The effects of in situ chemical oxidation on microbiological processes: A review

The effects of in situ chemical oxidation (ISCO) on biological processes, as reported in the literature, were researched to determine if coupling ISCO with in situ bioremediation could be achieved in field and laboratory experiments. Literature was compiled concerning the effect of ISCO on microbial communities following addition of a chemical oxidant at a range of concentrations designed to treat a variety of subsurface contaminants. The results indicate that although microbial communities may potentially be adversely affected by chemical oxidation in the short term, a rebound of microbial biomass and/or bioremediation activity can be expected. Successfully coupling ISCO with bioremediation in field applications may be a cost‐effective method of achieving risk‐based site remediation goals. © 2006 Wiley Periodicals, Inc.     

Enhancing Reductive Dechlorination With Nutrient Addition

Enhanced reductive bioremediation (ERB) of halogenated organics has become widely accepted and implemented to reduce risks to human health and the environment posed by these compounds. The family of chlorinated ethenes (e.g., trichloroethene) is among the most common contaminants treated using ERB techniques. The number of microbial species able to fully dechlorinate ethenes is small, and many times their population numbers are low. As a result, many ERB practitioners have turned to the injection of designer bacteria within treatment zones as a means to accelerate the process. Otherwise, many remediation projects encounter “cis‐stall,'' or the accumulation of the cis‐1,2‐dichloroethene and vinyl chloride intermediates. This article discusses three unique case studies where the addition of a balanced macro‐ and micro‐nutrient source substantially accelerated the chlorinated ethene ERB process, independent of the electron‐donor substrate applied. © 2013 Wiley Periodicals, Inc.     

石油污染土壤的微生物修复技术

介绍了石油污染土壤微生物修复技术的影响因素;概述了生物刺激、生物强化、固定化微生物、植物-微生物联合修复以及电动-微生物联合修复石油污染土壤的技术原理,分析了现阶段土壤修复过程中面临的难题,预测了微生物修复技术的研究方向。指出优化微生物的环境条件、培育新型高效的基因工程菌和开发经济高效的新型修复技术等将是未来微生物修复技术的发展趋势。     

Use of a low‐cost substrate in a continuous recirculation process to stimulate plumewide anaerobic dechlorination of chlorinated solvents

Over the past 20 years, significant time and money have been spent on better understanding and successfully applying bioremediation in the field. The results of these efforts provide a deeper un‐derstanding of aerobic and anaerobic microbial processes, the microbial species and environ‐mental conditions that are desirable for specific degradation pathways, and the limitations that may prevent full‐scale bioremediation from being successfully applied in heterogeneous subsur‐face environments. Numerous substrates have been identified as effective electron donors to stimulate anaerobic dechlorination of chlorinated ethenes, but methods of delivering these sub‐strates for in situ bioremediation (direct‐push injections, slug injections, high‐pressure injections, fracture wells, etc.) have yet to overcome the main limitation of achieving contact between these substrates and the contaminants. Therefore, although it is important (from a full‐scale remedia‐tion standpoint) to select an appropriate, low‐cost substrate that can be supplied in sufficient quantity to promote remediation of a large source area and its associated plume, it is equally im‐portant to ensure that the substrate can be delivered throughout the impacted plume zone. Failure to achieve substrate delivery and contact within the chlorinated solvent plume usually re‐sults in wasted money and limited remediation benefit. Bioremediation is a contact technology that cannot be effectively implemented on a large scale unless a method for rapidly delivering the low‐cost substrate across the entire source and plume areas is utilized. Unfortunately, many cur‐rent substrate delivery methods are not achieving sitewide distribution or treatment of the sorbed contaminant mass that exists in the organic fraction of a soil matrix. The following discussion sum‐marizes substrate delivery using an aggressive groundwater recirculation approach that can achieve plumewide contact between the contaminants and substrate, thus accelerating dechlori‐nation rates and shortening the overall remediation time frame. © 2006 Wiley Periodicals, Inc.     

