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An in vitro study of manure composition on the biochemical origins, composition, and accumulation of odorous compounds in cattle feedlots.

Very little is known about the biochemical origin of cattle feedlot odors and the environmental factors controlling their production. The tie between diet and manure composition is well established, but the effect of different manure compositions on odorous chemical production is unknown. This study describes the effect of starch, casein, and cellulose substrate additions to slurries of fresh (< 24 h) and aged cattle manure (> 1 d) on the anaerobic production of fermentation products and the consumption of substrates relative to no addition treatments. Aged cattle manure accumulated more VFA (245 to 290 mM) than the fresh manure (91 to 181 mM) irrespective of substrate additions (P < 0.001). In fresh manures, VFA concentrations were increased (P < 0.01) over no addition treatments when carbohydrate (starch or cellulose) was added, whereas starch and protein treatments to aged manure increased VFA content relative to no addition treatments (P < 0.001). Branched-chain VFA and aromatic compounds accumulated only in the aged manure (no addition and protein treatments), indicating that some protein fermentation occurred in those treatments. Based upon substrate loss, starch fermentation was the dominant process in both manures and all treatments with losses exceeding 18.6 g/L. Protein fermentation occurred only in the aged manure, specifically the no addition and protein treatments, when starch was no longer available. The production of odorous compounds from manure was controlled by substrate availability and pH, with pH related to lactate accumulation. We believe that calcareous soil and lactate-consuming microorganisms in the aged manure slurries minimized slurry acidification and resulted in greater accumulations of odorous products. Substrate additions had little effect on the overall accumulation of odor compounds in manure but had profound effects on odor compound composition. We propose that modifying cattle diets to limit starch and protein excretion would profoundly affect the production and accumulation of odor compounds in feedlots.

Animals↗

Degradation of organic pollutants by methane grown microbial consortia.

Microbial consortia were enriched from various environmental samples with methane as the sole carbon and energy source. Selected consortia that showed a capacity for co-oxidation of naphthalene were screened for their ability to degrade methyl-tert-butyl-ether (MTBE), phthalic acid esters (PAE), benzene, xylene and toluene (BTX). MTBE was not removed within 24 h by any of the consortia examined. One consortium enriched from activated sludge ("AAE-A2"), degraded PAE, including (butyl-benzyl)phthalate (BBP), and di-(butyl)phthalate (DBP). PAE have not previously been described as substrates for methanotrophic consortia. The apparent Km and Vmax for DBP degradation by AAE-A2 at 20 degrees C was 3.1 +/- 1.2 mg l(-1) and 8.7 +/- 1.1 mg DBP (g protein x h)(-1), respectively. AAE-A2 also showed fast degradation of BTX (230 +/- 30 nmol benzene (mg protein x h)(-1) at 20 degrees C). Additionally, AAE-A2 degraded benzene continuously for 2 weeks. In contrast, a pure culture of the methanotroph Methylosinus trichosporium OB3b ceased benzene degradation after only 2 days. Experiments with methane mono-oxygenase inhibitors or competitive substrates suggested that BTX degradation was carried out by methane-oxidizing bacteria in the consortium, whereas the degradation of PAE was carried out by non-methanotrophic bacteria co-existing with methanotrophs. The composition of the consortium (AAE-A2) based on polar lipid fatty acid (PLFA) profiles showed dominance of type II methanotrophs (83-92% of biomass). Phylogeny based on a 16S-rRNA gene clone library revealed that the dominating methanotrophs belonged to Methylosinus/Methylocystis spp. and that members of at least 4 different non-methanotrophic genera were present (Pseudomonas, Flavobacterium, Janthinobacterium and Rubivivax).

Bacteria↗

Solid waste treatment as a high-priority and low-cost alternative for greenhouse gas mitigation.

The increased concern about environmental problems caused by inadequate waste management, as well as the concern about global warming, promotes actions toward a sustainable management of the organic fraction of the waste. Landfills, the most common means to dispose of municipal solid waste (MSW), lead to the conversion of the organic waste to biogas, containing about 50% methane, a very active greenhouse gas (GHG). One unit of methane has a global warming potential of 21 computed for a 100-year horizon or 56 computed for 20 years. The waste sector in Israel contributes 13% of total greenhouse gases (GHG) emissions for a time horizon of 100 years (for a time horizon of 20 years, the waste sector contribution equals to more than 25% of total GHG emissions). The ultimate goal is to minimize the amount of methane (CH4) by converting it to CO2. This can be achieved by physicochemical means (e.g., landfill gas flare, incineration) or by biological processes (e.g., composting, anaerobic digestion). Since the waste in Israel has a high organic material content, it was found that the most cost-effective means to treat the degradable organic components is by aerobic composting (investment of less than US$ 10 to reduce emission of one ton CO2 equivalent per year). Another benefit of this technology is the ability to implement it within a short period. The suggested approach, which should be implemented especially in developing countries, could reduce a significant amount of GHG at relatively low cost and short time. The development of a national policy for proper waste treatment can be a significant means to abate GHG emissions in the short term, enabling a gain in time to develop other means for the long run. In addition, the use of CO2 quotas will credit the waste sector and will promote profitable proper waste management.

