Annalisa Onnis-Hayden

Mechanistic Toxicity Assessment of Nanomaterials by Whole-Cell-Array Stress Genes Expression Analysis

This study performed mechanistic toxicity assessment of nanosilver (nAg) and nanotitanium dioxide anatase (nTiO2_a) via toxicogenomic approach, employing a whole-cell-array library consisting of 91 recombinated Escherichia coli K12 strains with transcriptional GFP-fusions covering most known stress response genes. The results, for the first time, revealed more detailed transcriptional information on the toxic mechanism of nAg and nTiO2_a, and led to a better understanding of the mode of action (MOA) of metal and metal oxide nanomaterials (NMs). The detailed pathways network established for the oxidative stress system and for the SOS (DNA damage) repair system based on the temporal gene expression profiling data revealed the relationships and sequences of key genes involved in these toxin response systems. Both NMs were found to cause oxidative stress as well as cell membrane and transportation damage. Genotoxicity and DNA damage were also observed, although nTiO2_a induced SOS response via previously identified pathway and nAg seemed to induce DNA repair via a pathway different from SOS. We observed that the NMs at lower concentration tend to induce more chemical-specific toxicity response, while at higher concentrations, more general global stress response dominates. The information-rich real-time gene expression data allowed for identification of potential biomarkers that can be employed for specific toxin detection and biosensor developments. The concentration-dependent gene expression response led to the determination of the No Observed Transcriptional Effect Level (NOTEL) values, which can be potentially applied in the regulatory and risk assessment framework as an alternative toxicity assessment end point.

Implication of using different carbon sources for denitrification in wastewater treatments.

Application of external carbon sources for denitrification becomes necessary for wastewater treatment plants that have to meet very stringent effluent nitrogen limits (e.g., 3 to 5 mgTN/L). In this study, we evaluated and compared three carbon sources--MicroC (Environmental Operating Solutions, Bourne, Massachusetts), methanol, and acetate-in terms of their denitrification rates and kinetics, effect on overall nitrogen removal performance, and microbial community structure of carbon-specific denitrifying enrichments. Denitrification rates and kinetics were determined with both acclimated and non-acclimated biomass, obtained from laboratory-scale sequencing batch reactor systems or full-scale plants. The results demonstrate the feasibility of the use of MicroC for denitrification processes, with maximum denitrification rates (k(dmax)) of 6.4 mgN/gVSSh and an observed yield of 0.36 mgVSS/mgCOD. Comparable maximum nitrate uptake rates were found with methanol, while acetate showed a maximum denitrification rate nearly twice as high as the others. The maximum growth rates measured at 20 degrees C for MicroC and methanol were 3.7 and 1.2 day(-1), respectively. The implications resulting from the differences in the denitrification rates and kinetics of different carbon sources on the full-scale nitrogen removal performance, under various configurations and operational conditions, were assessed using Biowin (EnviroSim Associates, Ltd., Flamborough, Ontario, Canada) simulations for both pre- and post-denitrification systems. Examination of microbial population structures using Automated Ribosomal Intergenic Spacer Analysis (ARISA) throughout the study period showed dynamic temporal changes and distinct microbial community structures of different carbon-specific denitrifying cultures. The ability of a specific carbon-acclimated denitrifying population to instantly use other carbon source also was investigated, and the chemical-structure-associated behavior patterns observed suggested that the complex biochemical pathways/enzymes involved in the denitrification process depended on the carbon sources used.

Process optimization by decoupled control of key microbial populations: distribution of activity and abundance of polyphosphate-accumulating organisms and nitrifying populations in a full-scale IFAS-EBPR plant.

This study investigated the abundance and distribution of key functional microbial populations and their activities in a full-scale integrated fixed film activated sludge-enhanced biological phosphorus removal (IFAS-EBPR) process. Polyphosphate accumulating organisms (PAOs) including Accumulibacter and EBPR activities were predominately associated with the mixed liquor (>90%) whereas nitrifying populations and nitrification activity resided mostly (>70%) on the carrier media. Ammonia oxidizer bacteria (AOB) were members of the Nitrosomonas europaea/eutropha/halophila and the Nitrosomonas oligotropha lineages, while nitrite oxidizer bacteria (NOB) belonged to the Nitrospira genus. Addition of the carrier media in the hybrid activated sludge system increased the nitrification capacity and stability; this effect was much greater in the first IFAS stage than in the second one where the residual ammonia concentration becomes limiting. Our results show that IFAS-EBPR systems enable decoupling of solid residence time (SRT) control for nitrifiers and PAOs that require or prefer conflicting SRT values (e.g. >15 days required for nitrifiers and <5 days preferred for PAOs). Allowing the slow-growing nitrifiers to attach to the carrier media and the faster-growing phosphorus (P)-removing organisms (and other heterotrophs, e.g. denitrifiers) to be in the suspended mixed liquor (ML), the EBPR-IFAS system facilitates separate SRT controls and overall optimization for both N and P removal processes.

