Accumulation of organotin in Littorina littorea and Mya arenaria from Danish coastal waters

Organotin concentrations were determined in seawater, sediment and selected molluscs collected from the vicinity of the island of Fyn, Denmark. The extent of organotin contamination varied with the proximity of sampling sites to marinas and commercial shipping activities. Seasonal reductions in environmental TBT concentrations coincided with removal of pleasure craft from marinas during the winter season. At marina sites, concentration factors for TBT were 150-1500 for sediment and 500-10,000 for Littorina littorea (compared with seawater concentrations). Away from marina sites, Mya arenaria concentrated TBT by a factor of 57,000 to 220,000. The concentrations of TBT in M. arenaria from relatively clean sites were more than 10 times higher than found in L. littorea from heavily polluted marinas. M. arenaria were absent from marina sites and their immediate surroundings. Of the total organotin found in M. arenaria throughout the year, 80-90% was in the form of TBT. In contrast, in L. littorea approximately 60-70% of the total organotin was present as TBT in spring, but this proportion fell to 20-40% during autumn and winter. M. arenaria shows potential as a bioindicator organism of TBT pollution as it appears to have a very limited ability to metabolise and eliminate TBT, yet can tolerate quite high levels of TBT exposure. L. littorea might be used as a bioindicator of short-term fluctuations in TBT concentrations at heavily polluted sites. It is concluded that continued presence of TBT in the Danish coastal ecosystem may still pose an ecotoxicological threat.     

Tributyltin accumulation and effects in marine molluscs from West Greenland

The levels of the antifouling agent tributyltin (TBT) and its breakdown products in bivalves were investigated in 1999-2000 at six areas along the west coast of Greenland with focus on locations inside and outside harbours. In addition female gastropods were examined for the development of TBT-induced masculine characteristics in form of imposex or intersex. The highest TBT concentration, 254 ng x g(-1) ww, was found in the bivalve Mytilus edulis sampled inside Nuuk harbour, but significant TBT concentrations were also present in bivalves from the other harbour areas. Only low levels of TBT were detected in bivalves sampled outside the harbours and in several of the samples the TBT level was below the detection limit. The examination of neogastropods like Buccinum revealed that imposex development occurred in all the harbours. In contrast, imposex was not found in any neogastropods sampled outside the harbour areas. However, the value of marine neogastropods as indicators of TBT contamination in West Greenland seems limited, because of large species diversity and the difficulties in sampling enough specimens at least with the current sampling strategy. No effects, which could be related to TBT contamination, were found in the most abundant tidal gastropod in West Greenland, Littorina saxatilis.     

Effect of TBT on Ruditapes decussatus juveniles

The effects of sublethal concentrations of tributyltin (TBT) on growth of juvenile clams Ruditapes decussatus were determined during exposure to TBT concentrations of 50, 100 and 250 ng l(-1) (as Sn) for a period up to two years. Length and weight of clams increased continuously in all treatments throughout the experimental period, and, overall, rates were not significantly influenced by TBT exposure, although final length and weight were inversely related to increasing TBT concentration. Juvenile R. decussatus therefore appear to be less sensitive to TBT than larval stages. Some juveniles exposed to TBT developed abnormal shell growth, laterally, changing the typical flattened shape of clams into a more "rounded" form. This characteristic was more visible in the anterior margins of valves than posteriorly, and mainly observed in clams exposed to TBT at 50 ng l(-1) (as Sn).     

Experimental effects of tributyltin (TBT) contaminated sediment on a range of meiobenthic communities

Microcosm experiments have been carried out with whole natural meiobenthic communities to look at the effects of TBT sediment contamination on the community structure of the dominant nematode component of the meiobenthos. TBT has a high affinity for aquatic sediments, yet this is the first study of the effects of this contaminant in sediment on natural benthic communities. Three communities were studied from contrasting locations in south-west England: the intertidal of the Lynher estuary (muddy sediment) and the Exe estuary (sandy sediment) and the subtidal (50m depth) at Rame Head off Plymouth (muddy sand). Fresh sediment with natural meiobenthic communities was incubated for 2 months with TBT-contaminated sediment (three dose levels) in bottles. Nematodes were identified and enumerated and subjected to multivariate data analysis. The sandy Exe estuary fauna was significantly affected by TBT-contaminated sediment at all three doses (0.3, 0.6 and 0.9 microg g(-1) dry wt (as Sn) sediment), whereas the offshore fauna from Rame Head was significantly affected only at the highest dose. The muddy Lynher estuary meiofauna was affected (somewhat peculiarly) at the medium dose level only. Meiobenthic nematodes may not be as sensitive to TBT-contaminated sediment as other infaunal benthos but exhibited responses to levels of contamination still persisting in some UK estuaries and harbours. Comparing the effects of TBT with those of copper and zinc in the same laboratory experiments, our observations suggest that the relative impact of TBT on meiobenthic community structure is not as great as these contaminants in marine sediments. Although there are very few observations of TBT toxicity in sediment, it appears that TBT is toxic at much lower concentrations in seawater (ppb) than it is in sediment (ppm).     

