Abstracts of the Conference on Hazardous Waste Research, 1994


I. Abdel-Saheb and A.P. Schwab, Department of Agronomy, Kansas State University, Manhattan, KS 66506-5501

Vast areas of abandoned mine tailings in southeast Kansas are to be reclaimed by covering them with topsoil and revegetating. Although this procedure will reduce wind erosion and runoff of heavy metal contamination, the impact on leaching of metals is not known. A greenhouse experiment was initiated to measure the changes in metal mobility resulting from covering mine tailings with up to 60 cm of soil and establishing vegetation. Columns with different depths of topsoil over tailings were planted with a cool- season grass (tall fescue, Festuca arundinacea Schreb), and a warm-season grass (big bluestem Andropogon gerardii Vitm.). Duplicate columns for each treatment were prepared for unsaturated flow and leached with 0.001 M CaCl2 at a flow rate of 3.8 L/s. A vacuum (0.03 MPa) was applied to the bottom of the column, and effluents were collected every 24 h. The concentrations of cadmium, lead and zinc in the leachate were measured. After six months, plants were harvested and analyzed for heavy metals. Sequential extractions of column subsoil will be conducted to evaluate different metal forms labile for leaching.

Keyword(s): heavy metals, leaching, unsaturated flow.

Oral presentation in research track.


D.J.Adams, K.R. Gardner and B.E. Dinsdale, U.S. Bureau of Mines, Salt Lake City Research Center, 729 Arapeen Drive, Salt Lake City, UT 84108

The U.S. Bureau of Mines, Salt Lake City Research Center (SLRC), is investigating the biological reduction of selenate and selenite to develop a bioreactor for selenium removal from various processes and wastewaters. Indigenous selenium-reducing microorganisms vary somewhat from site to site and overgrowth of an established indigenous selenium-reducing microbial population by other indigenous nonselenium-reducing bacteria can pose a major bioreactor problem. In attempts to overcome this problem, the SLRC has used a consortium of selenium-reducing bacterial isolates as biofilms and immobilized in alginate beads to treat wastewaters containing 0.6 to 30.0 mg/L selenium. Reduction of both selenate and selenite to elemental selenium was obtained. Selenium reduction has been observed in a variety of bacteria under both aerobic and microaerophilic conditions. The SLRC has also used immobilized crude enzyme preparations to remove selenium from mining process solutions. The enzyme preparations reduced selenium in process solutions containing 100 mg/L cyanide; selenium- reducing bacteria are not cyanide tolerant. In laboratory tests, over 98% selenium removal has been attained using mining wastewaters.

Keyword(s): selenium reduction, bacteria, enzyme, immobilization.

Poster presentation.


O. Albinger, B.E. Logan, B. Biesemeyer and R.G. Arnold, Department of Chemical and Environmental Engineering, Arizona University, Tucson, AZ 85721

Predictions of bacterial transport/attenuation during passage through porous media are widely based on filtration theory. Cell-collector affinity, in this context the fraction of collisions between the biocolloid and collector material that result in bacterial sorption, is generally represented by a single value, a, that must be empirically determined. This is often accomplished through bench-scale simulations of field-scale situations. Using radiolabeled bacteria to measure biocolloid retention within a uniform bed of spherical borosilicate beads in a bench-scale reactor, we have shown that there is a wide range of affinities, or a values, within even monoclonal bacterial populations. A probability density function (PDF) representing the distribution of a values in a population of deep subsurface bacteria was mathematically deduced. That PDF was then used to test the importance of such variation in scaling from laboratory results for prediction of bacterial transport in large reactors or in the field. Prediction of far-field concentrations of residual microorganisms proved extremely sensitive to the representation of a values in the bacterial population.

Keyword(s): biocolloid transport, porous media, heterogeneities, modeling.

Poster presentation.


