Elevated nitrate enriches microbial functional genes for potential bioremediation of complexly contaminated sediments

Nitrate is an important nutrient and electron acceptor for microorganisms, having a key role in nitrogen (N) cycling and electron transfer in anoxic sediments. High-nitrate inputs into sediments could have a significant effect on N cycling and its associated microbial processes. However, few studies have been focused on the effect of nitrate addition on the functional diversity, composition, structure and dynamics of sediment microbial communities in contaminated aquatic ecosystems with persistent organic pollutants (POPs). Here we analyzed sediment microbial communities from a field-scale in situ bioremediation site, a creek in Pearl River Delta containing a variety of contaminants including polybrominated diphenyl ethers (PBDEs) and polycyclic aromatic hydrocarbons (PAHs), before and after nitrate injection using a comprehensive functional gene array (GeoChip 4.0). Our results showed that the sediment microbial community functional composition and structure were markedly altered, and that functional genes involved in N-, carbon (C)-, sulfur (S)-and phosphorus (P)- cycling processes were highly enriched after nitrate injection, especially those microorganisms with diverse metabolic capabilities, leading to potential in situ bioremediation of the contaminated sediment, such as PBDE and PAH reduction/degradation. This study provides new insights into our understanding of sediment microbial community responses to nitrate addition, suggesting that indigenous microorganisms could be successfully stimulated for in situ bioremediation of POPs in contaminated sediments with nitrate addition.     

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in vadose sediments

The unsaturated subsurface (vadose zone) receives significant amounts of hazardous chemicals, yet little is known about its microbial communities and their capacity to biodegrade pollutants. Trichloroethylene (TCE) biodegradation occurs readily in surface soils; however, the process usually requires enzyme induction by aromatic compounds, methane, or other cosubstrates. The aerobic biodegradation of toluene and TCE by indigenous microbial populations was measured in samples collected from the vadose zone at unpolluted and gasoline-contaminated sites. Incubation at field moisture levels showed little activity on either TCE or toluene, so samples were tested in soil suspensions. No degradation occurred in samples suspended in water or phosphate buffer solution; however, both toluene and TCE were degraded in samples suspended in mineral salts medium. TCE degradation depended on toluene degradation, and little loss occurred under sterile conditions. Studies with specific nutrients showed that addition of ammonium sulfate was essential for degradation, and addition of other mineral nutrients further enhanced the rate. Additional studies with vadose sediments amended with nutrients showed similar trends to those observed in sediment suspensions. Initial rates of biodegradation in suspensions were faster in uncontaminated samples than in gasolinecontaminated samples, but the same percentages of chemicals were degraded. Biodegradation was slower and less extensive in shallower samples than deeper samples from the uncontaminated site. Two toluene-degrading organisms isolated from a gasoline-contaminated sample were identified as Corynebacterium variabilis SVB74 and Acinetobacter radioresistens SVB65. Inoculation with 10⁶ cells of C. variabilis ml^–1 of soil solution did not enhance the rate of degradation above that of the indigenous population. These results indicate that mineral nutrients limited the rate of TCE and toluene degradation by indigenous populations and that no additional benefit was derived from inoculation with a toluene-degrading bacterial strain. Correspondence to: K.M. Scow     

Biodegradation of Trichloroethylene and Dichloromethane in Contaminated Soil and Groundwater

Soil column and serum bottle microcosm experiments were conducted to investigate the potential for in situ anaerobic bioremediation of trichloroethy lene (TCE) and dichloromethane (DCM) at the Pinellas site near Largo, Florida. Soil columns with continuous groundwater recycle were used to evaluate treatment with complex nutrients (casamino acids, methanol, lactate, sulfate); benzoate and sulfate; and methanol. The complex nutrients drove microbial dechlorination of TCE to ethene, whereas the benzoate/sulfate and methanol supported microbial dechlorination of TCE only to cis-1 ,2-dichloroethylene (cDCE). Microbial sulfate depletion in the benzoate/sulfate column allowed further dechlorination of cDCE to vinyl chloride. Serum bottle microcosms were used to investigate TCE dechlorination and DCM biodegradation in Pinellas soil slurries bioaugmented with liquid from the soil columns possessing TCE-dechlorinating activity and DCM biodegradation by indigenous microorganisms. Bioaugmented soil microcosms showed immediate TCE dechlorination in the microcosms with methanol or complex nutrients, but no dechlorination in the benzoate/sulfate microcosm. DCM biodegradation by indigenous microorganisms occurred in soil microcosms amended with either benzoate/sulfate or methanol, but not with complex nutrients. Bioaugmentation stimulated DCM biodegradation in both complex nutrient and methanol-amended microcosms, but appeared to inhibit DCM biodegradation in benzoate/sulfate-amended microcosms. TCE dechlorination occurred before DCM biodegradation in bioaugmented microcosms when both compounds were present.     

