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Transformation of copper oxychloride fungicide into copper oxalate by tolerant fungi and the effect of nitrogen source on tolerance.

Aspergillus niger and Penicillium chrysogenum were able to grow on Czapek Dox medium amended with elevated concentrations [up to 500 ppm active ingredient (ai)] of the fungicide copper oxychloride. Solubilization of the fungicide in solid medium was evident by the appearance of a clear (halo) zone underneath and around the growing colonies. The halo formed with A. niger, grown on fungicide-containing nitrate nitrogen medium, was found subsequently to enclose concentric rings of newly crystalline precipitate. These crystals were extracted, examined by scanning electron microscopy and IR, and identified as copper oxalate. The supplemented nitrogen source to the medium greatly affected both fungicide solubilization and fungal tolerance. Ratios of fungicide solubilization rate (R(S)) in relation to the colony growth rate (R(G)) were significantly higher on ammonium than on nitrate nitrogen medium for both fungal strains. Growth ratios (the colony extension rate in the presence of a given concentration of the fungicide in relation to the control colony growth rate) of A. niger were markedly lower on ammonium than on nitrate nitrogen medium. The cellular copper contents, taken up from the fungicide, and the medium titratable acidity were higher in ammonium than in nitrate medium for both fungi. These results suggested fungal possession of variable tolerance mechanisms to this fungicide by complexation and/or precipitation of copper in the medium. Additionally, this work emphasizes the activity of fungi in transformation of insoluble inorganic metal-containing fungicides into insoluble organic metal compounds, which has a potentiality in metal cycling in biogeochemical and environmental context.

Biodegradation, Environmental↗

[Hydrophobicity and microbial production. 1. Selection of oleophilic microorganisms and qualitative characterization by several ecologically and technologically relevant activities].

From samples of different origin microorganisms were selected using the property of oleophily. All oleophilic isolates (30 strains are characterized in more detail) proved to be Gram-positive, nonsporogenic bacteria, obviously belonging to the group of coryneforms. They are growing on a broad spectrum of substrates, 11 strains are facultatively methylotrophic. They are utilizing methanol as well as n-alkanes and glucose without any supplines. 6 strains are able to degrade and decolorize triphenylmethane dyes, for example crystal violet and malachite green. Because of the degradative capacities, combined with the hydrophobic properties of the cell surface, the group of the oleophilic bacteria seems to be very promising in relation to the development of environmental technologies and ecological investigations.

Actinomycetales↗

Factors influencing microbe insecticide interactions.

Microorganisms are present in almost all environmental components where insecticides are used and they will, therefore, encounter with the latter. In simplest terms, such interactions can be considered as (1) the action of microbes on insecticides and (2) the action of insecticides on microbes. These interactions can not be as clearly understood without understanding the influence of environmental factors that control the fate and activity of microorganisms. The proposed review will focus on two areas. The first is related to the test methods used to study the microbial insecticide interaction, the second area is concerned with the influence of environmental factors on the action of microbes and vice versa.

Bacteria↗

Metabolic engineering of bacteria for environmental applications: construction of Pseudomonas strains for biodegradation of 2-chlorotoluene.

In this article, we illustrate the challenges and bottlenecks in the metabolic engineering of bacteria destined for environmental bioremediation, by reporting current efforts to construct Pseudomonas strains genetically designed for degradation of the recalcitrant compound 2-chlorotoluene. The assembled pathway includes one catabolic segment encoding the toluene dioxygenase of the TOD system of Pseudomonas putida F1 (todC1C2BA), which affords the bioconversion of 2-chlorotoluene into 2-chlorobenzaldehyde by virtue of its residual methyl-monooxygenase activity on o-substituted substrates. A second catabolic segment encoded the entire upper TOL pathway from pWW0 plasmid of P. putida mt-2. The enzymes, benzyl alcohol dehydrogenase (encoded by xylB) and benzaldehyde dehydrogenase (xylC) of this segment accept o-chloro-substituted substrates all the way down to 2-chlorobenzoate. These TOL and TOD segments were assembled in separate mini-Tn5 transposon vectors, such that expression of the encoded genes was dependent on the toluene-responsive Pu promoter of the TOL plasmid and the cognate XylR regulator. Such gene cassettes (mini-Tn5 [UPP2] and mini-Tn5 [TOD2]) were inserted in the chromosome of the 2-chlorobenzoate degraders Pseudomonas aeruginosa PA142 and P. aeruginosa JB2. GC-MS analysis of the metabolic intermediates present in the culture media of the resulting strains verified that these possessed, not only the genetic information, but also the functional ability to mineralise 2-chlorotoluene. However, although these strains did convert the substrate into 2-chlorobenzoate, they failed to grow on 2-chlorotoluene as the only carbon source. These results pinpoint the rate of the metabolic fluxes, the non-productive spill of side-metabolites and the physiological control of degradative pathways as the real bottlenecks for degradation of certain pollutants, rather than the theoretical enzymatic and genetic fitness of the recombinant bacteria to the process. Choices to address this general problem are discussed.