Biodegradation, Environmental↗

Population dynamics of an introduced bacterium degrading chlorinated benzenes in a soil column and in sewage sludge.

The capacity of the beta-Proteobacterium Pseudomonas sp. strain P51, which degrades chlorinated benzenes, to metabolize 1,2,4-trichlorobenzene (TCB) under environmental conditions was tested by its release into two experimental systems. The first system consisted of laboratory scale microcosms which were operated with and without the addition of TCB and which were inoculated with sludge from a wastewater treatment plant. The second system consisted of a non sterile, water saturated soil column. We determined survival of strain P51 after its introduction and its ability to degrade TCB. The population dynamics was followed by selective plating and applying the polymerase chain reaction (PCR) to detect strain P51 and the chlorobenzene (tcb) genes on catabolic plasmid pP51. The results showed a completely different behaviour of strain P51 in the two habitats under the applied conditions. In the soil column the P51 bacteria inoculated the entire area and their population reached 2 x 10(6) cells/g soil. The population remained active since TCB was degraded to concentrations below the detection limit of 30 micrograms/l. In the sludge microcosms, the number of strain P51 cells immediately decreased from 4 x 10(7) cells/ml to 10(5) cells/ml over a period of 2 days after inoculation, and then the strain disappeared to levels below our detection limit (10(3)-10(4) cells/ml). In the reactor without TCB the population of P51 maintained a stable value of 10(5) cells/ml during 8 days but then also decreased to levels below the detection limit. In addition, no significant TCB degradation was found in the sludge reactors. The influence of presence of TCB on maintenance of strain P51 in the two habitats is discussed. This work demonstrates the possibility to successfully apply preselected strains to degrade otherwise poorly degradable substances in complex mixed microbial communities. However, survival and activity may depend strongly on the type of system into which the strain is introduced.

Base Sequence↗

In vitro water activity and pH dependence of mycelial growth and extracellular enzyme activities of Trichoderma strains with biocontrol potential.

AIMS: Water activity (aw) and pH are probably the most important environmental parameters affecting the activities of mycoparasitic Trichoderma strains. Therefore it is important to collect information on the effects of these factors on mycelial growth and on the in vitro activities of extracellular enzymes involved in nutrient competition (e.g. beta-glucosidase, cellobiohydrolase and beta-xylosidase) and mycoparasitism (e.g. N-acetyl-beta-glucosaminidase, trypsin-like protease and chymotrypsin-like protease) of Trichoderma strains with biocontrol potential. METHODS AND RESULTS: Water activity and pH dependence of the linear mycelial growth of five examined Trichoderma strains belonging to three different species groups was examined on yeast extract and soil extract media. Maximal growth rates were observed at aw 0.997 and pH 4.0 in the case of all strains. The activities of the examined extracellular enzymes at different aw and pH values were determined spectrophotometrically after incubation with chromogenic p-nitrophenyl and p-nitroaniline substrates. Maximal enzyme activities were measured at aw 0.950 for beta-glucosidase, trypsin-like protease and chymotrypsin-like protease, at 0.910 for cellobiohydrolase and at 0.993 for beta-xylosidase and N-acetyl-beta-glucosaminidase enzymes. Optimal pH values are suggested to be at 5.0 for beta-glucosidase, cellobiohydrolase and N-acetyl-beta-glucosaminidase, at 3.0 for beta-xylosidase, at 6.0 for trypsin-like protease and between 6.0 and 7.0 for chymotrypsin-like protease activities, respectively. CONCLUSIONS: Extracellular enzymes of the examined mycoparasitic Trichoderma strains are able to display activities under a wider range of aw and pH values than those allowing mycelial growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Data about the effects of aw and pH on mycelial growth and extracellular enzyme activities of Trichoderma reveal useful information about the applicability of biocontrol strains in agricultural soils with specific water and pH relations.

Biodegradation, Environmental↗

Degradation of benzo[a]pyrene by Mycobacterium vanbaalenii PYR-1.

