Water toxicity evaluations: Comparing genetically modified bioluminescent bacteria and CHO cells as biomonitoring tools.

The use of water for drinking and agriculture requires knowledge of its toxicity. In this study, we compared the use of genetically modified bioluminescent (GMB) bacteria whose luminescence increases in the presence of toxicants and Chinese Hamster Ovary (CHO) cells for the characterization of the toxicity of water samples collected from a lake and streams, hydroponic and aquaponic farms, and a wastewater treatment plant. GMB bacteria were used to probe genotoxicity, cytotoxicity and reactive oxygen species-induced effects in the whole water samples. Unlike GMB bacteria, the use of CHO cells requires XAD resin-based pre-concentration of toxic material present in water samples for the subsequent cytotoxicity assay. In addition to the examination of the toxicity of the water from the different sources, the GMB bacteria were also used to test the XAD extracts diluted to the concentrations causing 50% growth inhibition of the CHO cells. The two biomonitoring tools provided different results when they were used to test the above-mentioned diluted XAD extracts. A pre-concentration procedure based on adsorption by XAD resins with subsequent elution was not sufficient to represent the material responsible for the toxicity of the whole water samples toward the GMB bacteria. Therefore, the use of XAD resin extracts may lead to major underestimates of the toxicity of water samples. Although the toxicity findings obtained using the GMB bacteria and CHO cells may not correlate with each another, the GMB bacteria assay did provide a mechanism-specific biomonitoring tool to probe the toxicity of water samples without a need for the pre-concentration step.

Between source and sea: The role of wastewater treatment in reducing marine microplastics.

Wastewater treatment plants (WWTPs) are a focal point for the removal of microplastic (MP) particles before they are discharged into aquatic environments. WWTPs are capable of removing substantial quantities of larger MP particles but are inefficient in removing particles with any one dimension of less than 100 µm, with influents and effluents tending to have similar quantities of these smaller particles. As a single WWTP may release >100 billion MP particles annually, collectively WWTPs are significant contributors to the problem of MP pollution of global surface waters. Currently, there are no policies or regulations requiring the removal of MPs during wastewater treatment, but as concern about MP pollution grows, the potential for wastewater technologies to capture particles before they reach surface waters has begun to attract attention. There are promising technologies in various stages of development that may improve the removal of MP particles from wastewater. Better incentivization could speed up the research, development and adoption of innovative practices. This paper describes the current state of knowledge regarding MPs, wastewater and relevant policies that could influence the development and deployment of new technologies within WWTPs. We review existing technologies for capturing very small MP particles and examine new developments that may have the potential to overcome the shortcomings of existing methods. The types of collaborations needed to encourage and incentivize innovation within the wastewater sector are also discussed, specifically strong partnerships among scientific and engineering researchers, industry stakeholders, and policy decision makers.

Innovative ex-situ biological biogas upgrading using immobilized biomethanation bioreactor (IBBR).

Biogas, which typically consists of about 50-70% of methane gas, is produced by anaerobic digestion of organic waste and wastewater. Biogas is considered an important energy resource with much potential; however, its application is low due to its low quality. In this regard, upgrading it to natural gas quality (above 90% methane) will broaden its application. In this research, a novel ex-situ immobilized biomethanation bioreactor (IBBR) was developed for biologically upgrading biogas by reducing CO2 to CH4 using hydrogen gas as an electron donor. The developed process is based on immobilized microorganisms within a polymeric matrix enabling the application of high recirculation to increase the hydrogen bioavailability. This generates an increase in the consumption rate of hydrogen and the production rate of methane. This process was successfully demonstrated at laboratory-scale system, where the developed process led to a production of 80-89% methane with consumption of more than 93% of the fed hydrogen. However, a lower methane content was achieved in the bench-scale system, likely as a result of lower hydrogen consumption (63-90%). To conclude, the IBBRs show promising results with a potential for simple and effective biogas upgrading.

Abundance and diversity of anammox bacteria in a mainstream municipal wastewater treatment plant.

