Pyrene degradation in forest humus microcosms with or without pine and its mycorrhizal fungus

The mineralization potential of forest humus and the self-cleaning potential of a boreal coniferous forest environment for polycyclic aromatic hydrocarbon (PAH) compounds was studied using a model ecosystem of acid forest humus (pH = 3.6) and pyrene as the model compound. The matrix was natural humus or humus mixed with oil-polluted soil in the presence and absence of Scots pine (Pinus sylvestris L.) and its mycorrhizal fungus (Paxillus involutus). The rates of pyrene mineralization in the microcosms with humus implants (without pine) were initially insignificant but increased from Day 64 onward to 47 microg kg(-1) d(-1) and further to 144 microg kg(-1) d(-1) after Day 105. In the pine-planted humus microcosms the rate of mineralization also increased, reaching 28 microg kg(-1) d(-1) after Day 105. The 14CO2 emission was already considerable in nonplanted microcosms containing oily soil at Day 21 and the pyrene mineralization continued throughout the study. The pyrene was converted to CO2 at rates of 0.07 and 0.6 microg kg(-1) d(-1) in the oily-soil implanted microcosms with and without pine, respectively. When the probable assimilation of 14CO2 by the pine and ground vegetation was taken into account the most efficient microcosm mineralized 20% of the 91.2 mg kg(-1) pyrene in 180 d. The presence of pine and its mycorrhizal fungus had no statistically significant effect on mineralization yields. The rates of pyrene mineralization observed in this study for forest humus exceeded the total annual deposition rate of PAHs in southern Finland. This indicates that accumulation in forest soil is not to be expected.     

Effects of tree root-derived substrates and inorganic nutrients on pyrene mineralization in rhizosphere and bulk soil

This study investigated the effects of organic and inorganic nutrients on the microbial degradation of the common soil contaminant pyrene. The material used in this investigation was collected from potted trees that had been growing for over a year in a soil artificially contaminated with polycyclic aromatic hydrocarbons. Soil was removed from the nonroot (bulk) and root (rhizosphere) zones of these pots and used in mineralization studies that tracked microbial degradation of 14C-pyrene. The factors influencing degradation in these zones were then tested by amendment with essential inorganic nutrients or with root-derived materials. As expected, pyrene mineralization was greater in soil removed from the rhizosphere than in bulk soil. The rate of mineralization in rhizosphere soil was inhibited by inorganic nutrient amendment, whereas nutrients stimulated mineralization in the bulk soil. Pyrene mineralization in bulk soil was also increased by the addition of root extracts intended to mimic exudation by living roots. However, amendment with excised fine roots that were allowed to decay over time in soil initially inhibited mineralization. With time, the rate of mineralization increased, eventually exceeding that of unamended bulk soil. Combined, the initial inhibition and subsequent stimulation produced a zero net impact of decaying fine roots on bulk soil mineralization. Our results, in conjunction with known temporal patterns of fine root dynamics in natural systems, support the idea that seasonal variations in nutrient and substrate availability may influence the long-term effect of plants on organic degradation in soil, possibly reducing or negating the beneficial effects of vegetation that are often observed in short-term studies.     

Prediction of microbial accessibility of carbon-14-phenanthrene in soil in the presence of pyrene or benzo[a]pyrene using an aqueous cyclodextrin extraction technique

Traditionally, solvent extractions are routinely used in the assessment of contaminated land. However, vigorous solvent extractions only give total concentrations rather than that relating to the bioaccessible fraction. Recently, less harsh, aqueous-based extraction methods have been shown to be a better estimate of the microbial degradation of polycyclic aromatic hydrocarbons (PAHs). The aqueous-based hydroxypropyl-beta-cyclodextrin (HPCD) extraction technique was tested using 14C-PAHs in soils and compared against indigenous microbial mineralization (a measure of bioaccessibility) of 14C-phenanthrene in the presence of pyrene or benzo[a]pyrene (B[a]P) over a range of concentrations (0, 5, 10, or 50 mg kg(-1)) and aged for 0, 25, 50, and 100 d in four soils. At each time point, the total loss, extractability, and mineralization of 14C-phenanthrene was measured in each of the soils. The presence of the other PAHs had little effect on the behavior of 14C-phenanthrene in any of the soils. Comparisons between the amounts of 14C-phenanthrene extracted using HPCD and mineralized were made and showed that there was a correlation (1:1). This study demonstrates that HPCD extraction is able to predict the microbial accessibility fraction of 14C-phenanthrene in the presence of other PAHs in a range of soils, further supporting the applicability of this technique.     

