Accumulation of trinitrotoluene (TNT) in aquatic organisms: part 2--Bioconcentration in aquatic invertebrates and potential for trophic transfer to channel catfish (Ictalurus punctatus)

The potential of TNT to accumulate in aquatic organisms was assessed by determining bioconcentration factors for TNT and TNT biotransformation products using two benthic invertebrates (Chironomus tentans and Lumbriculus variegatus), and by determining the bioaccumulation factor of TNT and TNT biotransformation products due to TNT exposure via feeding for channel catfish (Ictalurus punctatus). In all three species, TNT was rapidly biotransformed resulting in minimal accumulation. The bioconcentration factors for parent TNT ranged from 3 to 4 ml g(-1) for the invertebrates studied, while the TNT bioaccumulation factor for catfish via oral exposure of food pellets was 2.4x10(-5) g g(-1) based on the concentration of TNT in the food pellet. As indicated by this small bioaccumulation factor, TNT accumulation in channel catfish through trophic transfer would be negligible compared to aqueous exposure (previously reported BCF of 0.79 ml g(-1)). TNT extractable biotransformation products accumulated to a greater degree than parent TNT for all three species. In addition, a large fraction of the radioactivity within all three species resisted solvent extraction. The highest bioconcentration factors occurred in L. variegatus with extractable radioactivity measuring 76 ml g(-1) and total radioactivity measuring 216 ml g(-1). Because the bioaccumulation of TNT is very low compared to the bioaccumulation of its biotransformation products, further research including identifying and determining the relative toxicities of these biotransformation products is necessary to fully evaluate the environmental risk posed by exposure to TNT.     

TNT and 4-amino-2,6-dinitrotoluene influence on germination and early seedling development of tall fescue

Cost-effective and environmentally acceptable methods are needed to remediate munitions-contaminated soil. Some perennial grass species are tolerant of soil contaminants and may promote remediation because of their high water use and extensive fibrous root systems. The effects of 2,4,6-trinitrotoluene (TNT) and its reduction product, 4-amino-2,6-dinitrotoluene (4ADNT), on germination and early seedling development of tall fescue (Festuca arundinacea Schreb.) were determined. Tall fescue seeds were germinated in nutrient-free agar containing 0-60 mg TNT litre(-1) or 0-15 mg 4ADNT litre(-1). Germination decreased linearly as TNT concentration increased but was not significantly affected by 4ADNT at these concentrations. Concentrations less than 30 mg TNT litre(-1) or 7.5 mg 4ADNT litre(-1) had little effect on seedling growth and development. Higher TNT or 4ADNT concentrations substantially delayed seedling development, caused abnormal radicle tissue development, and reduced secondary root and shoot growth. Seedling respiration rates decreased linearly with increasing TNT concentration. Experiments indicate that tall fescue may be grown in soils that maintain soil solution concentrations of 30 mg TNT litre(-1) or less.     

Do vesicle cells of the red alga Asparagopsis (Falkenbergia stage) play a role in bromocarbon production?

The Rhodophyceae (red algae) are an established source of volatile halocarbons in the marine environment. Some species in the Bonnemaisoniaceae have been reported to contain large amounts of halogens in structures referred to as vesicle cells, suggesting involvement of these specialised cells in the production of halocarbons. We have investigated the role of vesicle cells in the accumulation and metabolism of bromide in an isolate of the red macroalga Asparagopsis (Falkenbergia stage), a species known to release bromocarbons. Studies of laboratory-cultivated alga, using light microscopy, revealed a requirement of bromide for both the maintenance and formation of vesicle cells. Incubation of the alga in culture media with bromide concentrations below 64 mgl(-1) (the concentration of Br(-) in seawater) resulted in a decrease in the proportion of vesicle cells to pericentral cells. The abundance of vesicle cells was correlated with bromide concentration below this level. Induction of vesicle cell formation in cultures of Falkenbergia occurred at concentrations as low as 8 mgl(-1), with the abundance of vesicle cells increasing with bromide concentration up to around 100 mgl(-1). Further studies revealed a positive correlation between the abundance of vesicle cells and dibromomethane and bromoform production. Interestingly, however, whilst dibromomethane production was stimulated by the presence of bromide in the culture media, bromoform release remained unaffected suggesting that the two compounds are formed by different mechanisms.     

