Perchlorate reduction by a novel chemolithoautotrophic,hydrogen-oxidizing bacterium

Water treatment technologies are needed that can remove perchlorate from drinking water without introducing organic chemicals that stimulate bacterial growth in water distribution systems. Hydrogen is an ideal energy source for bacterial degradation of perchlorate as it leaves no organic residue and is sparingly soluble. We describe here the isolation of a perchlorate-respiring, hydrogen-oxidizing bacterium (Dechloromonas sp. strain HZ) that grows with carbon dioxide as sole carbon source. Strain HZ is a Gram-negative, rod-shaped facultative anaerobe that was isolated from a gas-phase anaerobic packed-bed biofilm reactor treating perchlorate-contaminated groundwater. The ability of strain HZ to grow autotrophically with carbon dioxide as the sole carbon source was confirmed by demonstrating that biomass carbon (100.9%) was derived from CO2. Chemolithotrophic growth with hydrogen was coupled with complete reduction of perchlorate (10 mM) to chloride with a maximum doubling time of 8.9 h. Strain HZ also grew using acetate as the electron donor and chlorate, nitrate, or oxygen (but not sulphate) as an electron acceptor. Phylogenetic analysis of the 16S rRNA sequence placed strain HZ in the genus Dechloromonas within the beta subgroup of the Proteobacteria. The study of this and other novel perchlorate-reducing bacteria may lead to new, safe technologies for removing perchlorate and other chemical pollutants from drinking water.     

Validation of the ion-exchange membrane bioreactor concept in a plate-and-frame module configuration

The ion exchange membrane bioreactor (IEMB) is a particular case of a membrane-supported biofilm reactor, in which oxy-anions, used as electron acceptors by an anoxic mixed microbial culture, are removed from a polluted water stream through an anion-exchange membrane. The opposite side of this membrane is used for the development of a biofilm, contacting a biocompartment, to which nutrients and chloride are fed as a source of “driving” counter-ion. The applicability of a plate-and-frame IEMB module configuration, consisting of a series of membranes, for the treatment of drinking water contaminated with nitrate and perchlorate, was evaluated. Permeation of carbon source across the membrane to the treated water stream was avoided by a dedicated start-up procedure involving a gradual increase of ethanol feeding to the IEMB biocompartment. It was demonstrated that the biocompartment pH must be controlled not only to guarantee a complete perchlorate removal, but also to avoid precipitation of struvite on the membrane surface, which provokes membrane scaling and decreases the availability of nutrients for the biofilm. Under these conditions, the IEMB was successfully operated maintaining both nitrate and perchlorate concentrations in the treated water below their recommended levels for drinking water supplies.     

Development of a Relative Source Contribution Factor for Drinking Water Criteria: The Case of Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)

The consideration of multiple or cumulative sources of exposure to a chemical is important for adequately protecting human health. This assessment demonstrates one way to consider multiple or cumulative sources through the development of a relative source contribution (RSC) factor for the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), using the Exposure Decision Tree approach (subtraction method) recommended by the U.S. Environmental Protection Agency. The RSC factor is used to ensure that the concentration of a chemical allowed by a regulatory criterion or multiple criteria, when combined with other identified sources of exposure common to the population of concern, will not result in unacceptable exposures. An exposure model was used to identify relevant potential sources for receptors. Potential exposure pathways include ingestion of soil, water, contaminated local crops and fish, and dermal contact with soil and water. These pathways are applicable only to areas that are in close proximity to current or former military bases where RDX may have been released into the environment. Given the physical/chemical properties and the available environmental occurrence data on RDX, there are adequate data to support a chemical-specific RSC factor for RDX of 50% for drinking water ingestion.     

