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.     

Recombinant luminescent bacterial sensors for the measurement of bioavailability of cadmium and lead in soils polluted by metal smelters

Environmental hazard of heavy metals in soils depends to a large extent on their bioavailability. The approach used in this study enables the determination of bioavailable metals in solid-phase samples. Two recombinant bacterial sensors, one responding specifically to cadmium and the other to lead and cadmium by increase of luminescence (firefly luciferase was used as a reporter) were used to determine the bioavailability of these metals in soil-water suspensions (a contact assay) and respective particle-free extracts. Fifty agricultural soils sampled near zinc and lead smelters in the Northern France containing up to (mg/kg) 20.1 of Cd, 1050 of Pb and 1390 of Zn were analysed. As the soil matrix interferes with the assay, recombinant luminescent control bacteria lacking the metal recognizing protein and corresponding promoter (thus, being not metal-inducible) but otherwise comparable to the sensor bacteria (the same host bacterium and plasmid encoding luciferase) were used in parallel to take into account the possible quenching and/or stimulating effects of the sample on the luminescence of the sensor bacteria. Both, chemical and sensor analysis showed that only microg/l levels of metals were extracted from the soil into the water phase (0.1% of the total Cd, 0.07% of Pb and 0.5% of Zn). However, 115-fold more Cd and 40-fold more Pb proved bioavailable if the sensor bacteria were incubated in soil suspensions (i.e., in the contact assay). The bioavailability of metals in different soils varied (depending probably on soil type) ranging from 0.5% to 56% for cadmium and from 0.2% to 8.6% for lead.     

Degradation of di-butyl-phthalate by soil bacteria

Twelve Gram-positive phthalate ester degraders were isolated from soil. Using Biolog GP2 plates, eight of them were identified as belonging to the Corynebacterium-Mycobacterium-Nocardia group, while the remaining four were unidentifiable. When cultured in the presence of di-butyl-phthalate (DBP) in basal salts solution, five of these isolates accomplished more than 90% of DBP degradation within 48 h (fast group), three were placed in the medium group, and the remaining four were placed in the slow group which caused less than 30% of DBP degradation within the same period of time. A 420 bp DNA fragment was amplified from six isolates and none of them fell within the slow group. When compared with the large subunit of phthalate dioxygenase gene (phtA) of Arthrobacter keyseri, 83% and 91% similarities were evident in the nucleotide and amino acid sequences, respectively. However, no correlation between cell surface hydrophobicity and phthalate degradation ability was evident. Six surfactants (Brij 30, Brij 35, Tergitoltype NP-10, Triton N-101, Triton X-100 and SDS) were tested for their abilities to increase degradation rate. When added at the critical micellar concentration (CMC), they all displayed strong growth inhibition against the three bacteria tested, with Brij 30 been the least toxic to isolates G2 and G11, and Brij 35 had the least inhibitory effect for G1. When half the CMC of Brij 30 was incorporated into the basal salts, the inhibitory effect on DBP degradation remained. Soil helped to minimize surfactant toxicity of surfactant and increase the degradation potential of some of the test bacteria. When DBP-amended soil had been aged for three months, decreases in bioavailability were observed but the effect varied tremendously between different organisms. For isolates G1, G2, G5, G7 and G17 the aging effects were almost non-exist. The present study indicates that selection of a suitable degrader may minimize the undesired effect of aging on bioremediation process.     

Role of loosely bound humic substances and humin in the bioavailability of phenanthrene aged in soil

A study was conducted to determine a possible role of loosely bound humic substances (i.e., humic and fulvic acids) in bioavailability of aged phenanthrene with time. In this study, long-term residence of phenanthrene in soil is defined as aging or sequestration, and the effect was determined by the declined bioavailability to bacteria of the polycyclic aromatic hydrocarbon with increased residence time. After 1, 7, and 100 days of aging of phenanthrene in Lima loam, about 90-93% of initial phenanthrene was recovered from the humin-mineral fraction of Lima loam whereas less than 12% was found in humic and fulvic acids of the same soil. Mineralization rates of phenanthrene aged in the humin-mineral fraction significantly decreased with time by the test bacterium P5-2. In terms of extents of mineralization, the difference with time was not appreciable, but still significant at P<0.05. Additional decreases in the rates and extents of mineralization were observed with the whole soil (i.e. Lima loam) to which phenanthrene had been aged. Data suggest that major sequestration sites for phenanthrene may reside in the humin-mineral fraction, and probably humic and fulvic acids may act as a physico-chemical barrier to bacterial degradation so that the compound's bioavailability may be limited.     