Differential responses of eubacterial, Mycobacterium, and Sphingomonas communities in polycyclic aromatic hydrocarbon (PAH)-contaminated soil to artificially induced changes in PAH profile

Recent reports suggest that Mycobacterium is better adapted to soils containing poorly bioavailable polycyclic aromatic hydrocarbons (PAHs) compared to Sphingomonas. To study this hypothesis, artificial conditions regarding PAH profile and PAH bioavailability were induced in two PAH-contaminated soils and the response of the eubacterial, Mycobacterium, and Sphingomonas communities to these changed conditions was monitored during laboratory incubation. Soil K3663 with a relatively high proportion of high molecular weight PAHs was amended with phenanthrene or pyrene to artificially change the soil into a soil with a relatively increased bioavailable PAH contamination. Soil AndE with a relatively high proportion of bioavailable low molecular weight PAHs was treated by a single-step Tenax extraction to remove the largest part of the easily bioavailable PAH contamination. In soil K3663, the added phenanthrene or pyrene compounds were rapidly degraded, concomitant with a significant increase in the number of phenanthrene and pyrene degraders, and minor and no changes in the Mycobacterium community and Sphingomonas community, respectively. However, a transient change in the eubacterial community related to the proliferation of several gamma-proteobacteria was noted in the phenanthrene-amended soil. In the extracted AndE soil, the Sphingomonas community initially developed into a more diverse community but finally decreased in size below the detection limit. Mycobacterium in that soil never increased to a detectable size, while the eubacterial community became dominated by a gamma-proteobacterial population. The results suggest that the relative bioavailability of PAH contamination in soil affects bacterial community structure but that the behavior of Mycobacterium and Sphingomonas in soil is more complex than prospected from studies on their ecology and physiology.     

Direct link between fluoranthene biodegradation and the mobility and sequestration of its residues during aging

The aim of this study was to assess the influence of the polycyclic aromatic hydrocarbons (PAH)-degrading activity in the fate of fluoranthene in soils. Three soil samples with different degrading activities (an industrial soil, the same industrial soil after biostimulation, and an agricultural soil) were spiked with 14C-fluoranthene and incubated for 6 mo with monitoring of biodegradation and mineralization. To follow the distribution of the 14C-fluoranthene residues (i.e., 14C-fluoranthene and its degradation products) among the soil compartments, we performed successively leaching, centrifugation (to collect intra-aggregate pore water), solvent extraction, and combustion of the soil columns. In the industrial soil, no mineralization of 14C-fluoranthene was observed, and only 3% of the initial 14C-activity was non-extractable (with acetone:dichloromethane) after 165 d of incubation. The biostimulation (addition of unlabeled polycyclic aromatic hydrocarbons) increased the degrading activity in this soil (59% of 14C-fluoranthene was mineralized) and increased the residues sequestration (13% of 14C-activity was non-extractable). The microflora of the agricultural soil mineralized 14C-fluoranthene more slowly and to a lesser extent (25%) than the biostimulated soil, but a higher amount of 14C-activity was sequestered (41%). Thus, the rate and extent of 14C-fluoranthene mineralization seemed to be related to the 14C-activity sequestration by controlling the accumulation of degradation products in the soil. 14C-Fluoranthene biodegradation enhanced the concentration of 14C-polar compounds in the intra-aggregate pore water. Our results point out the close link between fluoranthene biodegradation and two key aging processes, diffusion and sequestration, in soils. Biodegradation controls the mobility and sequestration of residues by transforming fluoranthene into more polar molecules that can diffuse into the intra-aggregate pore water and then might become bound to the matrix or entrapped in the microporosity.     

