil media to understand effects of the media and method of dosing on ENM toxicity. CuO ENMs were equally toxic to earthworms, or less toxic to plants than the dissolved Cu; whereas CdTe QD ENMs were more toxic than the micron-sized CdTe QDs. There was a coating effect in both, CuO and CdTe QD ENM experiments, the -COOH coated ENMs were most toxic in the fresh soil study, while -NH4+ coated ENMs were most toxic in the aged soil study. Despite the similarities in the toxicity ranking, the biological effects exerted were different between CuO and CdTe QD ENMs. In C. elegans exposures, the ENMs were more hazardous than dissolved Cu, but ranking of ENMs depended on the media and method of dosing. The results suggest the coating effect is determined by the reactivity of the coating in a given media, and it also depends on the core of the ENMs. As such, coating and ageing effects should be considered in the risk assessment of ENMs.

No description available.

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2011. / Please refer to full text for abstracts

Nanotechnology is an increasing market. Engineered nanoparticles (NPs), materials with at least one dimension between 1 and 100 nm, are produced on a large scale. NPs are vastly used in industrial processes and consumer products and they are most likely discharged into wastewater treatment plants after being used. Activated Sludge is one of the most applied biological wastewater treatment processes for the degradation of organic matter in sewage. Activated sludge produces an excess of sludge that is commonly treated and stabilized by anaerobic digestion. Recent studies have found that NPs accumulate in the activated sludge; thus, there is a potential for the concentrations of NPs to magnify as concentrated waste sludge is fed into the anaerobic digestion process. For this reason, it is important to study the possible toxic effects of NPs on the microorganisms involved in the anaerobic digestion process and the approaches to overcome toxicity if necessary. The present work evaluates the toxic effect of NPs on anaerobic wastewater treatment processes and also presents approaches for toxicity attenuation. The first objective of this dissertation (Chapter III) was to evaluate the toxicity of high concentrations (1, 500 mg L⁻¹) of Ag⁰, Al₂O₃, CeO₂, Cu⁰, CuO, Fe⁰, Fe₂O₃, Mn₂O₃, SiO₂, TiO₂, and ZnO NPs to acetoclastic and hydrogenotrophic methanogens and the effect of a dispersant on the NPs toxicity to methanogens. The findings indicated that only Cu⁰ and ZnO NPs caused severe toxicity to hydrogenotrophic methanogens and Cu⁰, CuO, and ZnO NPs to acetoclastic methanogens. The dispersant did not impact the NPs toxicity. The concentrations of Cu⁰ and ZnO causing 50% of inhibition (IC₅₀) to hydrogenotrophic methanogens were 68 and 250 mg L⁻¹, respectively. Whereas the IC₅₀ values for acetoclastic methanogens were 62, 68, and 179 for Cu⁰, ZnO, and CuO-Cu NPs respectively. These findings indicate that acetoclastic methanogens are more sensitive to NP toxicity compared to hydrogenotrophic methanogens and that Cu⁰ and ZnO NPs are highly toxic to both. Additionally, it was observed that the toxicity of any given metal was highly correlated with its final dissolved concentration in the assay irrespective of whether it was initially added as a NP or chloride salt, indicating that corrosion and dissolution of metals from NPs may have been responsible for the toxicity. The second objective of this dissertation (Chapter IV) was to evaluate the Cu⁰ NP toxicity to anaerobic microorganisms of wastewater treatment