Toxicity and bioaccumulation of reduced TNT metabolites in the earthworm Eisenia andrei exposed to amended forest soil

Soils contaminated with 2,4,6-trinitrotoluene (TNT) and TNT primary reduction products have been found to be toxic to certain soil invertebrates, such as earthworms. The mechanism of toxicity of TNT and of its by-products is still not known. To ascertain if one of the TNT reduction products underlies TNT toxicity, we tested the toxicity and bioaccumulation of TNT reduction products. 2-Amino-4,6-dinitrotoluene (2-ADNT), 4-amino-2,6-dinitrotoluene (4-ADNT), 2,4-diamino-6-nitrotoluene (2,4-DANT) and 2,6-diamino-4-nitrotoluene (2,6-DANT) were tested separately in adult earthworms (Eisenia andrei) following a 14-d exposure to amended sandy loam forest soil. TNT, 4-ADNT, and 2-ADNT were lethal to earthworms (14-d LC(50) were: 580, 531 and 1088 micromol kg(-1), or 132, 105 and 215 mgkg(-1) dry soil, respectively) and gave the following order of toxicity: 4-ADNT>TNT>2-ADNT. Exposure to 2,4-DANT and to 2,6-DANT caused no mortality at 600 micromol kg(-1) or 100 mgkg(-1) dry soil. We found that all four TNT reduction products accumulated in earthworm tissues and 2-ADNT reached the highest levels at 3.0+/-0.3 micromol g(-1) tissue. The 14-d bioaccumulation factors were 5.1, 6.4, 5.1 and 3.2 for 2-ADNT, 4-ADNT, 2,4-DANT and 2,6-DANT, respectively. Results also suggest that some TNT metabolites are at least as toxic as TNT and should be considered when evaluating the overall toxicity of TNT-contaminated soil to earthworms.     

Ozone-induced nitrate formation in needles and leaves of Picea abies, Fagus sylvatica and Quercus robur

Much attention has been paid to ozone as a major cause of novel forest decline in Europe. In combination with acidic mist, O(3) has been observed to increase ion leaching. Besides cations lake Mg(2+), Ca(2+), K(+), NH(4)(+), considerable amounts of nitrate were found to be leached by acidic mist from needles of Norway spruce. Controlled fumigation experiments, with 100, 300, and 600 microg O(3)m(-3) over 22 days continuously, have led to a nitrate accumulation of 94.1 +/- 14.8, 119.4 +/- 28.7 and 198.9 +/- 14.9 microg NO(3)(-1) g(-1) FW, respectively, in leaves of Quercus robur. Similar values were found in leaves of Fagus sylvatica and current and previous year needles of Picea abies. Nitrate levels of controls receiving charcoal filtered air were well below 40 microg NO(3)(-) g (-1) FW. Statistically significant elevated nitrate levels were observed after only 48 h of continuous fumigation with 600 microg O(3)m(-3), in all tree species tested, and after 144 h in the 100 microg O(3)m(-3) treatment. In another experiment, trees of Picea abies were kept in two charcoal (C) and two Purafil plus charcoal (P/C) ventilated chambers, and fumigated with O and 500 microg O(3)m(-3) in cabinets of each filter-type in order to eliminate NO(x) from chamber air. After 29 days of continuous ozone fumigation, NO(3)(-) accumulation in needles amounted to 102.0 +/- 37.7 and 137.4 +/- 40.5 microg g(-1) FW in P/C and C-filtered chambers, respectively. Nitrate contents of controls were below 30 microg NO(3)(-)g(-1) FW at the end of the experiment. No significant differences in NO(3)(-) accumulation between filter treatments were observed. Since NO(x) was reduced by more than 95% in the Purafil/charcoal versus the charcoal treatment, NO(3)(-) accumulation in needles can be attributed predominantly to the influence of ozone and not to direct NO(2) uptake of needles by the possible oxidation of NO to NO(2) in the presence of ozone.     

