Microbial Community Composition Near Depleted Uranium Impact Points

Military training activities can result in the deposition of depleted uranium (DU) into surface soils. Mechanisms of introduction include the generation of dust from firing and impact as well as the eventual corrosion of projectile fragments and unexploded ordnance. Microorganisms in surface soils have the potential to affect the transport of DU by direct binding of the metal to the cell surface, by altering near field soil chemistry that affects metal solubility, and by microbially influenced corrosion. We investigated the response (in terms of community composition) of a native soil microbiota to the presence of DU in an arid environment. Bacteria in soils outside of the test area were challenged in dilute media with “yellow cake” or U ₃ O ₈ . At concentrations of 200 to 20,000 mg L^{? 1} only species of Bacillus were identified. In situ characterizations (by PLFA analysis) of exposed site soils showed an enrichment in sulfate reducing bacterial (i17:1w7c up 39%) and Bacillus species (a15:0 up 35%) biomarkers. Three types of microbial communities were defined (as PLFA profiles) using exploratory statistics and related to three different levels of DU exposure. The community types were then statistically corresponded to site soil chemistry. Observed differences in site soil chemistry were attributed to munitions firing since enrichments (unexposed to exposed) were observed in the minerals magnesium (increase of ～ 18 mg kg^?1 ), potassium (increase of ～ 46 mg kg^?1 ) and sulfur (increase of ～ 12 mg kg^?1 ), all constituents of munitions residues. Increased concentrations of these minerals corresponded with a community type that was associated with an area of extensive DU round use.     

Utility of lipid biomarkers in support of bioremediation efforts at army sites

Lipid biomarker analysis has proven valuable in testing the hypothesis that attributes of the extant microbiota can directly reflect the occurrence of contaminant biodegradation. Two past research efforts have demonstrated this utility and are described here. A 4.5 m vertical core was obtained from a diesel fuel oil contamination plume. Core material was assayed for total petroleum hydrocarbons (TPH) and bacterial membrane phospholipids (PLFA) via a single solvent extraction. Microbial viable biomass and the relative abundance of Gram-negative bacterial PLFA biomarkers were found to be significantly correlated with TPH concentration. The core TPH profile also revealed two distinct areas where the average TPH level of 3,000 microg g(-1) fell to near detection limits. Both areas were characterized by a three-fold decrease in the hexadecane/pristane ratio, indicating alkane biodegradation, and a distinct PLFA profile that showed a close similarity to the uncontaminated surface soil. Low-order, incomplete detonations can deposit hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) into training range surface soils. Since surface soils are exposed to temporal and diurnal moisture cycles, we investigated the effect two very different soil moisture tensions had on the in situ microbiota and RDX biodegradation. Saturated soils were characterized by rapid RDX biodegradation, 4 day half-life, a decrease in number of species detected and increase in PLFA biomarkers for Gram-negative proteobacteria (n16:1omega7c, n18:1omega9c, and n18:1omega7c) and Gram-positive firmicutes (i15:0 and a15:0). Terminal restriction fragment length polymorphism (T-RFLP) profiles of endpoint microbial communities indicated a shift from 18 to 36% firmicutes, the loss of gamma-proteobacteria and the emergence of alpha-proteobacteria. These two past research efforts demonstrated the utility of the lipid biomarker analysis in identifying microbial community characteristics that were associated with two very different soil contaminants. Lipid biomarkers defined areas of TPH biodegradation and identified community shifts as a result of soil conditions that affected explosives fate. Information like this can be used to enhance the predictive power of ecological models such as the Army Training and Testing Area Carrying Capacity for munitions model [ATTACC].     