Persistence of testosterone and 17beta-estradiol in soils receiving swine manure or municipal biosolids

Natural and synthetic steroidal hormones can be carried to agricultural soil through fertilization with municipal biosolids, livestock manure, or poultry manure. The persistence and pathways of dissipation of [4-(14)C]-testosterone and of [4-(14)C]-17beta-estradiol in organic-amended soils were investigated using laboratory microcosms. Testosterone dissipation was investigated over a range of amendment concentrations, temperatures, and soil types. Under all conditions the parent compound and transformation products were dissipated within a few days. Addition of swine manure slurry to soil hastened the transformation of testosterone and 17beta-estradiol to the corresponding less hormonally active ketones, 4-androstene-3,17-dione and estrone. Two other testosterone transformation products, 5alpha-androstan-3,17-dione and 1,4-androstadiene-3,17-dione, were also detected. Experiments with sterilized soil and sterilized swine manure slurry suggested that the transformation of (14)C-labeled hormonal parent compounds was mainly caused by microorganisms in manure slurry, while mineralization of the hormones to (14)CO(2) required viable soil microorganisms. Organic amendments transiently inhibited the mineralization of [4-(14)C]-testosterone, perhaps by inhibiting soil microorganisms, or by enhancing sorption and reducing the bioavailability of testosterone or transformation products. Overall, organic amendments influenced the pathways and kinetics of testosterone and estradiol dissipation, but did not increase their persistence.     

大气颗粒物中多环芳烃污染特征及防治对策

介绍了大港油田大气颗粒物和多环芳烃的污染现状,并对大港油田地区大气特征构成及大气颗粒物中多环芳烃的污染特征作了初步分析,得出大港油田地区大气颗粒物中多环芳烃污染类型为燃油型,并据此提出了防治多环芳烃污染的相应对策。     

Integrating biodegradation and electroosmosis for the enhanced removal of polycyclic aromatic hydrocarbons from creosote-polluted soils

This paper presents a hybrid technology of soil remediation based on the integration of biodegradation and electroosmosis. We employed soils with different texture (clay soil and loamy sand) containing a mixture of polycyclic aromatic hydrocarbons (PAH) present in creosote, and inoculation with a representative soil bacterium able to degrade fluorene, phenanthrene, fluoranthene, pyrene, anthracene, and benzo[a]pyrene. Two different modes of treatment were prospected: (i) inducing in soil the simultaneous occurrence of biodegradation and electroosmosis in the presence of a biodegradable surfactant, and (ii) treating the soils sequentially with electrokinetics and bioremediation. Losses of PAH due to simultaneous biodegradation and electroosmosis (induced by a continuous electric field) were significantly higher than in control cells that contained the surfactant but no biological activity or no current. The method was especially successful with loamy sand. For example, benzo[a]pyrene decreased its concentration by 50% after 7 d, whereas 22 and 17% of the compound had disappeared as a result of electrokinetic flushing and bioremediation alone, respectively. The use of periodical changes in polarity and current pulses increased by 16% in the removal of total PAH and in up to 30% of specific compounds, including benzo[a]pyrene. With the aim of reaching lower residual levels through bioremediation, an electrokinetic pretreatment was also evaluated as a way to mobilize the less bioaccessible fraction of PAH. Residual concentrations of total biodegradable PAH, remaining after bioremediation in soil slurries, were twofold lower in electrokinetically pretreated soils than in untreated soils. The results indicate that biodegradation and electroosmosis can be successfully integrated to promote the removal of PAH from soil.     