Factors influencing organotin distribution in different marine environmental compartments, and their potential health risk

Tributyltin (TBT) and triphenyltin (TPT) in different marine environmental compartments such as seawater, sediments, and inshore fishes were investigated in 21 Taiwanese harbors between 2001 and 2004 in order to determine the major factors influencing their distribution. The existence of major input sources and the limited water exchange rate inside the harbors were indicated by higher TBT concentrations in seawater from inner harbor than from outer harbor areas. The levels of TBT in sediments were found to be mainly affected by their geographic distribution, water exchange rates and shipping activity. No significant correlations in TBT concentrations between water, sediment and fish suggested TBT accumulation by fish might not result from water and sediment, but from their food. TPT were detected in most fish samples, but found in few sediment samples and none in seawater, indicating fish could be as a target element for monitoring contaminated levels of TPT in the aquatic environment. Mean concentrations of TBT in fish muscle higher than tolerable average residue levels (TARLs), and mean hazard indices of TBT and TPT higher than 1 suggested consumption of fishes from Taiwanese harbor areas might have potential high risk to human health.     

TBT and TPhT persistence in a sludged soil

The persistence of tributyltin (TBT) and triphenyltin (TPhT) in soils was studied, taking into consideration the quantity of sewage sludge, TBT and TPhT concentrations in soil as well as the soil pH. The organotin compounds (OTC) were introduced into the soil via a spiked urban sludge, simulating agricultural practise. OTC speciation was achieved after acidic extraction of soil samples followed by gas chromatography–pulsed flame photometric analysis (GC–PFPD). Leaching tests conducted on a spiked sludge showed that more than 98% of TBT are sorbed on the sludge. TBT persistence in soil appeared to depend on its initial concentration in sludge. Thus, it was more important when concentration is over 1000 μg(Sn) kg⁻¹ of sludge. More than 50% of the initial TBT added into the soil were still present after 2 months, whatever the experimental conditions. The main degradation product appeared to be dibutyltin. About 90% of TPhT were initially sorbed on sludge, whatever the spiking concentration in sludge was. However, TPhT seemed to be quantitatively exchangeable at the solid/liquid interface, according to the leaching tests. It was also significantly degraded in sludged soil as only about 20% of TPhT remain present after 2 months, the monophenyltin being the main degradation product. pH had a significant positive effect on TBT and particularly TPhT persistence, according to the initial amounts introduced into the soil. Thus, at pH over 7 and triorganotin concentration over 100 μg(Sn) kg⁻¹, less than 10% of TBT but about 60% of TPhT were degraded. When the sludge was moderately contaminated by triorganotins (typically 50 μg(Sn) kg⁻¹ in our conditions) the pH had no effect on TBT and TPhT persistence.     

Tributyltin contamination around an oil terminal in Sullom Voe (Shetland)

This report provides clear evidence of tributyltin (TBT) contamination of coastal waters close to an oil terminal handling large tankers. Dogwhelk populations from Sullom Voe, Shetland, have been examined for the development by females of certain male sexual characteristics (e.g. a penis and vas deferens). This condition, termed imposex, is a response to exposure to tributylin compounds, and can be quantified by use of the relative penis size index (RPSI). Dogwhelk populations throughout Sullom Voe were strongly affected by TBT (RPSI 34 to 81%), and up to 90% of the females in the oil terminal area were sterile. Queen scallops collected in the Voe contained enhanced concentrations of tin in adductor muscle and gonad tissue (0.02 to 0.23 mg kg(-1) wet weight), but it is considered unlikely that their growth and development is adversely affected. Tin was rarely detected in scallops, queens, or dogwhelks outside the Voe, although the development of a vas deferens in many of the female dogwhelks indicated a degree of contamination of the open waters of Yell Sound. TBT was detectable (2 ng litre(-1) as tin) in only two sea water samples taken from the terminal area.     