Accumulation of trinitrotoluene (TNT) in aquatic organisms: part 1--Bioconcentration and distribution in channel catfish (Ictalurus punctatus)

Little is currently known regarding the toxicokinetics of TNT in fish. In the present study, the bioconcentration and distribution of trinitrotoluene (TNT) and TNT biotransformation products was investigated in juvenile channel catfish by exposing catfish to 14C-labeled TNT in water. Uptake experiments showed relatively fast rates (k(u)=10.1 ml g(-1) h(-1)) for TNT from the water; however, bioconcentration factors for TNT were low (0.79 ml g(-1)) due to rapid biotransformation and potential elimination of TNT. Accumulation of extractable radioactivity (TNT and all extractable biotransformation products) was much greater (BCF=10.5 ml g(-1)) than that for parent compound. TNT (parent compound) bioconcentrated to the greatest extent in the gills of the fish, while total radioactivity bioconcentrated to the greatest extent in the viscera. Residual portions of the fish that contained muscle and skin had lower concentrations of TNT than the whole fish, indicating that ingestion of fish fillets would result in decreased exposure to human consumers. Although the bioconcentration potential of TNT is very low, future research needs to be conducted to identify the biotransformation products that make up most of the radioactivity in exposed fish and evaluate their potential to promote toxicity.     

Treatment of trinitrotoluene by crude plant extracts

Crude plant extract solutions (spinach and parrotfeather) were prepared and spiked with 2,4,6-trinitrotoluene (TNT) (20 mgl(-1)). 90-h TNT removal by these solutions was compared to controls. Spinach and parrotfeather extract solutions removed 99% and 50% of the initial TNT, respectively; TNT was not eliminated in the controls or in extract solutions where removal activity was deactivated by boiling. A first-order removal constant of 0.052 h(-1) was estimated for spinach extract solutions treating 20 mgl(-1) TNT concentrations, which compared favorably to intact plant removal. Concentration variation was described by Michaelis-Menton kinetics. Detectable TNT degradation products represented only a fraction of the total TNT transformed, and the transformation favored the formation of 4-aminodinitrotoluene. The results indicated that crude plant extracts transform TNT, without the presence of the live plant.     

Comparative study of chromium biosorption by red, green and brown seaweed biomass

Dried biomass of the macroalgae Fucus vesiculosus and Fucus spiralis (brown), Ulva spp. (comprising Ulva linza, Ulva compressa and Ulva intestinalis) and Ulva lactuca (green), Palmaria palmata and Polysiphonia lanosa (red) were studied in terms of their chromium biosorption performance. Metal sorption was highly pH dependent with maximum Cr(III) and Cr(VI) sorption occurring at pH 4.5 and pH 2, respectively. Extended equilibrium times were required for Cr(VI) binding over Cr(III) binding (180 and 120min, respectively) thus indicating possible disparities in binding mechanism between chromium oxidation states. The red seaweed P. palmata revealed the highest removal efficiency for both Cr(III) and Cr(VI) at low initial concentrations. However, at high initial metal concentrations F. vesiculosus had the greatest removal efficiency for Cr(III) and performed almost identically to P. lanosa in terms of Cr(VI) removal. The Langmuir Isotherm mathematically described chromium binding to the seaweeds where F. vesiculosus had the largest q(max) for Cr(III) sorption (1.21mmol g(-1)) and P. lanosa had the largest Cr(VI) uptake (0.88mmol g(-1)). P. palmata had the highest affinity for both Cr(III) and Cr(VI) binding with b values of 4.94mM(-1) and 8.64mM(-1), respectively. Fourier transform infrared analysis revealed interactions of amino, carboxyl, sulphonate and hydroxyl groups in chromium binding to Ulva spp. The remaining seaweeds showed involvement of these groups to varying degrees as well as ether group participation in the brown seaweeds and for Cr(VI) binding to the red seaweeds.     