G. Amestoy, Montana Department of State Lands, Helena, MT 59620

Hard rock mining has been an important factor in the development of Montana as well as many western states for over one hundred years. As a result of these past mining operations that occurred prior to the passage of the mined land reclamation and other environmental statutes, mining was not regulated. Therefore many environmental degradation and reclamation problems exist at these "pre-law" mining sites that are abandoned or inactive today.

In an effort to eliminate the environmental degradation problems at these sites, the State of Montana, through the Montana Abandoned Mine Reclamation Bureau completed an inventory that identified some 6,000 sites, many of which require some form of reclamation/cleanup. It soon became apparent that the number of the abandoned and inactive mines needing cleanup far exceeded the Abandoned Mine Reclamation Bureau's ability to address this number of sites. Additionally, it was determined that many of these sites had multiple surface and mineral owners, including private, federal and state lands, as well as hazardous substances. These factors further complicate the issues of Responsible Party determination, responsibility for cleanup, and the cleanup standards that need to be met.

This presentation addresses the actual hands- on problems that need to be resolved when trying to coordinate and implement environmental cleanup and reclamation at these kinds of sites.

Keyword(s): hard rock mining, reclamation, inventory, hazardous substances, responsible parties.

Oral presentation in research track.


B. Anderson1 and B. Blackard2,1Superfund Basic Research and Training Program, National Institute of Environmental Health Sciences, P.O. Box 12233, Research Triangle Park, NC 27709; and 2Technology Planning and Management Corporation, Headquarters Park, Beta Building, Suite 220, 2222 Highway 54, Durham, NC 27713

The National Institute of Environmental Health Sciences (NIEHS) Superfund Basic Research and Training Program (SBRP) was established in 1986 by the Superfund Amendments and Reauthorization Act (SARA). This is a unique program of basic research and training grants directed towards understanding, assessing and attenuating the adverse effects on human health resulting from exposure to hazardous substances. Grants made under the SBRP are for coordinated, multicomponent, interdisciplinary studies. The program's primary goal is to provide an opportunity for researchers from the biomedical sciences, engineering, ecology, and the geosciences to explore the scope of the problems of uncontrolled hazardous waste and seek solutions. The program, now in its eighth year, is designed to provide a broader and more detailed body of scientific information to the field of environmental management. The research findings from this program are being used by state, local, and federal agencies, private organizations and industry in making decisions related to the management of hazardous substances.

Many innovative technologies for detecting, assessing and reducing toxic materials in the environment have been developed as a result of funding by the SBRP. To assist grantees, the SBRP has developed a technology transfer strategy designed to handle the unique problems associated with transferring multidisciplinary technology from basic research to applied research. By 1995 this will result in the validation of many of the research products being generated by this program as driven by technology demonstrations and commercialization. A number of these novel technologies are presented. The SBRP has actively pursued increasing the availability of these findings to the scientific community. Currently, the database detailing each program is available through the NIEHS Gopher server on the Internet. A description of how to access this system is given. To encourage the interdisciplinary nature of the program, the SBRP has strongly encouraged grantees’ participation in scientific conferences and workshops. A listing of conferences and workshops sponsored by the SBRP for the years of 1993-1994 will be presented.

Keyword(s): superfund, technology transfer, database.

Poster presentation.


D. Anderson1, L. Twidwell2 and G. Geesey3, 1Department of Biological Sciences, Montana Tech, Butte, MT 59701; 2Metallurgy Department, Montana Tech, Butte, MT 59701; and 3Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717

Biohydrometallurgy is the science and engineering concerned with the extraction of chemical elements from ores, concentrates and rocks through the solubilizing action of microorganisms. Acidophilic bacteria, especially Thiobacillus, are associated with this process and are used commercially in bioleaching (heap leaching.) These naturally occurring microorganisms also increase the rate at which acid mine (rock) drainage occurs. Traditional studies do not easily allow visualization of bioleaching. Flowthrough cell techniques adapted from biocorrosion/biofilm research used at Montana State University's Center for Biofilm Engineering will be presented and discussed. Comparisons between these new methods will be made with the more traditional batch/column studies, usually associated with biohydrometallurgical research.