An axenic plant culture system for optimal growth in long-term studies

The symbiotic co-evolution of plants and microbes leads to difficulties in understanding which of the two components is responsible for a given environmental response. Plant-microbe studies greatly benefit from the ability to grow plants in axenic (sterile) culture. Several studies have used axenic plant culture systems, but experimental procedures are often poorly documented, the plant growth environment is not optimal, and axenic conditions are not rigorously verified. We developed a unique axenic system using inert components that promotes plant health and can be kept sterile for at least 70 d. Crested wheatgrass (Agropyron cristatum cv. CDII) plants were grown in sand within flow-through glass columns that were positively pressured with filtered air. Plant health was optimized by regulating temperature, light level, CO2 concentration, humidity, and nutrients. The design incorporates several novel aspects, such as pretreatment of the sand with Fe, graduated sand layers to optimize the air-water balance of the root zone, and modification of a laminar flow hood to serve as a plant growth chamber. Adaptations of several sterile techniques were necessary for maintenance of axenic conditions. Axenic conditions were verified by plating and staining leachates as well as a rhizoplane stain. This system was designed to study nutrient and water stress effects on root exudates, but is useful for assessing a broad range of plant-microbe-environment interactions. Based on total organic C analysis, 74% of exudates was recovered in the leachate, 6% was recovered in the bulk sand, and 17% was recovered in the rhizosphere sand. Carbon in the leachate after 70 d reached 255 microg d(-1). Fumaric, malic, malonic, oxalic, and succinic acids were measured as components of the root exudates.     

搅拌棒吸附萃取气相色谱法测定地表水中多环芳烃

建立了新型的搅拌棒吸附萃取（SBSE）和热脱附系统（TDU）结合的气相色谱（GC）测定地表水中多环芳烃的方法。考察了萃取时间、搅拌条件及萃取温度对实验的影响,对7种多环芳烃（萘、荧蒽、苯并[b]荧蒽、苯并[k]荧蒽、苯并[ghi]苝、茚并[1,2,3-cd]芘和苯并[a]芘）的加标回收率为89.17%～99.38%,相对标准偏差（RSD）为1.6%～5.6%（n=3）。通过实际样品中PAHs的分析表明,该法快速、灵敏、简单,能满足痕量分析的需求。     

Degradation of polycyclic aromatic hydrocarbons by combined chemical pre-oxidation and bioremediation in creosote contaminated soil

The ability of pre-oxidation to overcome polycyclic aromatic hydrocarbons (PAH) recalcitrance to biodegradation was investigated in creosote contaminated soil. Sand and peat artificially spiked with creosote (quality WEI C) were used as model systems. Ozonation and Fenton-like treatment were proved to be feasible technologies for PAH degradation in soil. The efficiency of ozonation was strongly dependent on the water content of treated soil samples. The removal of PAH by Fenton-like treatment depended on the applied H2O2/soil weight ratio and ferrous ions addition. It was determined that the application of chemical oxidation in sand resulted in a higher PAH removal and required lower oxidant (ozone, hydrogen peroxide) doses. The enhancement of PAH biodegradability by different pre-treatment technologies also depended on the soil matrix. It was ascertained that combined chemical and biological treatment was more efficient in PAH elimination in creosote contaminated soil than either one alone. Thus, the combination of Fenton-like and the subsequent biological treatment resulted in the highest removal of PAH in creosote contaminated sand, and biodegradation with pre-ozonation was found to be the most effective technology for PAH elimination in peat.     

The potential of epiphytic hydrocarbon-utilizing bacteria on legume leaves for attenuation of atmospheric hydrocarbon pollutants

The leaves of two legumes, peas and beans, harbored on their surfaces up to 9×10? cells g?1 of oil-utilizing bacteria. Less numbers, up to 5×10? cells g?1 inhabited leaves of two nonlegume crops, namely tomato and sunflower. Older leaves accommodated more of such bacteria than younger ones. Plants raised in oily environments were colonized by much more oil-utilizing bacteria than those raised in pristine (oil-free) environments. Similar numbers were counted on the same media in which nitrogen salt was deleted, indicating that most phyllospheric bacteria were probably diazotrophic. Most dominant were Microbacterium spp. followed by Rhodococcus spp., Citrobacter freundii, in addition to several other minor species. The pure bacterial isolates could utilize leaf tissue hydrocarbons, and consume considerable proportions of crude oil, phenanthrene (an aromatic hydrocarbon) and n-octadecane (an alkane) in batch cultures. Bacterial consortia on fresh (but not on previously autoclaved) leaves of peas and beans could also consume substantial proportions of the surrounding volatile oil hydrocarbons in closed microcosms. It was concluded that phytoremediation through phyllosphere technology could be useful in remediating atmospheric hydrocarbon pollutants.     