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.     

Two-phase partitioning bioreactor for the biodegradation of high concentrations of pentachlorophenol using Sphingobium chlorophenolicum DSM 8671

An organic-aqueous two-liquid-phase partitioning system was developed to degrade high concentrations of pentachlorophenol (PCP). Dioctyl sebacate was selected among 12 non-aqueous phases as the most suitable solvent to control the delivery of PCP to the aqueous phase for being non-biodegradable and biocompatible. In shake-flask experiments, the two-phase system was able to support the removal of 1g PCP l(-1) of total liquid phase. The performance of the two-liquid phase partitioning system (TLPPS) in shake-flask was evaluated under different conditions. At the initial biomass concentrations of 7, 25, and 58 mg dry weight l(-1), the volumetric removal rates of PCP obtained were 25.7+/-0.5, 32.1+/-0.1, and 39.3+/-2.9 mg PCP l(-1)h(-1), respectively. Higher performance was observed at lower organic-aqueous phase ratios (16% and 28%) than higher ones (37% and 44%). In a 2-l TLPPS, the degradation of 10 g PCP was completed in less than 100 h at a total volumetric rate of 142 mg l(-1) h(-1). Kinetics study using Monod model showed that compared to monophasic systems, the biphasic system significantly enhanced the maximum specific growth rate and PCP removal rate. Results of this biphasic system showed no accumulation of unknown by-product(s) which has been reported for physical-pretreatment or high-performance biphasic systems of PCP degradation.     

Hydroxylated and methoxylated polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins in red alga and cyanobacteria living in the Baltic Sea

Hydroxylated polybrominated diphenyl ethers (OH-PBDEs) and methoxylated polybrominated diphenyl ethers (MeO-PBDEs) are present in the ecosystem of the Baltic Sea. OH-PBDEs are known to be both natural products from marine environments and metabolites of the anthropogenic polybrominated diphenyl ethers (PBDEs), whereas, MeO-PBDEs appear to be solely natural in origin. Polybrominated dibenzo-p-dioxins (PBDDs) are by-products formed in connection with the combustion of brominated flame retardants (BFRs), but are also indicated as natural products in a red alga (Ceramium tenuicorne) and blue mussels living in the Baltic Sea. The aims of the present investigation were to quantify the OH-PBDEs and MeO-PBDEs present in C. tenuicorne; to verify the identities of PBDDs detected previously in this species of red alga and to investigate whether cyanobacteria living in this same region of the Baltic Sea contain OH-PBDEs, MeO-PBDEs and/or PBDDs. The red alga was confirmed to contain tribromodibenzo-p-dioxins (triBDDs), by accurate mass determination and additional PBDD congeners were also detected in this sample. This is the first time that PBDDs have been identified in a red alga. The SigmaOH-PBDEs and SigmaMeO-PBDEs concentrations, present in C. tenuicorne were 150 and 4.6 ng g(-1) dry weight, respectively. In the cyanobacteria 6 OH-PBDEs, 6 MeO-PBDEs and 4 PBDDs were detected by mass spectrometry (electron capture negative ionization (ECNI)). The PBDDs and OH-PBDEs and MeO-PBDEs detected in the red alga and cyanobacteria are most likely of natural origin.     