Potential and Limitations of Microbial Reductive Dechlorination for Bioremediation Applications

Current knowledge and recent advances in the area of microbial reductive dechlorination of polychlorinated organic compounds are summarized. Factors which may limit the efficacy of the dechlorination process for the in situ bioremediation of contaminated soil and sediment systems are identified. Results of recent studies on the anaerobic biotransformation of soil-sorbed chlorinated ethenes and sediment-sorbed chlorinated benzenes are provided to illustrate how low contaminant bioavailability may control the rate and extent of dechlorination in subsurface systems, especially those with long-term contamination. Use of nonionic, polysorbate surfactants as the sole electron donors of a mixed, methanogenic culture supported the microbial sequential reductive dechlorination of either free or sediment-bound hexachlorobenzene (HCB) to primarily 1,3-dichlorobenzene, but did not enhance the bioavailability of sediment-bound HCB as compared to microcosms, which used glucose. Because current knowledge on the interactions of dechlorinating populations with other microbial populations in the presence of alternative terminal electron acceptors (e.g., nitrate, Fe³⁺ , Mn⁴⁺) is limited, such interactions and their effect on the dechlorination process in subsurface systems need to be further explored to improve our understanding of the reductive dechlorination process in complex environmental systems and lead to the development of more efficient in situ bioremediation technologies and strategies.     

土壤氰化物污染生物修复技术研究进展

综述了植物修复、微生物修复和生物联合修复等土壤氰化物污染生物修复技术的降解机理、降解途径及降解影响因素的研究进展,探讨了氰化物生物修复技术的发展趋势和应用前景。指出基于提高修复时效和针对土壤复合污染类型的多技术融合研究、基于提高微生物耐受性和降解效率的菌株固定化及菌根真菌-植物联合技术研究以及基于工程化应用为导向的现场试验研究是未来研究的重点领域,为土壤氰化物污染的综合治理和修复提出了新思路。     

Microbially Mediated In Situ Desorption Accelerates Remediation of Large Gasoline Product Plume

Remediation of a large separate‐phase hydrocarbon product and associated dissolved‐phase gasoline plume was accelerated by coupling multiphase extraction with in situ microbial stimulation. At the beginning of remediation activities, the separate‐phase hydrocarbon plume extended an estimated seven acres with product thickness measuring up to 2.1 feet thick. Within 18 months after beginning extraction, reduction of gasoline constituents in groundwater became asymptotic and measureable product disappeared from the upgradient source area. At that time, the remediation team initiated a program of limited in situ anaerobic bioremediation with the goal of stimulating production of natural surfactants from native microbes to release petroleum from the soil matrix. Groundwater concentrations of gasoline constituents increased gradually over the next three years, improving recovery without biofouling the pump‐and‐treat infrastructure. Using this approach, the groundwater component of the remedy was completed in less than five years, substantially less than the 10 years to 15 years predicted by modeling. This strategy demonstrated a more sustainable approach to remediation, reducing electrical usage by an estimated 800 megawatt hours, reducing infrastructure requirements, and preserving an estimated 150 million gallons of groundwater for this arid agricultural area. © 2014 Wiley Periodicals, Inc.     

Pilot‐scale evaluation using bioaugmentation to enhance PCE dissolution at dover AFB national test site

An Interstate Technology and Regulatory Council (ITRC) forum was recently held that focused on six case studies in which bioremediation of dense nonaqueous‐phase liquids (DNAPLs) was performed; the objective was to demonstrate that there is credible evidence for bioremediation as a viable environmental remediation technology. The first two case studies from the forum have been previously published; this third case study involves a pilot‐scale demonstration that investigated the effects of biological activity on enhancing dissolution of an emplaced tetrachloroethene (PCE) DNAPL source. It used a controlled‐release test cell with PCE as the primary DNAPL in a porous media groundwater system. Both laboratory tests and a field‐scale pilot test demonstrated that bioaugmentation can stimulate complete dechlorination to a nontoxic end product and that the mass flux from a source zone increases when biological dehalorespiration activity is enhanced through nutrient (electron donor) addition and bioaugmentation. All project goals were met. Important achievements include demonstrating the ability to degrade a PCE DNAPL source to ethene and obtaining significant information on the impacts to the microbial populations and corresponding isotope enrichments during biodegradation of a source area. © 2007 Wiley Periodicals, Inc.     

