Use of poultry litter for biodegradation of soil contaminated with 2,4- and 2,6-dinitrotoluene

Pseudomonas sp. and Pseudomonas putida can utilize dinitrotoluene (DNT) as N-source after the enzymatic removal of nitro groups from the aromatic ring. Addition of nutrients is known to stimulate the biodegradation process. Poultry litter has consortia of microorganisms (including Pseudomonas) along with many nutrients. The objective of this research was to study the biodegradation of 2,4- and 2,6-DNT contaminated soil (from Badger Army Ammunition Plant) using poultry litter. Complete biodegradation of both 2,4- and 2,6-DNT in the soil was observed after 1-day interaction with poultry litter. No degradation was observed using autoclaved litter.     

Use of clay mineral (montmorillonite) for reducing poultry litter leachate toxicity (EC50)

Poultry litter (PL) has useful nutrients and is therefore used as manure. In addition to N, P and K, PL also contains some heavy metals (As, Cd, Cu, Mn, Pb and Zn), antibiotics, antioxidants, mold inhibitors and other organic compounds. Poultry litter aqueous leachate (PLL) has been shown to be toxic to many organisms; PLL is more toxic than the aqueous leachate of other animal manures used on agricultural soils. Clayey soils are known to retain toxic heavy metals. The objective of this study was to measure the change in toxicity (EC50) of PLL on the addition of clay mineral--montmorillonite. A significant reduction (124%) in toxicity of the clay poultry litter leachate (CLL) after 7 days was observed compared to the toxicity of the PLL alone after 1 day. This indicates that some of the toxic components of the litter were adsorbed by the clay.     

Remediation of dinitrotoluene contaminated soils from former ammunition plants: soil washing efficiency and effective process monitoring in bioslurry reactors

A pilot-scale bioslurry system was used to test the treatment of soils highly contaminated with 2,4-dinitrotoluene (2,4-DNT) and 2,6-dinitrotoluene (2,6-DNT). The treatment scheme involved a soil-washing process followed by two sequential aerobic slurry reactors augmented with 2,4-DNT- and 2,6-DNT-mineralizing bacteria. Test soils were obtained from two former army ammunition plants, the Volunteer Army Ammunition Plant (VAAP, Chattanooga, TN) and the Badger Army Ammunition Plant (BAAP, Baraboo, WI). Soil washing was used to minimize operational problems in slurry reactors associated with large particulates. The Eimco slurry reactors were operated in a draw-and-fill mode for 3 months and were monitored for the biodegradation of 2,4-DNT and 2,6-DNT, nitrite production, NaOH consumption, and oxygen uptake rate. Results show that soil washing was very effective for the removal of sands and the recovery of soil fines containing 2,4-DNT and 2,6-DNT. Bioslurry reactors offered rapid and nearly complete degradation of both DNT isomers, but require real time monitoring to avoid long lag periods upon refeeding. Results found a significant discrepancy between the measured DNT concentrations and calculated DNT concentrations in the slurry reactors because of solids profiles in the slurry reactors and the presence of floating crystal of DNTs. Based on the actual amount of dinitrotoluene degradation, nitrite release, NaOH consumption, and oxygen uptake were close to the theoretical stoichiometric coefficients of complete DNT mineralization. Such stoichiometric relationships were not achieved if the calculation was based on the measured DNT concentrations due to the heterogeneity of DNT in the reactor. Results indicate that nitrite release, NaOH consumption, and oxygen uptake rates provide a fast assessment of 2,4-DNT degradation and microbial activity in a slurry reactor, but could not be extended to a second reactor in series where the degradation of a much lower concentration of 2,6-DNT degradation was achieved.     

Anaerobic biodegradation of an organic by-products leachate by interaction with different mine tailings

Deinking by-products from paper recycling have been used as cover materials on acid-producing tailings. Due to residual cellulose, anaerobic degradation leads to the production of an organic-rich leachate, which may percolate through the tailings. This study aims at describing the influence of the tailings’ characteristics on the attenuation of organic matter in leachate and at discussing the degradation mechanisms. To this end, leachate was mixed with different types of tailings, including three unoxidized tailings with varying acid generation potentials and one tailing in three states of oxidation. Regularly, selected biochemical parameters were analyzed to access the evolution of organic components. The results show that when leachate from deinking by-products was placed in contact with tailings, phase and acid–base balance reactions took place in the beginning. Subsequently, oxidation–reduction reactions dominated the chemistry of the system. The type, the constituents, and the state of oxidation of the mine tailings condition the mechanisms of biodegradation of organic components. Methanogenesis was predominant in the control sample (pure organic leachate) but was absent in all leachate-tailings mixtures. No biodegradation was observed in the liquid phase for oxidized tailings and the organic concentration remained constant for unoxidized tailings, independently of the acid-generation potential. The biodegradation efficiency was optimal when the leachate was in contact with a mixture of oxidized and unoxidized tailings due to sulfate-reduction.     