Solubilization of nitrotoluenes in micellar nonionic surfactant solutions

A key factor in selecting surfactants to enhance chemical or biological transformation or physical removal of an organic pollutant from contaminated soil is knowledge of the pollutant's solubility behavior in the surfactant solution. This study investigated the influence of nonionic surfactant structure on the solubility of 4-nitrotoluene (NT), 2,3-dinitrotoluene, 2,4-dinitrotoluene, 2,6-dinitrotoluene, and 2,4,6-trinitrotoluene (TNT) at room temperature. For a series of alkyl phenol ethoxylates (Tergitol NP-8 to NP-40), decreasing the ethoxylate chain length increased the solubility of these nitrotoluenes by a factor of two or less in 10 g l(-1) surfactant solutions, but did not significantly change their molar solubilization ratios (MSR, e.g. 0.02 for TNT) or their micelle-water partition coefficients (K(m), e.g. 3.4 for TNT). For Tergitol NP-8 solutions ranging from 1.0 to 12.4 g l(-1), no enhancement in NT solubility was found, suggesting that the cloud point was reached. The MSRs for Tween 80 were higher than those of Tween 20 and the MSRs of Brij-58 were higher than those for Brij-35. When comparing solutes, NT had the highest solubility and MSR (0.28-0.41), while TNT had the lowest solubility and MSR (0.02-0.03). A linear relationship between K(m) values and octanol-water partition coefficients based on Triton X-100 predicted the logK(m) values within 0.5 of their measured values. A linear solvation free energy correlation for K(m) suggested the importance of solute volume and effective hydrogen bond basicity in the partitioning process while implying that the nitrotoluenes are solubilized in a polar portion of the micelle.     

Growth and development of smooth bromegrass and tall fescue in TNT-contaminated soil

Plants can be used for effective and economical remediation of soil provided they are tolerant or resistant to contaminants. This study was conducted to determine effects of 2,4,6-trinitrotoluene (TNT) on growth and development of smooth bromegrass and tall fescue. Seeds of both species were grown in contaminated and non-contaminated soil mixed at ratios to obtain a range of concentrations and also in non-contaminated soil underlain by contaminated and non-contaminated soil mix. Germination, shoot and root dry weight, root length and area were measured. Germination and height of both species decreased with increasing TNT concentration. Shoot dry weight from tall fescue was 50% greater than smooth bromegrass at a given TNT concentration. Root length, area and dry weight of both species decreased with increasing TNT concentration. Root area and dry weight were greater for smooth bromegrass compared to tall fescue. This research indicates tall fescue and smooth bromegrass can germinate and grow in soils with concentrations less than 31 and 24 mg TNT l(-1), respectively.     

Ozone exposure-response relationships for biomass and root/shoot ratio of beech (Fagus sylvatica), ash (Fraxinus excelsior), Norway spruce (Picea abies) and Scots pine (Pinus sylvestris)

Current-year seedlings of beech, ash, Norway spruce and Scots pine were exposed during one growing season to different, but moderate, ozone (O(3)) scenarios representative for Switzerland (50, 85, 100% ambient, 50% ambient+30 nl l(-1)) in open-top chambers (OTCs) and to ambient O(3) concentrations in the field. Biomass significantly decreased with increasing O(3) dose in all species except for spruce. Losses of 25.5% (ash), 17.4% (beech), 9.9% (Scots pine) were found per 10 microl l(-1) h accumulated O(3) exposure over a threshold concentration of 40 nl l(-1) during daylight hours (AOT40). Ratios of root/shoot biomass (RSR) also significantly decreased with increasing AOT40 levels in beech and ash, but not in Norway spruce and Scots pine. The data show that the deciduous species beech and ash were more susceptible to O(3) with respect to RSR and biomass than the coniferous species Norway spruce and Scots pine.     

