Subhasis Ghoshal

Exposing culprit organic pollutants: A review

There is a continuing need for monitoring the health of the environment due to the presence of pollutants. Here, we review the development and attributes of biosensors by which bacteria have been genetically modified to express the luminescence genes, i.e. to glow, in a quantified manner, in response to pollutants. We have concentrated on the detection of organic hydrocarbon pollutants and discussed the molecular mechanisms by which some of these chemicals act as effector molecules on the respective regulatory systems. The future of environmental biosensors is predictably bright. As more knowledge is gathered on the sensing regulatory component, the possibility of developing targeted or pollutant-specific biosensors is promising. Moreover, the repertoire of biosensors for culprit organic pollutants is expected to be enlarged through advances in genomics technology and identification of new sensory or receptor molecules. The need for pollutant detection at concentrations in the parts per trillion range or biosensors configured in a nanoscale is anticipated.

Aggregation and deposition kinetics of carboxymethyl cellulose-modified zero-valent iron nanoparticles in porous media.

Transport and deposition of carboxymethyl cellulose (CMC)-modified nanoparticles of zero-valent iron (NZVI) were investigated in laboratory-scale sand packed columns. Aggregation resulted in a change in the particle size distribution (PSD) with time, and the changes in average particle size were determined by nanoparticle tracking analysis (NTA). The change in PSD over time was influenced by the CMC-NZVI concentration in suspension. A particle-particle attachment efficiency was evaluated by fitting an aggregation model with NTA data and subsequently used to predict changes in PSD over time. Changes in particle sizes over time led to corresponding changes in single-collector contact efficiencies, resulting in altered particle deposition rates over time. A coupled aggregation-colloid transport model was used to demonstrate how changes in PSD can reduce the transport of CMC-NZVI in column experiments. The effects of particle concentrations in the range of 0.07 g L(-1) to 0.725 g L(-1) on the transport in porous media were evaluated by comparing the elution profiles of CMC-NZVI from packed sand columns. Changes in PSD over time could reasonably account for a gradual increase in effluent concentration between 1 and 5 pore volumes (PVs). Processes such as detachment of deposited particles also likely contributed to the gradual increase in effluent concentrations. The particle-collector attachment efficiency increased with CMC-NZVI particle concentration due to a rise in dissolved Na(+) concentration with increased addition of Na-CMC. This inadvertent change in ionic strength led to decreased effluent concentrations at higher CMC-NZVI concentrations.

Assessment of transport of two polyelectrolyte-stabilized zero-valent iron nanoparticles in porous media

This study investigated the breakthrough patterns of carboxymethyl cellulose- and polyacrylic acid-stabilized zero-valent iron (Fe(0)) nanoparticles (NZVI) from packed sand columns under a range of pore water velocities of 0.02, 0.2 and 1 cm min(-1) and NZVI influent concentrations of 0.1, 0.5 and 3 g L(-1). The NZVI effluent relative concentrations of both types of particles decreased with slower flow velocities and increasing particle concentrations. PAA-NZVI exhibited slower elution from the columns than CMC-NZVI under identical experimental conditions, and this is attributed to more rapid aggregation kinetics of PAA-NZVI. The elution patterns of PAA-NZVI showed a stronger trend of gradually increasing effluent concentrations with flushing of additional pore volumes, especially at low flushing velocities and higher influent particle concentrations and this phenomenon too can be attributed to increasing aggregate sizes with time which caused decreases in the values of the single collector efficiency and thus the deposition rate constant. A 7 nm increase in CMC-NZVI aggregate size over 60 min was observed using nanoparticle tracking analysis. The reduction in colloidal stability due to aggregation of CMC- and PAA-NZVI was verified using sedimentation tests, and it was found that PAA-NZVI were less stable than CMC-NZVI. There were also notable inherent differences in the two NZVI particles. The CMC-NZVI were monodisperse with a mean diameter of 5.7 ± 0.9 nm, whereas PAA-NZVI had a bimodal particle size distribution with a small sub-population of particles with mean size of 30 ± 21 nm and a more abundant population of 4.6 ± 0.8 nm diameter particles. Furthermore, PAA-NZVI had a lower surface potential. These characteristics are also responsible for the different elution patterns CMC- and PAA-NZVI.