Life-Cycle Assessment of Advanced Nutrient Removal Technologies for Wastewater Treatment

Advanced nutrient removal processes, while improving the water quality of the receiving water body, can also produce indirect environmental and health impacts associated with increases in usage of energy, chemicals, and other material resources. The present study evaluated three levels of treatment for nutrient removal (N and P) using 27 representative treatment process configurations. Impacts were assessed across multiple environmental and health impacts using life-cycle assessment (LCA) following the Tool for the Reduction and Assessment of Chemical and Other Environmental Impacts (TRACI) impact-assessment method. Results show that advanced technologies that achieve high-level nutrient removal significantly decreased local eutrophication potential, while chemicals and electricity use for these advanced treatments, particularly multistage enhanced tertiary processes and reverse osmosis, simultaneously increased eutrophication indirectly and contributed to other potential environmental and health impacts including human and ecotoxicity, global warming potential, ozone depletion, and acidification. Average eutrophication potential can be reduced by about 70% when Level 2 (TN = 3 mg/L; TP = 0.1 mg/L) treatments are employed instead of Level 1 (TN = 8 mg/L; TP = 1 mg/L), but the implementation of more advanced tertiary processes for Level 3 (TN = 1 mg/L; TP = 0.01 mg/L) treatment may only lead to an additional 15% net reduction in life-cycle eutrophication potential.

Biotransformation of Two Pharmaceuticals by the Ammonia-Oxidizing Archaeon Nitrososphaera gargensis

The biotransformation of some micropollutants has previously been observed to be positively associated with ammonia oxidation activities and the transcript abundance of the archaeal ammonia monooxygenase gene (amoA) in nitrifying activated sludge. Given the increasing interest in and potential importance of ammonia-oxidizing archaea (AOA), we investigated the capabilities of an AOA pure culture, Nitrososphaera gargensis, to biotransform ten micropollutants belonging to three structurally similar groups (i.e., phenylureas, tertiary amides, and tertiary amines). N. gargensis was able to biotransform two of the tertiary amines, mianserin (MIA) and ranitidine (RAN), exhibiting similar compound specificity as two ammonia-oxidizing bacteria (AOB) strains that were tested for comparison. The same MIA and RAN biotransformation reactions were carried out by both the AOA and AOB strains. The major transformation product (TP) of MIA, α-oxo MIA was likely formed via a two-step oxidation reaction. The first hydroxylation step is typically catalyzed by monooxygenases. Three RAN TP candidates were identified from nontarget analysis. Their tentative structures and possible biotransformation pathways were proposed. The biotransformation of MIA and RAN only occurred when ammonia oxidation was active, suggesting cometabolic transformations. Consistently, a comparative proteomic analysis revealed no significant differential expression of any protein-encoding gene in N. gargensis grown on ammonium with MIA or RAN compared with standard cultivation on ammonium only. Taken together, this study provides first important insights regarding the roles played by AOA in micropollutant biotransformation.

Correlation of macroscopic aggregation behavior and microscopic adhesion properties of bacteria strains using a dimensionless Tabor's parameter.

Macroscopic adhesion-aggregation, floc formation, and subsequent transportation of microorganisms in porous media are closely related to the microscopic behavior and properties of individual cells. The classical Tabors parameter in colloidal science is modified to correlate the macroscopic aggregation and microscopic adhesion properties of microorganisms. Seven bacterial strains relevant to wastewater treatment and bioremediation were characterized in terms of their macroscopic aggregation index (AI) using an optical method, and their microscopic coupled adhesion and deformation properties using atomic force microscopy (AFM). Single cells were indented to measure the range and magnitude of the repulsive-attractive intersurface forces, elastic modulus, thickness and density of the cellular surface substances (CSS). The strong correlation suggests that cost and time effective microscopic AFM characterization is capable of making reliable prediction of macroscopic behavior.

Universal Quantifier Derived from AFM Analysis Links Cellular Mechanical Properties and Cell–Surface Integration Forces with Microbial Deposition and Transport Behavior

In this study, we employed AFM analysis combined with mathematical modeling for quantifying cell-surface contact mechanics and magnitude and range of cell-surface interaction forces for seven bacterial strains with a wide range of cell morphology, dimension, and surface characteristics. Comprehensive cell-surface characterization including surface charge, extracellular polymeric substance content, hydrophobicity, and cell-cell aggregation analyses were performed. Flow-through column tests were employed to determine the attachment efficiency and deposition-transport behavior of these bacterial strains. No statistically significant correlation between attachment efficiency and any single-cell surface property was identified. Single-cell characterization by atomic force microscopy (AFM) yielded the mechanical deformation and elastic modulus, penetration resistance to AFM probe penetration by cellular surface substances (CSS), range and magnitude of the repulsive-attractive intersurface forces, and geometry of each strain. We proposed and derived a universal dimensionless modified Tabors parameter to integrate all these properties that account for their collective behavior. Results showed that the Tabor parameter derived from AFM analysis correlated well with experimentally determined attachment efficiency (α), which therefore is able to link microscale cell-surface properties with macroscale bacterial transport behavior. Results suggested that the AFM tests performed between a single cell and a surface captured the key quantities of the interactions between the cell and the surface that dictate overall cell attachment behavior. Tabors parameter therefore can be potentially incorporated into the microbial transport model.