Butanol extraction to predict bioavailability of PAHs in soil

The feasibility of a mild-solvent extraction procedure to predict the bioavailability of individual polycyclic aromatic hydrocarbons (PAHs) in soil was assessed. The quantities that were degraded during the course of biodegradation of phenanthrene and pyrene in soil with or without plants correlated with the amounts extracted by n-butanol, with R2 values of 0.971 and 0.994, respectively. Six consecutive groups of earthworms removed ca. 70% of the pyrene remaining after extensive biodegradation, a value similar to the quantity extracted by n-butanol. The amount of chrysene aged in sterilized soil that was extracted by n-butanol was not statistically different from the quantities assimilated by earth-worms (Eisenia fetida) introduced into the soil. Such a mild extraction procedure may be useful as a means of predicting PAH bioavailability.     

Effect of Diuron on aquatic bacteria in laboratory-scale wastewater treatment ponds with special reference to Aeromonas species studied by colony hybridization

Six laboratory-scale wastewater treatment ponds were filled with sediment and water obtained from a reference pond (a wastewater treatment plant located in a rural environment at Montel-de-Gelat, Puy-de-D?me, France). They were kept at 20 degrees C, with alternative light and dark periods (12 h-12 h), and fed with raw effluent supplied weekly. Three of them were treated with Diuron (dissolved in DMSO) at a final concentration 10 mg/l, while the other three received only DMSO. Physico-chemical parameters, total bacteria, cultivable bacteria, and Aeromonas spp. were measured periodically until 41 days after the Diuron contamination. Total bacteria were treated with 4,6-diamidino 2-phenylindole (DAPI) and counted by epifluoroscence microscopy. The cultivable bacteria were quantified on plate count agar medium and Aeromonas spp. using colony hybridization. In the contaminated pilots, biochemical oxygen demand (BOD5), chemical oxygen demand (COD), suspended solids (SS), volatile suspended solids (VSS), ammonium, phosphorus, and bacteria increased, but dissolved oxygen decreased. The abundance of total bacteria, cultivable bacteria (multiplied by 30), and Aeromonas spp. increased for two weeks after Diuron introduction, reverting to initial values three weeks later. The percentage of cultivable bacteria relative to total bacteria was 0.2% in controls and 1.2% in treated pilots, while the percentage of Aeromonas spp. relative to cultivable bacteria decreased from 6-10% to 2%. Our results suggest that Diuron, which acts on the photosystem II of phototrophs, supports the development of cultivable bacteria through new carbon sources derived from the decomposition of photosynthetic micro-organisms, but does not specifically support Aeromonas spp.     

Bioavailability of dissolved organic nitrogen in treated effluents.

The research objective was to assess dissolved organic nitrogen (DON) bioavailability in wastewater effluents from a pilot-scale nitrification plant and a laboratory-scale total nitrogen (TN) removal plant. The DON bioavailability was assessed using a 14-day bioassay protocol containing bacterial and algal inocula. Nitrogen species, dissolved organic carbon, chlorophyll a, and biomass (as total suspended solids and culturable cell counts) concentrations were measured to assess DON bioavailability. The results showed an increase in algal chlorophyll a concentration, with a concurrent increase in algal biomass over time; increased bacterial counts and a decrease in DON concentration over time; and increased carbon-to-nitrogen ratio at the end of the 14-day bioassay, indicating effluent DON bioavailability to algae and bacteria. Approximately 18 to 61% of the initial DON in low-total-nitrogen wastewater effluent (TN = 4 to 5 mg/L) sample was bioavailable. The results show that bacteria and algae uptake and release DON during their growth.     

Effect of soil ageing on in vivo arsenic bioavailability in two dissimilar soils

Arsenic (As) bioavailability in spiked soils aged for up to 12 months was assessed using in vitro and in vivo methodologies. Ageing (natural attenuation) of spiked soils resulted in a decline in in vivo As bioavailability (swine assay) of over 75% in soil A (Red Ferrosol) but had no significant effect on in vivo As bioavailability even after 12 months of ageing in soil B (Brown Chromosol). Sequential fractionation, however, indicated that there was repartitioning of As within the soil fractions extracted during the time course investigated. In soil A, the As fraction associated with the more weakly bound soil fractions decreased while the residual fraction increased from 12% to 35%. In contrast, little repartitioning of As was observed in soil B indicating that natural attenuation may be only applicable for As in soils containing specific mineralogical properties.     

