Evaluation of tissue and cellular biomarkers to assess 2,4,6-trinitrotoluene (TNT) exposure in earthworms: effects-based assessment in laboratory studies using Eisenia andrei

The lysosomal neutral red retention time (NRRT) assay, a biomarker for lysosomal membrane stability, and the total immune activity (TIA) assay, a measure of non-specific immune system activity, were used in laboratory studies to assess the toxic effects of 2,4,6-trinitrotoluene (TNT) on earthworms (Eisenia andrei) in vivo. The results were compared with the concentration of TNT and its metabolites in earthworm tissue, as well as standard sublethal toxicity endpoints including growth (i.e. weight change) and reproduction effects from previously published studies. Filter paper experiments indicated a significant decrease in NRRT at ≥1.8 µg TNT cm^-2, whereas sublethal (weight loss) and lethal effects to earthworms were detected at ≥3.5 and 7.1 µg TNT cm^-2, respectively. Experiments in artificial soil showed that NRRT effects could be detected at lower TNT concentrations ( ≥55 mg TNT kg^-1 soil dry weight) compared with other sublethal endpoints (effects on growth and reproduction). The TIA biomarker did not significantly respond to TNT. Copper (as CuSO₄, filter paper contact tests) and 2-chloroacetamide (soil tests), which were used as reference toxicants, also decreased the NRRT. The use of the NRRT assay linked with tissue concentrations of TNT metabolites in earthworms was identified as a potentially appropriate biomarker approach for TNT exposure assessment under laboratory conditions and a novel tool for effects-based risk assessment.     

Lysosomal neutral red retention time as a biomarker of organophosphate exposure in the earthworm Aporrectodea caliginosa: laboratory and semi-field experiments

Aporrectodea caliginosa is the most common endogeic (topsoil) earthworm in New Zealand and, because of its habitat, is potentially vulnerable to surface-applied pesticides. Lysosomal damage to earthworms, which can be visualized by the use of the neutral red retention assay (NRRA) has been evaluated in this species as a biomarker of organophosphate exposure. Earthworms were exposed in the laboratory to sub-lethal concentrations of chlorpyrifos and diazinon. In a semi-field experiment, earthworms were placed in mesocosms in a field sprayed with these pesticides at the rate recommended for a vegetable crop. In the laboratory, the neutral red retention time (NRRT) was significantly reduced following exposure to both pesticides compared with controls. In the semi-field experiment, earthworm NRRT was significantly reduced by both pesticides. These experiments have shown that the NRRA is very sensitive to exposure to chlorpyrifos and diazinon even at field rates. It therefore shows promise as a potential biomarker of contamination of soil by organophosphates.     

Toxicity of A Novel Neonicotinoid Insecticide Paichongding to Earthworm Eisenia fetida

Traditional neonicotinoid insecticides are used worldwide. Paichongding (IPP), as a novel neonicotinoid pesticide, has been widely used in China. However, the ecotoxicity of IPP to non-target invertebrates in soil ecosystem has not been reported yet. In this study, acute toxicity of IPP to earthworm Eisenia fetida, as well as the antioxidant response after IPP exposure, was evaluated. In the filter paper contact test, the LC₅₀ at 24 hr and 48 hr for IPP were 14.98 μg/cm² and 7.59 μg/cm², respectively. In artificial soil test, the LC₅₀ (lethal concentration) at 14 days and 28 days for IPP were 541.07 mg/kg and 238.51 mg/kg, respectively. The LC₅₀ of IPP is much higher than that of traditional neonicotinoid insecticides. However, earthworm body weight assessment demonstrated that the growth of earthworm was inhibited by extended exposure to IPP at sublethal doses. The activities of antioxidative enzymes superoxide dismutase and catalase in earthworms were significantly induced after IPP exposure. Malondialdehyde, a biomarker of lipid peroxidation, was also increased after IPP exposure. Although the results indicated that IPP had potentially adverse effect on earthworms, its toxicity was much lower than traditional neonicotinoids.     