processes. Cu⁰ is known to be toxic to methanogens; nonetheless, little is known about its toxic effects on microorganisms of upper trophic levels of anaerobic digestion or other anaerobic process used for nitrogen removal. This specific objective evaluated Cu⁰ NP toxicity to glucose fermentation, syntrophic propionic oxidation, methanogenesis, denitrification and anaerobic ammonium oxidation (anammox). Chapter IV showed that anammox and glucose fermentation were the least and most inhibited processes with inhibition constants (K(i)) values of 0.324 and 0.004 mM of added Cu⁰ NPs, respectively. The Ki values obtained from the residual soluble concentration of the parallel experiments using CuCl₂ indicated that Cu⁰ NP toxicity is most likely caused by the release of soluble ions for each one of the microorganisms tested. The results taken as a whole demonstrate that Cu⁰ NPs are toxic to a variety of anaerobic microorganisms of wastewater treatment processes. The third objective of this document (Chapter V) was to study the role of biogenic sulfide in attenuating Cu⁰ and ZnO NP toxicity to acetoclastic methanogens. Previous literature results and research presented in this dissertation indicated that the release of soluble ions from Cu and ZnO NPs cause toxicity to methanogens. In the past, the application of sulfide to precipitate heavy metals as inert non-soluble sulfides was used to attenuate the toxicity of Cu and Zn salts. Building on this principle, Chapter V evaluated the toxicity of Cu⁰ and ZnO NPs in sulfate-containing (0.4 mM) and sulfate-free conditions. The results show that Cu⁰ and ZnO were 7 and 14x less toxic in sulfate-containing than in sulfate-free assays as indicated by the differences in K(i) values. The K(i) values obtained based on the residual metal concentration of the sulfate-free and sulfate-containing assays were very similar, indicating that the toxicity is well correlated with the release of soluble ions. Overall, this study demonstrated that biogenic sulfide is an effective attenuator of Cu⁰ and ZnO NP toxicity to acetoclastic methanogens. Finally, the last objective (Chapter VI) of this dissertation was to evaluate the effect of iron sulfide (FeS) on the attenuation of Cu⁰ and ZnO toxicity to acetoclastic methanogens. FeS is formed by the reaction of iron(II) and sulfide. This reaction is common in anaerobic sediments where the reduction of iron(III) to iron(II) and sulfate to sulfide occurs. FeS plays a key role controlling the soluble concentrations of heavy metals and thus their toxic effects in aquatic sediments. This study evaluated the application of FeS as an approach to attenuate Cu⁰ and ZnO NP toxicity and their salt analogs to acetoclastic methanogens. Two particle sizes, coarse FeS (FeS-c, 500-1200 µm) and fine FeS (FeS-f, 25-75 µm) were synthesized and used in this study. The results showed 2.5x less FeS-f than FeS-c was required to recover the methanogenic activity to the same extent from the exposure to highly inhibitory concentrations of CuCl₂ and ZnCl₂ (0.2 mM). The results also showed that a molar ratio of FeS-f/Cu⁰, FeS-f/ZnO, FeS-f/Zn Cl₂, and FeS-f/CuCl₂ of 3, 3, 6, and 12 respectively, was necessary to provide a high recovery of methanogenic activity (>75%). The excess of FeS needed to overcome the toxicity indicates that not all the sulfide in FeS was readily available to attenuate the toxicity. Overall, Chapter VI demonstrated that FeS is an effective attenuator of the toxicity of Cu⁰ NP and ZnO NPs and their salt analogs to methanogens, albeit molar excesses of FeS were required.