Keyword(s): biohydrometallurgy, bioleaching, biofilm, Thiobacillus, techniques.

Oral presentation in research track.


G.F. Andrews and S.G. Hansen, Idaho National Engineering Laboratory/EG&G Idaho, Inc., Office of Industrial Biotechnology and Process Engineering, P.O. Box 1625, Idaho Falls, ID 83415-2203

The differential soil bioreactor is a continuous- flow, laboratory treatability-study device in which groundwater, supplemented with nutrients, is recirculated through a disc of aquifer material at a rate that simulates actual groundwater flow. A high recycle ratio ensures that all bacteria in the disc are exposed to the same physiochemical environment, so rate and yield parameters needed for modeling in-situ bioremediation can, in principle, be derived directly from measurements of inlet and outlet concentrations of contaminants, nutrients and cells. Results are shown for the biodegradation of trichloroethylene by methanotrophic bacteria in sediments from the Savannah River site. The limitations of the technique for slow-flowing aquifers are discussed.

Keyword(s): in-situ, bioremediation, trichloroethylene, methanotrophs.

Oral presentation in research track.


J.S. Aronheim, T.H. Illangasekare, J.N. Ryan, G.L. Amy, R.W. Harvey, J.P. Loveland and A. Pieper, Department of Civil, Environmental and Architectural Engineering, University of Colorado at Boulder, Boulder, CO 80309; 303-492-6754; email: aronheim@uscu.colorado.edu

The transport of bacteriophages MS-2 and PRD-1 through three different saturated porous media formations were investigated in a two- dimensional pilot-scale flume (10m L, 0.05m W and 1.2 H), located in the Hydraulics Laboratory of the University of Colorado at Boulder. The migration of the virus-sized biocolloids co- injected with a bromide tracer were monitored in both space and time using up to fifteen sampling ports along the length of the flume. The three different simulated aquifer formations—a cross section of #30 sand (U.S. sieve size) homogeneously packed; a cross section of #30 sand, #8 sand (10 cm layer) and #30 sand; and a cross section of #30 sand, #30 sand coated with iron oxides (10 cm layer) and #30 sand—highlight the influences of physical and chemical aquifer media heterogeneities on the migration of virus- sized biocolloids. Included in this discussion is a description of the pilot-scale design and associated boundary conditions, methods for positioning injection and sampling points, techniques for packing and pre-experimental monitoring, approaches for analyzing and extrapolating information from breakthrough curves, and results from the experiments.

Keyword(s): biocolloid, transport, heterogeneities.

Oral presentation in research track.


G.A. Bala, C.P. Thomas, J.D. Jackson and R.A. McMillin, Idaho National Engineering Laboratory, EG&G Idaho Inc., Center for Industrial Bioprocess Engineering, Idaho Falls, ID 83415-2203

Processes have been developed for the restoration of environments contaminated with hydrocarbons and heavy organics. The intended product is a field deployable materials handling system and phase separation process ranging in size from 1 yd3/hr to 50 yd3/hr for commercial application to environmental problems associated with the exploration, production, refining and transport of petroleum, petroleum products and organic chemicals. Effluents from contaminated sites will be clean solids (classified by size if appropriate), and the concentrated contaminant.

The technology is based on biochemical solvation, liquid/liquid and liquid/solid extractions, materials classification, mechanical and hydraulic scrubbing, and phase separation of organic and aqueous phases. Fluid use is minimized through utilization of closed-loop (recycle) systems.

Contaminants that are removed from the solid material may be destroyed, disposed of using existing technologies, or used on-site for cogeneration of power for plant operations.

Additionally, if the contaminant is a valued product, the material may be recovered for application or sale. Clean solid material is not sterilized and may be returned to normal agricultural, commercial, residential or recreational use in most instances.

Keyword(s): hydrocarbon removal, restoration, petroleum contaminants.

Oral presentation in technology transfer track.