Single-step treatment of 2,4-dinitrotoluene via zero-valent metal reduction and chemical oxidation

Many nitroaromatic compounds (NACs) are considered toxic and potential carcinogens. The purpose of this study was to develop an integrated reductive/oxidative process for treating NACs contaminated waters. The process consists of the combination of zero-valent iron and an ozonation based treatment technique. Corrosion promoters are added to the contaminated water to minimize passivation of the metallic species. Water contaminated with 2,4-dinitrotoluene (DNT) was treated with the integrated process using a recirculated batch reactor. It was demonstrated that addition of corrosion promoters to the contaminated water enhances the reduction of 2,4-DNT with zero-valent iron. The addition of corrosion promoters resulted in 62% decrease in 2,4-DNT concentration to 2,4-diaminotoluene. The data shows that iron reduced the 2,4-DNT and ozone oxidized these products resulting in a 73% removal of TOC and a 96% decrease in 2,4-DNT concentration.     

Flexible single walled nanotube based chemical sensor for 2,4-dinitrotoluene sensing

In this work, we have attempted to fabricate flexible single walled carbon nanotube based sensor for detection of 2,4-dinitrotoluene (DNT) an explosive chemical. For analyte sensing study, the flexible sensor is fabricated by vacuum filtration method. These fabricated gas sensors are characterised by SEM and Raman spectroscopy. The sensor response is investigated toward the explosive chemicals which have NO₂ group in their molecular structure. The fabricated sensor is able to detect the traces of DNT at room temperature. The sensor gives 0.28–0.32% repeatable response to 0.22 ppm of DNT. The response of sensor increases with increase in the vapour concentration of the DNT vapours.     

Aerobic degradation of 2,4-dinitrotoluene by individual bacterial strains and defined mixed population in submerged cultures

The degradation efficiencies of isomeric mononitrotoluenes (2- and 4-NTs) and dinitrotoluenes (2,4-DNT and 2,6-DNT) by either individual bacterial strains (Bacillus cereus NDT4, Pseudomonas putida NDT1, Pseudomonas fluorescens NDT2, and Achromobacter sp. NDT3) or their mixture were compared in submerged batch cultivations. The mixed culture degraded 2,4-DNT nearly 50 times faster than any of the individual strains. The mixed culture also demonstrated significantly shorter lag periods in 2,4-DNT degradation, a lack of nitrite or organic intermediates accumulation in the liquid medium and the ability to degrade a broader spectrum of nitrotoluenes over a wider concentration range. The presence of both readily degradable 2-NT (or 4-NT) and poorly degradable 2,6-DNT in the medium negatively affected 2,4-DNT biodegradation. However, the mixed bacterial culture still effectively degraded 2,4-DNT with only slightly lower rates under these unfavorable conditions, thus showing potential for the remediation of 2,4-DNT contaminated sites.     

Adsorption of trace elements from poultry litter by montmorillonite clay

Poultry litter (PL) is used as fertilizer on agricultural lands because of its high nutrient content. However, the litter also contains trace elements such as As, Cd, Cu, Pb, and Zn. On land application of PL, these trace elements may be absorbed by crops, leach into groundwater, or enter the aquatic system as run-off. The objective of this research was to study the effect of the addition of montmorillonite clay-mineral (CM) in reducing the release of trace elements from PL. Cd, Cu, and Zn showed significant decreases of 29, 34, and 22%, respectively, in PL aqueous leachate (compared with the control-PL without CM) on mixing with 0.05 g CM but no change in As, Co, and Cr concentrations was observed. Lead showed a significant increase in PL aqueous leachate on mixing with 0.2 g CM but Pb concentration was two orders of magnitude less than in CM aqueous leachate alone. On washing, the settled precipitate (PL+CM) in the centrifuge tubes with water (desorption study) most of the adsorbed metals (Cd 85%, Cu 61%, and Zn 100%) were released. The results of this study show that the addition of CM resulted in significant adsorption of Cd and Cu from PL.     

Amperometric TNT biosensor based on the oriented immobilization of a nitroreductase maltose binding protein fusion.