A Long-Term Field Study of In Situ Bioremediation in a Fractured Conglomerate Trichloroethene Source Zone

ABSTRACT An 8-year bioremediation field study was conducted in a trichloroethene (TCE)-contaminated, highly indurated (i.e., hard), recharge-limited (i.e., contains little water) conglomerate where common remediation strategies, such as groundwater recirculation and direct push installation of a large well network, could not be used. A tracer test using isotopically distinct water from the Hetch Hetchy Reservoir indicated that remediation fluids mainly flowed through fractures and sand lenses in the conglomerate. This was confirmed during in situ bioremediation of the site, in which Dehalococcoides (from a bioaugmentation culture) and volatile fatty acids (from injection of lactate) were the most accurate indicators of transport between wells. Some contaminants were also displaced out of the area due to injection of tracer water. Despite these difficulties, dissolved contaminant mass decreased by an estimated 80% by the end of the test, reaching the lowest values ever recorded at this site. Furthermore, the persistence of ethene 4 years after bioaugmentation suggests that the dechlorinating capacity of the remaining microbial community is comparable to the matrix diffusion of TCE into the dissolved phase.     

Natural attenuation: what does the subsurface have in store?

Throughout the world, organic and inorganic substances leach intothe subsurface as a result of human activities and accidents. There, the chemicals pose director indirect threats to the environment and to increasingly scarce drinking water resources.At many contaminated sites the subsurface is able to attenuate pollutants which, potentially,lowers the costs of remediation. Natural attenuation comprises a wide range of processesof which the microbiological component, which is responsible for intrinsic bioremediation,can decrease the mass and toxicity of the contaminants and is, therefore, the mostimportant. Reliance on intrinsic bioremediation requires methods to monitor the process. Thesubject of this review is how knowledge of subsurface geology and hydrology, microbial ecologyand degradation processes is used and can be used to monitor the potential andcapacity for intrinsic bioremediation in the subsurface and to verify degradation in situ.As research on natural attenuation in the subsurface has been rather fragmented and limitedand often allows only conclusions to be drawn of the site under investigation, we providea concept based on Environmental Specimen Banking which will contribute to furtherunderstanding subsurface natural attenuation processes and will help to develop andimplement new monitoring techniques.     

Application of Pneumatic Fracturing to Enhance In Situ Bioremediation

A field pilot demonstration integrating pneumatic fracturing and in situ bioremediation was carried out in a gasoline-contaminated, low permeability soil formation. A pneumatic fracturing system was used to enhance subsurface air flow and transport rates, as well as to deliver soil amendments directly to the indigenous microbial populations. An in situ bioremediation zone was established and operated for a period of 50 weeks, which included periodic subsurface injections of phosphate, nitrate, and ammonium salts. Off-gas data indicated the formation of a series of aerobic, denitrifying, and methanogenic microbial degradation zones. Based on soil samples recovered from the site, 79% of soil-phase benzene, toluene, and xylenes (BTX) was removed by the integrated technology. From mass balance calculations, accounting for all physical losses, it was estimated that 85% of the total mass of BTX removed (based on mean concentration levels) was attributable to biodegradation.     

Growth ofAgrostis capillaris andDeschampsia flexuosa in relation to decomposition of and nutrient release by Sitka spruce litter

Summary Bags containing Sitka spruce litter (0, 15, 50 g) were placed in flower pots and covered with sand. Pots were watered at weekly intervals with nutrient solutions with and without nitrogen and with and without phosphorus. Decomposition was measured by carbon dioxide evolution from pots without plants. Neither added nitrogen nor added phosphorus had any marked effect on the rate of decomposition, which amounted to 14% loss of carbon in a year. The two grass species responded similarly to fertilizer; 72% of added nitrogen and 90% of added phosphorus were recovered in plant parts.A. capillaris captured nutrients more effectively from spruce litter than didD. flexuosa, recovering 13% of the phosphorus in the litter but only 5% of the nitrogen. Neither uptake of nitrogen nor uptake of phosphorus was enhanced in plants receiving fertilizer additions of the other nutrient.     