Alcohol Oxidoreductases↗

Disposable sensor for measuring the biochemical oxygen demand for nitrification and inhibition of nitrification in wastewater.

A disposable-type microbial sensor was developed for the determination of both the biochemical oxygen demand for nitrification (N-BOD) and inhibiting effects on nitrifying bacteria. The sensor was based on the respiratory activity of nitrifying bacteria immobilized on a miniature oxygen electrode. Typical response times for measuring N-BOD of ammonium standard solutions as well as of wastewater samples were in the range of 6-12 min. A dynamic evaluation of the signals after a measuring time of 120 s also resulted in good reproducibility and sensitivity. A daily profile of a municipal sewage plant was recorded, comparing the biosensor data with two standard methods. For the measurement of nitrification-inhibiting effects a 120-s dynamic signal evaluation was preferred to a steady-state method because of the long recovery times resulting from extended exposure to inhibitors. However, steady-state measurement techniques allowed allylthiourea detection with a ten times higher sensitivity. Because of the advantages of this miniaturized electrode, e.g. short response time, simple measuring procedure and low costs of production, this sensor system is considered to be suitable for commercial application in environmental analysis.

Bacteria↗

Uptake and degradation of DDT by hairy root cultures of Cichorium intybus and Brassica juncea.

Hairy root cultures of Cichorium intybus and Brassica juncea were used for their ability to uptake and degrade DDT (1,1,1-trichloro-2,2-bis-(4'-chlorophenyl)ethane). After 24 h of 14C DDT treatment, only 12-13% of the total applied radioactivity was detected in the culture media, indicating the efficient uptake of DDT by the hairy roots. The majority of the applied radioactivity was associated with the roots. DDT was degraded to various other products such as DDD, DDE and DDMU, along with some unknown compounds by hairy root cultures, which were detected by thin layer chromatography (TLC) and autoradiography. The rate of in situ degradation was found to be higher during the initial stages of culture and the residual 14C DDT in the roots was found to decrease from 77% to 61% over a period of 10-days. There was no spontaneous degradation of 14C DDT in media lacking hairy root cultures or in media with autoclaved hairy roots. This suggests that endogenous root enzymes play a role in the breakdown of 14C DDT. These results suggest the potential applicability and advantage of using these plant species for phytoremediation of persistent xenobiotics such as DDT in an eco-friendly and efficient manner for environmental clean up.

Biodegradation, Environmental↗

Urban environmental geochemistry of trace metals.

As the world's urban population continues to grow, it becomes increasingly imperative to understand the dynamic interactions between human activities and the urban environment. The development of urban environmental geochemistry has yielded a significant volume of scientific information about geochemical phenomena found uniquely in the urban environment, such as the distribution, dispersion, and geochemical characteristics of some toxic and potentially toxic trace metals. The aim of this paper is to provide an overview of the development of urban environmental geochemistry as a field of scientific study and highlight major transitions during the course of its development from its establishment to the major scientific interests in the field today. An extensive literature review is also conducted of trace metal contamination of the urban terrestrial environment, in particular of urban soils, in which the uniqueness of the urban environment and its influences on trace metal contamination are elaborated. Potential areas of future development in urban environmental geochemistry are identified and discussed.

Biodegradation, Environmental↗

Vortex-assisted liquid-liquid microextraction based on natural deep eutectic solvents for the determination of pyrethroid pesticides in urine.

A novel, facile, and environmentally friendly analytical method was developed based on vortex-assisted liquid-liquid microextraction and high-performance liquid chromatography with diode-array detection for detecting pyrethroid pesticides (PPs) in urine. Natural deep eutectic solvents (NADESs) were prepared using plant essential oil-derived monoterpenoids (thymol, carvacrol, and menthol) combined with aromatic primary alcohols (benzyl alcohol, phenethyl alcohol, and phenylpropyl alcohol) as hydrogen bond donors and acceptors. These solvents served as environmentally benign extraction media, thereby avoiding the use of conventional volatile, toxic organic solvents. NADESs are naturally derived, easy to prepare, biodegradable, and environmentally friendly solvents. Hydrophobic and π-π interactions between the NADESs and PPs may contribute to enhancing the affinity of PPs toward the NADESs phase. Vortex technology, accelerating mass transfer between the sample and extractant phases, enables fast extraction of PPs. Under optimized conditions, the method achieved a low detection limit (0.002 mg L-1), satisfactory precision with relative standard deviations (0.3%-2.4%), and acceptable recovery (80.7%-86.2%). The method demonstrated excellent performance in urine analysis and was feasible as a facile and green strategy for monitoring the content of PPs in biological matrices and assessing exposure risk.