Limitations to Natural Bioremediation of Perchlorate in a Contaminated Site

Perchlorate (ClO₄^-) has been detected in many drinking water supplies in the United States, including the Las Vegas Wash and Lake Mead, Nevada. These locations are highly contaminated and contribute perchlorate to Lake Mead and the Colorado River system. Essential elements for perchlorate bioremediation at these locations were examined, including the presence of perchlorate-reducing bacteria (PRB), sufficient electron donors, occurrence of competing electron acceptors, and ability of PRB to utilize a variety of electron donors. Enumeration of PRB was performed anoxically using most probable number (MPN). Values ranged from ≤20 to 230 PRB/100 ml or ≤20 to ≥ 1.6× 10⁵ PRB/g for Lake Mead water samples and Las Vegas Wash sediments, respectively. 16S rRNA sequences revealed that isolates were γ -proteobacteria, Aeromonas, Dechlorosoma, Rahnella and Shewanella. A screening of potential electron donors using BIOLOG^TM demonstrated that all isolates were capable of metabolic versatility. Measurements of total organic carbon (TOC), nitrate and dissolved oxygen (DO) indicated limited presence of electron donor at all sites, whereas the electron acceptors varied throughout the Wash and Lake Mead. The persistence of perchlorate in the sites is attributed to lack of available electron donor and/or the presence of competing electron acceptors. A location has been identified where perchlorate biodegradation could be implemented thereby halting the transport of perchlorate to Lake Mead and the Colorado River.     

Limitations to Natural Bioremediation of Perchlorate in a Contaminated Site

Perchlorate (ClO₄ ^?) has been detected in many drinking water supplies in the United States, including the Las Vegas Wash and Lake Mead, Nevada. These locations are highly contaminated and contribute perchlorate to Lake Mead and the Colorado River system. Essential elements for perchlorate bioremediation at these locations were examined, including the presence of perchlorate-reducing bacteria (PRB), sufficient electron donors, occurrence of competing electron acceptors, and ability of PRB to utilize a variety of electron donors. Enumeration of PRB was performed anoxically using most probable number (MPN). Values ranged from ≤20 to 230 PRB/100 ml or ≤20 to ≥ 1.6× 10⁵ PRB/g for Lake Mead water samples and Las Vegas Wash sediments, respectively. 16S rRNA sequences revealed that isolates were γ -proteobacteria, Aeromonas, Dechlorosoma, Rahnella and Shewanella. A screening of potential electron donors using BIOLOG^TM demonstrated that all isolates were capable of metabolic versatility. Measurements of total organic carbon (TOC), nitrate and dissolved oxygen (DO) indicated limited presence of electron donor at all sites, whereas the electron acceptors varied throughout the Wash and Lake Mead. The persistence of perchlorate in the sites is attributed to lack of available electron donor and/or the presence of competing electron acceptors. A location has been identified where perchlorate biodegradation could be implemented thereby halting the transport of perchlorate to Lake Mead and the Colorado River.     

Proteomic detection of proteins involved in perchlorate and chlorate metabolism

Mass spectrometry and a time-course cell lysis method were used to study proteins involved in perchlorate and chlorate metabolism in pure bacterial cultures and environmental samples. The bacterial cultures used included Dechlorosoma sp. KJ, Dechloromonas hortensis, Pseudomonas chloritidismutans ASK-1, and Pseudomonas stutzeri. The environmental samples included an anaerobic sludge enrichment culture from a sewage treatment plant, a sample of a biomass-covered activated carbon matrix from a bioreactor used for treating perchlorate-contaminated drinking water, and a waste water effluent sample from a paper mill. The approach focused on detection of perchlorate (and chlorate) reductase and chlorite dismutase proteins, which are the two central enzymes in the perchlorate (or chlorate) reduction pathways. In addition, acetate-metabolizing enzymes in pure bacterial samples and housekeeping proteins from perchlorate (or chlorate)-reducing microorganisms in environmental samples were also identified.     

Effects of iodine repletion on thyroid morphology in iodine and/or selenium deficient rat term fetuses, pups and mothers.

It has been suggested that selenium deficiency aggravates the iodine-induced thyroid inflammation and necrosis in iodine-deficient Wistar rats and possibly in man. Studies were carried out to determine whether large amounts of iodine given to iodine-deficient pregnant Sprague-Dawley rats with or without selenium deficiency would induce inflammation and necrosis in their term fetal thyroids. Iodine deficiency was induced in the dams by a low iodine diet or perchlorate in the drinking water and iodine excess was achieved by iodinated drinking water during pregnancy or daily subcutaneous injections of iodine from days 20 to 22 of pregnancy, 1 day after perchlorate was discontinued. Studies were also carried out in 30-day-old pups whose nursing mothers were iodine-deficient (perchlorate) with or without selenium deficiency from conception onward. The administration of iodine restored the morphologic changes in the thyroid induced by iodine deficiency, irrespective of selenium status, toward normal without inflammatory changes or necrosis. Possible explanations for these unexpected findings are discussed.     