Effects of earthworms and organic litter distribution on plant performance and aphid reproduction

Human management practices and large detritivores such as earthworms incorporate plant litter into the soil, thereby forming a heterogeneous soil environment from which plant roots extract nutrients. In a greenhouse experiment we investigated effects of earthworms and spatial distribution of ¹⁵N-labelled grass litter on plants of different functional groups [Lolium perenne (grass), Plantago lanceolata (forb), Trifolium repens (legume)]. Earthworms enhanced shoot and root growth in L. perenne and P. lanceolata and N uptake from organic litter and soil in all plant species. Litter concentrated in a patch (compared with litter mixed homogeneously into the soil) increased shoot biomass and ¹⁵N uptake from the litter in L. perenne and enhanced root proliferation in P. lanceolata when earthworms were present. Growth of clover (T. repens) was rather independent of the presence of earthworms and organic litter distribution: nevertheless, clover took up more nitrogen in the presence of earthworms and exploited more ¹⁵N from the added litter than the other plant species. The magnitude of the effects of earthworms and organic litter distribution differed between the plant species, indicating different responses of plants with contrasting root morphology. Aphid (Myzus persicae) reproduction was reduced on P. lanceolata in the presence of earthworms. We suggest that earthworm activity may indirectly alter plant chemistry and hence defence mechanisms against herbivores.     

Lysosomal neutral red retention time as a biomarker of organophosphate exposure in the earthworm Aporrectodea caliginosa: laboratory and semi-field experiments

Aporrectodea caliginosa is the most common endogeic (topsoil) earthworm in New Zealand and, because of its habitat, is potentially vulnerable to surface-applied pesticides. Lysosomal damage to earthworms, which can be visualized by the use of the neutral red retention assay (NRRA) has been evaluated in this species as a biomarker of organophosphate exposure. Earthworms were exposed in the laboratory to sub-lethal concentrations of chlorpyrifos and diazinon. In a semi-field experiment, earthworms were placed in mesocosms in a field sprayed with these pesticides at the rate recommended for a vegetable crop. In the laboratory, the neutral red retention time (NRRT) was significantly reduced following exposure to both pesticides compared with controls. In the semi-field experiment, earthworm NRRT was significantly reduced by both pesticides. These experiments have shown that the NRRA is very sensitive to exposure to chlorpyrifos and diazinon even at field rates. It therefore shows promise as a potential biomarker of contamination of soil by organophosphates.     

1H NMR and GC/MS metabolomics of earthworm responses to sub-lethal DDT and endosulfan exposure

The metabolic response of the earthworm Eisenia fetida to two pesticides, dichlorodiphenyltrichloroethane (DDT) and endosulfan, was characterized in contact tests using proton nuclear magnetic resonance (¹H NMR) and principal component analysis (PCA). PCA loading plots suggested that maltose, leucine and alanine were important metabolites contributing to the differences in dosed and control earthworms for both compounds at doses of 0.5, 1.0 and 2.0 μg/cm². Gas chromatography/mass spectrometry (GC/MS) was used to quantify the metabolites identified in E. fetida and determine if the changes in maltose, leucine and alanine following exposure to DDT and endosulfan (at 0.5 and 1.0 μg/cm²) were reproducible and greater than the natural variability. Quantification by GC/MS suggested that maltose was not a reliable biomarker since it both increased and decreased in earthworms exposed to DDT and increased by just 3% with exposure to endosulfan. Leucine was not stable with the GC/MS derivitization method used in this study and could not be confirmed as a reliable biomarker. However, alanine consistently increased for both DDT and endosulfan exposed E. fetida. Alanine showed considerable variability in control earthworms (±41.6%), yet the variability in alanine to glycine ratios was just ±10.5%. Increases in the alanine to glycine ratio were statistically significant at the P = 0.05 level for the 1.0 μg/cm² DDT dose and both the 0.5 and 1.0 μg/cm² endosulfan doses, suggesting that deviations from the normal homeostatic ratio of 1.5 for alanine to glycine is a potential biomarker of DDT and endosulfan exposure warranting further study. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Environmental Metabolomics Special Issue of Metabolomics.     