Toxicity and bioavailability to bacteria of particle-associated arsenite and mercury

The overall toxicity of soil, and the bioavailability and arsenite from soil were measured with the constructed constitutively luminescent strain Pseudomonas fluorescens OS8 (pNEP01) and with earlier published biosensor strains P. fluorescens OS8 (pTPT11) for mercury and P. fluorescens OS8 (pTPT31) for arsenite, respectively. Both spiked and authentic samples were studied. By combining bacterial assays enabled partial analysis of reasons for toxicity of environmental samples, some of which were highly toxic despite containing little or no heavy metals. The spiked soils were not toxic overall but the method of measuring concentration from water-extractable fraction or from soil-water slurry affected the results significantly. Mercury that was bound to clay even after water extraction was nevertheless found to be bioavailable to a high degree to the biosensor bacteria. Since induction of the luminescence genes takes place intracellularly the bacteria may able to apparently release mercury when in direct contact with clay particle. This type of biomobilisation was not observed with arsenite spiked soils. The same phenomenon was detected in one of the environmental samples.     

The effect of earthworms on the fractionation and bioavailability of heavy metals before and after soil remediation

The effect of two earthworm species, Lumbricus rubellus and Eisenia fetida, on the fractionation/bioavailability of Pb and Zn before and after soil leaching with EDTA was studied. Four leaching steps with total 12.5 mmol kg(-1) EDTA removed 39.8% and 6.1% of Pb and Zn, respectively. EDTA removed Pb from all soil fractions fairly uniformly (assessed using sequential extractions). Zn was mostly present in the chemically inert residual soil fraction, which explains its poor removal. Analysis of earthworm casts and the remainder of the soil indicated that L. rubellus and E. fetida actively regulated soil pH, but did not significantly change Pb and Zn fractionation in non-remediated and remediated soil. However, the bioavailability of Pb (assessed using Ruby's physiologically based extraction test) in E. fetida casts was significantly higher than in the bulk of the soil. In remediated soil the Pb bioavailability in the simulated stomach phase increased by 5.1 times.     

The influences of fly ash on stabilization for Cd in contaminated soils

Soil contaminated with potentially toxic metals (PTMs) has being a global environmental issue, which needs to be addressed on the priority basis. Fly ash (FA) is a kind of low-cost alkaline materials, which has been widely used in remediation of soil contaminated by PTMs, while the effects of FA on the stability for PTMs in contaminated farmland soil are still not clearly evaluated. In this study, cadmium (Cd) contaminated soil samples, collected from Shaanxi (SX), Hubei (HB), and Zhejiang (ZJ) province of China, were amended with FA addition (0, 1%, 2.5%, 5%, and 10% dose), and 1-year changes of Cd availability in soil samples were focused on. In addition, biological assessment method through pot culture was carried out to evaluate the reuse potential of Cd contaminated soils amended by FA. The result indicated that FA had a notable impact on decreasing the Cd mobility of SX soil (sand type), with 18.2~52.1% reduction in the DTPA extractable solution, followed by HB soil with 5.9~16.7% reduction, but no obvious effect of FA on ZJ soil (clay type) was observed. Furthermore, the results of pot experiment revealed that FA application could increase the biomass of Chinese cabbage. However, the DTPA extractable Cd in soils after planation and the Cd accumulation of plant increased. The results revealed that FA was not a promising soil stabilizer to immobilize HMs in Cd contaminated soil, and careful consideration should be given to Cd contaminated soils with FA restoration especially in their using for farmland productive due to the remaining risk of Cd bioavailability. These results also contributed to provide references for similar soil pollution remediation.

     

Phytoremediation of contaminated soils and groundwater: lessons from the field

Background, aim, and scope

The use of plants and associated microorganisms to remove, contain, inactivate, or degrade harmful environmental contaminants (generally termed phytoremediation) and to revitalize contaminated sites is gaining more and more attention. In this review, prerequisites for a successful remediation will be discussed. The performance of phytoremediation as an environmental remediation technology indeed depends on several factors including the extent of soil contamination, the availability and accessibility of contaminants for rhizosphere microorganisms and uptake into roots (bioavailability), and the ability of the plant and its associated microorganisms to intercept, absorb, accumulate, and/or degrade the contaminants. The main aim is to provide an overview of existing field experience in Europe concerning the use of plants and their associated microorganisms whether or not combined with amendments for the revitalization or remediation of contaminated soils and undeep groundwater. Contaminations with trace elements (except radionuclides) and organics will be considered. Because remediation with transgenic organisms is largely untested in the field, this topic is not covered in this review. Brief attention will be paid to the economical aspects, use, and processing of the biomass.

