Degradation of 2,4,6-trinitrotoluene by selected helophytes

Four emergent plants (helophytes, synonyms emersion macrophytes, marsh plants, etc.) Phragmites australis, Juncus glaucus, Carex gracillis and Typha latifolia were successfully used for degradation of TNT (2,4,6-trinitrotoluene) under in vitro conditions. The plants took up and transformed more than 90% of TNT from the medium within ten days of cultivation. The most efficient species was Ph. australis which took up 98% of TNT within ten days. The first stable degradation products 4-amino-2,6-dinitrotoluene (4-ADNT) and 2-amino-4,6-dinitrotoluene (2-ADNT) were identified and analysed during the cultivation period. [14C] TNT was used for the detection of TNT degradation products and their compartmentalization in plant tissues after two weeks of cultivation. Forty one percent of 14C was detected as insoluble or bound in cell structures: 34% in roots and 8% in the aerial parts. These results open the perspective of using the above-mentioned plants for the remediation of TNT contaminated waters.     

Enhanced uptake of As, Zn, and Cu by Vetiveria zizanioides and Zea mays using chelating agents

Vetiveria zizaniodes (vetiver) is commonly known for its effectiveness in soil and sediment erosion control. It can tolerate to extreme soil conditions and produce a high biomass even growing in contaminated areas. Zea mays (maize) can also produce a very high biomass with a fast growth rate and possesses some degree of metal tolerance. A greenhouse study was conducted to investigate the feasibility of using vetiver and maize for remediation of arsenic (As)-, zinc (Zn-), and copper (Cu)-amended soils and evaluate the effects of chelating agents on metal uptake by these plants. Vetiver had a better growth (dry weight yield of root and shoot) than maize under different treatment conditions. The effects of different chelating agents on As, Zn, and Cu extraction from soil to soil solution were studied. Among the nine chelating agents used, it was noted that 20 mmol NTA could maximize As and Zn bioavailability, while 20 mmol HEIDA could maximize Cu bioavailability in the soil solution. The surge time in maximizing metal uptake ranged from 16 to 20 days which indicated that timing on plant harvest was an important factor in enhanced metal accumulation. In general, vetiver was a more suitable plant species than maize in terms of phytoextraction of metals from metal-contaminated soil. Application of NTA in As-amended soil and HEIDA in Cu-amended soil at the rate of 20 mmol kg(-1) increased 3-4-fold of As and Cu in shoot of both plants, whereas application of NTA (20 mmol kg(-1)) increased 37- and 1.5-fold of Zn accumulation in shoot of vetiver and maize, respectively. The potential environmental risk of metal mobility caused by chelating agents used for phytoextraction should not be overlooked.     

大孔树脂吸附去除水中的2,4,6-三硝基甲苯

研究了2种大孔树脂XAD-4和NDA-804对水中的TNT的吸附行为。2种树脂对TNT的吸附等温线表明,温度的升高有利于吸附,在35℃条件下XAD-4树脂与NDA-804树脂对TNT的最高平衡吸附量分别达82.86 mg/g和94.26mg/g。采用Langmuir方程和Freundlich方程用于吸附等温线的解释,结果表明,吸附等温线更加符合Langmuir模型,相关系数均大于0.99。TNT在2种树脂上的吸附符合准二级动力学方程,TNT初始浓度越低,达到吸附平衡所需时间越短,在1 h内可达到吸附平衡。采用NDA-804处理对TNT废水,废水中TNT浓度由103.58 mg/L降至0.4 mg/L,去除率达99.6%,吸附后的树脂采用pH=2的乙醇和盐酸的混合液脱附可再生,高浓度再生溶液经蒸馏可回收TNT,实现了废水治理与资源化。     

Photocatalytic demethylation of 2,4,6-trinitrotoluene (TNT) by porphyrins

Illumination of tetraphenyl porphyrin sulfonate (TPPS), CuTPPs and FeTTPPS in solution with trinitrotoluene (TNT) at pH7 at room temperature using tungsten lamp illumination results in the degradation of TNT to yield trinitrobenzoic acid and trinitrobenzene. No other degradation products are observed. The rate of TNT degradation follows the series TPPS > FeTPPS > CuTPPS.     