Liquid Phase Microextraction↗

Degradation of recalcitrant compounds from stabilized landfill leachate using a combination of ozone-GAC adsorption treatment.

Laboratory experiments were undertaken to investigate the treatment performances of ozonation alone and/or its combination with granular activated carbon (GAC) adsorption for raw leachate from the NENT landfill (in Hong Kong). To improve its removal of recalcitrant contaminants from the leachate, the surface of GAC was oxidized with ozone prior to treatment. With respect to ozone dose and pH, the removal of COD and/or NH(3)-N from ozonation alone and combined ozone-GAC adsorption were evaluated and compared to those of other physico-chemical treatments in some reported studies. The removal mechanism of recalcitrant compounds by ozone-GAC adsorption treatment was presented. Among the various treatments studied, the combination of ozone-GAC adsorption using ozone-modified GAC had the highest removal for COD (86%) and/or NH(3)-N (92%) compared to ozonation alone (COD: 35%; NH(3)-N: 50%) at the same initial COD and/or NH(3)-N concentrations of 8000 and 2620 mg/L, respectively. Although the integrated treatment was more effective than ozonation alone for treating stabilized leachate, the results suggested that it could not generate treated effluent that complied with the COD limit of lower than 200 mg/L and the NH(3)-N discharge standard of less than 5 mg/L. Therefore, further biological treatments to complement the degradation of the leachate are still required to meet the environmental legislation.

Adsorption↗

Natural attenuation processes for remediation of arsenic contaminated soils and groundwater.

Arsenic (As) contamination presents a hazard in many countries. Natural attenuation (NA) of As-contaminated soils and groundwater may be a cost-effective in situ remedial option. It relies on the site intrinsic assimilative capacity and allows in-place cleanup. Sorption to solid phases is the principal mechanism immobilizing As in soils and removing it from groundwater. Hydroxides of iron, aluminum and manganese, clay and sulfide minerals, and natural organic matter are commonly associated with soils and aquifer sediments, and have been shown to be significant As adsorbents. The extent of sorption is influenced by As speciation and the site geochemical conditions such as pH, redox potential, and the co-occurring ions. Microbial activity may catalyze the transformation of As species, or mediate redox reactions thus influencing As mobility. Plants that are capable of hyperaccumulating As may translocate As from contaminated soils and groundwater to their tissues, providing the basis for phytoremediation. However, NA is subject to hydrological changes and may take substantial periods of time, thus requiring long-term monitoring. The current understanding of As NA processes remains limited. Sufficient site characterization is critical to the success of NA. Further research is required to develop conceptual and mathematical models to predict the fate and transport of As and to evaluate the site NA capacity. Engineering enhanced NA using environmentally benign products may be an effective alternative.

Arsenic↗

Alkylphenol ethoxylate degradation products in land-applied sewage sludge (biosolids).

Alkylphenol ethoxylates, widely used in commercial and household detergents in the United States, can degrade during the wastewater treatment process to more toxic, estrogenic, and lipophilic compounds. These include octylphenol (OP), nonylphenols (NPs), nonylphenol monoethoxylates (NP1EOs), and nonylphenol diethoxylates (NP2EOs). These compounds have received considerable attention due to their acute toxicity and ability to disrupt the endocrine system. In Europe, regulations have been established to control their impact on the environment. In this study, biosolids derived from all 11 U.S. wastewater treatment plants examined contained detectable levels of OP, NPs, NP1EOs, and NP2EOs. Nine exceeded the current Danish land application limit (30 mg/kg; sum of NPs, NP1EOs, and NP2EOs) by 6-33x. NPs were the major component, and their concentrations therein ranged from 5.4 to 887 mg/kg (dry weight). OP, reportedly 10-20x more estrogenic than NP, was detected in these same nine biosolids at levels up to 12.6 mg/kg. Three biosolids were also subjected to the U.S. Environmental Protection Agency Toxicity Characteristic Leaching Procedure Method 1311. NPs and NP1EOs were both detected in the leachate; the former at concentrations from 9.4 to 309 microg/L. On the basis of effect levels published in the literature, alkylphenol ethoxylate degradates in U.S. biosolids may cause adverse environmental impacts.