Conclusions and perspectives

It is clear that in spite of a growing public and commercial interest and the success of several pilot studies and field scale applications more fundamental research still is needed to better exploit the metabolic diversity of the plants themselves, but also to better understand the complex interactions between contaminants, soil, plant roots, and microorganisms (bacteria and mycorrhiza) in the rhizosphere. Further, more data are still needed to quantify the underlying economics, as a support for public acceptance and last but not least to convince policy makers and stakeholders (who are not very familiar with such techniques).     

Sorption and oxic degradation of the explosive CL-20 during transport in subsurface sediments

The abiotic sorption and oxic degradation processes that control the fate of the explosive CL-20, Hexanitrohexaazaisowurtzitane, in the subsurface environment were investigated to determine the potential for vadose and groundwater contamination. Sorption of aqueous CL-20 is relatively small (K(d) = 0.02-3.83 cm3 g(-1) for 7 sediments and 12 minerals), which results in only slight retardation relative to water movement. Thus, CL-20 could move quickly through unsaturated and saturated sediments of comparable composition to groundwater, similar to the subsurface behavior of RDX. CL-20 sorption was mainly to mineral surfaces of the sediments, and the resulting isotherm was nonlinear. CL-20 abiotically degrades in oxic environments at slow rates (i.e., 10s to 100s of hours) with a wide variety of minerals, but at fast rates (i.e., minutes) in the presence of 2:1 phyllosilicate clays (hectorite, montmorillonite, nontronite), micas (biotite, illite), and specific oxides (MnO2 and the ferrous-ferric iron oxide magnetite). High concentrations of surface ferrous iron in a dithionite reduced sediment degraded CL-20 the fastest (half-life < 0.05 h), but 2:1 clays containing no structural or adsorbed ferrous iron (hectorite) could also quickly degrade CL-20 (half-life < 0.2 h). CL-20 degradation rates were slower in natural sediments (half-life 3-800 h) compared to minerals. Sediments with slow degradation rates and small sorption would exhibit the highest potential for deep subsurface migration. Products of CL-20 oxic degradation included three high molecular weight compounds and anions (nitrite and formate). The 2-3.5 moles of nitrite produced suggest CL-20 nitro-groups are degraded, and the amount of formate produced (0.2-1.2 moles) suggests the CL-20 cage structure is broken in some sediments. Identification of further degradation products and CL-20 mineralization rates is needed to fully assess the impact of these CL-20 transformation rates on the risk of CL-20 (and degradation product) subsurface movement.     

Bioavailability and degradation of phenanthrene in compost amended soils

Bioavailability in soil of organic xenobiotics such as phenanthrene is limited by mechanisms of diffusion of the xenobiotics within soil micropores and organic matter. The agricultural utilization of compost may reduce the risk connected to organic xenobiotic contamination by means of: (i) a reduction of the bioavailable fraction through an increased adsorption and (ii) an enhanced degradation of the remaining bioavailable fraction through an inoculum of degrading microorganisms. Aim of this work is to test this hypothesis by assessing the effects of compost amendment on the bioavailability and degradation of phenanthrene in soil. Experiments were carried out in both sterilized and non-sterilized conditions, and chemical and microbiological analyses were carried out in order to determine the extent of degradation and bioavailability and to monitor the evolution of the soil micro flora in time. Bioavailability was assessed in sterilized microcosms, in order to assess the physical effects of compost on aging processes without the influence of microbial degradation. Results showed that bioavailability is significantly reduced in soils amended with compost, although no differences were found at the 2 doses of compost studied. In non-sterilized soils the amount of phenanthrene degraded was always higher in the amended soils than in the non-amended one. Microbiological analyses confirmed the presence of a higher number of phenanthrene degraders in the amended soils and in samples of compost alone. These results suggest that compost induces the degradation in soils of easily degradable compounds such as phenanthrene, when the proper bacteria are in the compost; more resistant xenobiotics may instead be trapped by the compost organic matter, thus becoming less available.     