Antimicrobial engineered nanomaterials (ENM) are increasingly incorporated into products despite limited understanding of the interactions between ENMs and bacteria that lead to toxic impacts. The hazard posed by increasing environmental release of antimicrobial ENMs is also poorly characterized. The overall objective of this thesis is to inform questions about the types of interactions that lead to an ENM inducing bacterial toxicity. Many antimicrobial ENMs are soluble, and the ion plays an important role in their toxicity. Some believe that, beyond release of ions, ENM toxicity is expected to derive from a nanoparticle (NP)-specific effect. This research compares bacterial responses to ENMs, their metal salts, and/or their transformed species within different experimental settings to improve our understanding of the interactions that enable ENM bacterial toxicity. The first objective is to characterize the potential hazard posed by pristine and transformed antimicrobial ENMs on microbial communities within a complex environmental system. One pair of ENMs (Ag0 and Ag2S) led to differential short-term impacts on surficial sediment microbial communities, while the other did not (CuO and CuS), showing that ENM transformation does not universally lead to distinct impacts. The metal ion (Cu2+) had a more profound microbial community impact than did any of the four ENMs. By 300 days the microbial community structure and composition re-converged, suggesting minimal long-term impacts of high pulse inputs of antimicrobial ENMs on microbial communities within complex environments. The second objective is to identify NP-specific effects of a common antimicrobial ENM on a model bacterium. Analysis of transcriptional responses identified NP-specific induction of a membrane stress responsive gene, providing evidence of a NP-specific effect. Otherwise, our results suggest that CuO NP toxicity triggers the same stress responses as does Cu2+, but at more moderate levels. Two ion treatments with the same total Cu input – one with pulse addition and one with gradual addition that was meant to better represent the slow dissolution of the CuO NP – led to temporally distinct responses. This calls for the use of more representative ion controls for comparison against soluble NP impacts in future nanotoxicity studies. The third objective is to investigate the potential use of CuO ENMs to reduce virulence and growth of an emerging bacterial pathogen. CuO NP exposure led to reduction in relative expression of three Staphylococcus aureus virulence factor genes, especially in methicillinresistant S. aureus (MRSA) clinical isolates. Growth was inhibited at high CuO NP concentrations for all four isolates, too. Comparison across all genes assayed showed isolatespecific transcriptional responses, but with NP- and ion-induced responses showing clear differences for each isolate, too. Altogether, this research contributes novel knowledge that will guide efforts to characterize potential hazard from release of ENMs into the environment and to apply ENMs for effective antibacterial treatment.

No description available.

The purpose of this dissertation was to broadly investigate the fate of antibiotic resistance genes (ARGs) and engineered nanomaterials (ENMs) as representative contaminants of emerging concern in wastewater treatment plants (WWTPs). WWTPs may have their performance impacted by ENMs and may also serve as a reservoir and point of release for both ENMs and ARGs into the environment. Of interest were potential adverse effects of ENMs, such as stimulation of antibiotic resistance in the WWTP, toxicity to microbial communities critical for WWTP performance, and toxicity to humans who may be exposed to effluents or aerosols containing ENMs and their transformation products. Response of nine representative ARGs encoding resistance to sulfonamide, erythromycin and tetracycline to various lab-scale sludge digestion processes were examined, and factors that drove the response of ARGs were discussed. Mesophilic anaerobic digestion significantly reduced sulI, sulII, tet(C), tet(G), and tet(X) with longer solids retention time (SRT) exhibiting a greater extent of removal. Thermophilic anaerobic digesters performed similarly to each other and provided more effective reduction of erm(B), erm(F), tet(O), and tet(W) compared to mesophilic digestion. Thermal hydrolysis pretreatment drastically reduced all ARGs, but they generally rebounded during subsequent anaerobic and aerobic digestion treatments. Bacterial community composition of the sludge digestion process, as controlled by the physical operating characteristics, was indicated to