Differential Tolerance of Cool- and W arm-Season Grasses to TNT -Contaminated Soil

Plants can be used for effective and economical remediation of soil provided they are tolerant or resistant to the contaminants. Greenhouse experiments were conducted to determine the tolerance of the cool-season grasses: smooth bromegrass (Bromus inermus Leyss.) and tall fescue (Festuca arundinaceae Schreb), and the warm-season grasses: big bluestem (Andropogon gerardii Vitman) and switchgrass (Panicum virgatum L.) to TNT (2,4,6-trinitrotoluene) in soil. TNT-contaminated soil was mixed with uncontaminated soil to obtain water-extractable TNT concentrations ranging from 71 to 435 mg kg^-1, corresponding to acetonitrile-extractable concentrations of 278 to 3115 mg kg^-1. Germination, shoot and root dry weight, and root area were measured in response to TNT concentrations in the soil mixtures. Germination and height of the warm-season grass species were more sensitive than the cool-season grass species to increasing TNT concentrations in soil. Significant reductions in shoot and root growth were observed in cool-season grasses at lower TNT concentrations in soil compared with warm-season grasses in the soil mixtures. Results indicated that the warm-season grasses can be established in soil containing less than 86 mg of water-extractable TNT kg^-1, based on 80% of measured growth in uncontaminated control soil.     

Bioavailability of 2, 4, 6-Trinitrotoluene (TNT) to Earthworms in Three Different Types of Soils in China

To study the effects of aging time (the length of time when contaminants are sequestered in soil) and soil properties on TNT bioavailability in soil, earthworms (Eisenia fetida) were exposed to three types of soils (fluvo-aquic soil, loessal soil, and black soil) contaminated by TNT for 7, 14, 21, 28, 35, and 42 days. The Earthworm-Soil Accumulation Factor (ESAF) of TNT and soil properties were analyzed. The ESAFs in black soil were significantly lower than those in fluvo-aquic soil and loessal soil (P < 0.05). In loessal soils, the ESAF increased with aging time, while that in black soils decreased. The ESAF of TNT had a significantly negative correlation with soil organic matter content, clay contents, and cation exchange capacity, which were the main factors affecting the TNT bioavailability in soils (P < 0.01). There was more quartz and feldspar in black soil, as well as more particles and micropores on the surface, which resulted in the easy adsorption and lower bioavailability of TNT. In conclusion, TNT bioavailability in soils is affected by aging time, soil physical and chemical properties, and mineral and surface properties, which must be considered when biotreatment for TNT in soils is applied.     

Toxicity and Bioaccumulation of Petroleum Mixtures with Alkyl PAHs in Earthworms

The toxicities of three oil products with boiling-point ranges representative of petroleum hydrocarbons were tested on earthworms (Eisenia fetida) to investigate the correlation between bioaccumulated concentrations of polycyclic aromatic hydrocarbons (PAHs) and toxicity. The toxicities to earthworms were in the sequence: kerosene > diesel > bunker-C. After 14 days, the LC50s of the soils contaminated with kerosene, diesel, and bunker-C were 1079, 9135, and 15,609 mg/kg, respectively. Analysis of the body residue concentrations of PAHs in the earthworms showed that the accumulation of alkyl PAHs predominated that of the 16 priority PAHs. Principal component analysis (PCA) identified 12 PAHs, including four alkylated naphthalenes, as the oil constituents that affected mortality in the kerosene-contaminated soil. For the diesel-contaminated soil, eight PAHs were identified, including dibenzothiophene. It was not clear which compounds affected mortality in the bunker-C soil. Across the three series, biota-to-soil accumulation factors (BSAFs) ranged from 10^–2.05 to 10^3.98, and generally increased as the hydrophobicity (K_ow) or molecular weight of the alkyl PAHs increased. The toxicity endpoints of each oil product can be used as reference values in the risk assessment of soils contaminated with petroleum, and individual PAHs screened out have implications for future toxicity assessment of petroleum hydrocarbons.     