Effect of Chemical Oxidation on Subsurface Microbiology and Trichloroethene (TCE) Biodegradation

Research was conducted to determine the effect of chemical oxidation on subsurface microbiology and cometabolic biodegradation capacity in a trichloroethene (TCE)/perchloroethene (PCE)-contaminated aquifer previously treated with Fenton's reagent. Groundwater pH declined from 5 to 2.4 immediately after the treatment, and subsequently rose to a range of 3.4 to 4.0 after 17 months. Limited microbial growth and TCE degradation were detected in the treated zone (pH 3.37 and TCE 5 to 21 mg/L) with carbon addition (i.e., methane and phenol). Methane addition resulted in the enrichment of yeast and fungi in microcosms at low pH. In contrast, methane addition to groundwater from the control well (pH 4.9 and TCE ca. 0.7 mg/L) stimulated methanotrophic growth, indicated by methane consumption, fluorescent antibody analysis, phospholipid-based markers, and rDNA probes. TCE degradation was measured in the control microcosms, but only when phenol was added. Although higher TCE concentrations in the treated zone might have inhibited TCE cometabolism, these results also indicate that low groundwater pH resulting from the chemical oxidation process (pH 3.37 versus 4.9) inhibited TCE degradation. Methanotrophic growth and TCE biodegradation may be possible as pH increases both in the treated zone and at the leading edge of plume, as long as the local soil is able to buffer the groundwater pH. Moreover, the Fenton's reagent process could be designed to operate at a higher pH (e.g., ≥ 4.5) and/or lower hydrogen peroxide concentration to minimize detrimental effects, providing an optimal environment to couple advanced oxidation processes with bioremediation technologies.     

Effect of Nitrogen Source on Growth and Trichloroethylene Degradation by Methane-Oxidizing Bacteria

The effect of nitrogen source on methane-oxidizing bacteria with respect to cellular growth and trichloroethylene (TCE) degradation ability were examined. One mixed chemostat culture and two pure type II methane-oxidizing strains, Methylosinus trichosporium OB3b and strain CAC-2, which was isolated from the chemostat culture, were used in this study. All cultures were able to grow with each of three different nitrogen sources: ammonia, nitrate, and molecular nitrogen. Both M. trichosporium OB3b and strain CAC-2 showed slightly lower net cellular growth rates and cell yields but exhibited higher methane uptake rates, levels of poly-β-hydroxybutyrate (PHB) production, and naphthalene oxidation rates when grown under nitrogen-fixing conditions. The TCE-degrading ability of each culture was measured in terms of initial TCE oxidation rates and TCE transformation capacities (mass of TCE degraded/biomass inactivated), measured both with and without external energy sources. Higher initial TCE oxidation rates and TCE transformation capacities were observed in nitrogen-fixing mixed, M. trichosporium OB3b, and CAC-2 cultures than in nitrate- or ammonia-supplied cells. TCE transformation capacities were found to correlate with cellular PHB content in all three cultures. The results of this study suggest that the nitrogen-fixing capabilities of methane-oxidizing bacteria can be used to select for high-activity TCE degraders for the enhancement of bioremediation in fixed-nitrogen-limited environments.