Uptake and metabolism of 2,4,6-trinitrotoluene in higher plants

The fate of the explosive 2,4,6-TNT in plants is of major interest. Therefore, a method was developed to analyse TNT and derivatives in plant tissue. The method was utilized to investigate the uptake and metabolism of TNT inMedicago sativa andAllium schoenoprasum grown in hydroponic cultures containing TNT levels of 0.1 to 10 mg/1. Detectable concentrations of nitrotoluenes were significantly higher inAllium schoenoprasum than inMedicago sativa. The uptake of TNT in plants was directly related to the initial TNT level. The principal nitroaromatic components in roots and shoots of both plant species were identified as 4-ADNT and 2-ADNT in equal amounts, with substantially less TNT.     

Enhanced bioavailability of sorbed 2,4,6-trinitrotoluene (TNT) by a bacterial consortium

Large quantities of trinitrotoluene (TNT) have been associated with past and present military activities worldwide. Because this contaminant is highly toxic and strongly sorbs to soil particles, bacteria that are able to transform it have had very little success, if any. This study was conducted to evaluate the bioavailability of 14C-labeled TNT in soil for microbial mineralization. Sorption-desorption experiments indicated that a Kendaia loam soil effectively adsorbs this explosive compound, with approximately 30-45% of the added TNT remaining sorbed to the soil after a total of 10 washings. A bacterial consortium isolated from explosive-contaminated sites was prepared in liquid medium and then tested in a TNT-spiked Kendaia loam soil. The concentration of TNT in the soil that was inoculated with the bacterial consortium was reduced by more than 30% of the initial concentration compared to the soil that did not contain the bacterial consortium within a period of 20 weeks. Nearly half of the TNT was mineralized as determined by the percentage of 14CO2 produced. Only one member of the consortium (i.e., Enterobacter sp.) significantly mineralized 25% of TNT although the extent of mineralization was significantly enhanced to 35% in the presence of the other two members of the consortium. The data suggest that some of the strongly adsorbed TNT may be accessible for metabolism if conditions for the right combination of microorganisms with specialized capabilities are optimized. The remaining sorbed fraction of substrate is presumably sequestered and thus unavailable to the microorganisms.     

The use of amino compounds for binding 2,4,6-trinitrotoluene in water

Sites polluted with 2,4,6-trinitrotoluene (TNT) constitute a worldwide problem. In this work, chemical reactions for binding TNT to amino-compounds are proposed as an initial step for developing new remediation techniques to clean-up groundwater and soils contaminated with TNT. Indeed, addition of aniline and an amino acid-like cysteine caused a decrease in free TNT of 86% and 68-100%, respectively. Using 13C-NMR spectroscopy, it was shown that TNT chemically forms a Meisenheimer complex with cysteine and aniline in 1/1 (by vol.) H2O/d6-acetone.     

Photolysis of nitroaromatics in aquatic systems. I. 2,4,6-trinitrotoluene

Photolysis of 2,4,6-trinitrotoluene (TNT) occurs rapidly in pure and natural waters irradiated with sunlight, with half-lives of less than a half-hour in some natural waters. Experiments to study the TNT photolysis products and to obtain evidence for complexation of TNT with natural substances are discussed.     

A study of 2,4,6-trinitrotoluene inhibition of benzo[a]pyrene uptake and activation in a microbial mutagenicity assay

A number of in vitro and in vivo studies have determined that binary and complex mixtures may interact to produce a toxicity that could not be predicted based on the individual chemicals. The present study was conducted with a binary mixture of model compounds to investigate possible interactions affecting their mutagenicity. The compounds included Benzo[a]pyrene (BAP), a polycyclic aromatic hydrocarbon that is an indirect-acting mutagen of great environmental concern, and 2,4,6-Trinitrotoluene (TNT), a nitro-aromatic compound that is a direct-acting mutagen frequently found as a soil contaminant at munitions sites. This study indicated that a binary mixture of BAP and TNT failed to induce the positive mutagenic response in Salmonella typhimurium strain TA98 characteristic of either compound alone. Spectrofluorometric analysis of BAP, and kinetic analyses of ³HBAP uptake in the presence or absence of TNT using TA98 cells that were treated or untreated with activated rat liver microsomes were performed. In cells preloaded with BAP, cellular BAP fluorescence was rapidly suppressed in the presence of TNT. Mass spectroscopy of BAP and TNT mixtures revealed a number of products, believed to be the result of complexation and nitration, that may account for the antagonistic action of TNT on BAP-induced mutagenicity in TA98 cells. Further, kinetic studies indicated that TNT inhibited the incorporation of BAP into cells.     