Butyltin compounds in sediment and molluscs from the shipping strait between Denmark and Sweden

The distribution of tributyltin (TBT) contamination in the subtidal zone of the waters between Denmark and Sweden was investigated in relation to major international shipping lanes. Sediment and different benthic molluscs were sampled in transects along and perpendicularly to the shipping lanes in the Sound (?resund) and the Kattegat/Skagerrak region. The samples were analysed for TBT and its degradation products, dibutyltin (DBT) and monobutyltin (MBT), using GC-PFPD. In sediments, the TBT concentration ranged from <1 to 19 ng g(-1) dry weight (dw) with a strong correlation between the TBT concentration and the organic fraction in sediment (r(2)=0.90) in the samples collected in the Sound, where the highest concentrations were found. This relationship was not observed in the samples from the Kattegat because the TBT concentration in most sediment samples was below the limit of detection. In the molluscs, TBT and its degradation products were detected in all samples from the entire area with concentrations ranging from 8.1 ng g(-1) dw in Buccinum undatum to 1316 ng g(-1) dw in Nuculana pernula. The deposit-feeding bivalve N. pernula was found to have a particularly high accumulation potential for TBT. In addition, a strong correlation between TBT concentrations in sediment and N. pernula was found. Therefore this species seems to be an ideal organism for monitoring sediment contamination. The TBT concentration in N. pernula was found to decrease gradually along the shipping lanes from the Sound, through the Kattegat and into the Skagerrak.     

Decreased TBT pollution and changing bioaccumulation pattern in gastropods imply butyltin desorption from sediments

Two monitoring surveys were repeated in Galicia (NW Spain): one in 2003 concerned the rocky shore gastropod Nucella lapillus (19 populations, the reference campaign in 1996), the other dealt in 2005 with the infaunal snail Nassarius reticulatus (25 sites, the previous one in 2000). Samples were subject to a standard protocol to determine the concentrations of butyltins (BTs) in tissues. Results show that pollution in most populations has considerably decreased over the last decade: for N. lapillus the mean descent ranged from 37% (for tributyltin -TBT-) to 66% (for monobutyltin -MBT-), and TBT concentrations were on average halved in N. reticulatus. However, derivatives generally increased in this latter species, to the extent that dibutyltin -DBT- in several 2005 samples exceeded the aggregate of all three BTs in 2000. As a consequence, a major change in the bioaccumulation patterns becomes evident, particularly when computing the butyltin degradation index [BDI: (DBT+MBT)/TBT]. This shift is most marked at sites where pollution has always been lowest, and it shows significant negative correlation between both gastropods. Since sources other than antifouling paints are not important in the area, it is proposed that observations are due to BT desorption from sediments through some interplay involving the different characteristics of the chemicals and the contrasting biology of the animals.     

Acute and chronic toxicities of tributyltin to various life stages of the marine polychaete Hydroides elegans

Experiments were performed to determine the acute and chronic toxicities of tributyltin (TBT) (nominal concentrations) to polychaete Hydroides elegans. Firstly, static tests were performed to examine the lethal effect of TBT on five life stages of H. elegans. The 48-h LC(50) (95% CI) for the eggs, 2-cells, trochophores, juveniles and adults was 0.18 (0.15-0.20), 0.97 (0.77-1.23), 2.36 (2.11-2.65), 2.86 (2.63-3.12) and 4.36 (4.04-4.71) microg TBT l(-1), respectively. Secondly, a 17-d static renewal chronic test was conducted to study sensitivity of the early development (egg to juvenile) to TBT. The survivorship, percent settlement and time to reach settlement were significantly reduced at 0.01 microg TBT l(-1). Effects of TBT on the juvenile growth and maturation were also studied through a 44-d chronic test. Lower survivorship and reduced tube growth were observed only at >or= 1 microg TBT l(-1), however, only H. elegans exposed to or= 0.1 microg TBT l(-1). Our results indicate that the early development of H. elegans is highly sensitive to TBT and this polychaete can be routinely employed as a test organism for both acute and chronic ecotoxicity bioassays in tropical and subtropical regions such as southern China.     