Among the factors that obstruct the application of anammox-based technology for nitrogen removal from mainstream municipal wastewater is the water's high organic loads. We hypothesized that some anammox species can adapt and grow in mainstream wastewater in which a minimal temperature of 13-15 °C is maintained. Using the AMX368F and AMX820R PCR-primers, anammox bacteria were detected in influent wastewater (COD/N ratio > 13) and in the anaerobic, anoxic, and aerobic chambers of a full-scale municipal wastewater treatment plant, reaching 107 copies/g VSS of the16S rRNA gene. Furthermore, anammox activity was demonstrated by 15N-isotopic tracing. The DNA sequences of clones randomly selected from a clone library were mainly clustered with Candidatus Brocadia flugida in addition to Ca. Brocadia sinica, Ca. Jettenia asiatica, and Ca. Anammoxoglobus propionicus. However, Ca. Brocadia was the only genus detected by high-throughput next-generation sequencing and denaturing gradient gel electrophoresis. The nitrite producers, ammonia-oxidizing archaea and bacteria, were both detected in the influent wastewater and the other chambers, while the nitrite consumers, Nitrospira nitrite oxidizers and the nirS-type denitrifiers, dominated all chambers. The results indicate the occurrence and potential activity of anammox bacteria in mainstream wastewater under certain conditions (proper temperature). The dominance of Brocadia flugida and Anammoxoglobus propionicus suggests a role for volatile fatty acids in selecting the anammox community in wastewater.

Mechanism of nanoplastics capture by jellyfish mucin and its potential as a sustainable water treatment technology.

The accumulation of nanoplastics (NPs) in the environment has raised concerns about their impact on human health and the biosphere. The main aim of this study is to understand the mechanism that governs the capture of NPs by jellyfish mucus extracted from the jellyfish Aurelia sp. (A.a.) and compare the capture/removal efficiency to that of conventional coagulants and mucus from other organisms. The efficacy of A.a mucus to capture polystyrene and acrylic NPs (∼100 nm) from spiked wastewater treatment plant (WWTP) effluent was evaluated. The mucus effect on capture kinetics and destabilization of NPs of different polymer compositions, sizes and concentrations was quantified by means of fluorescent NPs, dynamic light scattering and zeta potential measurements and visualized by scanning electron microscopy. A dosing of A.a. mucus equivalent to protein concentrations of ∼2-4 mg L-1 led to a rapid change in zeta potential from a baseline of -30 mV to values close to 0 mV, indicating a marked change from a stable to a non-stable dispersion leading to a rapid (<10 min) and significant removal of NPs (60 %-90 %) from a stable suspension. The A.a. mucus outperformed all other mucus types (0-37 %) and coagulants (0 %-32 % for ferric chloride; 23-40 % for poly aluminum chlorohydrate), highlighting the potential for jellyfish mucus to be used as bio-flocculant. The results indicate a mucus-particle interaction consisting of adsorption-bridging and "mesh" filtration. Further insight is provided by carbohydrate composition and protein disruption analysis. Total protein disruption resulted in a complete loss of the A.a. mucus capacity to capture NPs, while the breaking of disulfide bonds and protein unfolding resulted in improved capture capacity. The study demonstrates that natural jellyfish mucin can capture and remove NPs in water and wastewater treatment systems more efficiently than conventional coagulants, highlighting the potential for development of a new type of bio-flocculant.

Microplastic distributions in a domestic wastewater treatment plant: Removal efficiency, seasonal variation and influence of sampling technique.