Sublethal Toxicity Endpoints of Heavy Metals to the Nematode Caenorhabditis elegans

Caenorhabditis elegans, a free-living nematode, is commonly used as a model organism in ecotoxicological studies. The current literatures have provided useful insight into the relative sensitivity of several endpoints, but few direct comparisons of multiple endpoints under a common set of experimental conditions. The objective of this study was to determine appropriate sublethal endpoints to develop an ecotoxicity screening and monitoring system. C. elegans was applied to explore the sublethal toxicity of four heavy metals (copper, zinc, cadmium and chromium). Two physiological endpoints (growth and reproduction), three behavioral endpoints (head thrash frequency, body bend frequency and feeding) and two enzymatic endpoints (acetylcholine esterase [AChE] and superoxide dismutase [SOD]) were selected for the assessment of heavy metal toxicity. The squared correlation coefficients (R²) between the responses observed and fitted by Logit function were higher than 0.90 and the RMSE were lower than 0.10, indicating a good significance statistically. There was no significant difference among the half effect concentration (EC50) endpoints in physiological and behavioral effects of the four heavy metals, indicating similar sensitivity of physiological and behavioral effects. AChE enzyme was more sensitive to copper, zinc, and cadmium than to other physiological and behavioral effects, and SOD enzyme was most sensitive to chromium. The EC50 of copper, zinc, and cadmium, to the AChE enzyme in the nematodes were 0.68 mg/L, 2.76 mg/L, and 0.92 mg/L respectively and the EC50 of chromium to the SOD enzyme in the nematode was 1.58 mg/L. The results of this study showed that there was a good concentration-response relationship between all four heavy metals and the sublethal toxicity effects to C. elegans. Considering these sublethal endpoints in terms of simplicity, accuracy, repeatability and costs of the experiments, feeding is the relatively ideal sublethal toxicity endpoint of heavy metals to C. elegans.     

Long-Term Explosive Contamination in Soil: Effects on Soil Microbial Community and Bioremediation

Explosive contamination in soil is a great concern for environmental health. Following 50 years of munitions manufacturing and loading, soils from two different sites contained ≥ 6,435 mg 2,4,6-trinitrotoluene (TNT), 2,933 mg hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,135 mg octahydrol-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) kg^{? 1} soil. Extractable nitrate-N was as high as 315 and ammonium-N reached 150 mg N kg^{? 1} soil. Water leachates in the highly contaminated soils showed near saturation levels of TNT and RDX, suggesting great risk to water quality. The long-term contamination resulted in undetectable fungal populations and as low as 180 bacterial colony forming units (CFU) g^–1 soil. In the most severely contaminated soil, dehydrogenase activity was undetectable and microbial biomass carbon was very low (< 3.4 mg C _mic kg^–1 soil). The diminished biological activity was a consequence of long-term contamination because short-term (14 d) contamination of TNT at up to 5000 mg TNT kg^–1 soil did not cause a decline in the culturable bacterial population. Natural attenuation may not be a feasible remediation strategy in soils with long-term contamination by high concentrations of explosives.     

Progeny of the maize weevil,Sitophilus zeamais,is affected by parental exposure to clove and cinnamon essential oils

The interest in and utilization of botanical insecticides, particularly essential oils, has become increasingly relevant to the control of insect pests. However, the potential ecotoxicological risks or flaws (including sublethal effects on the targeted pest generation and its subsequent progeny) of this pest control tool have been neglected frequently. Here, we evaluated the effects of sublethal exposure to clove, Syzygium aromaticum (L.) Merrill & Perry (Myrtaceae), and cinnamon, Cinnamomum verum J. Presl (Lauraceae), essential oils on adult (F0) maize weevils, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and the physiology (e.g., body mass, respirometry, and grain consumption) and population dynamics (e.g., daily emergence and sex ratio) of their progeny. Longevities of the parents were negatively affected by the essential oils in a concentration‐dependent manner. Parental sublethal exposure to clove oil (0.17 μl cm^?2) accelerated offspring emergence but delayed the emergence of females compared to males. Parents that were sublethally exposed to clove (0.17 μl cm^?2) or cinnamon (0.35 μl cm^?2) essential oils produced heavier offspring. Parental sublethal exposure to cinnamon essential oil accelerated offspring emergence (at 0.70 μl cm^?2), delayed female emergence (at 0.17 μl cm^?2), and enhanced grain consumption (at 0.35 and 0.70 μl cm^?2) of the progeny. Thus, our findings indicate that sublethal exposure to clove and cinnamon essential oils is capable of promoting transgenerational effects in S. zeamais that can negatively impact the control efficacy of such products.     