Tributyltin (TBT) induces oxidative stress and modifies lipid profile in the filamentous fungus Cunninghamella elegans

To investigate the response of the tributyltin-degrading fungal strain Cunninghamella elegans to the organotin, a comparative lipidomics strategy was employed using an LC/MS-MS technique. A total of 49 lipid species were identified. Individual phospholipids were then quantified using a multiple reaction monitoring method. Tributyltin (TBT) caused a decline in the amounts of many molecular species of phosphatidylethanolamine or phosphatidylserine and an increase in the levels of phosphatidic acid, phosphatidylinositol and phosphatidylcholine. In the presence of TBT, it was observed that overall unsaturation was lower than in the control. Lipidome data were analyzed using principal component analysis, which confirmed the compositional changes in membrane lipids in response to TBT. Additionally, treatment of fungal biomass with butyltin led to a significant increase in lipid peroxidation. It is suggested that modification of the phospholipids profile and lipids peroxidation may reflect damage to mycelium caused by TBT.     

Occurrence of tributyltin (TBT)-resistant bacteria is not related to TBT pollution in Mekong River and coastal sediment: with a hypothesis of selective pressure from suspended solid

Tributyltin (TBT) is organotin compound that is toxic to aquatic life ranging from bacteria to mammals. This study examined the concentration of TBT in sediment from and near the Mekong River and the distribution of TBT-resistant bacteria. TBT concentrations ranged from <2.4 to 2.4 ng/g (dry wt) in river sediment and <2.4-15 ng g(-1) (dry wt) in harbor sediment. Viable count of total bacteria ranged from 2.0 x 10(4) to 1.4 x 10(7)cfu/g, and counts of TBT-resistant bacteria ranged <1.0 x 10(2) to 2.5 x 10(4)cfu/g. The estimated occurrence rate of TBT-resistant bacteria ranged from <0.01 to 34% and was highest in upstream sites in Cambodia. The occurrences of TBT in the sediment and of TBT-resistant bacteria were unrelated, and chemicals other than TBT might induce TBT resistance. TBT-resistant bacteria were more abundant in the dry season than in the rainy season. Differences in the selection process of TBT-resistant bacteria between dry and rainy seasons were examined using an advection-diffusion model of a suspended solid (SS) that conveys chemicals. The estimated dilution-diffusion time over a distance of 120 km downstream from a release site was 20 days during dry season and 5 days during rainy season, suggesting that bacteria at the sediment surface could be exposed to SS for longer periods during dry season.     

Levels of PCDDs and PCDFs in Korean river sediments and their detection by biomarkers

Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are a group of toxic halogenated aryl hydrocarbons inducing various physiological disorders against biological organisms. Here, we investigated their levels in sediment samples taken from 12 different rivers in Korea. The levels of PCDD/PCDFs in sediment samples were expressed as concentrations and international TEQ values. Among 17 PCDD/PCDFs selected as target compounds in this study, the 1,2,3,4,7,8-HxCDD and OCDD were found in all river sediments with significant variation in various congener profiles of PCDD/PCDFs in sediments. PCDD/PCDFs could be monitored by sensitive biomarkers using insect immune system. Out of 12 river sediment samples, the biomarkers reported four spots (up, middle, and down Singil sites and Ansan) as putative contamination areas. When comparing both chemical and biological monitoring results, two methods agreed three spots of Singil as contamination areas (above 10ppt levels) as well as six river sediment samples as relatively less-contaminated areas, but differed in the results in Ansan and Miho, probably due to relative non-specificity of biomarkers. Despite some disparity between bio- and chemical monitoring results, the biomarkers can be recommended as a device warning the contamination of dioxins in the environment because of a fast and inexpensive detection method.     

Imposex occurrence in marine whelks at a military facility in the high Arctic

Imposex was found in the Arctic whelk Buccinum finmarkianum at several marine stations off Thule Air Base, an US military facility in Northwest Greenland. This indicates a widespread contamination with the antifouling agents, tributyltin (TBT) or triphenyltin (TPhT) in the area, but such contamination was not supported by the organotin analyses in sediments, whelks and clams, which in general was below the analytical detection limit. Organotin concentrations above the detection limit were found only at one station close to a quay, where the highest frequency of imposex also occurred. This suggests that imposex in B. finmarkianum is a biomarker of TBT more sensitive than the detection limits, which the analytical chemistry could achieve in this study.     