Wastewater treatment plants (WWTPs) serve as an important route of microplastics (MPs) to the environment. Therefore, more effective MPs sampling and detection methodologies, as well as a better understanding of their influence on MPs occurrence and distributions in WWTP effluents, are needed for better removal and control. In this work, the efficiency of a municipal WWTP to remove MPs was assessed by collecting samples from raw to tertiary effluent during a 12-month sampling campaign (season-based) using different sampling methods (containers, 24-h composite and large grab samples). MPs retrieved from different treatment units within the WWTP were identified and quantified using plastic/non-plastic staining followed by optical microscopy, SEM and µ-Raman microscopy. Overall, the mean removal efficiency of MPs in the WWTP was 97%, with most MPs removed by the secondary stage and a mean effluent concentration of 1.97 MPs L-1 after sand filtration. The relative abundance of particles was lower than fibers in treated effluent compared with the raw wastewater, with MP fibers constituting 74% of the total MPs in raw wastewater and 91% in treated effluent. Taking seasonal variations into account is important as total MPs concentration in the effluent was notably higher in winter compared with the other seasons. Increasing the sampled volume using large samples or 24-h composite samples significantly reduced the variability between replicates. However, MPs concentration post the tertiary stage was significantly lower using morning sampling (9 am) by large grab sampling method (1.2 MPs L-1) compared to 24-h composite sampling (3.2 MPs L-1) possibly due to intra-daily changes. Using a finer mesh size (0.45 µm) to capture MPs beyond the size range typically studied (≥20 µm) effectively doubled the number of MPs detected in the tertiary effluent and highlights the importance of standardizing sampling procedures.

Controlling nitritation in a continuous split-feed/aeration biofilm nitrifying bioreactor.

This study explored the stability of partial ammonium oxidation at low feed concentration (50 g N/m3), suitable for anammox process, in continuous fixed bed up-flow biofilm reactors with external recirculation-aeration. The reactors, filled with crushed basalt, were fed with synthetic medium at 20-25 °C at constant flow-rate with limiting dissolved oxygen concentration controlled by the recirculation ratio (R). Successful nitritation was achieved at R ≅ 4-6 with approx. 50% of NH4+ oxidized to NO2- with <5% NO3-accumulation. q-PCR analysis along the reactor showed ammonia oxidizing bacteria being the prevalent nitrifiers over the three-fourths of the bed in the flow direction, negligible denitrifiers and absent ammonium oxidizing archaea. A numerical model for predicting the concentration of the nitrogen species and DO was formulated. The model successfully predicted the experimental results and displayed good sensitivity to intrinsic oxygen uptake parameters. The proposed numerical model can serve both as an operational and design tool.

Application of immobilized and granular dried anaerobic biomass for stabilizing and increasing anaerobic bio-systems tolerance for high organic loads and phenol shocks.

This study focuses on the stability and tolerance of continuous-flow bioreactors inoculated with anaerobic methanogens in three different configurations: (R1) dried granular biomass immobilized in PAC-enriched hydrophilic polyurethane foam, (R2) dried granular biomass, and (R3) wet granular biomass. These systems were tested under two different organic loading rates (OLR) of 6.25 and 10.94 (gCOD/(Lreactor∗d)), using a glucose-based synthetic mixture. The effect of an instantaneous shock load of phenol (5g/L for three days), and of phenol inclusion in the feed (0.5g/L) were also tested. At the lower OLR, all reactors performed similarly, however, increasing the OLR lead to a significant biomass washout and failure of R3. Biomass in R1 was more tolerant to phenol shock load than R2, though activity was recovered in both systems after about one month. PAC provided protection and shortened the adaptation time for 0.5g/L phenol that continuously was fed.

An independent prediction of the effect of dissolved organic matter on the transport of polycyclic aromatic hydrocarbons.

The transport of polycyclic aromatic hydrocarbons (PAH) in porous media in the presence of dissolved organic matter (DOM) was predicted with a transport bicontinuum model using independently obtained relationships to derive transport parameters for describing the effect of PAH binding to the DOM. The sorption constants of PAHs to soil and their binding constants to DOM were derived from basic correlations with K(ow) (indicator of hydrophobicity). The kinetic (rate) constants were derived from previously published correlations with K(p) (sorption constant). The independently obtained sorption and rate constants were corrected for binding to DOM and were used to predict the breakthrough curves (BTC) of contaminants in the presence and the absence of DOM. Column results confirmed the independently predicted BTC of PAHs in the presence of DOM that did not sorb to the solid phase, as well as the effect of DOM on the rate of the sorption and desorption processes. These findings confirm the ability to quantitatively describe how DOM facilitates transport of contaminants in the subsurface using independently derived parameters.

The effect of anaerobic biomass drying and exposure to air on their recovery and evolution.