A Comparative Toxicological Assessment of Perchlorate and Thiocyanate Based on Competitive Inhibition of Iodide Uptake as the Common Mode of Action

Thiocyanate and perchlorate are known to competitively inhibit thyroidal iodide uptake at the sodium-iodide symporter. Estimates of their relative potencies have recently been refined; thiocyanate is 15 times less potent than perchlorate on a serum concentration basis. Numerous studies have been published relating serum thiocyanate concentrations (or surrogate measures) with thyroid function in various populations including pregnant women and neonates in regions with varying degrees of iodine deficiency. Fifteen published studies were located that relate serum thiocyanate concentrations with thyroid function. In the absence of severe iodine deficiency or iodine excess, adverse thyroidal effects occur with chronic serum thiocyanate concentrations ≥ 200 μ mol/L whereas non-adverse effects are observed with concentrations in the range of 65–85 μ mol/L. No adverse or non-adverse effects are observed at serum concentrations below 50 μ mol/L, even among sensitive subpopulations. Recently, studies relating serum perchlorate concentrations with perchlorate dose have become available, thus making it possible to predict the perchlorate dose associated with a serum perchlorate concentration. Serum thiocyanate concentrations found to induce non-adverse or adverse thyroid effects can thereby be used to predict the perchlorate concentration and thus the perchlorate dose that would be expected to induce similar effects. To place a perspective on environmental perchlorate exposure, a serum thiocyanate concentration of 50 μ mol/L is equivalent to a serum perchlorate concentration of 3.3 μ mol/L in terms of iodine uptake inhibition. This serum perchlorate concentration would require a perchlorate dose of 0.27 mg/kg-day, or a drinking water equivalent level of 9 mg/L using standard default assumptions of a 70 kg adult drinking 2 liters of water daily.     

Changes in the structure and function of microbial communities in drinking water treatment bioreactors upon addition of phosphorus

Phosphorus was added as a nutrient to bench-scale and pilot-scale biologically active carbon (BAC) reactors operated for perchlorate and nitrate removal from contaminated groundwater. The two bioreactors responded similarly to phosphorus addition in terms of microbial community function (i.e., reactor performance), while drastically different responses in microbial community structure were detected. Improvement in reactor performance with respect to perchlorate and nitrate removal started within a few days after phosphorus addition for both reactors. Microbial community structures were evaluated using molecular techniques targeting 16S rRNA genes. Clone library results showed that the relative abundance of perchlorate-reducing bacteria (PRB) Dechloromonas and Azospira in the bench-scale reactor increased from 15.2% and 0.6% to 54.2% and 11.7% after phosphorus addition, respectively. Real-time quantitative PCR (qPCR) experiments revealed that these increases started within a few days after phosphorus addition. In contrast, after phosphorus addition, the relative abundance of Dechloromonas in the pilot-scale reactor decreased from 7.1 to 0.6%, while Zoogloea increased from 17.9 to 52.0%. The results of this study demonstrated that similar operating conditions for bench-scale and pilot-scale reactors resulted in similar contaminant removal performances, despite dramatically different responses from microbial communities. These findings suggest that it is important to evaluate the microbial community compositions inside bioreactors used for drinking water treatment, as they determine the microbial composition in the effluent and impact downstream treatment requirements for drinking water production. This information could be particularly relevant to drinking water safety, if pathogens or disinfectant-resistant bacteria are detected in the bioreactors.     

Perchlorate-reducing microorganisms isolated from contaminated sites

An extensive microcosm survey of perchlorate-contaminated sites was undertaken to assess the ability of indigenous microorganisms to degrade perchlorate. Samples from 12 contaminated sites and from one pristine location were analysed. Perchlorate was degraded to below detection limit in all electron donor-amended microcosms. Perchlorate-reducing microorganisms (PRMs) were numerous at most of these sites. Sixteen distinct PRMs were isolated that were phylogenetically related to either Dechloromonas in the Beta Proteobacteria (9/16 isolates) or to Azospirillum in the Alpha Proteobacteria (7/16 isolates). The majority of previously isolated PRMs are in the Beta Proteobacteria related to Dechloromonas or Dechlorosoma. This study indicates that PRMs of the genus Azospirillum may be more prevalent at contaminated sites than the current record of isolates suggests. Cell yields, electron donor to perchlorate ratios and maximum specific growth rates were similar among the isolates and similar to the few previously published values. However, the Monod half-saturation constants for perchlorate for the two Azospirillum isolates characterized were lower than those measured for other genera, suggesting that they may be more effective at low concentrations of perchlorate. These results extend the current understanding of PRMs from diverse environments and provide added confidence that microbial perchlorate reduction is ubiquitous, even at highly contaminated sites, and can be harnessed effectively for bioremediation.     