Long-Term Explosive Contamination in Soil: Effects on Soil Microbial Community and Bioremediation

Explosive contamination in soil is a great concern for environmental health. Following 50 years of munitions manufacturing and loading, soils from two different sites contained ≥ 6,435 mg 2,4,6-trinitrotoluene (TNT), 2,933 mg hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,135 mg octahydrol-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) kg^{? 1} soil. Extractable nitrate-N was as high as 315 and ammonium-N reached 150 mg N kg^{? 1} soil. Water leachates in the highly contaminated soils showed near saturation levels of TNT and RDX, suggesting great risk to water quality. The long-term contamination resulted in undetectable fungal populations and as low as 180 bacterial colony forming units (CFU) g^–1 soil. In the most severely contaminated soil, dehydrogenase activity was undetectable and microbial biomass carbon was very low (< 3.4 mg C _mic kg^–1 soil). The diminished biological activity was a consequence of long-term contamination because short-term (14 d) contamination of TNT at up to 5000 mg TNT kg^–1 soil did not cause a decline in the culturable bacterial population. Natural attenuation may not be a feasible remediation strategy in soils with long-term contamination by high concentrations of explosives.     

Effects of heavy metal contamination of soils on micronucleus induction in Tradescantia and on microbial enzyme activities: a comparative investigation

The aim of this study was to investigate correlation between genotoxic effects and changes of microbial parameters caused by metal contamination in soils. In total, 20 soils from nine locations were examined; metal contents and physicochemical soil parameters were measured with standard methods. In general, a pronounced induction of the frequency of micronuclei (MN) in the Tradescantia micronucleus (Trad-MN) assay was seen with increasing metal concentration in soils from identical locations. However, no correlations were found between metal contents and genotoxicity of soils from different locations. These discrepancies are probably due to differences of the physicochemical characteristics of the samples. Also, the microbial parameters depended on the metal content in soils from identical sampling locations. Inconsistent responses of the individual enzymes were seen in soils from different locations, indicating that it is not possible to define a specific marker enzyme for metal contamination. The most sensitive microbial parameters were dehydrogenase and arylsulfatase activity, biomass C, and biomass N. Statistical analyses showed an overall correlation between genotoxicity in Tradescantia on the one hand and dehydrogenase activity, biomass C, and the metabolic quotient on the other hand. In conclusion, the results of the present study show that the Trad-MN assay is suitable for the detection of genotoxic effects of metal contamination in soils and furthermore, that the DNA-damaging potential of soils from different origin cannot be predicted on the basis of chemical analyses of their metal concentrations.     

Enhanced transformation of TNT by Arabidopsis plants expressing an old yellow enzyme

2,4,6-Trinitrotoluene (TNT) is released in nature from manufacturing or demilitarization facilities, as well as after the firing or detonation of munitions or leakage from explosive remnants of war. Environmental contamination by TNT is associated with human health risks, necessitating the development of cost-effective remediation techniques. The lack of affordable and effective cleanup technologies for explosives contamination requires the development of better processes. In this study, we present a system for TNT phytoremediation by overexpressing the old yellow enzyme (OYE3) gene from Saccharomyces cerevisiae. The resulting transgenic Arabidopsis plants demonstrated significantly enhanced TNT tolerances and a strikingly higher capacity to remove TNT from their media. The current work indicates that S. cerevisiae OYE3 overexpression in Arabidopsis is an efficient method for the phytoremoval and degradation of TNT. Our findings have the potential to provide a suitable remediation strategy for sites contaminated by TNT.     

Sites of contamination and numbers of Erwinia carotovora present in stored seed potato stocks in Scotland

Forty-eight bulk and crate stored commercial seed potato stocks from the east of Scotland were examined for contamination by Erwinia carotovora in 1967 and 1968. Both E. carotovora var. carotovora and E. carotovora var. atroseptica were present, the latter, the blackleg pathogen, being detected four times more frequently. Most contamination was located on the tuber surface and in the lenticels; it was rare in the vascular ring and the organism was never detected in the cortex. The level of contamination of lenticels was only slightly affected after winter storage, which, however, substantially reduced surface contamination. More than 10³ cells were detected on the surface of tubers in over half the stocks and in half the stocks each lenticel sampled contained more than 10² bacteria.     