Optimized microwave extraction for trace detection of 2,4,6-trinitrotoluene in soil samples

An optimized microwave assisted extraction method for determination of trinitrotoluene (TNT) and related compounds in soil is presented. The new enhanced method exhibits improved extraction recovery and precision as well as sample handling time. For the separation and detection gas chromatography coupled to a thermoionic probe was used achieving TNT and dinitrotoluene detection limits per injection at the femtogram level. The generated extraction recovery and precision data are given for spiked and certified soil. Determined TNT and related compounds residues in soil collected from different parts of the world are presented.     

Germination and seedling development of switchgrass and smooth bromegrass exposed to 2,4,6-trinitrotoluene

It is estimated that explosives contaminate approximately 0.82 million cubic metres of soil at former military installations throughout the US; major contaminants often include 2,4,6-trinitrotoluene (TNT) and its degradation products. At some sites, phytoremediation may be a viable option to incineration or other costly remediation treatments. Grasses may be particularly suited for remediation because of their growth habit and adaptability to a wide range of soil and climate conditions. We characterized the effects of TNT on germination and early seedling development of switchgrass and smooth bromegrass to evaluate their potential use on contaminated sites. Switchgrass and smooth bromegrass seeds were germinated in nutrient-free agar containing 0 to 60 mg TNT litre(-1). Smooth bromegrass germination decreased as TNT concentration increased, while switchgrass germination was unaffected by TNT. Concentrations up to 15 mg TNT litre(-1) did not affect switchgrass root growth rate, but bromegrass root growth was reduced at TNT concentrations above 7.5 mg litre(-1). At 7.5 mg TNT litre(-1), however, shoot growth rate was reduced in both species. Examination at 20-fold magnification revealed switchgrass radicles were unaffected by TNT, while smooth bromegrass radicles appeared slightly swollen. Results indicate switchgrass is more tolerant of TNT than smooth bromegrass, but the establishment of both species may be limited to soil containing less than 50 mg kg(-1) of extractable TNT.     

Ecotoxicological evaluation of in situ bioremediation of soils contaminated by the explosive 2,4,6-trinitrotoluene (TNT)

To evaluate the environmental relevance of in situ bioremediation of contaminated soils, effective and reliable monitoring approaches are of special importance. The presented study was conducted as part of a research project investigating in situ bioremediation of topsoils contaminated by the explosive 2,4,6-trinitrotoluene (TNT). Changes in soil toxicity within different experimental fields at a former ordnance factory were evaluated using a battery of five bioassays (plant growth, Collembola reproduction, soil respiration, luminescent bacteria acute toxicity and mutagenicity test) in combination to chemical contaminant analysis. Resulting data reveal clear differences in sensitivities between methods with the luminescent bacteria assay performed with soil leachates as most sensitive toxicity indicator. Complete test battery results are presented in so-called soil toxicity profiles to visualise and facilitate the interpretation of data. Both biological and chemical monitoring results indicate a reduction of soil toxicity within 17 months of remediation.     

Coupling of 2,4,6-trinitrotoluene (TNT) metabolites onto humic monomers by a new laccase from Trametes modesta

During degradation of trinitrotoluene (TNT) by Trametes modesta, addition of humic monomers prevented the accumulation of all major stable TNT metabolites (aminodinitrotoluenes [AMDNT]) by at least 92% in the presence of 200 mM ferulic acid and guaiacol. Acute toxicity tests with individual TNT metabolites and in T. modesta cultures supplemented with 200 microM TNT demonstrated that the TNT biodegradation process lead to less toxic metabolites. Toxicity decreased in the order TNT>4-HADNT (4-hydroxylaminodinitrotoluene)>2-HADNT>2,6-DNT (2,6-dinitrotoluene)>2',2',6,6-azoxytetranitrotoluene>4-AMDNT>2-AMDNT>2,4-diamninonitrotoluene (2,4-DAMNT) while 2,4-DNT and 2,6-DAMNT were the least toxic. Ferulic acid is the best candidate for immobilization TNT biodegradation metabolites since it prevented the accumulation of AMDNTs in cultures during TNT biodegradation and its products were less toxic. All humic monomers were very effective in immobilizing 2-HADNT [100%], 4-HADNT [100%] and 2,2,6,6-azoxytetranitrotoluene [100%]. Two distinct laccase isoenzymes (LTM1 and LTM2) potentially involved in immobilization of TNT degradation products were purified to electrophoretic homogeneity. LTM1 and LTM2 have molecular weights of 77.6 and 52.5 kDa, are 18% and 24% glycosylated, have pI values of 3.6 and 4.2, respectively. Both enzymes oxidized all the typical laccase substrates tested. LTM1 showed highest kinetic constants (K(m)=0.03 microM; K(cat)=8.8 4x 10(7)s(-1)) with syringaldazine as substrate.     