Deposition of Carboxymethylcellulose-Coated Zero-Valent Iron Nanoparticles onto Silica: Roles of Solution Chemistry and Organic Molecules

Zero-valent iron nanoparticles (nZVI) used in the remediation of contaminated subsurface environments are commonly stabilized using polymer coatings. A bottom-up synthesis approach was used to synthesize carboxymethylcellulose (CMC)-coated nZVI particles with increased colloidal stability. The influence of water chemistry and selected environmental molecules, namely, fulvic acids and rhamnolipids, on the aggregate size and surface charge of the bare and CMC-coated nZVI particles was systematically examined using dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), and laser Doppler velocimetry. A quartz crystal microbalance with energy dissipation monitoring (QCM-D) was used to quantify the deposition rates of bare and CMC-coated nZVI particles onto a silica surface over a broad range of solution ionic strengths and in the presence of naturally occurring molecules. Nanoscale ZVI particle deposition was found to increase with IS for many of the conditions investigated. CMC acted as a better colloidal stabilizer when covalently bound to nZVI particles than when physisorbed onto the nanoparticle surface after particle synthesis. The lowest nanoparticle deposition rates were observed for CMC-coated nZVI in the presence of the rhamnolipid biosurfactant.

Biodegradation of semi- and non-volatile petroleum hydrocarbons in aged, contaminated soils from a sub-Arctic site: Laboratory pilot-scale experiments at site temperatures

This study evaluates the feasibility of landfarming biotreatment of petroleum-contaminated soils obtained from a sub-Arctic site at Resolution Island, Nunavut, Canada, and evaluates the changes in composition of the semi- and non-volatile petroleum hydrocarbon fractions during the biotreatment. Pilot-scale landfarming experiments were conducted in a laboratory in soil tanks under temperature profiles representative of the 3-year site air temperatures in July and August where temperature varied uniformly between 1 degrees C and 10 degrees C over 10 d. The site soils were acidic and N-deficient, but contained indigenous populations of hydrocarbon-degrading microorganisms. Biostimulation with nitrogen and phosphorus nutrient amendments to achieve C(TPH):N:P molar ratio of 100:9:1, and CaCO(3) amendment at 2000 mg Kg(-1) for maintaining neutral pH, and periodic 10-day tilling, reduced total petroleum hydrocarbon (TPH) concentrations by up to 64% over a 60-day period. The rate and extent of semi-volatile (F2: >C10-C16) and non-volatile (F3: >C16-C34) petroleum hydrocarbon fractions in the landfarms containing higher initial TPH levels ( approximately 2000 mg Kg(-1)) and lower TPH levels ( approximately 1000 mg Kg(-1)) were compared. Significant biodegradation of the F2 and F3 fractions occurred in both of those systems. First-order biodegradation rate constants of up to 0.019+/-0.001 d(-1) were determined for the F3 hydrocarbon fraction and were similar to the F2 fraction biodegradation rate constants of up to 0.024+/-0.005 d(-1). Biodegradation profiles of the C14, C16 and C18 alkanes revealed that at TPH concentrations above 1000 mg Kg(-1) these compounds are degraded concurrently, whereas below 1000 mg Kg(-1) the higher-molecular weight alkanes are preferentially degraded. After the 60-day treatment period, the TPH concentration was approximately 500 mg Kg(-1), and the residual TPH mass was largely associated with particles and aggregated particles with diameters of 0.6-2 mm, rather than the larger or finer particles and aggregates.

Rhamnolipid biosurfactant and soy protein act as effective stabilizers in the aggregation and transport of palladium-doped zerovalent iron nanoparticles in saturated porous media.

