In situ bioremediation of trichloroethylene-contaminated water by a resting-cell methanotrophic microbial filter

Abstract

An in situ microbial filter technology is being tested and developed for remediating migrating subsurface plumes contaminated with low concentrations of trichloroethylene (TCE). The current focus is the establishment of a replenishable bioactive zone (catalytic filter) along expanding plume boundaries by the injection of a representative methanotrophic bacterium, Methylosinus trichosporium OB3b. This microbial filter strategy has been successfully demonstrated using emplaced, attached resting cells (no methane additions) in a 1.1 m flow-through test bed loaded with water-saturated sand. Two separate 24 h pulses of TCE (109 ppb and 85 ppb), one week apart, were pumped through the system at a flow velocity of 15 mm h^?1; no TCE (< 0.5 ppb) was detected on the downstream side of the microbial filter. Subsequent excavation of the wet sand confirmed the existence of a TCE-bioactive zone 21 days after it had been created. An enhanced longevity of the cellular, soluble-form methane monooxygenase produced by this methanotroph is a result of the laboratory bioreactor culturing conditions. Additional experiments with cells in sealed vials and emplaced in the 1.1 m test bed yielded a high resting-cell finite TCE biotransformation capacity of about 0.25 mg per mg of bacteria; this is suitable for a planned sand-filled trench field demonstration at a Lawrence Livermore National Laboratory site.     

In situ determination of hafnium isotopes from rutile using LA-MC-ICP-MS

The hafnium isotopic analysis using laser ablation has been widely conducted on Hf-rich minerals(zircon/baddeleyite/ calzirtite/eudialyte), however, little work has been reported on Hf-poor(100 ppm) minerals. This work presents a detailed procedure of in situ hafnium isotopic analysis from rutile using laser ablation multiple collector inductively coupled plasma mass spectrometry(LA-MC-ICP-MS). The rutile U-Pb dating reference material JDX shows homogeneous hafnium isotopic ratios, with 176Hf/177Hf=0.281795±0.000015(2SD, n=33) and 176Lu/177Hf=0.000018±0.000004(2SD, n=17) that suggest the possibility of using JDX as a new reference material hafnium isotopic measurement. We also measure hafnium isotopic compositions of other rutile U-Pb dating reference material(R10, Sugluk-4 and PCA-S207) and the 176Hf/177 Hf values are similar to previously reported results, which confirms that we can acquire accurate and precise hafnium isotopic compositions using our developed analytical protocol. We analyzed hafnium isotopic compositions and U-Pb ages of rutile in high-temperature and ultrahigh-temperature granulites from various terrains of the Khondalite Belt from the North China Craton, combined with zircon results in the same area, suggesting that the metamorphic evolution history of the granulite is much more complicated than previously thought.     

In situ enzyme activities of filamentous scum bacteria in municipal activated sludge wastewater treatment plants

In situ phosphatase, esterase, lipase, and β‐glucuronidase activities were investigated in filamentous scum bacteria by ELF (enzyme labeled fluorescence)‐technology. Microthrix parvicella exhibited significant activities for all four enzymes with lipase activity being the highest. In situ activities were considerably higher in activated sludge as compared to scum indicating M. parvicella growth to occur mainly in the sludge fraction. “Nocardioform actinomycetes” showed significant activities for phosphatase, esterase, and β‐glucuronidase, lipase activity was only moderate. Activities revealed to be similar for activated sludge and scum. As population densities of enzyme‐active actinomycetes were noticeably higher in scum they presumably find good growth conditions in the scum layer. Enzyme activities in Nostocoida limicola morphotypes were only low with no lipase activity being detected. Due to their phylogenetic diversity activity assignment should be combined with fluorescence in situ hybridization (FISH). Numerous type 0041/0675 and type 1851 filaments showed all but lipase activity. However, some of their attached bacteria revealed to be lipase‐active. Only a few morphotype 0092 filaments revealed phosphatase, esterase, and β‐glucuronidase activity. ELF investigations proved suitable for monitoring in situ filamentous activity. Present imponderabilities of Eikelboom morphotype phylogenetic affiliation are discussed.     