Influence of ozonation on extractability of Pb and Zn from contaminated soils

The effect of soil ozonation on Pb and Zn extraction with EDTA, bioavailability (Ruby's Physiologically Based Extraction Test, PBET) and mobility (Toxicity Characteristic Leaching Procedure, TCLP) of Pb was studied on contaminated soils taken from 7 different locations in the Mezica Valley, Slovenia. EDTA extraction (40 mmol kg(-1)) removed from 27.4+/-1.5% to 64.8+/-1.5% of Pb, and from 1.9+/-0.2% to 22.4+/-2.0% of Zn from tested soils, and significantly reduced soil Pb bioavailability (PBET) and mobility (TCLP). Pretreatment of tested soils with ozone before EDTA extraction enhanced EDTA extractability of Pb for 11.0 to 28.9%, but had no effect on the extractability of Zn. In most of the soils, ozonation had no statistically significant effect on bioavailability and mobility of Pb, residual after EDTA extraction. Using linear regression analysis we found a significant increase (p<0.01) in EDTA extractability of Pb after soil ozonation in soils with a higher initial Pb content. EDTA extractability of Pb after soil ozonation was also significantly higher for soils with a lower Pb extractability when treated with EDTA alone. We found no correlation between soil organic matter content and the percentage of the Pb fraction bound to soil organic matter (where from 25.6+/-1.3% to 73.2+/-0.6% of Pb reside in tested soils) and Pb extractability with EDTA after soil ozonation.     

A pilot study of oral bioavailability of dioxins and furans from contaminated soils: Impact of differential hepatic enzyme activity and species differences

An in vivo pilot study of the oral bioavailability of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in two soils with distinct congener profiles (one dominated by PCDDs, the other by PCDFs) was conducted in rats and juvenile swine. The pilot study revealed potential confounding of relative bioavailability estimates compared to bioavailability in spiked corn oil gavage for tetrachlorodibenzofuran (TCDF) in the rat study due to differential EROD induction between groups receiving soil and those receiving spiked control PCDDs/PCDFs. A follow-up study in rats with the furan-contaminated soil was then conducted with reductions in the spiked control doses to 20%, 50% and 80% of the soil-feed dose in order to bracket hepatic enzyme induction levels in the soil group. When hepatic enzyme induction was matched between the soil and spiked control groups, the apparent relative bioavailability for TCDF was reduced significantly. Overall, after controlling for hepatic enzyme induction, estimates of relative bioavailability in rats and swine differed for the two soils. In the rat study, the relative bioavailability of the two soils were approximately 37% and 60% compared to corn oil administration for the PCDD- and PCDF- dominated soils, respectively, on a TEQ basis. In swine, both soils demonstrated relative bioavailability between 20% and 25% compared to administration in corn oil. These species differences and experimental design issues, such as controlling for differential enzyme induction between corn oil and soil-feed animals in a bioavailability study, are relevant to risk assessment efforts where relative bioavailability inputs are important for theoretical exposure and risk characterization.     

Microbe-induced changes in metal extractability from fly ash

A low cost and eco-friendly technology to bioremediate toxic metals associated with fly ash dumps that contaminate ground and surface water in and around fly ash settling ponds, was investigated. The impact of augmentation of fly ash tolerant bacterial strains, isolated from Typha latifolia growing naturally on fly ash dumps, was studied for metal extractability. It was observed that most of the bacterial strains either induced the bioavailability of Fe, Zn and Ni or immobilized Pb, Cr, Cu, Cd in the fly ash. However, there were few exceptions also. In case of Ni, eight strains enhanced metal mobility, while others caused metal immobilization. The findings also suggest that metal solublization and immobilization are specific to bacterial strains. While induced bioavailability of metals by bacteria may be used to accelerate the phytoextraction of metals from fly ash by hyper accumulator plants, immobilization of metals can check their migration to water reservoirs and reduce the human suffering in affected areas. Thus, bacteria serve the dual purpose and may result in the microbe- assisted phytoremediation of contaminated sites.     