Palladium-doped nanosized zerovalent iron (Pd-NZVI) particles can contribute to the transformation of chlorinated solvents and various other contaminants into innocuous products. To make Pd-NZVI an effective in situ subsurface remediation agent, these particles need to migrate through a targeted contaminated area. However, previous studies have reported very limited mobility of these particles in the groundwater environment and attributed it to rapid aggregation and subsequent pore plugging. In this study, we systematically investigated the influence of selected natural and nontoxic organic macromolecules (carboxymethyl cellulose, rhamnolipid biosurfactants, and soy protein) on the aggregation and transport behavior of bare and coated Pd-NZVI. Aggregation behavior was investigated using dynamic light scattering by monitoring the evolution of hydrodynamic diameter as a function of time, whereas transport behavior was investigated by conducting water-saturated sand-packed column experiments. While bare Pd-NZVI is prone to rapid aggregation, we observed good colloidal stability and concurrent enhanced transport of Pd-NZVI coated with carboxymethyl cellulose, rhamnolipid biosurfactants, and soy protein. Each surface modifier performed well at lower ionic strength (IS) (10 mM NaHCO3), and one of the rhamnolipid surface modifiers (JBR215) significantly enhanced transport of 150 mg/L Pd-NZVI at concentrations as low as 10 mg/L total organic carbon. However, an increase in the solution IS induced significant Pd-NZVI aggregation with a simultaneous decrease in the transport potential in accordance with the DLVO (Derjaguin, Landau, Verwey, and Overbeek) theory of colloidal stability. Nonetheless, at the highest IS (300 mM NaHCO3) investigated, the mobility of rhamnolipid-coated Pd-NZVI is significantly higher than that of Pd-NZVI coated with the other surface modifiers, suggesting that biosurfactants may be the most suitable surface modifiers in field application. Overall, this study emphasizes how stabilization of Pd-NZVI with natural macromolecules such as rhamnolipids can improve the transport potential of these reactive nanoparticles in subsurface remediation applications at concentrations significantly lower than those of other commonly used polymers.

A modified microbial adhesion to hydrocarbons assay to account for the presence of hydrocarbon droplets

The microbial adhesion to hydrocarbons (MATH) assay has been used widely to characterize microbial cell hydrophobicity and/or the extent of cell adhesion to hydrophobic liquids. The classical MATH assay involves spectrophotometric absorbance measurements of the initial and final cell concentrations in an aqueous cell suspension that has been contacted with a hydrocarbon liquid. In this study, microscopic examination of the aqueous cell suspension after contact with hexadecane or a hexadecane/toluene mixture revealed the presence of hydrocarbon droplets. The hydrocarbon droplets contributed to the absorbance values during spectrophotometric measurements and caused erroneous estimates of cell concentrations and extents of microbial adhesion. A modified MATH assay that avoids such artefacts is proposed here. In this modified assay, microscopic examination of the aqueous suspension and direct cell counts provides cell concentrations that are free of interference from hydrocarbon droplets. The presence of hydrocarbon droplets was noted in MATH assays performed with three bacterial strains, and two different hydrocarbons, at ionic strengths of 0.2 mM and 20 mM and pH 6. In these experiments, the formation of quasi-stable hydrocarbon droplets cannot be attributed to the presence of biosurfactants, or stabilization by biocolloids. The presence of surface potential at the hydrocarbon-water interface that was characterized by electrophoretic mobility of up to -1 and -2 microm cm/Vs, likely caused the formation of the quasi-stable hydrocarbon droplets that provided erroneous results using the classical MATH assay.

Biodegradation kinetics of naphthalene in nonaqueous phase liquid-water mixed batch systems: comparison of model predictions and experimental results.

A model is formulated to describe dissolution of naphthalene from an insoluble nonaqueous phase liquid (NAPL) and its subsequent biodegradation in the aqueous phase in completely mixed batch reactors. The physicochemical processes of equilibrium partitioning and mass transfer of naphthalene between the NAPL and aqueous phases were incorporated into the model. Biodegradation kinetics were described by Monods microbial growth kinetic model, modified to account for the inhibitory effects of 1,2-naphthoquinone formed during naphthalene degradation under certain conditions. System parameters and biokinetic coefficients pertinent to the NAPL-water systems were determined either by direct measurement or from nonlinear regression of the naphthalene mineralization profiles obtained from batch reactor tests with two-component NAPLs comprised of naphthalene and heptamethylnonane. The NAPLs contained substantial mass of naphthalene, and naphthalene biodegradation kinetics were evaluated over the time required for near complete depletion of naphthalene from the NAPL. Model predictions of naphthalene mineralization time profiles compared favorably to the general trends observed in the data obtained from laboratory experiments with the two-component NAPL, as well as with two coal tars obtained from the subsurface at contaminated sites and composed of many different PAHs (polycyclic aromatic hydrocarbon compounds). The effects of varying the NAPL mass and the naphthalene mole fractions in the NAPL are discussed. It was observed that the time to achieve a given percent removal of naphthalene does not change significantly with the initial mass of naphthalene in a fixed volume of the NAPL. Significant changes in the mineralization profiles are observed when the volume (and mass) of NAPL in the system is changed.