drive the distribution of ARGs present in the produced biosolids, more so than the influent ARG composition. Effects of silver (nanoAg), zero-valent iron (NZVI), titanium dioxide (nanoTiO2) and cerium dioxide (nanoCeO2) nanomaterials on nitrification function and microbial communities were examined in duplicate lab-scale nitrifying sequencing batch reactors (SBRs), relative to control SBRs received no materials or ionic/bulk analogs. Nitrification function was only inhibited by high load of 20 mg/L Ag+, but not by other nanomaterials or analogs. However, decrease of nitrifier gene abundances and distinct microbial communities were observed in SBRs receiving nanoAg, Ag+, nanoCeO2, and bulkCeO2. There was no apparent effect of nanoTiO2 or NZVI on nitrification, nitrifier gene abundances, or microbial community structure. A large portion of nanoAg remained dispersed in activated sludge and formed Ag-S complexes, while NZVI, nanoTiO2 and nanoCeO2 were mostly aggregated and chemically unmodified. Thus, the nanomaterials appeared to be generally stable in the activated sludge, which may limit their effect on nitrification function or microbial community structure. Considering an aerosol exposure scenario, cytotoxicity and genotoxicity of aqueous effluent and biosolids from SBRs dosed with nanoAg, NZVI, nanoTiO2 and nanoCeO2 to A549 human lung epithelial cells were examined, and the effects were compared relative to outputs from SBRs dosed with ionic/bulk analogs and undosed SBRs, as well as pristine ENMs. Although the pristine nanomaterials showed varying extents of cytotoxicity to A549 cells, and gentoxicity was observed for nanoAg, no significant cytotoxic or genotoxic effects of the SBR effluents or biosolids containing nanomaterials were observed. Studies presented in this dissertation provided new insights in the fate of ARGs in various sludge digestion processes and ENMs in nitrifying activated sludge system in lab-scale reactors. The study also yielded toxicity data of ENMs to biological wastewater treatment microbial communities and human lung cells indicated by a variety of toxicity markers. The results will aid in identifying appropriate management technologies for sludge containing ARGs and will inform microbial and human toxicity assessments of ENMs entering WWTPs. / Ph. D.

Metal-based engineered nanomaterials (ENMs) have potential to revolutionize diagnosis, drug delivery and manufactured products, leading to greater human ENM exposure. It is crucial to understand ENM pharmacokinetics and their association with biological barriers such as the blood-brain barrier (BBB). Physicochemical parameters such as size and surface modification of ENMs play an important role in ENM fate, including their brain association. Multifunctional ENMs showed advantages across the highly regulated BBB. There are limited reports on ENM distribution among the blood in the brain vasculature, the BBB, and brain parenchyma. In this study, ceria ENM was used to study the effect of size on its pharmacokinetics. Four sizes of ceria ENMs were studied. Five nm ceria showed a longer half-life in the blood and higher brain association compared with other sizes and 15 and 30 nm ceria had a higher blood cell association than 5 or 55 nm ceria. Because of the long circulation and high brain association of 5 nm ceria compared with other sizes, its distribution between the BBB and brain parenchyma was studied. The in situ brain perfusion technique showed 5 nm ceria (99%) on the luminal surface of the BBB rather than the brain parenchyma. For biomedical applications in the central nervous system (CNS), it is vital to develop stable and biocompatible ENMs and enhance their uptake by taking advantage of their unique properties. Cross-linked nanoassemblies entrapping iron oxide nanoparticles (CNA-IONPs) showed controlled particle size in biological conditions and less toxicity in comparison to Citrate-IONPs. CNA-IONPs considerably enhanced MRI T2 relaxivities and generated heat at mild hyperthermic temperatures (40 ~ 42°C) in the presence of alternating magnetic field (AMF). Numerous researchers showed mild whole body hyperthermia can increase BBB permeability for potential brain therapeutic application. Compared to conventional hyperthermia, AMF-induced hyperthermia increased BBB permeability with a shorter duration of hyperthermia and lower temperature, providing the potential to enhance IONP flux across the BBB with reduced toxicity. Overall, ENMs with optimized physicochemical properties can enhance their flux across the BBB into the brain with desirable pharmacokinetics, which provide great potential for diagnosis and therapy in the CNS.