Biotransformation of 2,4,6-trinitrotoluene (TNT) by enchytraeids (Enchytraeus albidus) in vivo and in vitro

2,4,6-Trinitrotoluene (TNT) is toxic to soil invertebrates, but little is known about its toxicokinetic behavior in soil. Tissue residue analysis was used to evaluate whether the presence of TNT and its reduced metabolites in soil invertebrates was due to uptake of these compounds from the soil into the organism, or due to microbial transformation of TNT associated with the organism followed by uptake. Adult white potworms (Enchytraeus albidus) were exposed to non-lethal concentrations of TNT in amended artificial soil for 21 d, or to TNT in solution for 20 h. Soil exposure studies confirmed earlier reports that TNT was transformed in enchytraeids in vivo to 2- and 4-aminodinitrotoluenes. However, enchytraeid exposure to TNT in solution led to the additional presence of 2,4-diaminonitrotoluene as well as 2- and 4- hydroxyamino-dinitrotoluenes and azoxy-compounds, suggesting that TNT can be metabolized in vivo in the absence of soil. Incubation of unexposed enchytraeid homogenates with TNT led to a protein-dependent appearance of these metabolites in vitro after > or =16 h incubation. Cellular fractionation studies indicated that most of this activity resided in the 8000 x g pellet, and was completely inhibited by broad-spectrum antibiotics. These studies demonstrate that enchytraeids can transform TNT in vivo and in vitro, at least in part, by bacteria associated with the host organism.     

Effect of photosensitizer riboflavin on the fate of 2,4,6-trinitrotoluene in a freshwater environment

The effect of riboflavin (1 microM) on the fate of TNT (20 mg/l) in a natural water environment was studied. The relative contribution of photolysis, microbial assemblages and freshwater matrix to TNT degradation was examined. The rates, extent and products of TNT and riboflavin transformation were compared under different experimental conditions. It was found that riboflavin significantly enhanced the degradation of TNT in natural water environment. Thus it is a potentially useful photosensitizing agent for the treatment of TNT-contaminated surface water. Furthermore, in the presence of riboflavin, two new intermediates with max. absorption wavelength of 230 nm were found, demonstrating that transformation of TNT in the presence of riboflavin undergoes different pathways.     

Effectiveness of urea in enhancing the extractability of 2,4,6-trinitrotoluene from chemically variant soils

One of the major challenges in developing an effective phytoremediation technology for 2,4,6-trinitrotoluene (TNT) contaminated soils is limited plant uptake resulting from low solubility of TNT. The effectiveness of urea as a solubilizing agent in increasing plant uptake of TNT in hydroponic systems has been documented. Our preliminary greenhouse experiments using urea were also very promising, but further characterization of the performance of urea in highly-complex soil-solution was necessary. The present study investigated the natural retention capacity of four chemically variant soils and optimized the factors influencing the effectiveness of urea in enhancing TNT solubility in the soil solutions. Results show that the extent of TNT sorption and desorption varies with the soil properties, and is mainly dependent on soil organic matter (SOM) content. Hysteretic desorption of TNT in all tested soils suggests irreversible sorption of TNT and indicates the need of using an extractant to increase the release of TNT in soil solutions. Urea significantly (p < 0.0001) enhanced TNT extraction from all soils, by increasing its solubility at the solid/liquid interface. Soil organic matter content and urea application rates showed significant effects, whereas pH did not exert any significant effect on urea catalysis of TNT extraction from soil. The optimum urea application rates (125 or 350 mg kg⁻¹) for maximizing TNT extraction were within the limits set by the agronomic fertilizer-N rates used for major agricultural crops. The data obtained from this batch study will facilitate the optimization of a chemically-catalyzed phytoremediation model for cleaning up TNT-contaminated soils.     