Optimal bioremediation conditions within contaminated aquifers are often found to be limited by the availability of nutrients, including nitrogen. Consequently, microorganisms that are capable of degrading contaminants as well as fixing molecular nitrogen as their sole nitrogen source could have a growth advantage in fixed-nitrogen-deficient environments that would be favorable for promoting in situ bioremediation.Trichloroethylene (TCE) is a major groundwater contaminant of concern in the United States due to its suspected carcinogenity and persistence in subsurface environments (31). However, a number of laboratory (1, 4, 13, 16, 18, 19, 22, 23, 26–28, 34) and field studies (3, 15, 24, 25) have shown that TCE can be cometabolically transformed into nontoxic end products (CO₂ and Cl⁻) by methane-oxidizing bacteria at the expense of reducing energy in the form of NADH. Many studies have also reported that some methane-oxidizing cultures (type II) are able to utilize different sources of nitrogen (N) for cellular growth (32, 33), including molecular nitrogen at reduced oxygen partial pressures (11, 12, 20, 33). The types of methanotrophs that are capable of nitrogen fixation also produce a type of oxygenase (i.e., soluble methane monooxygenase [sMMO]) which exhibits high activity with respect to the oxidation of TCE.Poly-β-hydroxybutyrate (PHB) is an internal reducing-energy storage polymer that can be used as an alternative reducing-energy source by a number of methane-oxidizing cultures under starvation conditions (9). Recently, a number of studies observed a correlation between TCE transformation capacities (T_c; mass of TCE transformed per mass of cells inactivated) and microbial PHB content (7, 16, 17), suggesting that PHB might be used as an alternative NADH source for TCE oxidation by methane-oxidizing bacteria in the absence of growth substrate. It has also been shown that the synthesis of PHB is stimulated in cells grown under nutrient-limited conditions, including nitrogen-fixing conditions (2, 9, 10, 21). As a result of the characteristics of methane-oxidizing microorganisms described above, it may be possible to select for nitrogen-fixing methane oxidizers in fixed-nitrogen-limited subsurface environments such that the burden of nutrient addition to the subsurface for the sustained growth of these contaminant degraders is diminished while contaminant degradation is enhanced during in situ bioremediation.A recent study conducted by us (7) explored the feasibility of using the nitrogen-fixing capabilities of methane oxidizers for the enhancement of bioremediation. Our results suggested that nitrogen-fixing mixed cultures were able to degrade TCE as effectively as nitrate-supplied cultures. Further, higher T_c and higher cellular PHB contents were observed in nitrogen-fixing cultures. Of particular interest were observations of lower TCE product toxicity, measured in terms of methane uptake rates following TCE exposure, for nitrogen-fixing cultures than for nitrate- or ammonia-supplied cultures. Since that study was conducted with mixed cultures, it was difficult to elucidate the reasons for the enhanced degradation performance of the nitrogen-fixing methane oxidizers. An understanding of the effects of nitrogen source on cell growth and TCE degradation ability will be particularly beneficial for designing, operating, and implementing in situ- or ex situ-engineered bioremediation systems. This study evaluates nitrogen source effects on methane-oxidizing bacteria, using two pure strains and one mixed chemostat culture. Nitrogen source effects are examined with regard to cellular growth, specific methane uptake rates, specific naphthalene oxidation rates, and TCE degradation ability.     

Comparison of TCE Transformation Abilities of Methane- and Propane-Utilizing Microorganisms

Subsurface microorganisms from McClellan Air Force Base (AFB) were grown in batch aquifer microcosms on methane, propane, and butane to evaluate the potential for aerobic trichloroethylene (TCE) cometabolism. Microorganisms stimulated on all three substrates indicated the existence of a subsurface microbial community capable of utilizing alkanes as growth substrates. Initial growth substrate utilization lag periods of 2 weeks for methane and 3 weeks for propane and butane were observed. Methane- and propane-utilizers were active toward TCE cometabolism, whereas butane-utilizers showed no ability to transform TCE. Gradually increasing TCE concentrations were effectively transformed with uniform additions of methane and propane for up to 1 year. TCE was transformed most rapidly during active methane utilization, and continued at a slower rate for approximately 1 week after methane was consumed. Propane microcosms maintained first-order TCE transformation for up to 4 weeks after propane was consumed. The microbial communities remained active toward primary substrate utilization as the TCE concentration was gradually increased. Both methane- and propane-utilizers showed positive correlations between TCE transformation rates and primary substrate utilization rates. Observed maximum TCE transformation yields were 0.068 g TCE/g methane and 0.048 g TCE/g propane. The methane-utilizers also transformed chloroform (CF) but not 1,1,1-trichloroethane (1,1,1-TCA). Propane-utilizers transformed both CF and 1,1,1-TCA, indicating they were better suited for cometabolizing chlorinated aliphatic hydrocarbon mixtures in the McClellan AFB subsurface.     