Spatial variability of in situ microbial activity: biotracer tests

Biotracer tests have been proposed as a means by which to characterize the in situ biodegradation potential for field-scale systems. In this study, field experiments were conducted at two sites to evaluate the utility of the biotracer method for characterizing the spatial variability of microbial activity. The first site is a mixed waste-contaminated surficial aquifer in Utah, and the second site is a chlorinated solvent-contaminated regional aquifer in Tucson, Arizona. Mass recovery of the biotracer decreased approximately linearly with increasing residence time for the Tucson site. Similar behavior was observed at the Utah site, except in the region adjacent to the injection zone, where percent recoveries were much lower than those predicted using a correlation determined using data collected downgradient of the injection zone. First-order biodegradation rate coefficients obtained from model calibration of the tracer data varied between 0.2 and 0.5/day for the Tucson site. For the Utah site, the values varied between 0.1 and 0.6/day downgradient of the injection wells, and between 0.7 and 2.6/day near the injection wells. Considering the large range over which biodegradation rate coefficients can vary, the rate coefficient exhibited relatively minimal spatial variability (factor of 2.5) for the Tucson site. Conversely, the spatial variability of the rate coefficient was an order of magnitude greater for the Utah site. These differences in variability are consistent with conditions associated with the respective sites. For example, the greater microbial activity observed in the vicinity of the injection wells for the Utah site is consistent with the biomass distribution determined from analysis of core samples, which shows larger bacterial cell densities for the region near the injection wells. These results illustrate the utility of biotracer tests for in situ characterization of microbial activity (e.g., biodegradation potential), including evaluation of potential spatial variability.     

Culture-dependent and independent analyses of subsurface microbial communities in oil-bearing strata of the Sagara oil reservoir

Abstract   Culture-dependent and independent methods were used to evaluate the microbial communities in cores collected at depths up to 200 m in oil-bearing and oil-free strata near the Sagara oil reservoir near Shizuoka, Japan. Direct microbial counts revealed much higher numbers (2.2 × 10⁴−7.9 × 10⁶/g) of microbes in the oil-bearing strata than in the oil-free zones, where counts were uniform at approximately 1.0 × 10⁴/g. Molecular taxonomic analyses via 16S rRNA gene sequence comparisons showed that the oil-free strata were dominated by members of the γ-Proteobacteria including Pseudomonas , Stenotrophomonas and Sphingomonas , whereas the oil-bearing strata were dominated by a single species closely related to Pseudomonas stutzeri . All archaeal clones were phylogenetically affiliated with the uncultured soil group in Crenarchaeota with the exception of a single phylotype that belonged to the genus Thermococcus . Culture-dependent analysis was carried out by most-probable-number culturing as well as direct plating to determine viable cell counts, using both complex organic substrates or native oil and autotrophic media. Both culture-dependent and independent methods revealed the abundant cultivable member was the aerobic oil-degrading bacterium Pseudomonas stutzeri and neither autotrophs nor anaerobic heterotrophs could be detected in the oil-bearing strata.     

In situ stress state in Mainland China

In situ stress measurements are one of the important methods for studying the recent tectonic stress field and stress state. In this paper, the stress state and variation of stresses with depth in Mainland China are presented and analyzed on the basis ofin situ stress data measured by both hydrofracturing and overcoring techniques.     

In situ bioventing of a diesel fuel spill

Abstract

An estimated 50 000 1 of diesel fuel contaminated soil over a depth of 1.5 to 3.5 m (approximately 1.5% by weight in the soil) following fuel recovery operations at the site of a diesel spill. Laboratory treatability identified oxygen supply treatment as having significant potential to enhance bioremediation of the soil in situ. A bioventing system was designed and tested on a quarter of the site. Venting alone over a period of six months reduced total hydrocarbon concentrations by 10 to 30% to a depth of 3 m. Venting with nutrient addition resulted in a further reduction of 30% over a subsequent 6 month period to the full depth of 3.5 m.     