Metabolism of the environmental pollutant benzo[a]pyrene in the bacterium Mycobacterium vanbaalenii PYR-1 was examined. This organism initially oxidized benzo[a]pyrene with dioxygenases and monooxygenases at C-4,5, C-9,10, and C-11,12. The metabolites were separated by reversed-phase high-performance liquid chromatography (HPLC) and characterized by UV-visible, mass, nuclear magnetic resonance, and circular dichroism spectral analyses. The major intermediates of benzo[a]pyrene metabolism that had accumulated in the culture media after 96 h of incubation were cis-4,5-dihydro-4,5-dihydroxybenzo[a]pyrene (benzo[a]pyrene cis-4,5-dihydrodiol), cis-11,12-dihydro-11,12-dihydroxybenzo[a]pyrene (benzo[a]pyrene cis-11,12-dihydrodiol), trans-11,12-dihydro-11,12-dihydroxybenzo[a]pyrene (benzo[a]pyrene trans-11,12-dihydrodiol), 10-oxabenzo[def]chrysen-9-one, and hydroxymethoxy and dimethoxy derivatives of benzo[a]pyrene. The ortho-ring fission products 4-formylchrysene-5-carboxylic acid and 4,5-chrysene-dicarboxylic acid and a monocarboxylated chrysene product were formed when replacement culture experiments were conducted with benzo[a]pyrene cis-4,5-dihydrodiol. Chiral stationary-phase HPLC analysis of the dihydrodiols indicated that benzo[a]pyrene cis-4,5-dihydrodiol had 30% 4S,5R and 70% 4R,5S absolute stereochemistry. Benzo[a]pyrene cis-11,12-dihydrodiol adopted an 11S,12R conformation with 100% optical purity. The enantiomeric composition of benzo[a]pyrene trans-11,12-dihydrodiol was an equal mixture of 11S,12S and 11R,12R molecules. The results of this study, in conjunction with those of previously reported studies, extend the pathways proposed for the bacterial metabolism of benzo[a]pyrene. Our study also provides evidence of the stereo- and regioselectivity of the oxygenases that catalyze the metabolism of benzo[a]pyrene in M. vanbaalenii PYR-1.

Benzopyrenes↗

Oil spill in the Rio de la Plata estuary, Argentina: 2-hydrocarbon disappearance rates in sediments and soils.

The 6-month assessment of the oil spill impact in the Rio de la Plata described in the preceding paper [Colombo, J.C., Barreda, A., Bilos, C., Cappelletti, N., Demichelis, S., Lombardi, P., Migoya, M.C., Skorupka, C., Suarez, G., 2004. Oil spill in the Rio de la Plata estuary, Argentina: 1 - biogeochemical assessment of waters, sediments, soils and biota. Environmental Pollution] was followed by a 13- and 42-month campaigns to evaluate the progress of hydrocarbon decay. Average sediment hydrocarbon concentrations in each sampling include high variability (85-260%) due to contrasting site conditions, but reflect a significant overall decrease after 3 years of the spill: 17 +/- 27, 18 +/- 39 to 0.54 +/- 1.4 microg g(-1) for aliphatics; 0.44 +/- 0.49, 0.99 +/- 1.6 to 0.04 +/- 0.03 microg g(-1) for aromatics at 6, 13 and 42 months, respectively. Average soil hydrocarbon levels are 100-1000 times higher and less variable (61-169%) than sediment values, but display a clear attenuation: 3678 +/- 2369, 1880 +/- 1141 to 6.0 +/- 10 microg g(-1) for aliphatics and 38 +/- 26, 49 +/- 32 to 0.06 +/- 0.04 microg g(-1) for aromatics. Hydrocarbon concentrations modeled to first-order rate equations yield average rate constants of total loss (biotic+abiotic) twice as higher in soils (k = 0.18-0.19 month(-1)) relative to sediments (0.08-0.10 month(-1)). Individual aliphatic rate constants decrease with increasing molecular weight from 0.21 +/- 0.07 month(-1) for isoprenoids and n-C27, similar to hopanes (0.10 +/- 0.05 month(-1)). Aromatics disappearance rates were more homogeneous with higher values for methylated relative to unsubstituted species (0.17 +/- 0.05 vs. 0.12 +/- 0.05 months(-1)). Continued hydrocarbon inputs, either from biogenic (algal n-C15,17; vascular plant n-C27,29) or combustion related sources (fluoranthene and pyrene), appear to contribute to reduced disappearance rate. According to the different loss rates, hydrocarbons showed clear compositional changes from 6-13 to 42 months. Aliphatics disappearance rates and compositional changes support an essentially microbiologically-mediated recovery of coastal sediments to pre-spill conditions in a 3-4 year period. The lower rates and more subtle compositional changes deduced for aromatic components, suggest a stronger incidence of physical removal processes.

Argentina↗

Differences in Michaelis-Menten kinetics for different cultivars of maize during cyanide removal.