Biodegradation, Environmental↗

Genetic engineering of bacteria from managed and natural habitats.

The genetic modification of bacteria from natural and managed habitats will impact on the management of agricultural and environmental settings. Potential applications include crop production and protection, degradation or sequestration of environmental pollutants, extraction of metals from ores, industrial fermentations, and productions of enzymes, diagnostics, and chemicals. Applications of this technology will ultimately include the release of beneficial agents in the environment. If safely deployed, genetically modified bacteria should be able to provide significant benefits in the management of environmental systems and in the development of new environmental control processes.

Agriculture↗

Quantification of chemotaxis to naphthalene by Pseudomonas putida G7.

The capillary assay was used to quantify the chemotactic response of Pseudomonas putida G7 to naphthalene. Experiments were conducted in which the cell concentration in the assay chamber, the naphthalene concentration in the capillary, or the incubation time was varied. Data from these experiments were evaluated with a model that accounted for the effect of diffusion on the distribution of substrate and the transport of cells from the chamber through the capillary orifice. By fitting a numerical solution of this model to the data, it was possible to determine the chemotactic sensitivity coefficient, chi0. The mean of the best-fit values for chi0 from the three types of experiments was 7.2 x 10(-5) cm2/s. A less computationally intensive model based on earlier approaches that ignore cell transport in the chamber resulted in chi0 values that were approximately three times higher. The models evaluated in the present study could simulate the results of capillary assays only at low chamber cell concentrations, for which the effect of consumption on the distribution of substrate was negligible. Results from this work suggest that it is possible to use the capillary assay to quantify taxis towards environmentally relevant chemoeffectors that have low aqueous solubility.

Biodegradation, Environmental↗

Stable hydrogen and carbon isotope fractionation during microbial toluene degradation: mechanistic and environmental aspects.

Primary features of hydrogen and carbon isotope fractionation during toluene degradation were studied to evaluate if analysis of isotope signatures can be used as a tool to monitor biodegradation in contaminated aquifers. D/H hydrogen isotope fractionation during microbial degradation of toluene was measured by gas chromatography. Per-deuterated toluene-d(8) and nonlabeled toluene were supplied in equal amounts as growth substrates, and kinetic isotope fractionation was calculated from the shift of the molar ratios of toluene-d(8) and nondeuterated toluene. The D/H isotope fractionation varied slightly for sulfate-reducing strain TRM1 (slope of curve [b] = -1.219), Desulfobacterium cetonicum (b = -1.196), Thauera aromatica (b = -0.816), and Geobacter metallireducens (b = -1.004) and was greater for the aerobic bacterium Pseudomonas putida mt-2 (b = -2.667). The D/H isotope fractionation was 3 orders of magnitude greater than the (13)C/(12)C carbon isotope fractionation reported previously. Hydrogen isotope fractionation with nonlabeled toluene was 1.7 and 6 times less than isotope fractionation with per-deuterated toluene-d(8) and nonlabeled toluene for sulfate-reducing strain TRM1 (b = -0.728) and D. cetonicum (b = -0.198), respectively. Carbon and hydrogen isotope fractionation during toluene degradation by D. cetonicum remained constant over a growth temperature range of 15 to 37 degrees C but varied slightly during degradation by P. putida mt-2, which showed maximum hydrogen isotope fractionation at 20 degrees C (b = -4.086) and minimum fractionation at 35 degrees C (b = -2.138). D/H isotope fractionation was observed only if the deuterium label was located at the methyl group of the toluene molecule which is the site of the initial enzymatic attack on the substrate by the bacterial strains investigated in this study. Use of ring-labeled toluene-d(5) in combination with nondeuterated toluene did not lead to significant D/H isotope fractionation. The activity of the first enzyme in the anaerobic toluene degradation pathway, benzylsuccinate synthase, was measured in cell extracts of D. cetonicum with an initial activity of 3.63 mU (mg of protein)(-1). The D/H isotope fractionation (b = -1.580) was 30% greater than that in growth experiments with D. cetonicum. Mass spectroscopic analysis of the product benzylsuccinate showed that H atoms abstracted from the toluene molecules by the enzyme were retained in the same molecules after the product was released. Our findings revealed that the use of deuterium-labeled toluene was appropriate for studying basic features of D/H isotope fractionation. Similar D/H fractionation factors for toluene degradation by anaerobic bacteria, the lack of significant temperature dependence, and the strong fractionation suggest that analysis of D/H fractionation can be used as a sensitive tool to assess degradation activities. Identification of the first enzyme reaction in the pathway as the major fractionating step provides a basis for linking observed isotope fractionation to biochemical reactions.

Anaerobiosis↗