Ecotoxicity of soils contaminated with industrial and domestic wastewater in western shenyang,China

Soil samples were collected from 7 sites in the up-, mid-and down-reach along and nearby the wastewater irrigation channel, western Shenyang of China. The concentrations of selected pollutants (mineral oil, PAHs - polycycle aromatic hydrocarbons and Cd) were determined by UV spectrometer, HPLC and AAS (atomic adsorption spectrometer) spectrometer, respectively. Toxicity effects of soils were evaluated by seedling emergence test with root length of wheat as the end-point and by earthworms test with the mortality rate and inhibition rates of body weight as endpoints. Results showed accumulation of pollutants for most soils with concentration of 200.2 mg.kg⁻¹∼1600 mg.kg⁻¹ for mineral oil, 0.33 mg.kg⁻¹∼1.81 mg.kg⁻¹ for Cd and 900.16 mg.kg⁻¹ ∼ 2737.91 mg.kg⁻¹ for PAHs. The inhibition rates of root elongation were from −20% up to 40 %, and mortality rates of earthworms ranged from 0%∼40% from the exposure period of two weeks to eight weeks by sampling interval of two weeks, the inhibition rates of earthworm growth were from −19.36% to 34.53%, showing effects of stimulation at 2 weeks to an increasing effects of inhibition at 4, 6 and 8 weeks, respectively. Mortality rates correlated with the loss of body weight of earthworms.

This study indicated the potential risk of pollutants of environmental low content in soil by the determination of selected chemicals combined with toxicity indexes.

     

Ecotoxicity of soils contaminated with industrial and domestic wastewater in western shenyang,China

Soil samples were collected from 7 sites in the up-, mid-and down-reach along and nearby the wastewater irrigation channel, western Shenyang of China. The concentrations of selected pollutants (mineral oil, PAHs - polycycle aromatic hydrocarbons and Cd) were determined by UV spectrometer, HPLC and AAS (atomic adsorption spectrometer) spectrometer, respectively. Toxicity effects of soils were evaluated by seedling emergence test with root length of wheat as the end-point and by earthworms test with the mortality rate and inhibition rates of body weight as endpoints. Results showed accumulation of pollutants for most soils with concentration of 200.2 mg.kg^?1～1600 mg.kg^?1 for mineral oil, 0.33 mg.kg^?1～1.81 mg.kg^?1 for Cd and 900.16 mg.kg^?1 ～ 2737.91 mg.kg^?1 for PAHs. The inhibition rates of root elongation were from ?20% up to 40 %, and mortality rates of earthworms ranged from 0%～40% from the exposure period of two weeks to eight weeks by sampling interval of two weeks, the inhibition rates of earthworm growth were from ?19.36% to 34.53%, showing effects of stimulation at 2 weeks to an increasing effects of inhibition at 4, 6 and 8 weeks, respectively. Mortality rates correlated with the loss of body weight of earthworms.This study indicated the potential risk of pollutants of environmental low content in soil by the determination of selected chemicals combined with toxicity indexes.     

Target Value of Intraabdominal Fat Area for Improving Coronary Heart Disease Risk Factors

Objective: The goal of this study was to determine an intraabdominal fat (IF) area target value for improving coronary heart disease (CHD) risk factors in response to weight reduction. Research Methods and Procedures: Subjects were 279 obese Japanese women, 21 to 66 years old, who were divided into diet‐alone and diet‐plus‐exercise groups and participated in a 14‐week weight reduction program. The IF area was measured by computerized tomography scans. Systolic blood pressure ≥ 140 mm Hg, diastolic blood pressure ≥ 90 mm Hg, total cholesterol ≥ 5.70 mM, triglycerides ≥ 1.70 mM, and fasting plasma glucose ≥ 6.99 mM were defined as CHD risk factors. Results: The best trade‐off between sensitivity (probability of correctly detecting true positive) and specificity (probability of correctly detecting true negative) was found at 100 cm² pretreatment in combined data of the two groups. At posttreatment, although a slight difference was found in the target value between the treatment groups (60 cm² for diet alone and 50 cm² for diet plus exercise), the combined data showed that the best trade‐off occurred at 60 cm² (sensitivity and specificity were 0.55 and 0.63, respectively). The percentage of subjects having no CHD risk factors was significantly lower in the group that had large IF areas (≥60 cm²) (46%) compared with the group that had normal IF areas (<60 cm²) (65%). However, the percentage of subjects having multiple CHD risk factors was significantly greater in the group that had large IF areas (16%) compared with the group with normal IF areas (7%) at posttreatment. Discussion: Our longitudinal data suggest that obese Japanese women should reduce their IF areas to <60 cm² through weight reduction to improve CHD risk factors independent of treatment.     