M.K. Banks1, A.P. Schwab2, R.S. Govindaraju1 and Zhi Chen1, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

A common environmental problem associated with the pumping and refining of crude oil is the disposal of petroleum sludge. This sludge is often treated by incorporation into the soil. If the soil is frequently tilled and fertilized, soil microorganisms will be stimulated and organic contaminants biodegraded. Unfortunately, the biodegradation rate of more recalcitrant and potentially toxic contaminants, such as the polynuclear aromatic hydrocarbons (PNAs), is rapid at first but declines quickly. Biodegradation of these compounds is limited by their strong adsorption potential and low solubility.

Recent research has suggested that vegetation may play an important role in the biodegradation of toxic organic chemicals, such as PNAs, in soil.

The establishment of vegetation on hazardous waste sites may be an economic, effective, low maintenance approach to waste remediation and stabilization. The use of plants for remediation may be especially well-suited for soils contaminated by organic chemicals to depths of less than 2 meters. The beneficial effects of vegetation on the biodegradation of hazardous organics are two-fold: organic contaminants may be taken up by the plant and accumulated, metabolized, or volatilized, and the rhizosphere microflora may accelerate biodegradation of the contaminants.

Completed greenhouse studies have indicated that vegetative remediation is a feasible method for clean-up of surface soil contaminated with petroleum products. However, a field demonstration is needed to exhibit this new technology to the industrial community. In this project, several petroleum contaminated field sites will be chosen in collaboration with three industrial partners. These sites will be thoroughly characterized for chemical properties, physical properties, and initial PNA concentrations. A variety of plant species will be established on the sites, including warm and cool season grasses and alfalfa Soil analyses for the target compounds over time will allow us to assess the efficiency and applicability of this remediation method.

Keyword(s): bioremediation, vegetation, petroleum, biodegradation, rhizosphere.

Poster presentation.


K.B. Barrett, J.M. Barnes and W.A. Apel, Industrial Biotechnology and Process Engineering, Idaho National Engineering Laboratory, PO Box 1625, Idaho Falls, ID 83415-2203

Nitrogen oxides (NOx) are primary air pollutants and, as such, there is considerable interest in the development of efficient, cost effective technologies to remediate NOx containing emissions. Biofiltration involves the venting of contaminated gas streams through biologically active material such as soil or compost. This technology has been used successfully to control odors as well as volatile organic compounds from a variety of industrial and public sources. The purpose of this study was to evaluate the feasibility of using biofiltration to convert NOx to nitrogen gas.

Bioreactor studies measuring nitric oxide (NO) removal by bacteria indigenous to coniferous wood compost were conducted. Vertical bioreactors (2 l volume) were constructed using glass process pipe (12 in. x 3 in.). Compaction of the compost in the bioreactors was minimized by the addition of wood chips (17% w/w) measuring approximately 1.2-1.5 cm in diameter. The compost/bark mixture was ten loaded (1.4 l) into the columns. A nitrogen gas stream containing various concentrations of NO (100-500 µl l-1) was purged (1 l min-1) through the compost under single pass continuous flow conditions. The effects of moisture, pH, temperature and exogenous carbon sources on nitric oxide removal in the biofilters were investigated.

Moisture measurements during nitric oxide removal experiments indicated that biological activity was optimum when the compost was moist but not saturated (9-12 centibars of soil suction). Studies demonstrated that pH control in the biofilter was a critical variable for maximum nitric oxide removal. Optimum denitrifying activity occurred at pH levels ranging between 6- 7. Nitric oxide removal rates were found to increase in biofilters treated with an external carbon and energy source. Biofilters maintained at 37şC and treated with lactate were shown capable of removing ł90% of the nitric oxide from an influent gas stream containing 500 ml/l NO.

Keyword(s): nitrogen oxides, biofiltration, denitrification.

Oral presentation in research track.