Growth of Plants on TBT-contaminated Harbour Sludge and Effect on TBT Removal (10 pp)

Background Worldwide, large amounts of sediments have to be dredged annually from waterways and harbours. These sediments are sometimes polluted with a variety of toxic compounds. In some countries, including Belgium, the load with the biocide tributyltin (TBT) from ship coatings prohibits the dumping of harbour sludge into the sea. Land-based dumping is a commonly used alternative. Objective This research investigated the feasibility to use land-deposited harbour sludge for plant production. In a field trial, the growth of 38 more or less salt-tolerant plant species on low and high TBT-contaminated sediments was studied. The elimination of TBT from sludge with and without vegetation was compared. The uptake of TBT and its degradation products di- and monobutyltin (DBT and MBT) into harvest products under field conditions was determined. - Experimental Set-up. Sediments dredged in May 2003 from the brackish waters of the port of Antwerp were analysed in the laboratory for soil texture, pH, electroconductivity, sodium, magnesium, potassium, calcium, ammonium, nitrate, total nitrogen, chloride, sulphur and the organotins TBT, DBT and MBT. The sediments were lagooned for one year to dewater, desalinate and improve their structure. Salt-tolerant domestic and wild plants were selected and sown in May 2004. In August 2004, plants were harvested and the produced biomass was determined. Samples were taken from vegetated and non-vegetated top and bottom sediments and from plants growing above soil and analysed for TBT, DBT and MBT. Results The fresh sediments showed a good supply with nutrients and a neutral pH, but were rather saline (EC 14 mS cm-1 of the saturated paste extract). The salinity decreased to 3.7 mS cm-1 during lagoonation. The high and the low contaminated sediment had initially 43 and 1.6 mg TBT kg-1 dry weight, respectively. Besides TBT, several other contaminants were present in the sediments at critical levels. The biomass production of the plant species from the field trial ranged from 0.2 to 13 tons dry mass per hectare. Plants performing excellently were barley, sorghum, rape seed, a clover/grass mix and reed. If at all, a positive influence of TBT on plant growth was seen. TBT was degraded significantly faster (>40%) below barley. The uptake of TBT, DBT and MBT into stem and leaves of reed, grass and clover was very low, but measurable and not related to concentrations in soil. No uptake of TBT or its metabolites into corn of barley was found. Discussion This study confirmed former results: the toxicity of TBT to higher plants is low, and even high levels in soils would not be a hindrance for crop production. The removal of TBT seemed to be increased by both lagooning and plant growth, although the target values for sea dumping in use in certain European countries were not reached. A plausible explanation for the faster degradation of TBT under vegetation is that oxygen is a limiting factor, and plants dewater the soil, thus aerating it. The uptake of the organotins TBT, DBT and MBT into harvest products is probably due to attached soil particles. Conclusions To summarize, barley was the optimal species: it grew very well despite the salinity of the dredged sediments, it had a significantly positive effect on TBT removal; it showed no measurable uptake of TBT or the other butyltins into the harvested product; and it is a cash crop well established in European agriculture. Outlook The amounts of dredged sediments are high, and good soils are becoming increasingly rare. The feasibility of using dredged sediments for non-food production, such as energy crops, should be investigated by a critical risk assessment.     

Horizontal distribution of butyltins in surface sediments from an enclosed bay system, Korea

Tributyltin (TBT), dibutyltin (DBT), and monobutyltin (MBT) compounds were quantitatively determined from surface sediment samples (total 59 stations) covering a whole basin where harbors, shipyards, and aquaculture farms were located. Butyltin compounds were detected from all the stations covering 640 km(2) of an enclosed bay system. TBT concentrations ranged from 4 to 382 ng/g as tin on a dry weight basis, and total butyltin concentrations, from 27 to 1763 ng/g. Horizontal distribution of TBT concentration showed apparent negative gradients from harbors and shipyards, indicating that its contamination was closely related to boating and dry-docking activities. However, TBT concentrations were decreased steeply from source areas. Elevated DBT and MBT levels in creeks imply the possible input of DBT from industrial wastewater. Total butyltin concentrations in sediments are significantly correlated with particulate organic carbon concentration for the subset of stations that are distant from source areas.     