Links between the detritivore and the herbivore system: effects of earthworms and Collembola on plant growth and aphid development

Effects of Collembola (Heteromurus nitidus and Onychiurus scotarius) and earthworms (Aporrectodea caliginosa and Octolasion tyrtaeum) on the growth of two plant species from different functional groups (Poa annua and Trifolium repens), and on the development of aphids (Myzus persicae) were investigated in a laboratory experiment lasting 20 weeks. Using soil from a fallow site which had been set aside for about 15 years, we expected that nitrogen would be of limited supply to plants and hypothesized that the soil animals studied, particularly earthworms, would increase nutrient availability to plants and thereby also modify aphid reproduction and development. Plant growth was modified strongly by the presence of soil animals. Earthworms caused a more than twofold increase in shoot and root mass of P. annua but increased that of T. repens by only 18% and 6%, respectively. However, earthworms neither affected plant shoot/root ratio nor the nitrogen concentration in plant tissue. In contrast, the presence of Collembola caused a reduction in plant biomass particularly that of P. annua roots, but plant tissue nitrogen concentration was increased, although only slightly. Aphid reproduction on T. repens was lowered in the presence of Collembola on average by 45% but on P. annua increased by a factor of about 3. It is concluded that Collembola decrease aphid reproduction on more palatable host plants like T. repens but increase that on less palatable ones like P. annua. Earthworm presence also affected aphid reproduction but the effect was less consistent than that of Collembola. In the presence of earthworms, aphid reproduction was in one experimental period increased by some 70%. Earthworms also modified the numbers of Collembola and their vertical distribution in experimental chambers. Exploitation of deeper soil layers by H. nitidus was increased but, generally, O. scotarius numbers were reduced whereas those of H. nitidus increased in earthworm treatments. The presence of Collembola also influenced earthworm body mass during the experiment. In general it declined, but in the presence of Collembola loss of body mass of A. caliginosa was more pronounced. We conclude that inhibiting effects between Collembola and earthworms resulted from the use of a common resource, litter material rich in nitrogen. This is supported by the higher C/N ratio of the litter material in the presence of earthworms and Collembola by the end of the experiment. Effects of soil invertebrates like Collembola and earthworms on plant performance and aphid development are assumed to be modified by complex direct and indirect interactions among soil animal groups. Received: 6 July 1998 / Accepted: 1 March 1999     

Chaotropic effects on 2,4,6-trinitrotoluene uptake by wheat (Triticum aestivum)

Previous research in our laboratory investigated the effectiveness of a common agrochemical, urea used as a chaotropic agent to facilitate 2,4,6-trinitrotoluene (TNT) removal by vetiver grass (Vetiveria zizanioides L.). Chaotropic agents disrupt water structure, increasing solubilization of hydrophobic compounds (TNT), and enhancing plant TNT uptake. Our findings showed that urea significantly enhanced TNT uptake kinetics by vetiver. We hypothesized that the beneficial effect of urea on the overall TNT uptake by vetiver grass was not plant-specific. We explored this hypothesis by testing the ability of wheat (Triticum aestivum L.) in removing TNT from aqueous media in the presence of urea. Results showed that untreated (no urea) wheat exhibited a slow, kinetically limited TNT uptake that was nearly half of the urea-treated wheat TNT capacity (250 mg kg⁻¹). Chaotropic effects of urea were illustrated by the significant (P < 0.001) increase in the TNT second-order reaction rate constants over those of the untreated (no urea) controls. Plant TNT speciation showed that TNT and several of its metabolites were detected in both root and shoot compartments of the plant, allowing for 110 and 36% recovery for the untreated and 0.1% urea treated plants. The lower % recovery of the urea-treated plants was attributed to a number of unknown polar TNT metabolites. Responsible Editor: Hans Lambers.     