The main goal of this study was to test the effect of various drying methods of granular anaerobic biomass on biomass survival, potential and rate of methane re-production, and structure. This may facilitate the development of drying methods to preserve excess anaerobic biomass in dry form for re-inoculation of existing digesters after process failure or wash out or for the start-up of new digesters. To that end, anaerobic granular biomass was collected from an up-flow anaerobic sludge blanket (UASB) reactor. The biomass was dried using two alternative methods: oven with air circulation at 50 °C for 24 h (DAO), and vacuum rotary evaporator at anaerobic conditions (DAN). For comparison, the control was a biomass with no drying (WET). Biomass samples were tested for specific methanogenic activity using synthetic wastewater. The microbial communities were also tested for viability using the LIVE/DEAD kit, and total biomass was initially quantified by qPCR targeting 16S rRNA genes. In addition, the mcrA functional gene was used s a target for the detection of the most abundant methanogens. Basic bacterial morphology classification was done by VIT(®) gene probe technology using a fluorescence microscope. Dried DAN and DAO biomasses required approximately four operational runs to recover their initial methanogenic activity compared to WET biomass. LIVE/DEAD results showed clear increases in the proportions of the viable biomass of the total bacterial communities over time, especially for the DAN and DAO samples. A comparison of the qPCR results of both DAN and DAO to the WET biomass showed that the methanogenic mcrA gene fraction of the total biomass population of 16S rRNA gene concentrations decreased moderately by about 17.2% in the samples of DAO and by approximately 6.7% in the samples of DAN over all runs after Run1.

Co-metabolic oxidation of pharmaceutical compounds by a nitrifying bacterial enrichment.

The biotransformation of five selected pharmaceuticals ibuprofen (IBP), ketoprofen (KTP), carbamazepine (CBZ), dexamethasone (DXM) and iopromide (IOP) by a stable nitrifying enrichment culture was investigated at concentrations ranging between 25 µg/L and 2mg/L. Complete biotransformation was observed only for IBP and KTP, although, an inverse correlation between transformation rate and concentration was found. The transformation pattern observed is consistent with ammonia monooxygenase (AMO) activity. The metabolic succession of the compounds according to the biotransformation rates was: IBP>KTP>DXM>CBZ>IOP. A linear correlation between the calculated diffusive flux of the model compounds across a bilayer membrane and their biotransformation rates was found. Our results support the concept that augmentation with nitrifying activity can enhance the removal of trace organic pollutants during effluent treatment. Furthermore, ammonia-oxidizing activity appears as a good indicator for estimation of potential of biodegradability of pharmaceuticals, especially at low concentrations.

Synergetic effect between photocatalytic degradation and adsorption processes on the removal of phenolic compounds from olive mill wastewater.

The photocatalytic degradation of two phenolic compounds, p-coumaric acid and caffeic acid, was performed with a suspended mixture of TiO(2) and powdered activated carbon (PAC) (at pH=3.4 and 8). Adsorption, direct photolysis and photocatalytic degradation were studied under different pH and UV light sources (sunlight vs. 365nm UV lamps). The potential for reusing this catalyst mixture in sequential photocatalytic runs was examined as well. Quantum yields for the direct photolysis of caffeic acid under solar and artificial 365nm light were calculated (for the first time) as 0.005 and 0.011, respectively. A higher removal rate of contaminants by either adsorption or photocatalysis was obtained at a low pH (pH 4). Furthermore, the addition of PAC increased the removal efficiency of the phenolic compounds. Fast removal of the pollutants from the solution over three sequential runs was achieved only when both TiO(2) and PAC were present. This suggests that at medium phenolic concentrations, the presence of PAC as a co-sorbent reduces surface poisoning of the TiO(2) catalyst and hence improves photocatalysis degradation of phenolic pollutants. The adsorption equilibrium of caffeic acid or p-coumaric acid on TiO(2), PAC and the combined mixture of TiO(2) and PAC follows the Langmuir isotherm model. Experiments with PAC TiO(2) mixture and olive mill wastewater (anaerobically treated and diluted by a factor of 10) showed higher removal of polyphenols than of chemical oxygen demand (COD). 87% removal of total polyphenols, compared to 58% of COD, was achieved after 24h of exposure to 365nm irradiation (7.6W/m(2)) in the presence of a suspended mixture of TiO(2) and PAC, indicating "self-selectivity" of polyphenols.