Application of a slow-release fertilizer for oil bioremediation in beach sediment

A 105-d field experiment was conducted to determine the potential of the slow-release fertilizer, Osmocote (Scotts, Marysville, OH), to stimulate the indigenous microbial biodegradation of petroleum hydrocarbons in an oil-spiked beach sediment on an intertidal foreshore in Singapore. Triplicate microcosms containing 80 kg of weathered sediment, spiked with 5% (w/w) Arabian light crude oil and 1.2% (w/w) Osmocote pellets, were established, together with control microcosms minus Osmocote. Relative to the control, the presence of the Osmocote sustained a significantly higher level of nutrients (NH(4)(+)-N, NO(3)(-)-N, and PO(4)(3-)-P) in the sediment pore water over the duration of the experiment. The metabolic activity of the indigenous microbial biomass, as measured using an intracellular dehydrogenase enzyme assay, was also significantly enhanced over the duration of the experiment in amended sediments. The loss of total recoverable petroleum hydrocarbons (TRPH) and biodegradation of total n-alkanes (C(10)-C(33)), branched alkanes (pristane and phytane), as well as total target polycyclic aromatic hydrocarbons (PAHs) (two- to six-ring), in both the control and Osmocote-amended sediments, followed a first-order biodegradation model. The first-order loss rate of total recoverable petroleum hydrocarbons was 2.57 times greater than that of the control. The hopane-normalized rate constants for total n-alkane, branched alkane, and total target PAH biodegradation in the Osmocote-treated sediments were 3.95-, 5.50-, and 2.45-fold higher than the control, respectively. Overall, the presence of Osmocote was able to significantly enhance and accelerate the biodegradation of aliphatics and PAHs in oil-contaminated sediments under natural field conditions in an intertidal foreshore environment.     

Solubilization of mixed polycyclic aromatic hydrocarbons through a rhamnolipid biosurfactant

The solubilization of phenanthrene (PHE) and pyrene (PYR) by rhamnolipid biosurfactant was systematically investigated. The solubilities of both polycyclic aromatic hydrocarbons (PAHs) were increased linearly with the biosurfactant concentration at above critical micelle concentration. A competitive effect was observed between PHE and PYR. The solubility of PHE in a mixed system was lower than that in a single PAH system, whereas the solubility of PYR in a mixed system was enhanced. This is because the hydrophobicity of PYR is higher than that of PHE, so PYR is favored in the competitive solubilization. The combined effect of biosurfactant and dissolved organic matter (DOM) on PAH solubilization was also examined. Two kinds of DOM (derived from soil and from compost) were used. There was an obvious enhancement of solubility for PHE and PYR in systems with concurrence of DOM and biosurfacrant compared with systems with only DOM or biosurfactant; however, the enhancement in the mixed system was less than their additive. This could be explained as the formation of a DOM-biosurfactant complex. In addition, the solubility enhancement of PAHs in a compost-DOM system was higher than that in a soil-DOM system. This could be explained as functional group differences of two DOM types.     

Phenanthrene degradation in soils co-inoculated with phenanthrene-degrading and biosurfactant-producing bacteria