Knowledge of the kinetic parameters, the half-saturation constant (K(m)) and the maximum metabolic capacity (v(max)), is very useful for the characterization of enzymes and biochemical processes. Little is known about rates of which vegetation metabolizes environmental chemicals. It is known, however, that vascular plants possess an enzyme system that detoxifies cyanide by converting it into the amino acid asparagine. This study investigated the differences in Michaelis-Menten kinetics of cyanide removal by different cultivars of maize. Detached leaves (1.0 g fresh weight) of seven different cultivars of maize (Zea mays L.) were kept in glass vessels with 100mL of aqueous solution spiked with potassium cyanide at 25+/-0.5 degrees C for 28 h. Four treatment concentrations of cyanide were used, ranging from 0.43 to 7.67 mgCNL(-1). The disappearance of cyanide from the aqueous solution was analyzed spectrophotometrically. Realistic values of K(m) and v(max) were estimated by a computer program using non-linear regression treatment. Lineweaver-Burk plots were also used to estimate the kinetic parameters for comparison. Using non-linear regression treatments, values of v(max) and K(m) were found to be between 10.80 and 22.80 mgCNkg(-1)h(-1), and 2.57 and 7.09 mgCNL(-1), respectively. The highest v(max) was achieved by the cultivars HengFen 1, followed by NongDa 108. The lowest v(max) was demonstrated by JingKe 8. The highest K(m) was found in NongDa 108, followed by HengFen 1. The lowest K(m) was associated with JingKe 8. Results from this study indicated that significant removal of cyanide from an aqueous solution was observed in the presence of plant materials without apparent phytotoxicity, even at the high concentration of cyanide used in this study. All maize cultivars used in this study were able to metabolize cyanide efficiently, although with different metabolic capacities. Results also showed a small variation of metabolic rates between the different cultivars. This leads to the conclusion that plants can be used as a phytoremediation agent in the detoxification of cyanide, presenting a feasible option for cleaning up soils and water contaminated with cyanide.

Biodegradation, Environmental↗

Lead accumulation by tall fescue (Festuca arundinacea Schreb.) grown on a lead-contaminated soil.

Phytoextraction is gaining acceptance as a cost-effective and environmentally friendly phytoremediation strategy for reducing toxic metal levels from contaminated soils. Cognizant of the potential of this phytoremediation technique as an alternative to expensive engineering-based remediation technologies, experiments were conducted to evaluate the suitability of some plants as phytoextraction species. From one of our preliminary studies, we found that tall fescue (Festuca arundinacea Schreb. cv. Spirit) can tolerate and accumulate significant amounts of lead (Pb) in its shoots when grown in Pb-amended sand. To further evaluate the suitability of tall fescue as one of the potential crop rotation species for phytoextraction, a study was conducted to determine whether the addition of ethylenediaminetetraacetic acid (EDTA) alone or in combination with acetic acid can further enhance the shoot uptake of Pb. Seeds were planted in 3.8 L plastic pots containing top soil, peat, and sand (4:2:1, v:v:v) spiked with various levels (0,1000, 2000 mg Pb/kg dry soil) of lead. At six weeks after planting, aqueous solutions (0, 5 mmol/kg dry soil) of EDTA and acetic acid (5 mmol/kg dry soil) were applied to the root zone, and all plants were harvested a week later. Results revealed that tall fescue was relatively tolerant to moderate levels of Pb as shown by non-significant differences in root and shoot biomass among treatments. An exception to this trend however, was the slight reduction in root and shoot biomass of plants exposed to the highest Pb level in combination with the two chelates. Root Pb concentration increased with increasing level of soil-applied Pb. Further increases in root Pb concentrations were attributed to chelate amendments. Translocation index, which is a measure of the partitioning of the metal to the shoots, was significantly enhanced with chelate addition especially when both EDTA and acetic acid were used. Chelate-induced increases in translocation indices correspondingly led to higher shoot Pb concentrations.

Biodegradation, Environmental↗

Haloalkane hydrolysis by Rhodococcus erythropolis cells: comparison of conventional aqueous phase dehalogenation and nonconventional gas phase dehalogenation.