Enhanced anaerobic bioremediation of a TCE source at the Tarheel Army Missile Plant using EOS

EOS, or emulsified oil substrate, was used to stimulate anaerobic biodegradation of trichloroethene (TCE) and tetrachloroethene (PCE) at a former Army‐owned manufacturing facility located in the Piedmont area of North Carolina. Previous use of chlorinated solvents at the facility resulted in soil and groundwater impacts. Ten years of active remediation utilizing soil vacuum extraction and air sparging (SVE/AS) were largely ineffective in reducing the TCE/PCE plume. In 2002, the Army authorized preparation of an amended Remedial Action Plan (RAP) to evaluate in situ bioremediation methods to remediate TCE in groundwater. The RAP evaluated eight groundwater remediation technologies and recommended EOS as the preferred bioremediation alternative for the site. Eight wells were drilled within the 100 × 100 feet area believed to be the primary source area for the TCE plume. In a first injection phase, dilute EOS emulsion was injected into half of the wells. Distribution of the carbon substrate through the treatment zone was enhanced by pumping the four wells that were not injected and recirculating the extracted water through the injection wells. The process was repeated in a second phase that reversed the injection/extraction well pairs. Overall, 18,480 pounds of EOS were injected and 163,000 gallons of water were recirculated through the source area. Anaerobic groundwater conditions were observed shortly after injection with a corresponding decrease in both PCE and TCE concentrations. Dissolved oxygen, oxidation‐reduction potential, and sulfate concentrations also decreased after injection, while TCE‐degradation products, ferrous iron, and methane concentrations increased. The reduction in TCE allowed the Army to meet the groundwater remediation goals for the site. Approximately 18 months after injection, eight wells were innoculated with a commercially prepared dechlorinating culture (KB‐1) in an attempt to address lingering cis‐1,2‐dichloroethene (cis‐DCE) and vinyl chloride (VC) that continued to be observed in some wells. Dehalococcoides populations increased slightly post‐bioaugmentation. Both cis‐DCE and VC continue to slowly decrease. © 2007 Wiley Periodicals, Inc.     

In-situ bioremediation of hanford groundwater

The U.S. Department of Energy has generated liquid wastes containing radioactive and hazardous chemicals throughout the more than forty years of operation at its Hanford site in Washington State. Many of the waste components, including nitrate and carbon tetrachloride (CCl₄), have been detected in the Hanford groundwater. In-situ bioremediation of CCl₄ and nitrate is being considered to clean the aquifer. Preliminary estimates indicate that this technology should cost significantly less than ex-situ bioremediation and about the same as air stripping/granular activated carbon. In-situ bioremediation has the advantage of providing ultimate destruction of the contaminant and requires significantly less remediation time. Currently, a test site is under development. A computer-aided design tool is being used to design optimal remediation conditions by linking subsurface transport predictions, site characterization data, and microbial growth and contaminant destruction kinetics.     

The use of enhanced bioremediation at the Savannah River Site to remediate pesticides and PCBs

Enhanced bioremediation is quickly developing into an economical and viable technology for the remediation of contaminated soils. Until recently, chlorinated organic compounds have proven difficult to bioremediate. Environmentally recalcitrant compounds, such as polychlorinated biphenyls (PCBs) and persistent organic pesticides (POPs) such as dichlorodiphenyl trichloroethane (DDT) have shown to be especially arduous to bioremediate. Recent advances in field‐scale bioremedial applications have indicated that biodegradation of these compounds may be possible. Engineers and scientists at the Savannah River Site (SRS), a major DOE installation near Aiken, South Carolina, are using enhanced bioremediation to remediate soils contaminated with pesticides (DDT and its metabolites, heptachlor epoxide, dieldrin, and endrin) and PCBs. This article reviews the ongoing remediation occurring at the Chemicals, Metals, and Pesticides (CMP) Pits using windrow turners to facilitate microbial degradation of certain pesticides and PCBs. © 2003 Wiley Periodicals, Inc.     