Ceria (cerium oxide) nanomaterials, or nanoceria, have commercial catalysis and energy storage applications. The cerium atoms on the surface of nanoceria can store or release oxygen, cycling between Ce3+ and Ce4+, and can therefore act as a therapeutic to relieve oxidative stress within living systems. Nanoceria dissolution is present in acidic environments in vivo. In order to accurately define the fate of nanoceria in vivo, nanoceria dissolution or stabilization is observed in vitro using acidic aqueous environments. Nanoceria stabilization is a known problem even during its synthesis; in fact, a carboxylic acid, citric acid, is used in many synthesis protocols. Citric acid adsorbs onto nanoceria surfaces, capping particle formation and creating stable dispersions with extended shelf lives. Nanoceria was shown to agglomerate in the presence of some carboxylic acids over a time scale of up to 30 weeks, and degraded in others, at pH 4.5 (representing that of phagolysosomes). Sixteen carboxylic acids were tested: citric, glutaric, tricarballylic, α-hydroxybutyric, β-hydroxybutyric, adipic, malic, acetic, pimelic, succinic, lactic, tartronic, isocitric, tartaric, dihydroxymalonic, and glyceric acid. Each acid was introduced as 0.11 M, into pH 4.5 iso-osmotic solutions. Controls such as ammonium nitrate, sodium nitrate, and water were also tested to assess their effects on nanoceria dissolution and stabilization. To further test stability, nanoceria suspensions were subject to light and dark milieu, simulating plant environments and biological systems, respectively. Light induced nanoceria agglomeration in some, but not all ligands, and is likely to be a result of UV irradiation. Light initiates free radicals generated from the ceria nanoparticles. Some of the ligands completely dissolved the nanoceria when exposed to light. Citric and malic acids form coordination complexes with cerium on the surface of the ceria nanoparticle that can inhibit agglomeration. This approach identifies key functional groups required to prevent nanoceria agglomeration. The impact of each ligand on nanoceria was analyzed and will ultimately describe the fate of nanoceria in vivo. In addition, simulated biological fluid (SBF) exposure can change nanoceria’s surface properties and biological activity. The citrate-coated nanoceria physicochemical properties such as size, morphology, crystallinity, surface elemental composition, and charge were determined before and after exposure to simulated lung, gastric, and intestinal fluids. SBF exposure resulted in either loss or overcoating of nanoceria’s surface citrate by some of the SBF components, greater nanoceria agglomeration, and small changes in the zeta potential. Nanocomposites are comprised of a polymer matrix embedded with nanoparticles. These nanoparticles can alter material and optical properties of the polymer. SR-399 (dipentaerythritol pentaacrylate) is a fast cure, low skin irritant monomer that contains five carbon-carbon double bonds (C=C). It is a hard, flexible polymer, and also resistant to abrasion. It can be used as a sealant, binder, coating, and as a paint additive. In this case, metal oxide nanoparticles were added to the monomer prior to polymerization. Titania nanoparticles are known to absorb UV light due to their photocatalytic nature. Titania nanoparticles were chosen due to their high stability, non-toxicity, and are relatively quick, easy, and inexpensive to manufacture. Channels in thin monomer films were created using a ferrofluid manipulated by magnetic fields. The mechanical properties of a microfluidic device by rapid photopolymerization is dependent on the crosslinking gradient observed throughout the depth of the film. Quantitative information regarding the degree of polymerization of thin film polymers polymerized by free radical polymerization through the application of UV light is crucial to estimate material properties. In general, less cure leads to more flexibility, and more cure leads to brittleness. The objective was to quantify the degree of polymerization to approximate the C=C concentration and directly relate it to the mechanical properties of the polymer. Polymerization of C=C groups was conducted using a photoinitiator and an UV light source from one surface of a thin film of a multifunctional monomer. The C=C fraction in the film was found to vary with film depth and UV light intensity. The extents of conversion and crosslinking estimates were compared to local mechanical moduli and optical properties. A mathematical model linking the mechanical properties to the degree of polymerization, C=C composition, as a function of film depth and light intensity was then developed. For a given amount of light energy, one can predict the hardness and modulus of elasticity. The correlation between the photopolymerization and the mechanical properties can be used to optimize the mechanical properties of thin films within the manufacturing and energy constraints, and should be scalable to other multifunctional monomer systems.