In situ cosmogenic Be in olivines and pyroxenes

This study proposes an efficient new cleaning procedure for measuring in situ cosmogenic ¹⁰Be in olivines and pyroxenes. This chemical routine is specially designed to decontaminate the abundant meteoric ¹⁰Be from these minerals. The method was tested on mafic minerals from basaltic flows of Mt. Etna volcano and from Hawaiian flows and moraines. A sequential dissolution test shows that ¹⁰Be concentrations decrease with the number of cleaning steps until reaching a constant value. This is a necessary condition to demonstrate the efficiency of the method in properly decontaminating samples of meteoric ¹⁰Be. Moreover, cross-calibration with cosmogenic ³He measured within the same samples yielded a sea level high-latitude production rate of 4.5±0.4 at g⁻¹ a⁻¹ for cosmogenic ¹⁰Be in mafic minerals. This rate is within 1σ uncertainty of empirically or model-derived rates for ¹⁰Be on the same targets. Such concordance supports the consistency of the new method.     

In situ disdrometer calibration using multiple DSD moments

In situ calibration is a proposed strategy for continuous as well as initial calibration of an impact disdrometer. In previous work, a collocated tipping bucket had been utilized to provide a rainfall rate based ～11/3 moment reference to an impact disdrometer’s signal processing system for implementation of adaptive calibration. Using rainfall rate only, transformation of impulse amplitude to a drop volume based on a simple power law was used to define an error surface in the model’s parameter space. By incorporating optical extinction second moment measurements with rainfall rate data, an improved in situ disdrometer calibration algorithm results due to utilization of multiple (two or more) independent moments of the drop size distribution in the error function definition. The resulting improvement in calibration performance can be quantified by detailed examination of the parameter space error surface using simulation as well as real data.     

In situ local shock speed and transit shock speed

A useful index for estimating the transit speeds was derived by analyzing interplanetary shock observations. This index is the ratio of the in situ local shock speed and the transit speed; it is 0.6–0.9 for most observed shocks. The local shock speed and the transit speed calculated for the results of the magnetohydrodynamic simulation show good agreement with the observations. The relation expressed by the index is well explained by a simplified propagation model assuming a blast wave. For several shocks the ratio is approximately 1.2, implying that these shocks accelerated during propagation in slow-speed solar wind. This ratio is similar to that for the background solar wind acceleration.     

越冬浮游植物光合作用活性的原位研究

水体中浮游植物在越冬期间数量少,关于湖泊中越冬浮游植物如蓝藻、绿藻、硅藻等浮游植物的光合作用活性情况少见报道.本实验通过Phyto-PAM(Pulse Amplitude Modulation)对最大光量子产量Fv/Fm(可变荧光和最大荧光之比)和快速光曲线的测定,结合显微镜镜检结果,并在室内进行了硫酸铜处理以及鲁哥氏液处理实验,对太湖、巢湖和玄武湖处于越冬期间的浮游植物进行了原位光合作用活性的研究.显微镜镜检结果表明太湖和玄武湖水体中以绿藻和硅藻为主,而蓝藻很少,巢湖水体中有大量蓝藻、绿藻和硅藻.Phyto-PAM在对玄武湖和太湖水样原位测定时未检测出蓝藻,而绿藻和硅藻有光合作用活性,其Fv/Fm在0.5左右;对巢湖的水体检测到的蓝藻、绿藻、硅藻都具有光合作用活性,其中绿藻和硅藻的Fv/Fm在0.5左右,蓝藻的Fv/Fm在0.3左右;用硫酸铜处理实验在巢湖水样中发现水体仅有硅藻活性(Fv/Fm下降至0.2左右),而用鲁哥氏液处理后无任何藻类光合作用活性.上述实验结果表明,即使处于越冬期间,以较多数量出现在水体中的蓝藻、绿藻和硅藻仍然具有较高的光合作用活性.     

越冬浮游植物光合作用活性的原位研究

对太湖北岸贡湖和梅梁湾湖滨带(水-陆交错区)观测场中的主要水生植物群落的种类组成、生物量进行了测定,定量分析了其生长型和群落特征的季节变化,比较分析了上述两个湖滨带观测场的水质变化情况,并且在此基础上进一步探讨了影响湖滨带沉水植物生长的关键因素.结果表明:(1)贡湖观测场的荇菜(Nymphoides peltatum) 马来眼子菜(Potamogeton malaianus)群落由3种浮叶植物和3种沉水植物构成,共5种水生植物生长型.而梅梁湾观测场的菹草群落为单优群落,只有一种沉水植物菹草(Potamogeton crispus),其生长型为大眼子菜型.以上两个湖滨带观测场水生植物群落特征与人为活动的干扰(如水质污染、人工堤岸建设等)密切相关;(2)以上两湖湾的主要污染指标为氮、磷营养物,均已达到富营养水平,但贡湖观测场的水质(除透明度外)明显好于梅梁湾观测场;(3)透明度、水深和风浪可能是影响贡湖和梅梁湾湖滨带沉水植物种群数量、分布和格局的重要因素.     