Contaminant sorption within the soil matrix frequently limits biodegradation. However, contaminant bioavailability can be species-specific. This study investigated bioavailability of phenanthrene (PHE) to two PHE-degrading bacteria (Pseudomonas strain R and isolate P5-2) in the presence of rhamnolipid biosurfactant and/or a biosurfactant-producing bacterium, Pseudomonas aeruginosa ATCC 9027. Pseudomonas strain R mineralized more soil-sorbed PHE than strain P5-2, but in aqueous cultures the rate and extent of PHE mineralization by P5-2 exceeded that by P. strain R. In Fallsington sandy loam (fine-loamy, mixed, active, mesic Typic Endoaquult) (high PHE-sorption capacity) the addition of rhamnolipid increased PHE mineralization by P. strain R. Phenanthrene mineralization in soils inoculated with P5-2 was minimal and no enhancement in PHE degradation was observed when biosurfactant was added. Co-inoculation of Fallsington sandy loam with the biosurfactant producer did not affect PHE mineralization by isolate P5-2, but significantly enhanced PHE mineralization by P. strain R. The enhancement of PHE mineralization could not be explained by P. aeruginosa-mediated PHE degradation. The addition of rhamnolipid at concentrations above the critical micelle concentration (CMC) resulted in enhanced PHE release from test soils. These results suggest that the PHE-degrading strains were able to access different pools of PHE and that the biosurfactant-enhanced release of PHE from soils did not result in enhanced biodegradation. The results also demonstrated that bacteria with the catabolic potential to degrade sorbed hydrophobic contaminants could interact commensally with surfactant-producing strains by an unknown mechanism to hasten the biodegradation of aromatic hydrocarbons. Thus, understanding interactions among microbes may provide opportunities to further enhance biodegradation of soil-bound organic contaminants.     

Gaseous contaminant emissions as affected by burning scrap tires in cement manufacturing

We studied the environmental impact (gaseous emissions) of using scrap tires as a fuel substitute at a cement plant that produces one million tons of cement per year. Using a combination of tires and coal as opposed to only coal caused variations in the pollutant emission rate. The study recorded a 37% increase in the rate of emission for CO, a 24% increase for SO2, an 11% decrease for NOx, and a 48% increase for HCl when tires were included. The rate of emission for metals increased 61% for Fe, 33% for Al, 487% for Zn, 127% for Pb, 339% for Cr, 100% for Mn, and 74% for Cu, and decreased 22% for Hg. On the other hand, the emission rate of organic compounds dropped by 14% for polycyclic aromatic hydrocarbons, 8% in naphthalene, 37% in chlorobenzene, and 45% in dioxins and furans. We used a Gaussian model of atmospheric dispersion to calculate the average pollutant concentration (1-h, 24-h, and annual concentrations) in the ambient air at ground level with the help of the ISC-ST2 software program developed by the USEPA. When tires were used, we observed (i) a 12 to 24% increase in particulate matter, this range considering the concentration variation depending on the average used (1-h, 24-h, and annual basis), 31 to 52% in CO, 22 to 34% in SO2, 39 to 52% in HCl, 12 to 27% in Fe, -3 to 8% in Al, 30 to 37% in Zn, and 270 to 885% in Pb; (ii) a decrease of 8 to 13% in NOx, 9 to 13% in polycyclic aromatic hydrocarbons, 6 to 7% in naphthalene, 32 to 39% in chlorobenzene, and 32 to 45% in dioxins and furans. The results obtained showed that the maximum ground-level concentrations were well within the environmental standards (for operation with only coal as well as for operation with a combination of coal and tires).     

Distribution behavior of pyrene to adsorbed humic acids on kaolin

The distribution behavior of pyrene on humic acid (HA)-kaolin complexes, prepared by adsorbing HA on kaolin, was investigated by batch experiments. The distribution coefficient (Kd) of pyrene on the HA-kaolin complex increased with the fraction (f(oc)) of organic carbon adsorbed to the surface of the kaolin. This can be attributed to hydrophobic interactions between pyrene and the adsorbed HAs. The effects of adsorbed HAs were quantitatively evaluated by calculating the distribution coefficient (K(oc)) and affinity constant (K(oc)ads) for pyrene to the adsorbed HAs. A fluorescence quenching method was employed to determine the affinity constant (K(oc)aq) of pyrene to HAs dissolved in an aqueous solution. When the K(oc) values were compared with the K(oc)aq values, the K(oc) values were found to be 4 to 11 times larger than the K(oc)aq values. On the other hand, the K(oc)ads values were 4 to 9 times larger than the K(oc)aq values. These indicate that the affinity for pyrene is enhanced by the adsorption of HAs to kaolin. In addition, the K(oc) values increased with increasing average molecular weights of the HAs. These results demonstrate that HAs, when they are adsorbed to clay minerals, play an important role in the deposition of polycyclic aromatic hydrocarbons (PAHs) in a soil environment.     