Adsorption of RDX to Soil with Low Organic Carbon: Laboratory Results,Field Observations,Remedial Implications

The purpose of this article is to review both laboratory and field observations of RDX adsorption to soils and to use those results to estimate the effects of a planned remedial action. Adsorption isotherms for RDX are generally observed to be linear and reversible. Statistical tests were performed to determine the relationship between K_d and various soil characteristics. A linear relationship between Kd and soil organic carbon was observed, as expected, but regression of K_d to organic carbon content indicated a non-zero intercept, suggesting that other sorbents may also be significant at low OC (e.g., > 0.5 %). No other soil properties were significantly related to Kd so the mechanism of adsorption at low organic carbon was not determined. These results were used to interpret observations of RDX in the vadose zone at Milan Army Ammunition Plant (MAAP), TN. MAAP exhibits widespread soil contamination by RDX. Depth to groundwater ranges from 40 to 80?ft. Unsaturated soils are fine grained near the surface, and sandy near the water table. RDX is concentrated in the upper 2?ft, where concentrations in some places exceed 1 %. Subsurface concentrations are generally less than 50?mg/kg. The distribution of RDX in soil, soil moisture and groundwater, and soil physical testing data were interpreted using simple models. The distribution of RDX is consistent with the following conceptual model: ??Water containing RDX was dis charged to the land surface (prior to 1983); ??Crystalline RDX remains in surface soil (remedial activities are ongo ing); ??Infiltrating rainwater leaches RDX from surface soils; ??This leachate carries RDX through the deeper vadose zone, resulting in significant soil contamination through out the full thickness of the vadose zone; these soils can generate leachate and adversely affect ground- water quality for many years to come. Field results were consistent with the adsorption studies. Simple models consistent with the field and laboratory observations indicate that deeper soils that are not planned to be remediated may continue to leach unacceptable concentrations to groundwater for approximately 180 years. The Army intends to evaluate whether it will be most cost-effective to address this continuing source by treating soils or groundwater.     

Proposal for an integrated approach to microbial environmental monitoring in cultural heritage: experience at the Correggio exhibition in Parma

Microbial environmental monitoring represents one of the most useful methods to assess potential risks related to the integrity of cultural heritage and people’s health. The monitoring plan described in the present work is based on standardized techniques for measuring microbial air and surface contamination. Air contamination is assessed through both active and passive samplings, measuring the concentration of microbes in air (in colony forming units per cubic metre, CFU/m³) and the rate at which microorganisms settle on surfaces (expressed by the Index of Microbial Air Contamination, IMA, CFU/dm²/h). For surface contamination, two parameters are measured using nitrocellulose membranes: the Microbial Buildup (MB, the total number of microorganisms accumulated on a surface in an unknown period of time prior to the sampling) and the Hourly Microbial Fallout (HMF, the number of microorganisms that settle on a specific surface during 1 h). The monitoring plan was implemented at the Pilotta Palace in Parma, Italy, during the Correggio exhibition in 2009. Samplings were taken before and during opening times. Some microbial contamination was already detected before the arrival of visitors: air contamination mean values of 99.1 CFU/m³ and 5.2 CFU/dm²/h were recorded, while MB and HMF mean values for surfaces were 92 and 7 CFU/dm², respectively. A significant increase was recorded in air contamination during opening times, with mean values of 323.7 CFU/m³ and 19.4 CFU/dm²/h; surface contamination values increased as well. This monitoring plan represents a contribution towards the definition of a much needed standardized methodology.     

Natural rhizobial populations and nodulation status of woody legumes growing in diverse Kenyan conditions

The rhizobial populations and nodulation status of both indigenous (mainly Acacia species) and some introduced woody legume species were assessed under glasshouse conditions in soils collected from 12 sites located in different ecological zones of Kenya. The populations among the sites, as estimated by the MPN technique, varied from <3.6 to>2.3×10⁵ cells g^-1 of soil. There were some intrasite variations in population estimates depending on the trap host species, date of soil collection and the method used in sampling the soils. Nodulation in whole soil also varied across the sites with test species frequently showing higher nodulation ability in native soils. Sesbania sesban (L.) Merr. was the most prolific nodulating species while Acacia tortilis (Forsskal) Hayne was very erratic in nodulation. Nodulation of most species showed interplant and intraspecific variability within a single soil source.     