Degradation of 2,4,6-trinitrotoluene in water and soil slurry utilizing a calcium peroxide compound

The degradation of 2,4,6-trinitrotoluene was examined in pure water and contaminated soil slurry using calcium peroxide as a source of solid hydrogen peroxide and oxygen. The extent of TNT oxidation was compared with that obtained by using hydrated lime, which is normally generated by slurrying CaO2 in water and contained in CaO2 technical formulation (approximately 50%, w/w). Complete TNT degradation occurred between 280 min, 0.1% CaO2/Ca(OH)2 and 20 min, 1% CaO2/Ca(OH)2. A large part of the generated oxidation products, 80-90%, were absorbed on the solid calcium hydroxide, whereas the remaining 10-20% was detected in solution until 48 h. Removal of nitro groups was extremely effective in CaO2 slurry, where all the nitrogen (3 mol per mol of TNT) was removed from TNT within 240 min. Respect to calcium hydroxide, the peroxy compound liberated H2O2 in solution, 370 mg l-1 at 0.2% CaO2, w/v, which then decomposed within 480 min. Most of the 14C-TNT was retained more strongly on the calcium hydroxide generated by slurrying CaO2. This pool remained adsorbed on the solid until pH dropped below 5.8. The treatment of a contaminated soil slurry, 700 mg TNT kg-1, reduced CH3CN extractable TNT below 20 mg kg-1 at very low concentration of CaO2/Ca(OH)2, approximately 0.2%, w/w. Both oxidants do not lead to soil sterilization as the phosphorus added to neutralize the pH serves as a source of nutrient for the soil biomass.     

Aniline and 2,4,6-trinitrotoluene associate preferentially to low molecular weight fractions of dissolved soil organic matter

Aniline and 2,4,6-trinitrotoluene (TNT) were equilibrated with particulate (POM) and dissolved organic matter (DOM) from an organic soil at different compositions of adsorbed major cations (Na, Al) and pH (aniline: 3.7–5.1, TNT: 4.8–5.0). After separation of POM, concentrations of ¹⁴C-labelled aniline and TNT* (including TNT degradation products) were determined in DOM size fractions using size-exclusion chromatography (SEC) and UV-detection. Concentrations in the <3.5 kDa size fraction were 2.8–6.0 and 8.5–9.5 times higher for aniline and TNT*, respectively, as compared to the >40 kDa fraction. Thus, both aniline and TNT* were preferentially associated to the smallest DOM size fraction. The significant binding to DOM (similar extent as to POM) and the fact that the <3.5 kDa DOM fraction was less susceptible to flocculation by major metals suggests that the mobility of aniline and TNT is highly affected by the solubility of soil organic matter.     

Growth and lead accumulation by the grasses Vetiveria zizanioides and Thysanolaena maxima in lead-contaminated soil amended with pig manure and fertilizer: a glasshouse study

Bo Ngam lead mine soils contain high concentrations of lead (up 1% total Pb) and low amounts of organic matter and major nutrients (N, P, K). A glasshouse study was conducted to compare growth performance, metal tolerance and metal uptake by two grasses, Thysanolaena maxima (Roxb.) O. Kuntze and four ecotypes of Vetiveria zizanioides (L.) Nash, syn. Chrysopogon zizanioides (L.) Roberty (three from Thailand: Surat Thani, Songkhla and Kamphaeng Phet, and one from Sri Lanka) and to study the effects of pig manure (20% and 40% w/w) and inorganic fertilizer (75 and 150 mg kg(-1)) amendments to this lead mine soil. The results showed that both T. maxima and V. zizanioides (Surat Thani and Songkhla) could tolerate high Pb concentrations in soil (10750 mg kg(-1)) and had very good growth performance. Application of pig manure increased electrical conductivity (EC) and reduced DTPA-extractable Pb concentration in the soils. Pig manure application improved the growth of vetiver, especially at 20%, application dosage. Vetiver had the highest biomass. T. maxima could not tolerate high EC values. The uptake by roots and transport of Pb to shoots of both species was reduced when soils were amended with pig manure. Application of inorganic fertilizer did not improve growth of vetiver but did improve that of T. maxima. Fertilizer application did not have any great influence on the Pb uptake in vetiver while T. maxima took up more Pb as a result of the fertilizer enhancing its biomass yield. Both species transported low Pb concentrations to shoots (8.3-179 mg kg(-1)) and accumulated higher concentrations in roots (107-911 mg kg(-1)). In summary, both species may be species well suited for phytostabilization in tropical lead mine areas.