The preparation and characterization of an amperometric 2,4,6-trinitrotoluene (TNT) biosensor based on the surface immobilization of a maltose binding protein (MBP) nitroreductase (NR) fusion (MBP-NR) onto an electrode modified with an electropolymerized film of N-(3-pyrrol-1-ylpropyl)-4,4'-bipyridine (PPB) are described. The MBP domain of MBP-NR exhibits a high and specific affinity toward electropolymerized films of PPB with the immobilized enzyme retaining virtually all of its enzymatic activity. Under similar conditions, the wild-type NR enzyme (i.e., without the MBP domain) loses most of its enzymatic activity. The kinetics of the catalytic reaction between the biosensor and TNT and 2,4-dinitrotoluene (DNT) were characterized using rotated disk electrode and cyclic voltammetry techniques, and values of 1.4 x 10(4) and 7.1 x 10(4) M(-1) s(-1) were obtained for TNT and DNT, respectively. The apparent Michaelis-Menten constants (KM) for MBP-NR in solution and on the surface, using TNT as substrate, were determined to be 27 and 95 microM, respectively. The corresponding value for "wild-type" NR in solution containing TNT was 78 microM, which is very close to the value obtained for MBP-NR on the surface. The limits of detection for both TNT and DNT were estimated to be 2 microM, and the sensitivities were determined to be 205 and 222 nA/microM, respectively.     

Stability and 2,4-dinitrotoluene response of organic field effect transistors based on π-conjugated thiophene oligomers

This paper reports on organic field effect transistors (OFETs) based on two π-conjugated oligomers derived from thiophenes and their use as sensors for the detection of 2,4-dinitrotoluene (DNT). The detection mechanism relies on donor-acceptor interactions between the π-conjugated system (donor) and the nitrated molecule (acceptor). An important feature of sensors is the stability under operation, so, a large part of this work will be dealing with the behavior of OFETs under bias stress experiments as well as with the influence of temperature during operation. Most of results reported here are concerning hexyl capped tetra Thienylene–Vinylene (denominated 4-TV). Some preliminary results on the promising hexyl capped quinquethiophene derived from 3,4-ethylenedioxythiophene (denominated TETET) are also reported. Under a DNT contaminated air atmosphere ( 7 ppm), 4-TV based OFETs exhibit an increase of the drain current when DNT is present in the atmosphere as expected.     

Chlorophenols in leachates originating from different landfills and aerobic composting plants

Both type and concentration of organic contaminants in landfill leachates show great variation depending on many factors, such as type of wastes, rate of water application, moisture content, landfill design and operation age. In this paper, highly toxic chlorophenol derivatives, poorly biodegradable, carcinogenic existence and recalcitrant properties are determined by solid phase microextraction (SPME)-GC/FID in different leachates from landfill and composting plant in Istanbul. Leachates originated from acidogenic, methanogenic phases of Odayeri sanitary landfill (OSL) and from an aerobic composting plant are considered for different chlorophenol types. It is observed that acidogenic leachate from Odayeri landfill includes 2,4-dichlorophenol, 2,6-dichlorophenol, 2,3,4-trichlorophenol, 2,3,4,5-tetrachlorophenol and 2,3,4,6-tetrachlorophenol at concentration ranges, 15-130, 18-65, 8-40, 5-20 and 10-25 microg/l, respectively. Whereas, only 2,4-dichlorophenol at a concentration range 8-40 microg/l is determined in the methanogenic leachate of the landfill, which can be considered as an indication of reductive dechlorination. There is no chlorophenol derivative in aerobic composting leachate. It is determined that acidogenic leachate from Odayeri landfill includes more species of chlorinated phenols at higher concentration.     

Recovery of nitrotoluenes in wastewater by solvent extraction

Toluene extraction was utilized to recover 2,4-dinitrotoluene (DNT), 2,6-DNT, and 2,4,6-trinitrotoluene (TNT) from wastewater of toluene nitration process. The batch-wise experiments were performed to elucidate the influence of various operating variables on the extracting behavior, including extracting temperature, volume ratios of solvent versus wastewater, agitation time, acidity of wastewater, and extraction stages. It was found that recovery of total organic compounds (TOC) was significantly elevated with increasing extraction temperature. Besides, high volume ratio of toluene/wastewater (2.0) and wastewater acidified to lower pH value enhanced the recovery percentage of TOC, in which extractable tendency was as follows: 2,6-DNT>2,4-DNT>2,4,6-TNT. It is worth noting that the nitrotoluenes in wastewater would be almost completely recovered using three sequential stages toluene extraction at the agitation time of 12min and pH 3.0. It is apparent that this established method is promising for the treatment of wastewater from toluene nitration processed industrially.     