Kinetics and mechanism of TNT degradation in TiO2 photocatalysis

The photocatalytic degradation of TNT in a circular photocatalytic reactor, using a UV lamp as a light source and TiO(2) as a photocatalyst, was investigated. The effects of various parameters such as the initial TNT concentration, and the initial pH on the TNT degradation rate of TiO(2) photocatalysis were examined. In the presence of both UV light illumination and TiO(2) catalyst, TNT was more effectively degraded than with either UV or TiO(2) alone. The reaction rate was found to obey pseudo first-order kinetics represented by the Langmuir-Hinshelwood model. In the mineralization study, TNT (30 mg/l) photocatalytic degradation resulted in an approximately 80% TOC decrease after 150 min, and 10% of acetate and 57% of formate were produced as the organic intermediates, and were further degraded. NO(-)(3) NO(-)(2), and NH(+)(4) were detected as the nitrogen byproducts from photocatalysis and photolysis, and more than 50% of the total nitrogen was converted mainly to NO(-)(3)in the photocatalysis. However, NO(-)(3) did not adsorbed on the TiO(2) surface. TNT showed higher photocatalytic degradation efficiency at neutral and basic pH.     

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.     

2,4,6-Trinitrotoluene mineralization and bacterial production rates of natural microbial assemblages from coastal sediments

The nitrogenous energetic constituent, 2,4,6-Trinitrotoluene (TNT), is widely reported to be resistant to bacterial mineralization (conversion to CO₂); however, these studies primarily involve bacterial isolates from freshwater where bacterial production is typically limited by phosphorus. This study involved six surveys of coastal waters adjacent to three biome types: temperate broadleaf, northern coniferous, and tropical. Capacity to catabolize and mineralize TNT ring carbon to CO₂ was a common feature of natural sediment assemblages from these coastal environments (ranging to 270+/−38 μg C kg⁻¹ d⁻¹). More importantly, these mineralization rates comprised a significant proportion of total heterotrophic production. The finding that most natural assemblages surveyed from these ecosystems can mineralize TNT ring carbon to CO₂ is consistent with recent reports that assemblage components can incorporate TNT ring carbon into bacterial biomass. These data counter the widely held contention that TNT is recalcitrant to bacterial catabolism of the ring carbon in natural environments.     

Aqueous uptake and sublethal toxicity of p,p'-DDE in non-feeding larval stages of Antarctic krill (Euphausia superba)

This study evaluated the toxicological sensitivity of non-feeding larval stages of a key Antarctic species (Antarctic krill, Euphausia superba) to p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE) exposure. The aqueous uptake clearance rate of 84 mL g(-1) preserved weight (p.w.) h(-1) determined for p,p'-DDE in Antarctic krill larvae is comparable to previous findings for small cold water crustaceans and five times slower than the rates reported for an amphipod inhabiting warmer waters. Natural variations in larval physiology appear to influence contaminant uptake and larval krill behavioural responses, strongly highlighting the importance of time of measurement for ecotoxicological testing. Sublethal narcosis (immobility) was observed in larval Antarctic krill from p,p'-DDE body residues of 0.2 mmol/kg p.w., which is in agreement with findings for adult krill and temperate aquatic species. The finding of comparable body residue-based toxicity of p,p'-DDE between polar and temperate species supports the tissue residue approach for environmental risk assessment of polar ecosystems.     

The effect of solvents on metal ion adsorption by the alga Chlorella vulgaris

The dead dried alga, Chlorella vulgaris, was used for metal ion sequestering. The uptake of each of Cu(II), Cd(II), Fe(III), and Sn(IV) from their aqueous solutions decreased upon reuse of the biomass. Introducing mixed ethanol/water (50% v/v) metal ion solutions in batch systems enhanced the metal uptake of the exhausted biomass by 90% for iron, 40% for tin and only 14% for cadmium. In the column system, 20% v/v ethanol/water proved to be superior to acetone/water and isopropanol/water metal solutions in enhancing metal uptake. This increase in uptake offers a new method for regeneration of the algal capacity to remove metal ions from their solutions. Mossbauer results of iron and tin loaded algae from aqueous and mixed ethanol water solutions exhibited the characteristic doublet for (FeCl4-) at 0.350-0.565 mms(-1) respectively and the major doublet for inorganic Sn(IV) moieties at -0.217 to -0.365 mms(-1) respectively, excluding the possibility of microprecipitation of iron and tin, which is expected at such high concentrations of metals in the alga.     

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.