Biofiltration of air polluted by volatile organic compounds is now recognized by the industrial and research communities as an effective and viable alternative to standard environmental technologies. Whereas many studies have focused on solid/liquid/gas biofilters, there have been fewer reports on waste air treatment using other biological processes, especially in a solid/gas biofilter. In this study, a comparison was made of the hydrolysis of halogenated compounds (such as 1-chlorobutane) by lyophilized Rhodococcus erythropolis cells in a novel solid/gas biofilter and in the aqueous phase. We first determined the culture conditions for the production of R. erythropolis cells with a strong dehalogenase activity. Four different media were studied and the amount of 1-chlorobutane was optimized. Next, we report the possibility to use R. erythropolis cells in a solid/gas biofilter in order to transform halogenated compounds in corresponding alcohols. The effect of experimental parameters (total flow into the biofilter, thermodynamic activity of the substrates, temperature, carbon chain length of halogenated substrates) on the activity and stability of lyophilized cells in the gas phase was determined. A critical water thermodynamic activity (a(w)) of 0.4 is necessary for the enzyme to become active and optimal dehalogenase activity for the lyophilized cells is obtained for an a(w) of 0.9. A temperature of reaction of 40 degrees C represents the best compromise between stability and activity. Activation energy of the reaction was determined and found equal to 59.5 KJ/mol. The pH effect on the dehalogenase activity of R. erythropolis cells was also studied in the gas phase and in the aqueous phase. It was observed that pH 9.0 provided the best activity in both systems. We observed that in the aqueous phase R. erythropolis cells were less sensitive to the variation in pH than R. erythropolis cells in the gas phase. Finally, the addition of volatile Lewis base (triethylamine) in the gaseous phase and the action of the lysozyme in order to permeabilize the cells was found to be highly beneficial to the effectiveness of the biofilter.

Air Pollutants↗

Environmental biochemistry of arsenic.

Microorganisms are involved in the redistribution and global cycling of arsenic. Arsenic can accumulate and can be subject to various biotransformations including reduction, oxidation, and methylation. Bacterial methylation of inorganic arsenic is coupled to the methane biosynthetic pathway in methanogenic bacteria under anaerobic conditions and may be a mechanism for arsenic detoxification. The pathway proceeds by reduction of arsenate to arsenite followed by methylation to dimethylarsine. Fungi are also able to transform inorganic and organic arsenic compounds into volatile methylarsines. The pathway proceeds aerobically by arsenate reduction to arsenite followed by several methylation steps producing trimethylarsine. Volatile arsine gases are very toxic to mammals because they destroy red blood cells (LD50 in rats; 3.0 mg kg-1). Further studies are needed on dimethylarsine and trimethylarsine toxicity tests through inhalation of target animals. Marine algae transform arsenate into non-volatile methylated arsenic compounds (methanearsonic and dimethylarsinic acids) in seawater. This is considered to be a beneficial step not only to the primary producers, but also to the higher trophic levels, since non-volatile methylated arsenic is much less toxic to marine invertebrates. Freshwater algae like marine algae synthesize lipid-soluble arsenic compounds and do not produce volatile methylarsines. Aquatic plants also synthesize similar lipid-soluble arsenic compounds. In terrestrial plants, arsenate is preferentially taken up 3 to 4 times the rate of arsenite. In the presence of phosphate, arsenate uptake is inhibited while in the presence of arsenate, phosphate uptake is only slightly inhibited. There is a competitive interaction between arsenate and phosphate for the same uptake system in terrestrial plants. The mode of toxicity of arsenate is to partially block protein synthesis and interfere with protein phosphorylation but the presence of phosphate prevents this mode of action. There appears to be a higher affinity for phosphate than arsenate with a discriminate ratio of 4:1. It is estimated that as much as 210 x 10(5) kg of arsenic is lost to the atmosphere in the vapor state annually from the land surface. The continental vapor flux is about 8 times that of the continental dust flux indicating that the biogenic contribution may play a significant role in cycling of arsenic. It has not been established whether volatile arsenic can be released by plants. Further studies are needed to determine mass balances in the rate of transfer (fluxes) of arsenic in the environment.

Animals↗

Dual system to reinforce biological containment of recombinant bacteria designed for rhizoremediation.

Active biological containment (ABC) systems have been designed to control at will the survival or death of a bacterial population. These systems are based on the use of a killing gene, e.g., a porin-inducing protein such as the one encoded by the Escherichia coli gef gene, and a regulatory circuit that controls expression of the killing gene in response to the presence or absence of environmental signals. An ABC system for recombinant microorganisms that degrade a model pollutant was designed on the basis of the Pseudomonas putida TOL plasmid meta-cleavage regulatory circuit. The system consists of a fusion of the Pm promoter to lacI, whose expression is controlled by XylS with 3-methylbenzoate, and a fusion of a synthetic P(lac) promoter to gef. In the presence of the model pollutant, bacterial cells survived and degraded the target compound, whereas in the absence of the aromatic carboxylic acid cell death was induced. The system had two main drawbacks: (i) the slow death of the bacterial cells in soil versus the fast killing rate in liquid cultures in laboratory assays, and (ii) the appearance of mutants, at a rate of about 10(-8) per cell and generation, that did not die after the pollutant had been exhausted. We reinforced the ABC system by including it in a Deltaasd P. putida background. A P. putida Deltaasd mutant is viable only in complex medium supplemented with diaminopimelic acid, methionine, lysine, and threonine. We constructed a P. putida Deltaasd strain, called MCR7, with a Pm::asd fusion in the host chromosome. This strain was viable in the presence of 3-methylbenzoate because synthesis of the essential metabolites was achieved through XylS-dependent induction. In the P. putida MCR7 strain, an ABC system (Pm::lacI, xylS, P(lac)::gef) was incorporated into the host chromosome to yield strain MCR8. The number of MCR8 mutants that escaped killing was below our detection limit (<10(-9) mutants per cell and generation). The MCR8 strain survived and colonized rhizosphere soil with 3-methylbenzoate at a level similar to that of the wild-type strain. However, it disappeared in less than 20 to 25 days in soils without the pollutant, whereas an asd(+), biologically contained counterpart such as P. putida CMC4 was still detectable in soils after 100 days.

