Biodegradation of 2,4-dinitrotoluene using poultry litter leachate

Various micro-organisms are known to degrade 2,4-dinitrotoluene (DNT) through pathways involving intermediates. Addition of nutrients stimulates the biodegradation process. Poultry litter has a consortia of micro-organisms along with many nutrients. The objective was to study the DNT biodegradation using poultry litter in an aqueous medium. Complete biodegradation of 10-50 ppm 2,4-DNT solutions was observed after two days interaction with poultry litter leachate without the formation of any intermediates. No degradation was observed using autoclaved litter leachate.     

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.     

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.     

Separation and detection of explosives on a microchip using micellar electrokinetic chromatography and indirect laser-induced fluorescence

A new approach for sensitive detection on a microfabricated chip is presented. Indirect laser-induced-fluorescence (IDLIF) was used to detect explosive compounds after separation by micellar electrokinetic chromatography (MEKC). The detection setup was used in an epifluorescence configuration with excitation provided by a near-IR diode laser operating at 750 nm. To achieve indirect detection, a low concentration of a dye (5 microM Cy7) was added to the running buffer as a visualizing agent. Using this methodology, a sample containing 14 explosives (EPA 8330 mixture) was examined. Concentrations of 1 ppm of trinitrobenzene (TNB), trinitrotoluene (TNT), dinitrobenzene (DNB), tetryl, and 2,4-dinitrotoluene (2,4-DNT) could be detected with S/N ratios between 3 and 10. Analyses showing 10 peaks, with plate numbers on the order of 60000, were completed within 60 s using a 65 mm long separation channel. The three isomers of nitrotoluene (2-, 3-, and 4-nitrotoluene) were not resolved. Additionally, the two nitramines (HMX and RDX) could only be detected at much higher concentrations, likely due to the low fluorescence quenching efficiencies of these compounds. The analysis method was also used to separate and detect nitroaromatic compounds in extracts from spiked soil samples. The presence of 1 ppm (1 microg of analyte/1 g of soil) of TNB, DNB, TNT, tetryl, 2,4-DNT, 2,6-DNT, 2-NH2-4,6-DNT, and 4-NH2-2,6-DNT could readily be detected. In the interest of increasing the sensitivity of the analysis, various on-chip injection schemes were evaluated. It was found that a 250 microm double-T injector gave a 35% increase in peak signal compared to a straight-cross injector, which is less than expected based on injected volume.     

Enhanced electrokinetic dissolution of naphthalene and 2,4-DNT from contaminated soils

Electrokinetic soil remediation has been proven to remove heavy metals and polar organics from low hydraulic conductivity subsurface environment. In this study, carboxymethyl-beta-cyclodextrin (CMCD) was used as a carrier to assist electrokinetic treatment for removal of low polarity organic contaminants from soils (2.2% organic carbon content). Naphthalene and 2,4-dinitrotoluene (2,4-DNT) were selected as the test compounds. The results from columns experiments showed that 46 and 43% of naphthalene and 2,4-DNT, respectively, were removed using 0.01 N NaNO(3) flushing solution with 40 V electrical potential while 70 and 72% of naphthalene and 2,4-DNT were removed using 2 g/L CMCD solution. Naphthalene and 2,4-DNT removal was enhanced to 83 and 89%, respectively, by using 2 g/L CMCD with 40 V electrical potential. Findings from this study also demonstrated that cyclodextrin assisted electrokinetics can enhance the removal rate of naphthalene and 2,4-DNT. Electric potential applied has more influence on the contaminant removal than the amount of CMCD used. Higher voltage application caused increase in the removal rate of naphthalene and 2,4-DNT, and appeared to be one of the critical factors in obtaining higher contaminant removal while increasing CMCD solution concentration above 2 g/L appeared to have little effect on the contaminant removal.     

Application of Box-Wilson experimental design method for 2,4-dinitrotoluene treatment in a sequential anaerobic migrating blanket reactor (AMBR)/aerobic completely stirred tank reactor (CSTR) system

A sequential aerobic completely stirred tank reactor (CSTR) following the anaerobic migrating blanket reactor (AMBR) was used to treat a synthetic wastewater containing 2,4-dinitrotoluene (2,4-DNT). A Box-Wilson statistical experiment design was used to determine the effects of 2,4-DNT and the hydraulic retention times (HRTs) on 2,4-DNT and COD removal efficiencies in the AMBR reactor. The 2,4-DNT concentrations in the feed (0-280 mg/L) and the HRT (0.5-10 days) were considered as the independent variables while the 2,4-DNT and chemical oxygen demand (COD) removal efficiencies, total and methane gas productions, methane gas percentage, pH, total volatile fatty acid (TVFA) and total volatile fatty acid/bicarbonate alkalinity (TVFA/Bic.Alk.) ratio were considered as the objective functions in the Box-Wilson statistical experiment design in the AMBR. The predicted data for the parameters given above were determined from the response functions by regression analysis of the experimental data and exhibited excellent agreement with the experimental results. The optimum HRT which gave the maximum COD (97.00%) and 2,4-DNT removal (99.90%) efficiencies was between 5 and 10 days at influent 2,4-DNT concentrations 1-280 mg/L in the AMBR. The aerobic CSTR was used for removals of residual COD remaining from the AMBR, and for metabolites of 2,4-DNT. The maximum COD removal efficiency was 99% at an HRT of 1.89 days at a 2,4-DNT concentration of 239 mg/L in the aerobic CSTR. It was found that 280 mg/L 2,4-DNT transformed to 2,4-diaminotoluene (2,4-DAT) via 2-amino-4-nitrotoluene (2-A-4-NT) and 4-amino-2-nitrotoluene (4-A-2-NT) in the AMBR. The maximum 2,4-DAT removal was 82% at an HRT of 8.61 days in the aerobic CSTR. The maximum total COD and 2,4-DNT removal efficiencies were 99.00% and 99.99%, respectively, at an influent 2,4-DNT concentration of 239 mg/L and at 1.89 days of HRT in the sequential AMBR/CSTR.     