Harnessing microbial activities for environmental cleanup

Human activities have released large amounts of toxic organic and inorganic chemicals into the environment. Toxic waste streams threaten dwindling drinking water supplies and impact terrestrial, estuarine and marine ecosystems. Cleanup is technically challenging and the costs based on traditional technologies are exceeding the economic capabilities of even the richest countries. Recent advances in our understanding of the microbiology contributing to contaminant transformation and detoxification has led to successful field demonstrations. Hence, harnessing the activity of naturally occurring bacteria, particularly the power of anaerobic reductive processes, is a promising approach to restore contaminated subsurface environments, protect drinking water reservoirs and to safeguard ecosystem health.     

Modelling salinity inhibition effects during biodegradation of perchlorate

AIMS: To determine the mathematical kinetic rates and mechanisms of acclimated perchlorate (ClO)-reducing microbial cultures by incorporating a term to relate the inhibitory effect of high salinity during biological reduction of concentrated perchlorate solutions. METHODS AND RESULTS: Salt toxicity associated with the biodegradation of concentrated perchlorate (200, 500, 1100, 1700 and 2400 mg l(-1) as ClO) was investigated using two microbial cultures isolated from a domestic wastewater treatment plant [return activated sludge (RAS) and anaerobic digester sludge (ADS)]. Experiments were performed in wastewaters containing various sodium chloride concentrations, ranging from 0% to 4.0% (w/v) NaCl (ionic strength: 0.14-0.82 mol l(-1), total dissolved solids: 5.3-42.6 g l(-1)) at near-neutral values of pH (6.7-7.8). Perchlorate biodegradation was stimulated through stepwise acclimation to high salinity. The ADS culture was capable of reducing perchlorate at salinities up to 4% NaCl, while the RAS culture exhibited complete inhibition of perchlorate degradation at 4% NaCl, probably resulting from either a toxic effect or enzyme inactivation of the perchlorate-reducing microbes. Further, a kinetic growth model was developed based on experimental data in order to express an inhibition function to relate specific growth rate and salinity. CONCLUSIONS: Biological reduction of concentrated perchlorate wastewaters using either acclimated RAS or ADS cultures is feasible up to 3% or 4% NaCl, respectively. In addition, the kinetic model including a salinity inhibition term should be effective in many practical applications such as improving reactor design and management, furthering the understanding of high salinity inhibition, and enhancing bioremediation under high salinity loading conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Applications of these findings in water treatment practice where ion exchange or membrane technologies are used to remove perchlorate from water can have the potential to increase the overall attractiveness of these processes by eliminating the need to dispose of a concentrated perchlorate solution.     

Direct measurement of perchlorate exposure biomarkers in a highly exposed population: a pilot study

Exposure to perchlorate is ubiquitous in the United States and has been found to be widespread in food and drinking water. People living in the lower Colorado River region may have perchlorate exposure because of perchlorate in ground water and locally-grown produce. Relatively high doses of perchlorate can inhibit iodine uptake and impair thyroid function, and thus could impair neurological development in utero. We examined human exposures to perchlorate in the Imperial Valley among individuals consuming locally grown produce and compared perchlorate exposure doses to state and federal reference doses. We collected 24-hour urine specimen from a convenience sample of 31 individuals and measured urinary excretion rates of perchlorate, thiocyanate, nitrate, and iodide. In addition, drinking water and local produce were also sampled for perchlorate. All but two of the water samples tested negative for perchlorate. Perchlorate levels in 79 produce samples ranged from non-detect to 1816 ppb. Estimated perchlorate doses ranged from 0.02 to 0.51 μg/kg of body weight/day. Perchlorate dose increased with the number of servings of dairy products consumed and with estimated perchlorate levels in produce consumed. The geometric mean perchlorate dose was 70% higher than for the NHANES reference population. Our sample of 31 Imperial Valley residents had higher perchlorate dose levels compared with national reference ranges. Although none of our exposure estimates exceeded the U. S. EPA reference dose, three participants exceeded the acceptable daily dose as defined by bench mark dose methods used by the California Office of Environmental Health Hazard Assessment.     