Aspartic Acid↗

Natural formation and degradation of chloroacetic acids and volatile organochlorines in forest soil--challenges to understanding.

GOAL, SCOPE AND BACKGROUND: The anthropogenic environmental emissions of chloroacetic acids and volatile organochlorines have been under scrutiny in recent years because the two compound groups are suspected to contribute to forest dieback and stratospheric ozone destruction, respectively. The two organochlorine groups are linked because the atmospheric photochemical oxidation of some volatile organochlorine compounds is one source of phytotoxic chloroacetic acids in the environment. Moreover, both groups are produced in higher amounts by natural chlorination of organic matter, e.g. by soil microorganisms, marine macroalgae and salt lake bacteria, and show similar metabolism pathways. Elucidating the origin and fate of these organohalogens is necessary to implement actions to counteract environmental problems caused by these compounds. MAIN FEATURES: While the anthropogenic sources of chloroacetic acids and volatile organochlorines are relatively well-known and within human control, knowledge of relevant natural processes is scarce and fragmented. This article reviews current knowledge on natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soils, with particular emphasis on processes in the rhizosphere, and discusses future studies necessary to understand the role of forest soils in the formation and degradation of these compounds. RESULTS AND DISCUSSION: Reviewing the present knowledge of the natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soil has revealed gaps in knowledge regarding the actual mechanisms behind these processes. In particular, there remains insufficient quantification of reliable budgets and rates of formation and degradation of chloroacetic acids and volatile organochlorines in forest soil (both biotic and abiotic processes) to evaluate the strength of forest ecosystems regarding the emission and uptake of chloroacetic acids and volatile organochlorines, both on a regional scale and on a global scale. CONCLUSION: It is concluded that the overall role of forest soil as a source and/or sink for chloroacetic acids and volatile organochlorines is still unclear; the available laboratory and field data reveal only bits of the puzzle. Detailed knowledge of the natural degradation and formation processes in forest soil is important to evaluate the strength of forest ecosystems for the emission and uptake of chloroacetic acids and volatile organochlorines, both on a regional scale and on a global scale. RECOMMENDATION AND PERSPECTIVE: As the natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soil can be influenced by human activities, evaluation of the extent of this influence will help to identify what future actions are needed to reduce human influences and thus prevent further damage to the environment and to human health caused by these compounds.

Acetates↗

Evaluation of major constraints to revegetation of lead/zinc mine tailings using bioassay techniques.

The residues from the extraction of lead/zinc (Pb/Zn) ores of most Pb/Zn mines are permanently stored in tailings ponds, which require revegetation to reduce their environmental impact. This can only be done if the main constraints on plant establishment are evaluated. This can readily be done by field and greenhouse studies. To test this, the properties of different tailings from Lechang Pb/Zn mine located at the north of Guangdong Province in southern China have been studied. Physical and chemical properties including concentrations of metals (Pb, Zn, Cd and Cu) in the tailings and soils collected from different sites have been measured. The results showed that tailings contain low nitrogen (0.016-0.075%), low-organic matter (0.58-1.78%), high salt (3.55-13.85 dS/m), and high total and diethylene-tetramine-pentaacetic acid (DTPA)-extractable metal concentrations (total: 1,019-1,642 microg g(-1) Pb, 3,078-6,773 microg g(-1) Zn, 8-23 microg g(-1) Cd, and 85-192 microg g(-1) Cu; DTPA-extractable: 59-178 microig g(-1) Pb, 21-200 microg g(-1) Zn, 0.30-1.5 mcirog g(-1) Cd, and 4.3-12 microg g(-1) Cu). Aqueous extracts of tailings/soils (10%, 20% and 30%, w/v) from different sites were prepared for testing their effects on seed germination and root elongation of a vegetable crop Brassica chinensis and a grass species Cynodon dactylon. It was found that root elongation provided a better evaluation of toxicity than seed germination. The ranking of toxicity using root elongation was: high-sulfur tailings > tailing dam > sparsely vegetated tailings > densely vegetated tailings > mountain soil for both plants. This order was consistent with DTPA-extractable Pb contents in the tailings and soils. B. chinensis seedlings were then grown in the mixtures of different proportions of tailings and farm soil for 4 weeks, and the results (dry weights of seedlings) were in line with the root elongation test. All these demonstrated that heavy metal toxicity, especially available Pb, low content of nutrient, and poor physical structure were major constraints on plant establishment and colonization on the Pb/Zn mine tailings.