Comparison of visible and near-infrared Raman cross-sections of explosives in solution and in the solid state

Raman cross-sections of explosives in solution and in the solid state have been measured using visible and near-infrared excitation via secondary calibration. These measurements are valuable for both fundamental scientific purposes and applications in the standoff detection of explosives. The explosive compounds RDX, HMX, TNT, 2,4-DNT, 2,6-DNT, and ammonium nitrate were measured using discrete excitation wavelengths ranging from 532 nm to 785 nm. A comparison of the spectral features and cross-sections between the solid state and solution was performed. Comparison is also made to cross-sections measured with deep ultraviolet excitation.     

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-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.     

Photolysis of 2,4-dinitrotoluene in various water solutions: effect of dissolved species

This work investigates the photolysis of 2,4-dinitrotoluene (2,4-DNT) in the presence of different dissolved species. Initial experiments revealed that the direct photolysis of 2,4-DNT in deionized water solutions under sunlight and artificial light followed a pseudo-first order kinetic. Humic acids (HA) appeared to act as sensitizers in the aqueous photolysis of 2,4-DNT and the calculated half life was found to be approximately 2h, which is faster than the half life calculated in the case of deionized water ( approximately 4h). The presence of salt (NaCl) in the deionized water solutions was found to have a more pronounced sensitizing effect upon the photolysis of 2,4-DNT, yielding half lives of the order of 1h. Investigations on seawater and groundwater spiked with 2,4-DNT, revealed that photolysis is enhanced in the order seawater > groundwater approximately deionized water.     

Treatment of metal cyanide bearing wastewater by simultaneous adsorption and biodegradation (SAB)

This paper presents process review and comparative study of biodegradation and adsorption alone with simultaneous adsorption and biodegradation (SAB) process using Pseudomonas fluorescens. Ferrocyanide solution was used for all studies with initial CN(-) concentrations of 50, 100, 200 and 300mg/L, and initial pH of 6. Pseudomonas fluorescens used ferrocyanide as sole source of nitrogen and biodegradation efficiency was observed as 96.4, 94.1, 86.2 and 69.3%, respectively after 60h of agitation. Whereas in adsorption process with granular activated carbon (GAC) as adsorbent, CN(-) removal efficiency was found to be 85.6, 80.1, 70.2 and 50.2%, respectively. But in SAB process the removal efficiency could be more than 70% for all concentrations only at 36h of agitation and achieved removal efficiency of 99.9% for 50 and 100mgCN(-)/L. It was found that SAB process is more effective than biodegradation and adsorption alone.     

Electrochemical destruction of dinitrotoluene isomers and 2,4,6-trinitrotoluene in spent acid from toluene nitration process

Mineralization of dinitrotoluene (DNT) isomers and 2,4,6-trinitrotoluene (TNT) in spent acid was conducted by in situ electrogenerated hydrogen peroxide. The electrolytic experiments were carried out to elucidate the influence of various operating parameters on the performance of mineralization of total organic compounds (TOC) in spent acid, including electrode potential, reaction temperature, oxygen dosage and concentration of sulfuric acid. It is worth noting that organic compounds could be completely mineralized by hydrogen peroxide obtained from cathodic reduction of oxygen, which was mainly supplied by anodic oxidation of water. Based on the spectra identified by gas chromatograph/mass spectrometer (GC/MS), it is proposed that oxidative degradation of 2,4-DNT and/or 2,6-DNT, 2,4,6-TNT results in o-mononitrotoluene (MNT) and 1,3,5-trinitrobenzene, respectively. Due to the removal of TOC and some amount of water, the electrolytic method established is promising for industrial application to regeneration of spent acid from toluene nitration process.     

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.     