PAH Degradation Capacity of Soil Microbial Communities—Does It Depend on PAH Exposure?

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants of the environment. But is their microbial degradation equally wide in distribution? We estimated the PAH degradation capacity of 13 soils ranging from pristine locations (total PAHs ≈ 0.1 mg kg^?1) to heavily polluted industrial sites (total PAHs ≈ 400 mg kg^?1). The size of the pyrene- and phenanthrene-degrading bacterial populations was determined by most probable number (MPN) enumeration. Densities of phenanthrene degraders reflected previous PAH exposure, whereas pyrene degraders were detected only in the most polluted soils. The potentials for phenanthrene and pyrene degradation were measured as the mineralization of ¹⁴C-labeled spikes. The time to 10% mineralization of added ¹⁴C phenanthrene and ¹⁴C pyrene was inversely correlated with the PAH content of the soils. Substantial ¹⁴C phenanthrene mineralization in all soils tested, including seven unpolluted soils, demonstrated that phenanthrene is not a suitable model compound for predicting PAH degradation in soils. ¹⁴C pyrene was mineralized by all Danish soil samples tested, regardless of whether they were from contaminated sites or not, suggesting that in industrialized areas the background level of pyrene is sufficient to maintain pyrene degradation traits in the gene pool of soil microorganisms. In contrast, two pristine forest soils from northern Norway and Ghana mineralized little ¹⁴C pyrene within the 140-day test period. Mineralization of phenanthrene and pyrene by all Danish soils suggests that soil microbial communities of inhabited areas possess a sufficiently high PAH degradation capacity to question the value of bioaugmentation with specific PAH degraders for bioremediation.     

Fate and Transport of High Explosives in a Sandy Soil: Adsorption and Desorption

Several areas of the Massachusetts Military Reservation (MMR) have soils with significant levels of high explosives (HE) contamination because of a long history of training and range activities (such as open burning, open detonation, disposal, and artillery and mortar firing). Site-specific transport and attenuation mechanisms were assessed in sandy soils for three contaminants of concern: the nitramine hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and the nitroaromatics 2,4-dinitrotolune (2,4-DNT) and 2,4,6-trinitrotoluene (TNT). For all three contaminants, linear distribution coefficients (K_d) were dependent on the fraction of organic carbon in soil. The nitroaromatics sorbed much more strongly than RDX in both soils. Over 120 hours, the desorption rate of RDX from field contaminated surface soil was much slower than its sorption rate, with the desorption K_d (1.5 L/kg) much higher than K_d for sorption (0.37 L/kg). Desorption of 2,4-DNT was negligible over 120 hours. Thus, applying sorption-derived K_d values for transport modeling may significantly overestimate the flux of explosives from MMR soils. Based on multiple component column transport tests, RDX will be the most mobile of these contaminants in MMR soils. In saturated columns packed with uncontaminated soil, RDX broke through rapidly, whereas the nitroaromatics were significantly attenuated by irreversible sorption or abiotic transformations.     

三亚市果蔬地重金属空间分布及其污染评价

通过野外调查采样,采用标准对比法和地累积指数法,评价了三亚市果蔬地表层土壤重金属的污染状况。结果表明:土壤中Hg、Cd、Cr、Pb和As5种重金属的积累不显著,其平均值均未超过国家环境质量二级标准(GB15618-1995);但Hg、Cd和Pb的平均值高于海南岛农用地的自然背景值,表明存在区域轻度污染;Cr、Hg、Pb和As4种重金属元素的变异系数均>1,表明土壤中Cr、Hg、Pb和As的分布极不均匀。同时,利用地理信息系统(GIS)Krging插值方法得到了5种重金属的空间分布图,显示出不同重金属元素的空间分异特征,为三亚市果蔬地的可持续开发利用提供指导。     

Evaluation of Soil Contamination by Polycyclic Aromatic Hydrocarbons in Gipuzkoa (Northern Spain)