Biodegradation, Environmental↗

Combined sewer overflows to surface waters detected by the anthropogenic marker caffeine.

Continuous progress in wastewater treatment technology and the growing number of households connected to wastewater treatment plants (WWTPs) have generally resulted in decreased environmental loading of many pollutants. Nonetheless, further reduction of pollutant inputs is required to improve the quality of surface waters in densely populated areas. In this context, the relative contribution of combined sewer overflows as sources of wastewater-derived contaminants has attracted more and more attention, but the quantitative importance of these overflows has barely been investigated. In this study, caffeine was successfully used as a chemical marker to estimate the fraction of sewer overflows in the catchment area of lake Greifensee, Switzerland. Caffeine is a ubiquitous compound in raw, domestic wastewater with typical per capita loads of approximately 16 mg person(-1) d(-1). In WWTPs of the Greifensee region, caffeine is largely eliminated (>99%), resulting in much smaller loads of < or = 0.15 mg person(-1) d(-1) in treated wastewater. However, in receiving streams as in the inflows to Greifensee, caffeine loads (0.1-1.6 mg person(-1) d(-1)) were higher than those in WWTP effluents, indicating additional sources. As the loads in the streams correlated with precipitation during sampling, it was concluded that combined sewer overflows were the most likely source of caffeine. Using a mass balance approach, it was possible to determine the fraction of wastewater (in dry weather equivalents) discharged untreated to the receiving streams (up to 10%, annual mean, approximately 2-3%). The concept of caffeine as a marker for combined sewer overflows was then applied to estimate phosphorus inputs to Greifensee with untreated and treated wastewater (approximately 1.5 and 2.0 t P y(-1), respectively), which corresponded well with P inputs determined in a separate study based on hydraulic considerations. For compounds with high elimination in WWTPs such as phosphorus (96-98% in the Greifensee area), inputs from combined sewer overflows are thus of similar magnitude as inputs from treated wastewater. The study demonstrated that caffeine is a suitable marker for untreated wastewater (from combined sewer overflows, direct discharges, etc.), but its sensitivity depends on regional conditions and decreases with decreasing elimination efficiency in WWTPs.

Biodegradation, Environmental↗

Metal bioremediation through growing cells.

Heavy-metal pollution represents an important environmental problem due to the toxic effects of metals, and their accumulation throughout the food chain leads to serious ecological and health problems. Metal remediation through common physico-chemical techniques is expensive and unsuitable in case of voluminous effluents containing complexing organic matter and low metal contamination. Biotechnological approaches that are designed to cover such niches have, therefore, received great deal of attention in the recent years. Biosorption studies involving low-cost and often dead/pretreated biomass have dominated the literature and, subsequently, extensive reviews focusing on equilibrium and kinetics of metal biosorption have also come up. However, the low binding capacity of biomass for certain recalcitrant metals such as Ni and failure to effectively remove metals from real industrial effluents due to presence of organic or inorganic ligands limit this approach. At times, when pure biosorptive metal removal is not feasible, application of a judicious consortium of growing metal-resistant cells can ensure better removal through a combination of bioprecipitation, biosorption and continuous metabolic uptake of metals after physical adsorption. Such approach may lead to simultaneous removal of toxic metals, organic loads and other inorganic impurities, as well as allow optimization through development of resistant species. However, sensitivity of living cells to extremes of pH or high metal concentration and need to furnish metabolic energy are some of the major constraints of employing growing cells for bioremediation. The efforts to meet such challenges via isolation of metal-resistant bacterial/fungal strains and exploitation of organic wastes as carbon substrates have began. Recent studies show that the strains (bacteria, yeast and fungi) isolated from contaminated sites possess excellent capability of metal scavenging. Some bacterial strains possess high tolerance to various metals and may be potential candidates for their simultaneous removal from wastes. Evidently, the stage has already been set for the application of metal-resistant growing microbial cells for metal harvesting. This review focuses on the applicability of growing bacterial/fungal/algal cells for metal removal and the efforts directed towards cell/process development to make this option technically/economically viable for the comprehensive treatment of metal-rich effluents.

Adsorption↗

Potential tree species for use in the restoration of unsanitary landfills.