Determination of vapor pressure of low-volatility compounds using a method to obtain saturated vapor with coated capillary columns

The vapor pressures of O-ethyl S-2-diisopropylaminoethyl methylphosphonothiolate (VX), O-isobutyl S-2-diethylaminoethyl methylphosphonothiolate (RVX), and 2,4-dinitrotoluene (2,4-DNT) were determined with the gas saturation method in temperatures ranging from -12 to 103 degrees C. The saturated vapor was generated using a fused-silica column coated with the compound. This column was placed in a gas chromatograph, and the vapor pressure was determined directly from the detector signal or by sampling on Tenax tubes that were subsequently analyzed. From the linear relationships obtained by plotting log P vs 1/T, the enthalpies of vaporization (deltaHvap) and the vapor pressures at selected temperatures were determined. The vapor pressure of VX at 25 degrees C was 0.110 Pa and the deltaHvap 77.9 kJ x mol(-1). The corresponding results for RVX were 0.082 Pa and 76.6 kJ x mol(-1). The vapor pressure of 2,4-DNT at 72 degrees C (melting point) was determined to 6.0 Pa, and the enthalpies of the solid and the liquid state were 94.2 and 75.3 kJ x mol(-1), respectively. Using capillary columns to generate saturated vapors has three major advantages: short equilibrium time, low consumption of sample, and safe handling of toxic compounds.     

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.     

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.     

Degradation characteristics of waste lubricants under different nutrient conditions

We investigated the limits and extent of lubricants biodegradation at different nutrient conditions and evaluated several soil biological activities with regard to their usefulness for monitoring the bioremediation process in a soil contaminated with lubricants. To examine the effects of nutrient addition on lubricants biodegradation, a bench-scale investigation was conducted under different nutrient conditions for over 105 days testing period. When nutrients were added to contaminated soil with aged lubricant, great stimulation was occurred in fertilized soil for hydrocarbon degradation activity compared to non-fertilized soil. At the end of the experiment (105 days after), the initial level of contamination (9320+/-343 mg/kg) was reduced by 42-51% in the fertilized soil, whereas, only 18% of the hydrocarbon was eliminated in the non-fertilized soil. The effect of biostimulation of indigenous soil microorganisms declined with time, apparently 42% of the initial concentration of hydrocarbon remained at the end of experiment. Lubricants biodegradation process could be monitored well by soil biological parameters. In fertilized soil, biological parameters (number of HUB, soil respiration, dehydrogenase and catalase activities) were significantly enhanced and correlated with each other, as well as the residual lubricant concentration.     

Enhanced detection of nitroaromatic explosive vapors combining solid-phase extraction-air sampling,supercritical fluid extraction,and large-volume injection-GC

A complete method for sampling and analyzing of energetic compounds in the atmosphere is described. The method consists of the hyphenation of several techniques: active air sampling using a solid-phase extraction cartridge to collect the analytes, extraction of the sorbed analytes by toluene/methyl tert-butyl ether modified supercritical fluid extraction (SFE), and analysis of the extract by large-volume injection GC-nitrogen/phosphorus detection. The GC system is equipped with a loop-type injection interface with an early solvent vapor exit, a utilizing concurrent solvent evaporation technique. Chemometric approaches, based on a Plackett-Burman screening design and a central composite design for response surface modeling, were used to determine the optimum SFE conditions. The relative standard deviations of the optimized method were determined to be 4.3 to 7.7%, giving raise to method detection limits ranging from 0.06 to 0.36 ng in the sampling cartridge, equivalent to 6.2-36.4 pg/L in the atmosphere, standard sampling volume 10 L. The analytical method was applied to characterize headspace composition above military grade trinitrotoluene (TNT). Results confirm that 2,4-dinitrotoluene (DNT) and 1,3-dinitrobenzene (DNB) constitute the largest vapor flux, but TNT, 2,6-DNT, and trinitrobenzene TNB were also consistently detected in all the samples.     

Effect of soil organic matter chemistry on sorption of trinitrotoluene and 2,4-dinitrotoluene

The sorption of organic contaminants in soil is mainly attributed to the soil organic matter (SOM) content. However, recent studies have highlighted the fact that it is not the total carbon content of the organic matter, but its chemical structure which have a profound effect on the sorption of organic contaminants. In the present study sorption of two nitroaromatic contaminants viz. trinitrotoluene (TNT) and 2,4-dinitrotoluene (2,4-DNT) was studied in different SOM fractions viz. a commercial humic acid, commercial lignin and humic acid and humin extracted from a compost. ¹³C-DP/MAS NMR studies indicated that the structural composition of the organic carbon in different SOM fractions was different. The order of sorption of the nitroaromatics in the different sorbents was: humic acid-commercial > humic acid-compost > humin ～ lignin. Among the aliphatic and aromatic carbon fractions (representing bulk of SOM matrix), adsorption parameter K_f(1/n) for nitroaromatics sorption correlated well with the aliphatic carbon (r = 0.791 for TNT and 0.829 for 2,4-DNT) than the aromatic carbon (r = 0.634 for TNT and r = 0.616 for 2,4-DNT). However, among carbon containing functional groups, carbonyl carbon showed strong positive correlation with sorption of TNT (r = 0.991) and 2,4-DNT (r = 0.967) while O-alkyl carbon showed negative correlation (r = 0.832 for TNT and r = 0.828 for 2,4-DNT). The study indicates that aliphatic domains in the SOM significantly affect the non-specific sorption of both the nitroaromatic contaminants.