E.A. Betterton1, K.D. Warren2 and R.G. Arnold2, 1Department of Physics and Atmospheric Chemistry and 2Department of Chemical and Environmental Engineering, Arizona University, Tucson, AZ 85721

Elemental iron and zinc have been used to reductively convert part-per-thousand levels of aqueous-phase tetrachloromethane to trichloromethane in a few hours. Products include free metal irons, chloride ion and hydrogen gas.

Hydrogen ions are consumed. Process kinetics are dependent on solution pH, surface area of the elemental metal, tetrachloromethane concentration, and buffer selection. In mixed solvent experiments, conversion kinetics and process stoichiometry were dependent on the alcohol that was added to water. When isopropanol was added, acetone was among the suite of products. Dehalogenation reactions using elemental metals as reductants offer promise as a means for initiating the destruction of heavily halogenated aliphatic compounds. However, models of process mechanism, and therefore factors that will influence remediation designs, are incomplete. The results of recent experiments will be described.

Keyword(s): reductive dehalogenation, elemental metals, groundwater remediation, tetrachloromethane.

Poster presentation.


Ron K. Bhada1, Abbas Ghassemi1, Ricardo Jacquez1 and Dave Kauffman2, 1New Mexico State University, Box 30001, Dept. WERC, Las Cruces, NM 88003-8001; 2College of Engineering, University of New Mexico, Farris Engineering Center, Albuquerque, NM 87131

In 1990, the Department of Energy approved a cooperative agreement for a Waste-management Education and Research Consortium (WERC) program. This consortium includes as its members the New Mexico State University (NMSU), the University of New Mexico (UNM), the New Mexico Institute of Mining and Technology (NMIMT), the Navajo Community College, the Los Alamos National Laboratory, and the Sandia National Laboratories working with industrial affiliates.

The formative years have conclusively demonstrated that the partnership of universities, national laboratories and industry developed by WERC is an effective tool for education, technology development and technology transfer, with the education process also playing a critical role in technology transfer.

The university/national laboratory/industry partnership of WERC has resulted in unique solutions to technology issues. These have attained the demonstration stage at DOE and industry sites within the short period of three years after the start of the program. Further, the program has resulted in students with conventional course type education as well as experience on practical development projects at the leading edge of technology, thus forming a base for technology transfer as these students flow into government and industry jobs. Several of our past students occupy responsible positions and are already technology transfer agents.

Several of the unique technologies developed via WERC are successfully demonstrated via application at national laboratories and industrial sites. Examples are listed below.

Remediation of soil contaminated with plutonium using a polymer capture process has been applied at a DOE waste site.

Subsurface mapping of buried waste has been applied at a DOE integrated demonstration (ID) site.

A pipeline detection system has been applied to leak detection from storage tanks and pipelines in the oil industry.

This paper will present how the WERC partnership has been successful because of its unique structure and method of operation.

Keyword(s): technology transfer, partnerships, education.

Oral presentation in technology transfer track.


T. Boronina and K.J. Klabunde, Department of Chemistry, Kansas State University, Manhattan, KS 66506

The kinetics of the transformation of volatile chlorocarbons in water solution in the presence of the zero-valence metal particles was studied using Gas Chromatography and Gas Chromatography/Mass spectroscopy. Metals including magnesium, zinc, tin were exposed to a low concentration (100 to 1000 ul L-l) of carbon tetrachloride in water. Tin caused the degradation of carbon tetrachloride and appearance of carbon dioxide, chloroform and tin oxide. The transformation of chlorocarbons was found to depend on the surface properties of the metal particles.

Keyword(s): metal particles, chlorocarbons, water, degradation.

Oral presentation in research track.


S. Bradley, M. Roberts and R. Crawford, Center for Hazardous Waste Remediation Research, FRC 105, University of Idaho, Moscow, ID 83843

2,4-D is a widely used systemic herbicide for the control of broadleaf weeds in grain crops, field corn, pastures, and other crop and non-crop land.

This herbicide also has a large market in home and garden use. Under aerobic conditions, 2,4-D is rapidly degraded by microbes in soil.