Recent applications of phytoremediation technologies

Although the application of microbe biotechnology has been successful with petroleum-based constituents, microbial digestion has met with limited success for widespread residual organic and metal pollutants located above the potentiometric surface. Vegetation-based remediation, on the other hand, shows potential for accumulating, immobilizing, and transforming low levels of persistent contamination from the subsurface. Agricultural bioremediation, called geobotany or phytoremediation, relies on the remediating abilities of contaminant-accumulating plants to remove contamination from soil or groundwater. In natural ecosystems, plants act to filter and metabolize substances generated by nature. Phytoremediation affirmatively applies this process to help clean up contamination created by artificial means. Plants have proven effective at remediating areas contaminated with organic chemical wastes such as petroleum products, solvents, wood preservatives, pesticides, and metals. Phytoremediation is not the best technology for every site but has shown success with lead, cadmium, zinc, and radionuclides. The phytoremediation process takes much longer than conventional methods to clean a site and is dependent upon the type and degree of contamination. Concentrations must be within a narrow range of tolerable levels and the presence of the contamination must be at the appropriate depth. Nevertheless, phytoremediation offers an effective alternative to conventional, engineered remedial plans that usually involve costly activities like excavation, treatment, and disposal of soil or pump-and-treat technologies for groundwater. Phytoremediation also seems to be a promising new technology for the treatment of stormwater, industrial wastewater, and sewage. The relative low costs of capital for start-up together with negligible operations and maintenance costs provide a strong incentive for further investigation and development of phytoremediation projects.     

A universal design approach for in situ bioremediation developed from multiple project sites

This article describes a design approach that has been developed for bioremediation of chlorinated volatile organic compound–impacted groundwater that is based upon experience gained during the past 17 years. The projects described in the article generally involve large‐scale enhanced anaerobic dechlorination (EAD) and combined aerobic/anaerobic bioremediation techniques. Our design approach is based on three primary objectives: (1) selecting and distributing the proper additives (including bioaugmentation) within the targeted treatment zone; (2) maintaining a neutral pH (and adding alkalinity when needed); and (3) sustaining the desired conditions for a sufficient period of time for the bioremediation process to be fully completed. This design approach can be applied to both anaerobic and aerobic bioremediation systems. Site‐specific conditions of hydraulic permeability, groundwater velocity, contaminant type and concentrations, and regulatory constraints will dictate the best remedial approach and design parameters for in situ bioremediation at each site. The biggest challenges to implementing anaerobic bioremediation processes are generally the selection and delivery of a suitable electron donor and the proper distribution of the donor throughout the targeted treatment zone. For aerobic bioremediation processes, complete distribution of adequate concentrations of a suitable electron acceptor, typically oxygen or oxygen‐yielding compounds such as hydrogen peroxide, is critical. These design approaches were developed based on understanding the biological processes involved and the mechanics of groundwater flow. They have evolved based on actual applications and results from numerous sites. An EAD treatment system, based on our current design approach, typically uses alcohol as a substrate, employs groundwater recirculation to distribute additives, and has an operational period of two to four years. An aerobic in situ treatment system based on our current design approach typically uses pure oxygen or hydrogen peroxide as an electron acceptor, may involve enhancements to groundwater flow for better distribution, and generally has an operational period of one to four years. These design concepts and specific project examples are presented for 17 sites. © 2012 Wiley Periodicals, Inc.     

Integrated approach to PCE‐impacted site characterization,site management,and enhanced bioremediation

Tetrachloroethene (PCE)‐ and trichloroethene (TCE)‐impacted sites pose significant challenges even when site characterization activities indicate that biodegradation has occurred naturally. Although site‐specific, regulatory, and economic factors play roles in the remedy‐selection process, the application of molecular biological tools to the bioremediation field has streamlined the assessment of remedial alternatives and allowed for detailed evaluation of the chosen remedial technology. The case study described here was performed at a PCE‐impacted site at which reductive dechlorination of PCE and TCE had led to accumulation of cis‐dichlorethene (cis‐DCE) with concentrations ranging from approximately 10 to 100 mg/L. Bio‐Trap® samplers and quantitative polymerase chain reaction (qPCR) enumeration of Dehalococcoides spp. were used to evaluate three remedial options: monitored natural attenuation, biostimulation with HRC®, and biostimulation with HRC‐S®. Dehalococcoides populations in HRC‐S‐amended Bio‐Traps deployed in impacted wells were on the order of 10³ to 10⁴ cells/bead but were below detection limits in most unamended and HRC‐amended Bio‐Traps. Thus the in situ Bio‐Trap study identified biostimulation with HRC‐S as the recommended approach, which was further evaluated with a pilot study. After the pilot HRC‐S injection, Dehalococcoides populations increased to 10⁶ to 10⁷ cells/bead, and concentrations of cis‐DCE and vinyl chloride decreased with concurrent ethene production. Based on these results, a full‐scale HRC‐S injection was designed and implemented at the site. As with the pilot study, full‐scale HRC‐S injection promoted growth of Dehalococcoides spp. and stimulated reductive dechlorination of the daughter products cis‐DCE and vinyl chloride. © 2008 Wiley Periodicals, Inc.     