Medium composition affects the degree and pattern of cadmium inhibition of naphthalene biodegradation

Metals have been reported to inhibit organic pollutant biodegradation; however, widely varying degrees and patterns of inhibition have been reported. To investigate the roles of medium composition and metal bioavailability on these different degrees and patterns of inhibition, we assessed the impact of cadmium on naphthalene biodegradation by a newly isolated strain of Comamonas testosteroni in three chemically-defined minimal salts media (MSM): Tris-buffered MSM, PIPES-buffered MSM, and Bushnell-Haas medium. Cadmium (total concentrations of 100 and 500 microM) inhibited biodegradation in each medium. Degrees of inhibition were different in each medium. Cadmium was most inhibitory in PIPES-buffered MSM and least inhibitory in Bushnell-Haas. For example, in Bushnell-Haas medium, 100 microM cadmium reduced the cell yield more than 4-fold compared to controls not containing cadmium. The same concentration of cadmium completely inhibited growth in PIPES-buffered MSM. No difference in inhibition was observed in any medium when cadmium was added 24 h before inoculation rather than when added within one minute of inoculation. Two patterns of inhibition were observed. Inhibition occurred in a dose dependent pattern in Tris- and PIPES-buffered MSM and in a non-dose dependent pattern in Bushnell-Haas. Specifically, in Bushnell-Haas, 100 microM total cadmium extended the lag phase by 23+/-8.66 h, whereas 500 microM did not extend the lag phase. Soluble, ionic cadmium (Cd2+) concentrations were measured and modeled in each medium to assess cadmium bioavailability. In media containing 500 microM total cadmium, bioavailability was highest in Tris- and PIPES-buffered MSM and lowest in Bushnell-Haas. In Bushnell-Haas, cadmium bioavailability was initially higher in the 500 microM treatments (196+/-21.2 microM) than in the 100 microM treatments (78.2+/-2.04 microM); however, after 12 h, bioavailability was higher in the 100 microM treatments (56.4+/-24.8 micro) than the 500 microM treatments (13.3+/-1.2 microM). These data suggest that the type of medium determines the degrees and patterns by which metals inhibit biodegradation and emphasize the importance of coupling metal toxicity and bioavailability data.     

Bioremediation of tributyltin contaminated sediment: degradation enhancement and improvement of bioavailability to promote treatment processes

Bioremediation of tributyltin (TBT) contaminated sediment was studied and degradation enhancement and improvement of bioavailability were also investigated. In TBT spiked sediment, the half-life of TBT in the control sample, representing natural attenuation, was 578 d indicating its persistence. In the stimulated sample (pH 7.5, aeration and incubated at 28 °C), the half-life was significantly reduced to 11 d. Further stimulation by nutrient addition (succinate, glycerol and l-arginine) or inoculation with Enterobacter cloacae (∼10⁷ viable cells g⁻¹ of sediment) resulted in half-life reduction to 9 and 10 d, respectively. In non-spiked sediment, the indigenous microorganisms were able to degrade aged TBT, but the extended period of contamination decreased the degradation efficiency. To improve bioavailability, addition of surfactant, adjustment of salinity and sonication were studied. The highest percentage solubilisation of TBT in water was obtained by adjusting salinity to 20 psu, which increased the solubility of TBT from 13% to 33%. Half-lives after bioavailability was improved were 5, 4 and 4 d for stimulation, stimulation w/nutrient addition and stimulation w/inoculation, respectively. However, natural attenuation in the control sample was not enhanced. The results show that providing suitable conditions is important in enhancing TBT biodegradation, and bioavailability improvement additionally increased the rate and degraded amount of TBT. Unfortunately, nutrient addition and inoculation of the degrader did not enhance the degradation appreciably.     

Eichhornia crassipes capability to remove naphthalene from wastewater in the absence of bacteria

The aim of the current study was to investigate the potential of an aquatic plant, the water hyacinth (Eichhornia crassipes) devoid rhizospheric bacteria, to reduce naphthalene (a polyaromatic hydrocarbon) present in wastewater and wetlands.The capability of sterile water hyacinth plants to remove naphthalene from water and wastewater was studied in batch systems. Water hyacinths enhance the removal of pollutants through their consumption as nutrients and also through microbial activity of their rhizospheric bacteria.Experimental kinetics of naphthalene removal by water hyacinth coupled with natural rhizospheric bacteria was 100% after 9 d. Plants, decoupled of rhizospheric bacteria, reduced naphthalene concentration up to 45% during 7 d. Additionally, naphthalene uptake by water hyacinth revealed a biphasic behavior: a rapid first phase completed after 2.5 h, and a second, considerably slower rate, phase (2.5-225 h). In conclusion, water hyacinth devoid rhizospheric bacteria reduced significantly naphthalene concentration in water, revealing a considerable plant contribution in the biodegradation process of this pollutant.