Groundwater contamination is limited due to the pesticide’s high rate of microbial degradation and uptake by plants. Groundwater is susceptible to contamination at point sources where 2,4-D is formulated, loaded onto application equipment, and at clean-up areas where large amounts of rinseates can be produced. We have developed an aerobic biological process to degrade 2,4-D found in these rinseates and other water which may have contamination. This process involves establishing an environment in which a known 2,4-D degrading bacterium can utilize the herbicide as a carbon source and achieve mineralization to carbon dioxide and water. The environment conducive to 2,4-D degradation is produced by using an inexpensive nitrogen and phosphate source such as agricultural grade diammoniumphosphate and monoammoniumphosphate producing a buffer.

Oxygen is provided by vigorous aeration of the water with compressed air. The bacterium used is Alcaligenes eutrophus JMP134 (pJP4). The organism is produced in large amounts at relatively low cost by fermentation. The organism is air dried on peat and has exhibited good survival and degradation capabilities when stored at room temperature in a desiccated form for at least 60 days. This process has been successfully tested on rinseates produced by the cleaning of commercial spray rigs applying the herbicide Curtail®. Optimum physical parameters for the process such as pH, temperature, inoculam amounts, and buffer concentrations have been determined.

Keyword(s): 2,4-D, degradation, rinseates.

Poster presentation.


F. Brockman, W. Payne, D. Workman, A. Soong, S. Manley, W. Sun and A. Ogram, Pacific Northwest Laboratory, Richland, WA 99352; and Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164

The field site was manipulated with injection of air (control experiment), 1% methane (in air), pulsing of air only and 4% methane, and pulsing of 4% methane supplemented with gaseous forms of nitrogen and phosphorous. Gases were injected through a horizontal well into the aquifer and a vacuum was established in a second horizontal well in the vadose zone. Following each injection regime, sediment samples from the contaminated region (100-140 feet below ground surface) were analyzed. Analyses included most-probable- number enrichments for physiological groups known or suspected to degrade trichloroethylene (TCE) and per-chloroethylene (PCE), TCE and PCE removal from enrichments, and DNA extraction and hybridization with various gene probes corresponding to enzymes known to degrade TCE or TCE metabolites.

Culturable populations in sediments following methane pulsing plus nitrogen and phosphorous (+ N + P) compared to pre-methane sediments showed methanotrophs increased 0 to >3 orders of magnitude (to >2400/g), propane-oxidizers increased 2 to >3 orders of mag (to >2400/g), and ammonia-oxidizers increased 2 to >4 orders of magnitude (to >24000/g). These three microbial trophic groups are known to degrade TCE.

Culturable methylotroph populations increased 1 to >2 orders of magnitude (to >2400/g) and culturable anaerobes showed little to no increase.

Following methane pulsing + N + P, TCE and PCE removal occurred in methanotrophic enrichments for most sediments with 1 mg of inoculum and 100 mg of inoculum, respectively.

In contrast, TCE and PCE removal in methanotrophic enrichments from pre-methane sediments rarely occurred with 1 g of inoculum. A messenger RNA probe corresponding to the soluble methane monooxygenase gene showed that the cultural methods underestimated the density of the gene in sediments by several orders of magnitude. The underestimation occurred during all injection regimes but was most severe following the 1% methane injection. Populations of TCE-degrading microorganisms corresponded with increases in contaminant removal in vadose zone piezometers and groundwater, strongly suggesting that microorganisms were responsible for much of the TCE and PCE removal.

Keyword(s): bioremediation, trichloroethylene, nucleic acid probes, methanotrophs.

Oral presentation in research track.

This work was supported by the U.S. Department of Energy as part of Pacific Northwest Laboratory’s Laboratory Directed Research and Development. Pacific Northwest Laboratory is operated for the U.S. Department of Energy by Battelle Memorial Institute under contract DE-AC06-76RLO 1830.