Phosphorus Fractionation and Removal in Wastewater Treatment - Implications for Minimizing Effluent Phosphorus

To address increasing water quality problems associated with eutrophication, state environmental agencies and the United States Environmental Protection Agency (EPA) are requiring more stringent WWTP effluent total phosphorus concentrations that are below 0.10 mg/L and more recently values as low as 0.01 to 0.06 mg/L have been considered. These represent a large decrease in effluent phosphorus permit concentration, which has challenged the POTWs to investigate and evaluate treatment alternatives to achieve additional phosphorus removal by tertiary treatment and to determine the most minimum concentrations possible. Previous studies indicated that as more advanced tertiary treatment processes are applied to eliminate nearly all the soluble reactive P in order to meet the extremely low TP limits, other non-reactive phosphorus species – such as organic P and acid-hydrolysable P – become a larger fraction of the effluent P and their behavior under different treatment methods needs to be understood. The fractional composition of effluent phosphorus and its variations between different WWTP processes have previously received little attention. Therefore, an urgent need exists to gain further fundamental understanding of effluent P chemical species in order to interpret and improve technologies for phosphorus removal to very low limits. The objective of this study is to conduct a survey of P fractions and their fate and susceptibility to a range of different P removal processes, especially tertiary P removal processes. This will help us gain insights into the removal efficiency and mechanism of different P fractions through various treatment technologies. This title belongs to WERF Research Report Series . ISBN: 9781780406893 (eBook)

Protocol to Evaluate Alternative External Carbon Sources for Denitrification at Full-Scale Wastewater Treatment Plants

Addition of external carbon to biological nutrient removal processes to enhance denitrification to meet more stringent effluent nitrogen limits (e.g., total nitrogen < 3 to 6 mg/L) has become more common for publicly owned treatment works. The objective of this report is to establish a framework for a comprehensive and practice-orientated standardized methodology and a procedure to assess the efficiency and feasibility of an alternative carbon source for enhancing nitrogen removal at full-scale wastewater treatment facilities. The report includes a comprehensive literature review that summarizes various types of external carbon sources that can be applied for supporting denitrification. The summary included denitrification kinetics rates and coefficients. A road map is developed that depicts the overall framework and identifies the key components required for a comprehensive and systematic assessment of an alternative carbon source for denitrification. A procedure for pre-screening carbon alternatives is established and an evaluation matrix is provided. The document presents a list of basic parameters that should be obtained or measured to determine the potential of a carbon source. These parameters are often available from the supplier. These basic parameters are used to determine the carbon-to-nitrogen demand ratio and process performance monitoring varies for field testing different nitrogen removal processes. Therefore, the guidance document first provides protocols for evaluating denitrification performance in a general anoxic denitrification reactor. Then, more configuration-specific examples are given for the most commonly used denitrification processes, including the modified Ludzack-Ettinger (MLE) process and 4-stage Bardenpho, either as a suspended-growth system or a fixed-film system. The final selection of the external carbon source is based on a combination of literature review information, bench test parameters, and full-scale testing results. This title belongs to WERF Research Report Series . ISBN: 9781843396185 (eBook)

Comparative Life Cycle Assessment of Advanced Wastewater Treatment Processes for Removal of Chemicals of Emerging Concern

The potential health effects associated with contaminants of emerging concern (CECs) have motivated regulatory initiatives and deployment of energy- and chemical-intensive advanced treatment processes for their removal. This study evaluates life cycle environmental and health impacts associated with advanced CEC removal processes, encompassing both the benefits of improved effluent quality as well as emissions from upstream activities. A total of 64 treatment configurations were designed and modeled for treating typical U.S. medium-strength wastewater, covering three policy-relevant representative levels of carbon and nutrient removal, with and without additional tertiary CEC removal. The USEtox model was used to calculate characterization factors of several CECs with missing values. Stochastic uncertainty analysis considered variability in influent water quality and uncertainty in CEC toxicity and associated characterization factors. Results show that advanced tertiary treatment can simultaneously reduce nutrients and CECs in effluents to specified limits, but these direct water quality benefits were outweighed by even greater increases in indirect impacts for the toxicity-related metrics, even when considering order-of-magnitude uncertainties for CEC characterization factors. Future work should consider water quality aspects not currently captured in life cycle impact assessment, such as endocrine disruption, in order to evaluate the full policy implications of the CEC removal.