A Novel Approach to Biological Treatment of Contaminated Water

Over the past few years, the focus of our research has been to respond to the recognized needs for novel biological processes that are capable of destroying a wide range of biodegradable pollutants and providing the perfect environment for complex interspecies interactions required for the degradation of environmental contaminants. A new biotechnology process called Biological Permeable Barrier (BPB) was developed to provide high microbial density, stable environmental conditions, and protective measures for microbial activities for in‐situ bioremediation of contaminated groundwater. This patented technology (U.S. Patent 6,337,019 ) is based on the creation of a structured matrix, or Bio‐beads, that provides the perfect environment for organic‐degrading microorganisms to establish biofilms capable of destroying the contaminants in water with remarkable stability and control. For over 240 days, the viability and performance of the BPB (Bio‐beads) system were shown for biodegradation of a targeted contaminant, 2,4,6‐ trichlorophenol (TCP), under a variety of operating and stress conditions (Razavi‐Shirazi, 1997 ). Extensive batch experiments were also conducted to obtain necessary data to determine the rate of TCP diffusion into the Bio‐beads, adsorption properties of the Bio‐beads, and substrate‐use rate of the mixed bioculture as free cells and as immobilized cells (Bio‐beads). A simulated model of BPB was also characterized with its porosity, permeability, and compressibility or deformation under typical field conditions. Our extensive research showed that BPB takes advantage of a controlled biotechnology process to overcome the disadvantages and uncertainties associated with conventional biological processes. A summary of our investigation is presented here. © 2002 Wiley Periodicals, Inc.     

Spatial Distribution of Geobacteraceae and Sulfate‐Reducing Bacteria During In Situ Bioremediation of Uranium‐Contaminated Groundwater

Analysis of the physiological status of subsurface microbial communities generally relies on the study of unattached microorganisms in the groundwater. These approaches have been employed in studies on bioremediation of uranium‐contaminated groundwater at a study site in Rifle, Colorado, in which Geobacter species typically account for over 90 percent of the microbial community in the groundwater during active uranium reduction. However, to develop efficient in situ bioremediation strategies it is necessary to know the status of sediment‐associated microorganisms as well. In order to evaluate the distribution of the natural community of Geobacter during bioremediation of uranium, subsurface sediments were packed into either passive flux meters (PFMs) or sediment columns deployed in groundwater monitoring wells prior to acetate injection during in situ biostimulation field trials. The trials were performed at the Department of Energy's (DOE's) Rifle Integrated Field Research Challenge site. Sediment samples were removed either during the peak of Fe(III) reduction or the peak of sulfate reduction over the course of two separate field experiments and preserved for microscopy. Direct cell counts using fluorescence in situ hybridization (FISH) probes targeting Geobacter species indicated that the majority of Geobacter cells were unattached during Fe(III) reduction, which typically tracks with elevated rates of uranium reduction. Similar measurements conducted during the sulfate‐reducing phase revealed the majority of Geobacter to be attached following exhaustion of more readily bioavailable forms of iron minerals. Laboratory sediment column studies confirmed observations made with sediment samples collected during field trials and indicated that during Fe(III) reduction, Geobacter species are primarily unattached (90 percent), whereas the majority of sulfate‐reducing bacteria and Geobacter species are attached to sediment surfaces when sulfate reduction is the predominant form of metabolism (75 percent and 77 percent, respectively). In addition, artificial sediment experiments showed that pure cultures of Geobacter uraniireducens, isolated from the Rifle site, were primarily unattached once Fe(III) became scarce. These results demonstrate that, although Geobacter species must directly contact Fe(III) oxides in order to reduce them, cells do not firmly attach to the sediments, which is likely an adaptive response to sparsely and heterogeneously dispersed Fe(III) minerals in the subsurface. © 2013 Wiley Periodicals, Inc.     