B. Bugbee1 and W.J. Doucette2, 1Utah Agricultural Experiment Station and 2Utah Water Research Laboratory, Utah State University, Logan, UT 84322

Plants have profound effects on physical, chemical and biological processes in soils and can potentially accelerate the rate of degradation of hazardous organic chemicals. Quantitative information describing the effects of vegetation on the fate of organic chemicals in soils is needed to increase our general understanding of the fate process and evaluate the potential use of plants in enhancing the remediation of contaminated soils.

This study was designed to examine rates and extent of degradation by the plant/soil system, as well as plant uptake, accumulation and metabolism, for several representative organic chemicals. A flow-through system, consisting of six modules, was designed to accommodate rapid flow rates through the plant growth chamber. Rapid flow rates prevent condensation in the chamber and also prevent the unnatural accumulation of volatile compounds in the chamber air. Each system module consisted of planted or unplanted soil covered with a glass bell jar, through which passed a continuous stream of air. Bell jars stood in a growth chamber, and plants (or soil controls) were kept at 23 ± 1 C, with a 16 hr photoperiod. Pre- and post-chamber flow rates were carefully balanced to minimize pressure gradients and thus minimize air leaks. Six such modules ran simultaneously, three each for the planted and unplanted systems.

Radiolabeled (14C) and non-radiolabeled aromatic hydrocarbons, dissolved in ethanol, were added to the soil to yield a concentration of approximately 100 mg/g dry weight of soil. Plants were started from seed at the beginning of a trial.

Concentrations of the 14C labeled test compounds in the vapor, plant and soil phases were determined, after appropriate extraction and/or concentration techniques, by liquid scintillation counting. Preliminary results, using the system and procedures described above, show that mineralization of the test aromatic hydrocarbons is significantly greater in the rhizosphere soil than in the unplanted soil after two weeks.

Keyword(s): plants, biodegradation, organic contaminants.

Oral presentation in research track.


John A. Bumpus, C.W. Chang and Matthew Tatarko, Department of Chemistry and Biochemistry, Department of Biological Sciences and the Center for Bioengineering and Pollution Control, University of Notre Dame, Notre Dame, IN 46556

The wood rotting Basidiomycete Phanerochaete chrysosporium is able to degrade a wide variety of environmentally persistent organic pollutants to carbon dioxide. The unique biodegradative abilities of this fungus are due, in part, to lignin peroxidases, oxidative enzymes that are secreted in response to nutrient deprivation. Lignin peroxidases catalyze the initial oxidation of many of the organic pollutants that are degraded by this fungus. Lignin peroxidases mediate the initial oxidation of N,N,N',N',N",N"- hexamethylpararosaniline, several azo dyes and certain polycyclic aromatic hydrocarbons. Lignin peroxidases also mediate oxidative dechlorination.

For example, lignin peroxidases oxidize pentachlorophenol to 2,3,5,6-tetrachloro- 2,5cyclohexadiene-1,4-dione. Similarly these enzymes mediate oxidative oligomerization of 4- chloroaniline resulting in production of several dimers, trimers and tetramers and net dechlorination of the aromatic ring. Interestingly, many, but not all, of the reactions mediated by lignin peroxidases are also mediated by other plant, fungal and mammalian peroxidases. In some instances the major role of lignin peroxidases is the oxidation an intermediate. In the case of phenanthrene degradation lignin peroxidases do not mediate the initial oxidation.

However, these enzymes do mediate the oxidation of 9-Phenanthrol, forming Phenanthrene-9,10- dione. This is of interest because 9-Phenanthrol is a major metabolite formed under nonligninolytic conditions while Phenanthrene-9,10-dione is a major metabolite formed in ligninolytic cultures.

Thus, in this case, lignin peroxidases appear to link the pathway occurring under nonligninolytic conditions with the pathway that predominates in ligninolytic cultures. (Supported by NIEHS grant ESO 4492.)

Keyword(s): Phanerochaete chrysosporium, lignin peroxidase, biodegradation.

Poster presentation.