Effect of nitrogen and phosphorus addition on phenanthrene biodegradation in four soils

Phenanthrene mineralization rates were found to vary widely among four soils; differences in soil nutrient levels was one hypothesis to explain this variation. To test this hypothesis, phenanthrene mineralization rates were measured in these soils with, and without, added nitrogen and phosphorus. Mineralization rates either remained unchanged or were depressed by the addition of nitrogen and phosphorus. Phenanthrene degradation rates remained unchanged in the soil which had the highest indigenous levels of nitrogen and phosphorus and which showed the largest increase in phosphorus levels after nutrients were added. The soils in which degradation rates were depressed had lower initial phosphorus concentrations and showed much smaller or no measurable increase in phosphorus levels after nutrients were added to the soils. To understand the response of phenanthrene degradation rates to added nitrogen and phosphorus, it may be necessary to consider the bioavailability of added nutrients and nutrient induced changes in microbial metabolism and ecology.     

Dilution bioassays: Their application to assessments of nutrient limitation in

Hypereutrophic waters, which are characterized by nutrient inputs exceeding phytoplankton nutrient requirements, are often sites of chronic nuisance algal blooms and associated water quality deterioration problems. In order to restore such systems to acceptable water quality standards, identification of growth-limiting nutrients is of central importance. Conventional nutrient addition bioassay techniques are often ineffective in identifying potentially limiting nutrients, due to persistent nutrient excesses in hypereutrophic systems. Accordingly, we have developed a nutrient dilution bioassay, in which stepwise dilutions of phytoplankton nutrients (nitrogen, phosphorus, iron, trace metals) with a nutrient-free major ion solution are capable of; 1) identifying those nutrients potentially most limiting, and 2) establishing magnitudes of respective nutrient input cutbacks required to bring about nutrient-limited control of phytoplankton growth. In situ deployment of dilution bioassays should help establish criteria governing minimal nutrient inputs required to arrest undesirable impacts of hypereutrophy. We have evaluated the field applicability of dilution bioassays, during a 2 year trial in the periodically hypereutrophic Neuse River, North Carolina.     

The response of microbial biomass and hydrolytic enzymes to a decade of nitrogen, phosphorus, and potassium addition in a lowland tropical rain forest

Nutrient availability is widely considered to constrain primary productivity in lowland tropical forests, yet there is little comparable information for the soil microbial biomass. We assessed microbial nutrient limitation by quantifying soil microbial biomass and hydrolytic enzyme activities in a long-term nutrient addition experiment in lowland tropical rain forest in central Panama. Multiple measurements were made over an annual cycle in plots that had received a decade of nitrogen, phosphorus, potassium, and micronutrient addition. Phosphorus addition increased soil microbial carbon (13 %), nitrogen (21 %), and phosphorus (49 %), decreased phosphatase activity by ~65 % and N-acetyl β-glucosaminidase activity by 24 %, but did not affect β-glucosidase activity. In contrast, addition of nitrogen, potassium, or micronutrients did not significantly affect microbial biomass or the activity of any enzyme. Microbial nutrients and hydrolytic enzyme activities all declined markedly in the dry season, with the change in microbial biomass equivalent to or greater than the annual nutrient flux in fine litter fall. Although multiple nutrients limit tree productivity at this site, we conclude that phosphorus limits microbial biomass in this strongly-weathered lowland tropical forest soil. This finding indicates that efforts to include enzymes in biogeochemical models must account for the disproportionate microbial investment in phosphorus acquisition in strongly-weathered soils.     

培养法和免培养法联合检测油藏环境烃降解菌和产甲烷菌群多样性

烃降解菌和产甲烷菌是油藏环境微生物生态系统中重要的功能菌群, 采用DGGE和FISH方法分析了不同油藏样品中两类菌群的多样性和产甲烷活性。DGGE结果表明, 不同水样的alkB基因多样性相差较大, 而且注水井条带明显多于采油井。FISH结果表明, 油藏水样中产甲烷菌含量明显高于烃降解菌, 且两者空间分布的位置较近; 说明油藏环境中烃降解菌和产甲烷菌结成一定的相互关系。富集培养表明, 胜利油田产出液接种物培养130 d后, 石油烃降解率达到50%以上, 产甲烷的最大速率达到1.57×10^?2 mmol/(L?d)。利用分子生物学方法分析油藏环境功能菌群的多样性, 可以为开展微生物采油技术的应用提供有用信息。     