Genotoxicity is unrelated to total concentration of priority carcinogenic polycyclic aromatic hydrocarbons in soils undergoing biological treatment

A solid-phase microbiological assay was used to determine the changes in genotoxicity associated with sequestration or biodegradation of carcinogenic compounds in contaminated soils. The concentration of six carcinogenic polycyclic aromatic hydrocarbons (PAHs) did not change in 59 d in sterile soil, but the genotoxicity declined markedly. In a soil undergoing bioremediation in the field for 147 d or biodegradation in the laboratory for 180 d, the concentrations either changed little or declined at different rates, but the genotoxicity increased followed by a decline. The genotoxicity of a second soil declined as a result of biological treatment. The data show that genotoxicity of contaminated soils may be unrelated to the concentration of carcinogenic PAHs because of aging or new mutagens formed during biological treatment.     

Responses to iron limitation in Hordeum vulgare L. as affected by the atmospheric CO2 concentration

Elevated atmospheric CO2 treatments stimulated biomass production in Fe-sufficient and Fe-deficient barley plants, both in hydroponics and in soil culture. Root/shoot biomass ratio was increased in severely Fe-deficient plants grown in hydroponics but not under moderate Fe limitation in soil culture. Significantly increased biomass production in high CO2 treatments, even under severe Fe deficiency in hydroponic culture, indicates an improved internal Fe utilization. Iron deficiency-induced secretion of PS in 0.5 to 2.5 cm sub-apical root zones was increased by 74% in response to elevated CO2 treatments of barley plants in hydroponics but no PS were detectable in root exudates collected from soil-grown plants. This may be attributed to suppression of PS release by internal Fe concentrations above the critical level for Fe deficiency, determined at final harvest for soil-grown barley plants, even without additional Fe supply. However, extremely low concentrations of easily plant-available Fe in the investigated soil and low Fe seed reserves suggest a contribution of PS-mediated Fe mobilization from sparingly soluble Fe sources to Fe acquisition of the soil-grown barley plants during the preceding culture period. Higher Fe contents in shoots (+52%) of plants grown in soil culture without Fe supply under elevated atmospheric CO2 concentrations may indicate an increased efficiency for Fe acquisition. No significant influence on diversity and function of rhizosphere-bacterial communities was detectable in the outer rhizosphere soil (0-3 mm distance from the root surface) by DGGE of 16S rRNA gene fragments and analysis of marker enzyme activities for C-, N-, and P-cycles.     

Validation of procedures to quantify nonextractable polycyclic aromatic hydrocarbon residues in soil

This study was conducted to optimize butanol solvent shake extraction, dichloromethane soxtec extraction, and methanolic saponification extraction for the selective extraction of aged polycyclic aromatic hydrocarbons from soil. Extraction kinetics for these methods was established to determine the optimal time necessary to achieve exhaustive compound extraction. This resulted in times of 12, 6, and 5 h, respectively, for butanol, dichloromethane, and saponification, to extract polycyclic aromatic hydrocarbons from previously spiked, then aged soil. Increasing the soil mass to butanol volume ratio reduced the proportion of polycyclic aromatic hydrocarbon extracted by butanol, highlighting the importance of determining and maintaining a constant soil to solvent ratio for comparative purposes. Drying soil samples before dichloromethane soxtec extraction reduced by 30 to 76% the amount of polycyclic aromatic hydrocarbons extracted. The effect of sample drying is discussed with relevance to enhancing the formation of nonextractable compounds in soil and compound losses previously assumed by volatilization. The optimized extraction procedures provided low variability with relative standard deviations < or = 5.2% for analysis of multiple replicates. The results obtained by the optimized procedures provided equivalent or improved reproducibility to those obtained by other methods reported in the literature.