Given that they represent the most economical option for disposing of refuse, waste landfills are widespread in urban areas. However, landfills generate air and water pollution and require restoration for landscape development. A number of unsanitary waste landfills have caused severe environmental problems in developing countries. This study aimed to investigate the colonization status of different tree species on waste landfills to assess their potential for restoring unsanitary landfills in South Korea. Plot surveys were conducted using 10 x 10-m quadrats at seven waste landfill sites: Bunsuri, Dugiri, Hasanundong, Gomaeri, Kyongseodong, Mojeonri, and Shindaedong. We determined the height, diameter at breast height (DBH), and number of tree species in the plots, and enumerated all saplings < or =1 m high. Because black locust, Robinia pseudoacacia, was the dominant tree species in the waste landfills, we measured the distance from the presumed mother plant (i.e., the tallest black locust in a patch), height, and DBH of all individuals in black locust patches to determine patch structure. Robinia pseudoacacia, Salix koreensis, and Populus sieboldii formed canopy layers in the waste landfills. The basal area of black locust was 1.51 m(2)/ha, and this species had the highest number of saplings among all tree species. The diameter of the black locust patches ranged from 3.71 to 11.29 m. As the patch diameter increased, the number of regenerated saplings also tended to increase, albeit not significantly. Black locust invaded via bud banks and spread clonally in a concentric pattern across the landfills. This species grew well in the dry habitat of the landfills, and its growth rate was very high. Furthermore, black locust has the ability to fix nitrogen symbiotically; it is therefore considered a well-adapted species for waste landfills. Eleven woody species were selected for screening: Acer palmatum, Albizzia julibrissin, Buxus microphylla var. koreana, Ginkgo biloba, Hibiscus syriacus, Koelreuteria paniculata, Ligustrum obtusifolium, Liriodendron tulipifera, Pinus koraiensis, Pinus thunbergii, and Sophora japonica. As a result of a comparison of the total ratio (sum of shoot extension and diameter growth at the landfill relative to a reference site) and mortality, six species (Liriodendron tulipifera, Albizzia julibrissin, Ligustrum obtusifolium, Buxus microphylla var. koreana, Hibiscus syriacus, and Sophora japonica), which had a total ratio >1 and experienced low mortality, are recommended as potentially suitable species for waste landfill remediation. We suggest that mixed plantations of ubiquitous adaptable species and naturally occurring black locust will enhance the landscape through synergistic effects.

Adaptation, Physiological↗

Stability of 1-nitropyrene and 1,6-dinitropyrene in environmental water samples and soil suspensions.

This study examined the stability of mutagenic 1-nitropyrene (1-NP) and (1,6-dinitropyrene (1,6-diNP) in environmental water samples and various soil suspensions containing 0.1% peptone and in water samples containing no peptone. The water samples or the soil suspensions were mixed with NPs and incubated at 30 degrees C. The stability of NPs was expressed as mutagenic activity remaining in the test solutions. The mutagenicity decreased rapidly when 1-NP or 1,6-diNP was incubated in unautoclaved test solutions containing 0.1% peptone but not when incubated in autoclaved test solutions. The mutagenicity in the soil suspensions, especially in the sludge, decreased faster than in the water samples. This was due to the large number of colony-forming units (CFU) in the soil suspensions. In the water samples containing 0.1% peptone, the mutagenicity of NPs in the polluted Tamiya River water decreased faster than in the unpolluted Yoshino River water. The rate of decrease was dependent on the number of CFU in the water samples. A large number of CFU decreased the mutagenicity more rapidly than did a small number of CFU in samples. The disappearance of mutagenicity was dependent on the initial concentrations of NPs. The periods required for a 50% decrease in the mutagenicity of 1-NP at the low concentration (0.2 microgram/ml) was shorter than that at the high concentration (3 micrograms/ml). 1-Aminopyrene was detected in the 1-NP test solution after incubation when it was analyzed by high-pressure liquid chromatography. In the water samples containing no peptone, the mutagenicity of 1-NP (0.2 microgram/ml) decreased gradually during 30 days of incubation. After incubation for 1540 days, the remaining mutagenicity of 1-NP in the water samples was almost the same as that in autoclaved water samples. On the other hand, the mutagenicity of 1,6-diNP (10 ng/ml) decreased and the remaining mutagenicity, except in the Yoshino River water, was less than 20% after 30 days of incubation and was completely lost during the 1540-day incubation. However, the mutagenicity of 1,6-diNP in autoclaved water samples was very stable and almost all mutagenicity, except in sea water, remained after 1540 days of incubation at 30 degrees C. These results suggest that the microflora in the environment plays an important role in the primary degradation and decontamination of relatively low concentrations of NPs.

Biodegradation, Environmental↗