Single cell protein production from digested and undigested poultry litter by Candida utilis: optimization of process parameters using response surface methodology

In this study, production of single cell protein (SCP) was carried out using Candida utilis from undigested poultry litter (UPL) and digested poultry litter (DPL). UPL and DPL were hydrolyzed by acid hydrolysis initially and the resulting crude hydrolysates were used for SCP production. Fermentation variables of the production medium were optimized by single variables optimization methodology. By this conventional approach, SCP production from UPL and DPL has increased to 25 and 20 %, respectively. The proportional analysis of these results revealed that UPL was the suitable substrate than DPL for the production of SCP by C. utilis. Further, 2³ central composite design was applied to determine the optimal process parameters for SCP production using UPL as substrate. The analysis of treatment combinations showed that a second-order polynomial regression model was in good agreement with experimental results at R ² value ?0.98 (P < 0.05). These results suggest that UPL concentration was the most influential factor affecting the SCP production with a P value of 2.22E-09 followed by yeast extract and pH at 0.0339 and 0.0439, respectively. Based on contour plots, the optimum conditions for SCP production were found to be the UPL concentration—5.6 % w/v, yeast extract concentration—2.3 % w/v, and pH—7. After statistical optimization, SCP production by C. utilis strain has increased 29 %. It is understood from this study that UPL could be used effectively as substrate for the production of animal feed supplement with benefit of reducing feed cost as well as mitigating environmental problems associated with poultry litter.     

The fate and transport of RDX, HMX, TNT and DNT in the volcanic soils of Hawaii: a laboratory and modeling study

The adsorption and degradation behavior of RDX, HMX, TNT and DNT and the impact of pH, ionic strength and dissolved organic matter on sorption were examined for two volcanic soils of a former military training area on Hawaii Island, Hawaii, USA. The transport of these chemicals in the soil was also studied in small packed columns and simulated using a water-flow and solute-transport model, HYDRUS_1D. The results show that HMX and RDX are both significantly more mobile than TNT and DNT. The adsorbability of the four chemicals was ranked as: RDXRDX>DNT>TNT. No significant trend was observed for the effect of ionic strength, pH and dissolved organic carbon (DOC) on the adsorption of explosive compounds within the concentrations and pH ranges evaluated. The simulation results show that TNT and DNT would not leach beyond a depth of 30cm soil profile whereas a significant amount of HMX and RDX would pass the 30cm depth. It seems that the risk for contamination of groundwater is much higher for both HMX and RDX than for DNT and TNT as the substratum in this area consists of highly permeable lavas.     

Spectroscopic ultra-trace detection of nitroaromatic gas vapor on rationally designed two-dimensional nanoparticle cluster arrays

Nanoparticle cluster arrays (NCAs) are engineered two-dimensional plasmonic arrays that provide high signal enhancements for critical sensing applications using surface enhanced Raman spectroscopy (SERS). In this work we demonstrate that rationally designed NCAs are capable of detecting ultra-traces of 2,4-dinitrotoluene (DNT) vapor. NCAs functionalized with a thin film of an aqueous NaOH solution facilitated the detection of DNT vapor at a concentration of at least 10 ppt, even in the presence of an excess of potential interferents, including Diesel fuel, fertilizers, and pesticides. Both in the presence and in the absence of this complex background the SERS signal intensity of the NO(2) stretching mode showed a continuous, concentration dependent response over the entire monitored concentration range (10 ppt-100 ppb). The small size, superb sensitivity, and selectivity, as well as the fast response time of <5 min, make NCAs a valuable photonic sensor platform for ultra-trace nitroaromatic gas vapor detection with potential applications in landmine removal and homeland security.     

Acclimated mixed microbial responses to organic species in industrial landfill leachate

Acclimated mixed microbial cultures utilize organic carbon and other nutrients solely from leachate and exhibit a diauxic type of growth, implying the presence of two groups of organic “substrates” in the leachate under study. Carbon balance calculations provide clear evidence that the loss of dissolved organic carbon is due to biological oxidation and not to sorption, stripping or evaporation. Substrate inhibition and low sludge yield are observed. The oxygen requirements of the mixed culture are nominal. It is possible to treat high-strength hazardous wastewater at an overall organic carbon removal of about 80%. Application of reverse osmosis improves the quality of effluents from biological degradation.     