Applications of mulch biowalls—three case studies

Mulch biowalls are proving to be an effective means of generating reducing conditions for the in situ anaerobic reduction of contaminants in groundwater that are amenable to the reduction process. Mulch is an inexpensive and readily available substrate that provides a long‐lasting carbon and electron donor source for the stimulation of the anaerobic reduction process in groundwater. Examples of contaminants that are amenable to the biotic anaerobic reduction process include: chlorinated alkenes and alkanes, explosives, perchlorate, some metals, and petroleum hydrocarbons. The microbial degradation of cellulose fibers (mulch) is arguably the oldest reduction process known and is evident anywhere that plant material, soil, and water are present together. This article presents three case studies discussing three different uses of mulch biowalls to stimulate the anaerobic bioremediation of contaminants in shallow soils and groundwater. © 2009 Wiley Periodicals, Inc.     

Rapid full‐scale bioremediation of perchlorate in soil at a large brownfields site

The former Bermite site north of Los Angeles, California, was used to manufacture various explosives and related products containing energetic compounds, including perchlorate. Remediation of perchlorate in site soil and groundwater is being conducted to meet regulatory requirements and allow planned redevelopment activities to proceed. The general approach to perchlorate remediation of shallow soil at the site includes excavation of affected soils followed by ex situ bioremediation. Glycerin was chosen for use as an electron donor because of its stability, safety, low cost, and regulatory acceptance. However, full‐scale bioremediation operation with glycerin initially resulted in inconsistent results despite consistent perchlorate biodegradation observed in treatability study microcosms. To eliminate the inconsistency and optimize the biotreatment process, additional studies were performed in the field on parallel tracks to determine crucial factor(s) that influenced inconsistent breakdown of perchlorate in site soils. Total Kjeldahl nitrogen (TKN) was determined to be a significant factor limiting perchlorate biodegradation. The addition of di‐ammonium phosphate (DAP) resulted in the consistent and complete perchlorate removal, generally within two weeks of incubation with a median destruction rate of about 200 μg/kg/day. Soil processing rates were gradually increased over the year, and, by the summer, approximately 2,000 to 2,500 tons of soil were being processed per day with a total of approximately 160,000 tons processed by the end of July. The total unit treatment cost for the process is about approximately $35/ton. The glycerin‐DAP process is playing a major role in the remediation of this 1,000‐acre former industrial site. © 2008 Wiley Periodicals, Inc.     

Bioremediation of Dissolved and Free Phase Oil in Wastewater from Equipment Maintenance Activities

Land treatment facilities can provide effective treatment of secondary oily wastewater from maintenance operations, particularly in arid climates. Soil and underlying groundwater from a land treatment facility, which has been operating for eight years, were analyzed to determine the effectiveness of using bioremediation for the treatment of dissolved and free‐phase oil in maintenance wastewater. The study was conducted at a mining site in Western Australia. The facility was capable of treating 140 kiloliters (kL) of oily wastewater per day. The average petroleum hydrocarbon content of the wastewater was 2 percent weight per volume (w/v) based on data available for the first five years. The soil data indicate that the land treatment process has been operating efficiently even at high wastewater loadings with maximum degradation rates of 10–242 mg/kg per day. Based on the soil data, there is no evidence of accumulation of any metal or polycyclic aromatic hydrocarbon (PAH) compounds. The land treatment facility has led to only low levels of TPH (total petroleum hydrocarbons) contamination (<4 ppm) in the underlying groundwater. However, nitrate concentrations in the groundwater were shown to increase over the first five years of the facility's operation. This article reports and discusses the operational data from the land treatment process, illustrating its effectiveness in treating oily wastewater. © 2001 John Wiley & Sons, Inc.