Arsenic toxicity,mutagenesis, and carcinogenesis – a health risk assessment and management approach

A comprehensive analysis of published data indicates that arsenic exposure induces cardiovascular diseases, developmental abnormalities, neurologic and neurobehavioral disorders, diabetes, hearing loss, hematologic disorders, and various types of cancer. Although exposure may occur via the dermal, and parenteral routes, the main pathways of exposure include ingestion, and inhalation. The severity of adverse health effects is related to the chemical form of arsenic, and is also time- and dose-dependent. Recent reports have pointed out that arsenic poisoning appears to be one of the major public health problems of pandemic nature. Acute and chronic exposure to arsenic has been reported in several countries of the world where a large proportion of drinking water (groundwater) is contaminated with high concentrations of arsenic. Research has also pointed significantly higher standardized mortality rates for cancers of the bladder, kidney, skin, liver, and colon in many areas of arsenic pollution. There is therefore a great need for developing a comprehensive health risk assessment (RA) concept that should be used by public health officials and environmental managers for an effective management of the health effects associated with arsenic exposure. With a special emphasis on arsenic toxicity, mutagenesis, and carcinogenesis, this paper is aimed at using the National Academy of Science's RA framework as a guide, for developing a RA paradigm for arsenic based on a comprehensive analysis of the currently available scientific information on its physical and chemical properties, production and use, fate and transport, toxicokinetics, systemic and carcinogenic health effects, regulatory and health guidelines, analytical guidelines and treatment technologies.     

Neonatal thyroid-stimulating hormone level and perchlorate in drinking water

BACKGROUND: The effect of perchlorate in drinking water on neonatal blood thyroid-stimulating hormone (thyrotropin; TSH) levels was examined for Las Vegas and Reno, Nevada. METHODS: The neonatal blood TSH levels in Las Vegas (with up to 15 microg/L (ppb) perchlorate in drinking water) and in Reno (with no perchlorate detected in the drinking water) from December 1998 to October 1999 were analyzed and compared. The study samples were from newborns in their first month of life (excluding the first day of life) with birth weights of 2, 500-4,500 g. A multivariate analysis of logarithmically transformed TSH levels was used to compare the mean TSH levels between Las Vegas and Reno newborns, with age and sex being controlled as potential confounders. RESULTS: This study of neonatal TSH levels in the first month of life found no effect from living in the areas with environmental perchlorate exposures of 相似文献   

Dissimilatory perchlorate reduction: a review

In the United States anthropogenic activities are mainly responsible for the wide spread perchlorate contamination of drinking water, surface water, groundwater, and soil. Even at microgram levels, perchlorate causes toxicity to flora and fauna and affects growth, metabolism and reproduction in humans and animals. Reports of antithyroid effects of perchlorate and its detection in common food items have raised serious public health concerns, leading to extensive decontamination efforts in recent years. Several physico-chemical removal and biological decontamination processes are being developed. Although promising, ion exchange is a non-selective and incomplete process as it merely transfers perchlorate from water to the resin. The perchlorate-laden spent resins (perchlorate 200-500 mg L⁻¹) require regeneration resulting in production of concentrated brine (6-12% NaCl) or caustic waste streams. On the contrary, biological reduction completely degrades perchlorate into O₂ and innocuous Cl⁻. High reduction potential of ClO₄⁻/Cl⁻ (E° = ∼1.28 V) and ClO₃⁻/Cl⁻ pairs (E° = 1.03 V) makes these contaminants thermodynamically ideal e⁻ acceptors for microbial reduction. In recent years unique dissimilatory perchlorate reducing bacteria have been isolated and detailed studies pertaining to their microbiological, biochemical, genetics and phylogenetic aspects have been undertaken which is the subject of this review article while the various physico-chemical removal and biological reduction processes have been reviewed by others.     