Biodegradation of petroleum hydrocarbons in contaminated clayey soils from a sub-arctic site: The role of aggregate size and microstructure

This study investigates the extent of biodegradation of non-volatile petroleum hydrocarbons (C16-C34) and the associated microbial activity in predominant aggregate sizes during a pilot-scale biopile experiment conducted at 15 °C, with a clayey soil, from a crude oil-impacted site in northern Canada. The in situ aggregate microstructure was characterized by N2 adsorption and X-ray CT scanning. The soils in the nutrient (N)-amended and unamended biopile tanks were comprised of macroaggregates (>2 mm) and mesoaggregates (0.25-2 mm). Nutrient addition significantly enhanced petroleum hydrocarbon biodegradation in macroaggregates, but not in mesoaggregates. At the end of 65-d biopile experiment, 42% of the C16-C34 hydrocarbons were degraded in the nutrient-amended macroaggregates, compared to 13% in the mesoaggregates. Higher microbial activity in the macroaggregates of the nutrient amended biopile was inferred from a larger increase in extractable protein concentrations, compared to the other aggregates. Terminal Restriction Fragment Length Polymorphism (T-RFLP) of 16S rRNA genes showed that there was no selection of bacterial populations in any of the aggregates during biopile treatment, suggesting that the enhanced biodegradation in nutrient-amended macroaggregates was likely due to metabolic stimulation. X-ray micro CT scanning revealed that the number of pores wider than 4 μm, which would be easily accessible by bacteria, were an order of magnitude higher in macroaggregates. Also, N2 adsorption analyses showed that pore surface areas and pore volumes per unit weight were four to five-times larger, compared to the mesoaggregates. Thus the higher porosity microstructure in macroaggregates allowed greater hydrocarbon degradation upon biostimulation by nutrient addition and aeration.

Pilot-scale bioremediation of a petroleum hydrocarbon-contaminated clayey soil from a sub-Arctic site.

Bioremediation is a potentially cost-effective solution for petroleum contamination in cold region sites. This study investigates the extent of biodegradation of petroleum hydrocarbons (C16-C34) in a pilot-scale biopile experiment conducted at 15°C for periods up to 385 days, with a clayey soil, from a crude oil-impacted site in northern Canada. Although several studies on bioremediation of petroleum hydrocarbon-contaminated soils from cold region sites have been reported for coarse-textured, sandy soils, there are limited studies of bioremediation of petroleum contamination in fine-textured, clayey soils. Our results indicate that aeration and moisture addition was sufficient for achieving 47% biodegradation and an endpoint of 530 mg/kg for non-volatile (C16-C34) petroleum hydrocarbons. Nutrient amendment with 95 mg-N/kg showed no significant effect on biodegradation compared to a control system without nutrient but similar moisture content. In contrast, in a biopile amended with 1340 mg-N/kg, no statistically significant biodegradation of non-volatile fraction was detected. Terminal Restriction Fragment Length Polymorphism (T-RFLP) analyses of alkB and 16S rRNA genes revealed that inhibition of hydrocarbon biodegradation was associated with a lack of change in microbial community composition. Overall, our data suggests that biopiles are feasible for attaining the bioremediation endpoint in clayey soils. Despite the significantly lower biodegradation rate of 0.009 day(-1) in biopile tank compared to 0.11 day(-1) in slurry bioreactors for C16-C34 hydrocarbons, the biodegradation extents for this fraction were comparable in these two systems.