Acceleration of tributyltin chloride (TBT) degradation in liquid cultures of the filamentous fungus Cunninghamella elegans

In this study, we have examined the effects of synthetic medium ingredients and culture incubation conditions on growth and tributyltin chloride (TBT) degradation activity of the fungus Cunninghamella elegans. The best efficiency of TBT conversion to less toxic derivatives: dibutyltin and monobutyltin was noticed on media which contained glucose, NH(4)Cl, K(2)HPO(4) and MgSO(4). Next, the constructed M3 medium (with the above components) ensured vigorous growth of C. elegans and allowed the reduction of 80% of the initial TBT content (10 mg l(-1)), after 3d of biodegradation. The further acceleration of the biocide utilization by C. elegans was achieved by additional oxygen supply (pO(2) >or = 20%) to the growing fungus (89% after 2d of incubation in the BioFlo II bioreactor). The efficient xenobiotic biodegradation was related to the intensity of fungal growth. The obtained results suggest a cometabolic nature of TBT utilization by C. elegans.     

Organotins and imposex in the rock shell, Thais clavigera, from oyster mariculture areas in Taiwan

The amounts of organotin compounds such as butyltins [sigma BTs: tributyl- (TBT), dibutyl- (DBT), monobutyl- (MBT)] and phenyltins [sigma PhTs: triphenyl- (TPhT), diphenyl- (DPhT) and monophenyl- (MPhT)] in rock shells, Thais clavigera, from the northern (Shiangsan), central (Lukang) and southern (Chiku) Taiwan oyster mariculture areas varied with season and location. High values of sigma BTs (mainly TBT) and low values of sigma PhTs (mainly TPhT) were found during winter (January 1999) at Shiangsan, while high amounts of sigma PhTs (mainly TPhT) and low sigma BTs (mainly TBT) were found during summer (August 1998) at Lukang. The snails were also affected by imposex. Imposex at Shiangsan, Lukang and Chiku increased from 67.1, 59.3 and 36.7% in summer to 100, 100 and 80% in winter, respectively. Relative penis size indices similar trend at Chiku site. No male displayed rock shells were found at Shiangsan and Lukang in winter. Linear correlations between pseudopenis length versus TBT (r = 0.7655, P < 0.001), DBT (r = 0.4253, P < 0.05), MBT (r = 0.5865, P < 0.01) and TPhT (r = -0.6160, P < 0.01) were obtained significant. Among 200 samples, significant positive correlations between length (pseudopenis/shell) ratio of female versus TBT (r = 0.6944, P < 0.005) and sigma BTs (r = 0.6413, r < 0.01) were also observed. The weak correlations between length (pseudopenis/shell) ratio of female versus DBT (r = 0.3085) and MBT (r = 0.4240, P < 0.1) and TPhT (-0.4917, P < 0.05; negative correlation) could indicate that DBT, MBT and TPhT have little or no effect on the development of imposex in rock shells.     

Soil microbial parameters and luminescent bacteria assays as indicators for in situ bioremediation of TNT-contaminated soils

In situ bioremediation is increasingly being discussed as a useful strategy for cleaning up contaminated soils. Compared to established ex situ procedures, meaningful and reliable approaches for monitoring the remediation processes and their efficiency are of special importance. The subject of this study was the significance of two bioassays for monitoring purposes. The work was performed within the scope of a research project on the in situ bioremediation of topsoil contaminated with 2,4,6-trinitrotoluene (TNT). To evaluate changes within different experimental fields during a 17-month remediation period, the results of soil microbial assays and luminescent bacteria assays were compared with chemical monitoring data. The luminescent bacteria assays showed a significant reduction of the water-soluble soil toxicants in the treated fields. This bioassay proved to be a sensitive screening indicator of toxicity and may effectively aid the ecotoxicological interpretation of chemical monitoring data. Microbial biomass (C(mic)), the metabolic quotient (qCO2), and the ratio of microbial to organic carbon (C(mic)/C(org)) showed a highly significant correlation with total concentrations of TNT in the soil. But, in contrast to luminescent bacteria assays, this approach did not reveal any recovery of the soil at the end of the remediation period. There is clear evidence for persistent adverse effects of chronic TNT contamination on the site-specific microbial community and the local carbon cycle in the soil. The study clearly exhibits the differences between, as well as the complementary value of both bioassay approaches for monitoring short-term and long-term effects of soil contamination and the efficiency of remediation.