Representative polycyclic aromatic hydrocarbons (PAHs) of low-medium molecular weight were determined using headspace solid-phase microextraction and gas chromatography with a flame ionization detector (HS-SPME-GC-FID) in ten surface soil samples from Gipuzkoa (Northern Spain). The sum of the PAHs ranged from 0.21 to 136.26 mg kg^?1. Pyrene and chrysene were the most abundant detected PAHs with an average concentration around 3.1 mg kg^?1. Pearson's correlation and PAH isomer ratios were applied to study the different origins of contamination. The results indicated that the PAH contamination in the studied area was a mixed pattern of pyrolytic and petrogenic inputs. Multivariate exploratory techniques, principal component analysis (PCA), and cluster analysis (CA) were also applied corroborating the PAH compounds patterns in the soils.     

Microbiological Contamination of Hospital Air: I. Quantitative Studies

The levels of airborne contamination in various areas of two hospitals were determined during a 15-month sampling period, using Casella and Andersen volumetric samplers. Based on nearly 5,000 samples, the mean count per ft³ ranged from 4.5 in obstetric-gynecology delivery rooms to 72.4 in waste-handling areas. The mean count for the entire hospital environment was on the order of 20 contaminants per ft³; 48% were associated with particles >5 μ diam, 30% with particles between 2 and 6 μ diam, and 22% with particles <2 μ diam. The airborne contamination was influenced by traffic, activity, ventilation considerations, and gross surface contamination, but not markedly by seasonal changes. When suitable control measures were implemented, the level of contamination could be diminished and kept low.     

Changes in soil Acidobacteria communities after 2,4,6-trinitrotoluene contamination

Despite their widespread occurrence in soils, the ecology of Acidobacteria and their response to environmental perturbations due to human activities remain very poorly documented. This study was aimed at assessing changes in the diversity and abundance of Acidobacteria in soils contaminated with 2,4,6-trinitrotoluene (TNT) compared with nonpolluted soils. The analysis of Acidobacteria communities at two sites with long-term and short-term contamination revealed that TNT has a drastic impact on the relative abundance of Acidobacteria in soil bacterial 16S rRNA gene libraries. The disappearance of most Acidobacteria from these soils was concomitant with a shift in Acidobacteria community composition and a loss of diversity, although the extent of diversity erosion depended on the sampling site.     

Dissemination and survival of non-indigenous bacterial genomes in pristine Antarctic environments

Continental Antarctic is perceived as a largely pristine environment, although certain localized regions (e.g., parts of the Ross Dependency Dry Valleys) are relatively heavy impacted by human activities. The procedures imposed on Antarctic field parties for the handling and disposal of both solid and liquid wastes are designed to minimise eutrofication and contamination (particularly by human enteric bacteria). However, little consideration has been given to the significance, if any, of less obvious forms of microbial contamination resulting from periodic human activities in Antarctica. The predominant commensal microorganism on human skin, Staphylococcus epidermidis, could be detected by PCR, in Dry Valley mineral soils collected from heavily impacted areas, but could not be detected in Dry Valley mineral soils collected from low impact and pristine areas. Cell viability of this non-enteric human commensal is rapidly lost in Dry Valley mineral soil. However, S. epidermidis can persist for long periods in Dry Valley mineral soil as non-viable cells and/or naked DNA.     

Composite Sampling of a Bacillus anthracis Surrogate with Cellulose Sponge Surface Samplers from a Nonporous Surface