Nutrient Input and Carbon and Microbial Dynamics in an Ombrotrophic Bog

Slow rates of plant production and decomposition in ombrotrophic bogs are believed to be partially the result of low nutrient availability. To test the effect of nutrient availability on decomposition, carbon dioxide (CO₂) flux dynamics, microbial biomass, and nutrients, we added nitrogen (N) with phosphorus (P) and potassium (K), to prevent limitation of the latter 2 nutrients, over 2 growing seasons to plots at Mer Bleue peatland, Ontario, Canada. After the first growing season, increasing N fertilization (with constant P and K) decreased in vitro CO₂ production potential and increased microbial biomass measured with a chloroform fumigation-extraction technique in the upper peat profile, while by the end of the second season, CO₂ production potential was increased in response to N plus PK treatment, presumably due to more easily decomposable newly formed plant material. In situ CO₂ fluxes measured using chamber-techniques over the second year corroborated this presumption, with greater photosynthetic CO₂ uptake and ecosystem respiration (ER) during high N plus PK treatments. The more efficient microbial community, with slower CO₂ production potential and larger biomass, after the first year was characterized by larger fungal biomass measured with signature phospholipid fatty acids. The majority of N was likely quickly sequestered by the vegetation and transferred to dissolved organic forms and microbial biomass in the upper parts of the peat profile, while additional P relative to controls was distributed throughout the profile, implying that the vegetation at the site was N limited. However, in situ CO₂ flux data suggested the possibility of P or NPK limitation. We hypothesize that nutrient deposition may lead to enhanced C uptake by altering the microbial community and decomposition, however this pattern disappears through subsequent changes in the vegetation and production of more readily decomposable plant tissues.     

Nutrient-limited biodegradation of PAH in various soil strata at a creosote contaminated site

The effects of nutrient addition on the in situ biodegradation of polycyclic aromatic hydrocarbons in creosote contaminated soil were studied in soil columns taken from various soil strata at a wood preserving plant in Norway. Three samples were used: one from the topsoil (0–0.5 m), one from an organic rich layer (2–2.5 m) and one from the sandy aquifer (4.5–5 m). The addition of inorganic nitrogen and phosphorous stimulated the degradation of polycyclic aromatic hydrocarbons (PAHs) in the top soil and the aquifer sand. These two soils, which differed strongly in contamination levels, responded similarly to nutrient addition with the corresponding degradation of 4-ring PAHs. The ratio between available nitrogen (N) and phosphorous (P) might explain the degree of degradation observed for the 4-ring PAHs. However, the degree of degradation of 3-ring PAHs did not significantly increase after nutrient addition. An increase in the respiration rate, after nutrient addition, could only be observed in the topsoil. In the aquifer sand, 4-ring PAH degradation was not accompanied by an increase in the respiration rate or the number of heterotrophic micro-organisms. PAH degradation in the organic layer did not respond to nutrient addition. This was probably due to the low availability of the contaminants for micro-organisms, as a result of sorption to the soil organic matter. Our data illustrate the need for a better understanding of the role of nutrients in the degradation of high molecular weight hydrocarbons for the successful application of bioremediation at PAH contaminated sites.     

辽西北沙地苹果-大豆间作对土壤养分和微生物量分布的影响

为了解辽西北沙地果农间作系统中土壤养分及微生物量分布特征,选取研究区具有代表性的苹果(Malus pumila)-大豆(Glycine max)间作系统为研究对象,对间作系统0~60 cm土层、0~300 cm水平距离范围内的土壤养分和微生物量进行了测定,并与大豆单作、苹果单作进行对比。结果表明:辽西北沙地苹果与大豆间作系统中土壤养分十分匮乏,全氮、碱解氮、有效磷极缺乏;在水平方向上,随距苹果树距离的增加,间作系统中土壤有机质、碱解氮、全磷、有效磷总体上呈现先降低再升高后趋于平稳的变化规律;各养分竞争激烈区位于果树带区[0,100 cm);在垂直方向上,苹果与大豆间作系统各土壤养分均表现出表聚性,表土层(0~20 cm)土壤养分含量显著高于深土层(20~60 cm);土壤微生物量碳、氮在果树带区及表土层含量高,且在该区域细菌数量高;间作与单作各土壤养分、微生物量在表土层差异较大,差异性随土层加深而缩小,间作系统除碱解氮外对土壤养分表现为负效应,不能满足间作系统对土壤养分的需求,尤其对土壤氮素、磷素竞争需求量较大。综上所述,辽西北沙地苹果-大豆间作系统中存在明显的竞争关系,竞争激烈区域位于果树带区,应在此处加强氮肥和磷肥施加。     