Aerobic Gram-Positive and Gram-Negative Bacteria Exhibit Differential Sensitivity to and Transformation of 2,4,6-Trinitrotoluene (TNT)

A systematic evaluation of the ability of different bacterial genera to transform 2,4,6-trinitrotoluene (TNT), and grow in its presence, was conducted. Aerobic Gram-negative organisms degraded TNT and evidenced net consumption of reduced metabolites when cultured in molasses medium. Some Gram-negative isolates transformed all the initial TNT to undetectable metabolites, with no adsorption of TNT or metabolites to cells. Growth and TNT transformation capacity of Gram-positive bacteria both exhibited 50% reductions in the presence of approximately 10 μg TNT ml⁻¹. Most non-sporeforming Gram-positive organisms incubated in molasses media amended with 80 μg TNT ml⁻¹ became unculturable, whereas all strains tested remained culturable when incubated in mineral media amended with 98 μg TNT ml⁻¹, indicating that TNT sensitivity is linked to metabolic activity. These results indicate that the microbial ecology of soil may be severely impacted by TNT contamination. Received: 2 December 1996 / Accepted: 3 February 1997     

Mass Balance Studies with 14C-Labeled 2,4,6-Trinitrotoluene (TNT) Mediated by an Anaerobic Desulfovibrio Species and an Aerobic Serratia Species

Investigations were carried out to evaluate the level of incorporation of radiolabeled 2,4,6-trinitrotoluene (TNT) and metabolites into the bacterial biomass of two different bacterial species after cometabolically mediated TNT transformation. Biotransformation experiments with ¹⁴C-TNT indicated that TNT was not mineralized; however, carbon derived from TNT became associated with the cells. It was found that more than 42% of the initially applied radiolabel was associated with the cell biomass after cometabolic ¹⁴C-TNT transformation with the strictly anerobic Desulfovibrio species strain SHV, whereas with the strictly aerobic Serratia plymuthica species strain B7, 32% of cell-associated ¹⁴C activity was measured. The remainder of the radiolabel was present in the supernatants of the liquid cultures in the form of different TNT metabolites. Under anoxic conditions with the Desulfovibrio species, TNT was ultimately transformed to 2,4,6-triaminotoluene (TAT) and both diaminonitrotoluene isomers, whereas under oxic conditions with the Serratia species, TNT was converted to hydroxylaminodinitrotoluenes and aminodinitrotoluenes, with 4-amino-2,6-dinitrotoluene (4ADNT) being the major end product. In both culture supernatants, small amounts of very polar, radiolabeled, but unidentified metabolites were detected. At the end of the experiments approximately 92% and 96% of the originally applied radioactivity was recovered in the studies with the Serratia and Desulfovibrio species, respectively. Received: 21 May 1998 / Accepted: 6 July 1998     

溶酶体检测在海洋污染监测中的应用研究进展

溶酶体(lysosome)是真核细胞内重要的细胞器.近年来随着对溶酶体结构和功能研究的深入,溶酶体被认为是亚细胞水平上的有毒物质的靶点,在国外已广泛应用于海洋污染监测.本文概述了溶酶体标志酶、溶酶体 自噬系统和溶酶体膜的生物学特性,在此基础上介绍了利用溶酶体检测技术进行海洋污染监测的原理和方法.双壳贝类消化腺和鱼类肝脏最适于作为溶酶体检测的敏感器官;采用溶酶体膜稳定性测定(LMS)、溶酶体中性红保留时间测定(NRRT)、溶酶体形态测量(MM)、溶酶体标志酶免疫组化测定(Ih)和电镜(EM)观察等技术,能够指示海洋污染状况,因此溶酶体可作为生物标志物监测海洋环境污染.文中还分析了溶酶体检测的优缺点以及应注意问题,对其应用前景进行了展望.