A series of experiments was conducted to explore the utility of composite-based collection of surface samples for the detection of a Bacillus anthracis surrogate using cellulose sponge samplers on a nonporous stainless steel surface. Two composite-based collection approaches were evaluated over a surface area of 3716 cm² (four separate 929 cm² areas), larger than the 645 cm² prescribed by the standard Centers for Disease Control (CDC) and Prevention cellulose sponge sampling protocol for use on nonporous surfaces. The CDC method was also compared to a modified protocol where only one surface of the sponge sampler was used for each of the four areas composited. Differences in collection efficiency compared to positive controls and the potential for contaminant transfer for each protocol were assessed. The impact of the loss of wetting buffer from the sponge sampler onto additional surface areas sampled was evaluated. Statistical tests of the results using ANOVA indicate that the collection of composite samples using the modified sampling protocol is comparable to the collection of composite samples using the standard CDC protocol (p  =  0.261). Most of the surface-bound spores are collected on the first sampling pass, suggesting that multiple passes with the sponge sampler over the same surface may be unnecessary. The effect of moisture loss from the sponge sampler on collection efficiency was not significant (p  =  0.720) for both methods. Contaminant transfer occurs with both sampling protocols, but the magnitude of transfer is significantly greater when using the standard protocol than when the modified protocol is used (p<0.001). The results of this study suggest that composite surface sampling, by either method presented here, could successfully be used to increase the surface area sampled per sponge sampler, resulting in reduced sampling times in the field and decreased laboratory processing cost and turn-around times.     

Identification of a Brevibacterium marker gene specific to poultry litter and development of a quantitative PCR assay

Aim: To identify a DNA sequence specific to a bacterium found in poultry litter that was indicative of faecal contamination by poultry sources. Methods and Results: Faecally contaminated poultry litter and soils were used as source material for the development of a quantitative polymerase chain reaction (qPCR) method targeting the 16S rRNA gene of a Brevibacterium sp. The identified sequence had 98% nucleotide identity to the 16S rRNA gene of Brevibacterium avium. The qPCR method was tested on 17 soiled litter samples; 40 chicken faecal samples; and 116 nontarget faecal samples from cattle, swine, ducks, geese, and human sewage collected across the United States. The 571‐bp product was detected in 76% of poultry‐associated samples, but not in 93% of faecal samples from other sources. Marker concentrations were 10⁷–10⁹ gene copies per gram in soiled litter, up to 10⁵ gene copies per gram in spread‐site soils, and 10⁷ gene copies per litre in field run‐off water. Results were corroborated by a blinded study conducted by a second laboratory. Conclusion: The poultry‐specific PCR product is a useful marker gene for assessing the impact of faecal contamination as a result of land‐applied poultry litter. Significance and Impact of the Study: This study describes the first quantitative, sensitive and specific microbial source tracking method for the detection of poultry litter contamination.     

城市土壤质量演变及其生态环境效应

城市化是人类活动影响下自然生态系统向人工生态系统的急剧变化形式,它引起一系列生态环境条件的改变。城市对土壤资源的数量和质量产生深刻的影响,包括土壤地表封闭引起的土壤生态功能彻底消失,土壤物理性质恶化,土壤形态学特征和演变过程深受人为作用的影响。短程变异增加而总体多样性降低,土壤污染加剧,土壤养分富集等多方面,城市过程中土地利用变化对土壤产生一系列生态环境效应,如养分径流输出增加,热缓冲能力下降,污染转移危险性上升等,有必要迅速开展城市化过程中土壤质量演变及其生态环境效应的系统研究,为我国城市可持续发展提供理论依据。     

Lime Treatment of Explosives-Contaminated Soil from Munitions Plants and Firing Ranges

Microcosms were prepared using soils from munitions plants and active firing ranges and treated with hydrated lime. The presence of particulate explosives and co-contaminants, and the concentration of soil total organic carbon (TOC) on the alkaline hydrolysis reaction were studied. Trinitrobenzene (TNB) and dinitrobenzene (DNB) were sensitive to alkaline hydrolysis under these experimental conditions. The TNT metabolites, 2A- and 4A-DNT, were also removed, although more slowly than the parent compound, and the reaction required a higher pH (>12). RDX retention in the soil was proportional to the TOC content. The degradation intermediates of the alkaline hydrolysis reaction partitioned in the soil matrix in a manner similar to the parent. Solid particles of explosives are also degraded by alkaline hydrolysis. RDX and HMX exhibited 74 and 57% removal, respectively, in 21 days. TNT, as whole and broken grains, showed 83 and 99.9% removal in 21 days, respectively. The propellants, 2,4- and 2,6-DNT, were insensitive to alkaline hydrolysis. Alkaline hydrolysis is an inexpensive and effective means of reducing the varied explosives contamination.