Intrinsic bioremediation in a solvent-contaminated alluvial groundwater

An industrial site contaminated with a mixture of volatile organic compounds in its subsurface differed from previously reported locations in that the contamination consisted of a mixture of chlorinated, brominated, and non-halogenated aromatic and aliphatic solvents in an alluvial aquifer. The source area was adjacent to a river. Of the contaminants present in the aquifer, benzene, toluene, and chlorobenzene (BTC) were of primary concern. Studies of the physical, chemical, and microbiological characteristics of site groundwater were conducted. The studies concentrated on BTC, but also addressed the fate of the other aquifer VOCs. Gas chromatographic analyses performed on laboratory microcosms demonstrated that subsurface microorganisms were capable of BTC degradation. Mineralization of BTC was demonstrated by the release of ¹⁴CO₂ from radiolabelled BTC. In the field, distribution patterns of nutrients and electron acceptors were consistent with expression of in situ microbial metabolic activity: methane, conductivity, salinity and o-phosphate concentrations were all positively correlated with contaminant concentration; while oxidation-reduction potential, nitrate, dissolved oxygen and sulfate concentrations were negatively correlated. Total aerobes, aerotolerant anaerobes, BTC-specific degraders, and acridine orange direct microscopic microorganism counts were strongly and positively correlated with field contaminant concentrations. The relative concentrations of benzene and toluene were lower away from the core of the plume compared to the less readily metabolized compound, chlorobenzene. Hydrodynamic modeling of electron-acceptor depletion conservatively estimated that 450 kg of contaminant have been removed from the subsurface yearly. Models lacking a biodegradation term predicted that 360 kg of contaminant would reach the river annually, which would result in measurable contaminant concentrations. River surveillance, however, has only rarely detected these compounds in the sediment and then only at trace concentrations. Thus, the combination of field modeling, laboratory studies, and site surveillance data confirm that significant in situ biodegradation of the contaminants has occurred. These studies establish the presence of intrinsic bioremediation of groundwater contaminants in this unusual industrial site subsurface habitat. Received 01 December 1995/ Accepted in revised form 27 July 1996     

Bacterial community analysis of shallow groundwater undergoing sequential anaerobic and aerobic chloroethene biotransformation

At Department of Energy Site 300, beneficial hydrocarbon cocontaminants and favorable subsurface conditions facilitate sequential reductive dechlorination of trichloroethene (TCE) and rapid oxidation of the resultant cis-dichloroethene (cis-DCE) upon periodic oxygen influx. We assessed the geochemistry and microbial community of groundwater from across the site. Removal of cis-DCE was shown to coincide with oxygen influx in hydrocarbon-containing groundwater near the source area. Principal component analysis of contaminants and inorganic compounds showed that monitoring wells could be differentiated based upon concentrations of TCE, cis-DCE, and nitrate. Structurally similar communities were detected in groundwater from wells containing cis-DCE, high TCE, and low nitrate levels. Bacteria identified by sequencing 16S rRNA genes belonged to seven phylogenetic groups, including Alpha-, Beta-, Gamma- and Deltaproteobacteria, Nitrospira, Firmicutes and Cytophaga-Flexibacter-Bacteroidetes (CFB). Whereas members of the Burkholderiales and CFB group were abundant in all wells (10(4)-10(9) 16S rRNA gene copies L(-1)), quantitative PCR showed that Alphaproteobacteria were elevated (>10(6) L(-1)) only in wells containing hydrocarbon cocontaminants. The study shows that bacterial community structure is related to groundwater geochemistry and that Alphaproteobacteria are enriched in locales where cis-DCE removal occurs.