Modeling the effect of immobilization of microorganisms on the rate of biodegradation of phenol under inhibitory conditions.

The main objective of this research is to model the effect of biodegradation process of phenol at high initial concentrations using a well known immobilization technique of the biomass. This work focused on testing the effect of activated carbon and clay while considering the diffusive internal mass transfer limitations. Biodegradation of phenol was performed by using enriched microorganisms from a compost of agricultural wastes. The average phenol biodegradation rate (uptake) of free biomass system was 235.3 mg g(-1) h(-1) at initial concentration range of 212-260 mg/L. However, the values for the systems of immobilized biomass in alginate and activated carbon (1 mm), alginate, activated carbon (4 mm), alginate, activated carbon and clay (1 mm) and alginate, activated carbon and clay (4 mm) were 64.9, 27.6, 27.5, and 8 mg g(-1) h(-1) respectively. The effective diffusion factors in different matrix were obtained using an intra-particle diffusion-based mathematical model. Diffusion limitation was observed when the matrix contained clay in addition to activated carbon. The diffusion coefficient was decreased from 1.6 × 10(-8) to 1.2 × 10(-9) cm(2)/s when clay was added to the matrix of 1 mm of alginate and activated carbon. Also, slight differences between the diffusion factors were observed for larger beads. The combination of clay and AC contributes to better mineralization of phenol at high concentrations. This could be attributed to the synergism of both additives.

Immobilization of soot particles in a silica matrix: A sorbent-carrier system for studying organic chemical sorption.

A new method for studying sorption with diesel and hexane sootwas developed, tested, and applied. A commercial silica-based chromatography medium was used as an inert inorganic carrier for immobilization (entrapment) of soot particles and their aggregates, thus creating a combined sorbent for sorption of hydrophobic organic chemicals (HOCs). After precombustion to remove potential organic carbon contaminants, the silica particles and soot samples were mixed under dry conditions that allowed the soot to be incorporated within the pore structure of the much larger (> 180 microm) carrier particles. Unincorporated soot was removed by multiple rinses with Milli-Q water. Sorption rate and equilibrium experiments were conducted, using phenanthrene as a probe HOC. Strong nonlinear sorption of phenanthrene was observed, in agreement with results previously obtained using air-bridge and flocculation-based methods. Batch kinetic studies suggested that 60 d of prewetting is required to obtain full water saturation, as perhaps needed for proper assessment of phenanthrene uptake rate by soot in aqueous systems. Forthe determination of equilibrium phenanthrene sorption, however, 1-d prewetting is sufficient so long as final equilibration is for at least 60 d. The new method is a practical approach to sorption measurement that may prove especially useful for study of strongly sorbing chemicals.

Sorption nonlinearity for organic contaminants with diesel soot: method development and isotherm interpretation.

An experimentally practical and precise flocculation-based method was developed, tested, and applied to determine phenanthrene and 1,2,4-trichlorobenzene sorption with NIST SRM 2975 diesel particulate matter. Following an initial equilibration period, polyaluminum chloride (PACI) solution was added to the sorption tubes in order to facilitate the formation of flocculated aggregates of soot particles. After separation of the solids through centrifugation, supernatant concentrations were determined as with conventional batch methods. The flocculation-based method was tested on three kinds of soot and then used to evaluate sorption kinetics and equilibrium with SRM 2975. Kinetic results showed that wetting of the soot required more than 20 days, but that 60 days was sufficient to achieve equilibration with both water and phenanthrene. Sixty-day isotherms for both phenanthrene and 1,2,4-trichlorobenzene were strongly nonlinear. At approximate 10(-3) of solubility, carbon-normalized distribution coefficients (Koc) were 10-20 times higher than those for absorption to sediment organic matter. Measurements at closer to solubility indicated much lower Koc, suggesting a total sorption capacity at aqueous solubility that is of similar magnitude to that in sediment organic matter. Independent analysis of extractable hydrocarbons suggests that absorption into a native hydrocarbon phase was not a major component of sorption.