Perchlorate Remediation by Electrokinetic Extraction and Electrokinetic Injection of Substrates

Perchlorate (ClO4?) contamination of groundwater has recently become a major concern across the nation. Electrokinetic (EK) extraction with the simultaneous EK injection of organic material to promote degradation could allow for the efficient removal of perchlorate while simultaneously promoting degradation of perchlorate. Column experiments were conducted to evaluate the technology. Lactate and glycine served as organic substrates to promote degradation after injection into the columns as well as maintaining the pH near neutral. Removal of perchlorate from contaminated materials kaolin, sand, and a natural soil historically contaminated by perchlorate was controlled by the ionic flux of perchlorate and not by transport from the osmotic flux which was only significant for kaolin experiments. Perchlorate was removed from contaminated sand and clay below our detection limits (5 ppb). Both lactic acid and glycine were successfully injected into clay and a sand matrix. Results from a contaminated site soil indicate that the Chemical Oxygen Demand was increased after electrokinetic injection of glycine and lactate. Experiments using soil from a contaminated site confirmed that EK can be used to both remove perchlorate and stimulate bioremediation by the injection of lactate or glycine. The use of EK technology to both remove and provide for continued source removal by bioremediation offers a potential new tool to treat low permeability systems.     

Direct enrichment of perchlorate-reducing microbial community for efficient electroactive perchlorate reduction in biocathodes

Biological reduction of perchlorate (ClO₄ ^?) has emerged as a promising solution for the removal of perchlorate in contaminated water and soils. In this work, we demonstrate a simple process to enrich perchlorate-reducing microbial communities separately using acetate as electron donor and the municipal aerobic membrane bioreactor sludge as inoculum. Inoculation of cathodes in microbial fuel cells (MFCs) with these enrichments, and further electrochemical enrichment at constant resistance operation of the MFCs, led to perchlorate-reducing biocathodes with peak reduction rates of 0.095 mM/day (2 mg/m²/day). Analysis of the microbial diversity of perchlorate-reducing biocathodes using PCR-DGGE revealed unique community profiles when compared to the denitrifying biocathode communities. More importantly, the total time taken for enrichment of the electroactive communities was reduced from several months reported previously in literature to less than a month in this work.     

Review of Campylobacter spp. in drinking and environmental waters

Consumption of contaminated drinking water is a significant cause of Campylobacter infections. Drinking water contamination is known to result from septic seepage and wastewater intrusion into non-disinfected sources of groundwater and occasionally from cross-connection into drinking water distribution systems. Wastewater effluents, farm animals and wild birds are the primary sources contributing human-infectious Campylobacters in environmental waters, impacting on recreational activities and drinking water sources. Culturing of Campylobacter entails time-consuming steps that often provide qualitative or semi-quantitative results. Viable but non-culturable forms due to environmental stress are not detected, and thus may result in false-negative assessments of Campylobacter risks from drinking and environmental waters. Molecular methods, especially quantitative PCR applications, are therefore important to use in the detection of environmental Campylobacter spp. Processing large volumes of water may be required to reach the desired sensitivity for either culture or molecular detection methods. In the future, applications of novel molecular techniques such as isothermal amplification and high-throughput sequencing applications are awaited to develop and become more affordable and practical in environmental Campylobacter research. The new technologies may change the knowledge on the prevalence and pathogenicity of the different Campylobacter species in the water environment.     

Perchlorate Remediation by Electrokinetic Extraction and Electrokinetic Injection of Substrates

Perchlorate (ClO₄^-) contamination of groundwater has recently become a major concern across the nation. Electrokinetic (EK) extraction with the simultaneous EK injection of organic material to promote degradation could allow for the efficient removal of perchlorate while simultaneously promoting degradation of perchlorate. Column experiments were conducted to evaluate the technology. Lactate and glycine served as organic substrates to promote degradation after injection into the columns as well as maintaining the pH near neutral. Removal of perchlorate from contaminated materials kaolin, sand, and a natural soil historically contaminated by perchlorate was controlled by the ionic flux of perchlorate and not by transport from the osmotic flux which was only significant for kaolin experiments. Perchlorate was removed from contaminated sand and clay below our detection limits (5 ppb). Both lactic acid and glycine were successfully injected into clay and a sand matrix. Results from a contaminated site soil indicate that the Chemical Oxygen Demand was increased after electrokinetic injection of glycine and lactate. Experiments using soil from a contaminated site confirmed that EK can be used to both remove perchlorate and stimulate bioremediation by the injection of lactate or glycine. The use of EK technology to both remove and provide for continued source removal by bioremediation offers a potential new tool to treat low permeability systems.