In situ tests simulating traffic-load-induced settlement of alluvial silt subsoil

In China, The Yellow River delta is the youngest large river delta, and the low liquid limit alluvial silt is widely distributed there. The silt is easy to liquefy so that the silt subsoil shows large settlement under traffic load. At present, few in situ model tests were conducted to study the traffic-load-induced settlement of silt subsoil. Therefore, a falling-weight simulation equipment of traffic load was developed. By adjusting the technical parameters such as the falling height of the weight, different types of traffic loads can be well simulated. With the equipment, in situ tests were carried out to study cumulative settlement of silt subsoil in the Yellow River delta. Tests indicate that the settlement and excess pore water pressures rapidly grow initially and then tend to be stable with increasing the number of load cycles, and they also increase with the magnitude of the traffic load. When the load attains a threshold value, liquefaction takes place in the silt subsoil. After terminating loading, the pore water pressure rapidly decreases and the settlement increases simultaneously, while after one hour they tend to stabilize. Based on Chai–Miura cumulative deformation model of soil, the traffic-load-induced cumulative settlement of silt subsoil was calculated and compared with the test results. The calculated cumulative settlement with increasing number of load cycles agrees well with the test results, except the initial phase of cyclic loading where the settlement observed in the situ tests is overestimated. This is mainly because Chai–Miura model assumes undrained conditions while the subsoil under traffic loads is partly drained.     

In situ rates of sulfate reduction in response to geochemical perturbations

Rates of in situ microbial sulfate reduction in response to geochemical perturbations were determined using Native Organism Geochemical Experimentation Enclosures (NOGEEs), a new in situ technique developed to facilitate evaluation of controls on microbial reaction rates. NOGEEs function by first trapping a native microbial community in situ and then subjecting it to geochemical perturbations through the introduction of various test solutions. On three occasions, NOGEEs were used at the Norman Landfill research site in Norman, Oklahoma, to evaluate sulfate-reduction rates in wetland sediments impacted by landfill leachate. The initial experiment, in May 2007, consisted of five introductions of a sulfate test solution over 11 d. Each test stimulated sulfate reduction with rates increasing until an apparent maximum was achieved. Two subsequent experiments, conducted in October 2007 and February 2008, evaluated the effects of concentration on sulfate-reduction rates. Results from these experiments showed that faster sulfate-reduction rates were associated with increased sulfate concentrations. Understanding variability in sulfate-reduction rates in response to perturbations may be an important factor in predicting rates of natural attenuation and bioremediation of contaminants in systems not at biogeochemical equilibrium.     

In situ experimental study of the evolution and recolonization of polluted sediments

When perturbing conditions influencing a benthic ecosystem improve, an extremely polluted sediment may very rapidly be colonized by a structured assemblage. This study was made after the implantation of modules containing chemically very contaminated sediment from a polluted biotope, in a lesser contaminated site. The colonization and dynamic processes of the studied mollusc populations do not depend on the concentrations of the solid phase pollutants but are related to the release of these pollutants in the interstitial water of sediments, which in turn is induced by ambiant condition variations and particularly by salinity.     

In situ uptake of mercury by the intertidal mussel,Mytilus californianus

The in situ uptake of mercury in tissues of Mytilus californianus was studied by exposing control specimens to waters at four depths near a submarine wastewater discharge for a period of 24 weeks. The response patterns observed for digestive gland, adductor muscle and gonadal tissues were strikingly different. Digestive gland concentrations increased rapidly to the heightened levels found for indigenous specimens. On the other hand, changes in the mercury content of adductor muscle were not statistically significant, and gonadal tissues generally exhibited a low rate of increase. No correlation with depth was observed.