Bioremediation of contaminated soil beds and groundwater — A simulation study

Pollution has reached levels which demand immediate attention and scientific and technological solutions are required on an urgent basis. We are concerned in this paper with bioremediation of soil and groundwater, i.e. the use of indigenous micro-organisms to clean up soil beds and groundwater contaminated with organic pollutants. To achieve managedin situ bioremediation in practice, treated water is recycled with added nutrients into the ground so that oxygen and nitrogen are carried with the water to the subsurface regions. Sorption, convective-dispersive flow and chemical and biological transformations are the chief processes involved that have to be modelled. Here we discuss a simulation model developed to aid in designing an efficient system that maximizes the rate of biodegradation. Simulation models are a must in this case since laboratory experiments take time periods of the order of months. An unusual feature of this simulation model is that it is governed by coupled partial and ordinary differential equations. Partial differential equations (PDEs) model the diffusion and biodegradation processes occurring in the micropores of soil aggregates while ordinary differential equations (ODEs) describe the bioremediation in the interstitial spaces between soil aggregates, both partial and ordinary differential equations being nonlinear. The model is applied to the case of high initial contaminant concentrations. This work is part of a joint project with Regional Research Laboratory, Jorhat and has been carried out in close cooperation with N N Dutta. Discussions with K S Yajnik have been very useful.     

Pyrene-modified quartz crystal microbalance for the detection of polynitroaromatic compounds

The synthesis of a dithiol-functionalized pyrene derivative is reported, together with studies of interactions between this receptor (and other related pyrenes) and nitroaromatic compounds (NACs), in both solution and in the solid state. Spectroscopic analysis in solution and X-ray crystallographic analysis of cocrystals of pyrene and NACs in the solid state indicate that supramolecular interactions lead to the formation of defined π-π stacked complexes. The dithiol-functionalized pyrene derivative can be used to modify the surface of a gold quartz crystal microbalance (QCM) to create a unique π-electron rich surface, which is able to interact with electron poor aromatic compounds. For example, exposure of the modified QCM surface to the nitroaromatic compound 2,4-dinitrotoluene (DNT) in solution results in a reduction in the resonant frequency of the QCM as a result of supramolecular interactions between the electron-rich pyrenyl surface layer and the electron-poor DNT molecules. These results suggest the potential use of such modified QCM surfaces for the detection of explosive NACs.     

Potential for biodegradation of polychlorinated biphenyls (PCBs) by Sinorhizobium meliloti

Resting cell assay and soil microcosms were set up to investigate the biodegradation capability and metabolic intermediate of polychlorinated biphenyls (PCBs) by a rhizobial strain Sinorhizobium meliloti. Biodegradation was observed immediately after 2,4,4'-TCB was supplied as a sole source of carbon and energy in liquid cultures. After 6 days, the percent biodegradation of 2,4,4'-TCB was 77.4% compared with the control. The main intermediate was identified as 2-hydroxy-6-oxo-6-phenylhex-2,4-dienoic acid (HOPDA) for 2,4,4'-TCB as determined by gas chromatography-mass spectrometry (GC-MS). Inoculation with S. meliloti greatly enhanced the degradation of target PCB mixtures in the soil. Moreover, soil culturable bacteria, fungi and biphenyl degrading bacteria counts showed significant increase after inoculation of S. meliloti. This study suggests that S. meliloti is promising in PCB bioremediation.     

The use of 2D non-uniform electric field to enhance in situ bioremediation of 2,4-dichlorophenol-contaminated soil

In situ bioremediation is a safe and cost-effective technology for the cleanup of organic-contaminated soil, but its remediation rate is usually very slow, which results primarily from limited mass transfer of pollutants to the degrading bacteria in soil media. This study investigated the feasibility of adopting 2D non-uniform electric field to enhance in situ bioremediation process by promoting the mass transfer of organics to degrading bacteria under in situ conditions. For this purpose, a 2D non-uniform electrokinetic system was designed and tested at bench-scale with a sandy loam as the model soil and 2,4-dichlorophenol (2,4-DCP) as the model organic pollutant at two common operation modes (bidirectional and rotational). Periodically, the electric field reverses its direction at bidirectional mode and revolves a given angle at rotational mode. The results demonstrated that the non-uniform electric field could effectively stimulate the desorption and the movement of 2,4-DCP in the soil. The 2,4-DCP was mobilized through soil media towards the anode at a rate of about 1.0 cmd(-1)V(-1). The results also showed that in situ biodegradation of 2,4-DCP in the soil was greatly enhanced by the applied 2D electric field upon operational mode. At the bidirectional mode, an average 2,4-DCP removal of 73.4% was achieved in 15 days, and the in situ biodegradation of 2,4-DCP was increased by about three times as compared with that uncoupled with electric field, whereas, 34.8% of 2,4-DCP was removed on average in the same time period at the rotational mode. In terms of maintaining remediation uniformity in soil, the rotational operation remarkably excelled the bidirectional operation. In the hexagonal treatment area, the 2,4-DCP removal efficiency adversely increase with the distance to the central electrode at the bidirectional mode, while the rotational mode generated almost uniform removal in soil bed.