S.R. Burckhard1, A.P.Schwab2 and M.K. Banks1, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

A laboratory study was conducted to determine whether the concentration of heavy metals, zinc (Zn), lead (Pb), and cadmium (Cd), in leachate from mine tailings could be affected by the presence of organic acids exuded by plant roots and microbes in the rhizosphere. Geochemical modeling predicted that some organic ligands found in the rhizosphere have the capability to complex and increase the solubility of Zn. The leachate was analyzed for Zn, Pb and Cd content and for organic acid concentration. These results were confirmed by batch and column studies in which mine tailings were exposed to 0 to 10,000 mM organic acids.

Keyword(s): heavy metals, organic acids, rhizosphere, plants.

Oral presentation in research track.


Karl E. Burgher1 and Abbas Ghassemi2, 1Director, MWTPP Training and Education, Montana Tech of the University of Montana, Butte, MT; and 2Director, WERC Special Programs, New Mexico State University, Las Cruces, NM

The Mine Waste Technology Pilot Program (per published material) is involved with testing technologies to clean up mine and mineral waste and with the education and training of people who are or will be involved with mine waste issues.

The education component is a part of the larger MWTPP, sponsored by The Environmental Protection Agency through The Department of Energy, and administered by MSE, Inc., in Butte, Montana. Montana Tech's education role is to implement the following four point program: Master of Science degree with a mine and mineral waste emphasis, providing students with enhanced exposure to the interdisciplinary field of mine waste processing under existing engineering/science graduate programs; awards fellowships, graduate research assistantships, and graduate teaching assistantships; educational and training opportunities for industry professionals through specialized workshops; and educational opportunities for students and teachers, from kindergarten through high school.

In 1990, the Department of Energy approved a cooperative agreement for a Waste-Management Education and Research Consortium.. This consortium includes as its members the New Mexico State University, the University of Mexico, the New Mexico Institute of Mining and Technology, the Navajo Community College, the Los Alamos National Laboratory, and the Sandia National Laboratories. The model five year program was assigned the mission of demonstrating that a university/national laboratory partnership can effectively expand the nation’s capability to address the issues related to management of all types of waste via education and technology development. This partnership of universities, national laboratories, and industry developed by WERC is an effective tool for education, technology development, and technology transfer, with the education process playing a critical role in technology transfer. The WERC program is available to over 3000 students in academic institutions with a minority population of 25-95%.

Keyword(s): mine, hazardous, waste, education, university.

Oral presentation in research track.


J.G. Burken and J.L. Schnoor, Department of Civil and Environmental Engineering, University of Iowa, Iowa City, IA 52242

The study reported here investigated the overall effect poplar trees exert on the fate and transport of atrazine in a variety of soil types. The research was conducted in batch reactors.

Reactors utilizing small poplar cuttings, reactors quantifying sorption/desorbtion phenomenon, and reactors dosed with surrogate and actual poplar root exudate were prepared containing a silica sand or a silt-loam soil. The atrazine fate was monitored by the use of atrazine labeled with carbon-14 isotopes. The extent of uptake in the poplars was then quantified through oxidation of the poplar biomass followed by liquid scintillation techniques. Experimental results indicate that virtually all of the atrazine in sand can be taken up by the poplars in a reasonably short amount of time, with little adverse effects on the trees. In organic soils, the poplars can take up only mobil atrazine which is not bound to the soil. Results also indicate that the poplars have either a positive effect or no detrimental effect on all fate and transport mechanisms studied. This would indicate that poplar trees may have a promising future in the role of remediation of pesticide contaminated waters.

Keyword(s): atrazine, pesticides, poplars, remediation.

Oral presentation in research track.


M.C. Canty and T.F. McIntyre, Mine Waste Technology Pilot Program (MWTPP), MSE, Inc., Butte, MT 59701

A field application using sulfate-reducing bacteria (SRB) to treat acidic mine water discharging from the Lilly/Orphan Boy Mine near Elliston, Montana, is described. The field application is a project under the MWTPP implemented by MSE, Inc., funded by the U.S. Environmental Protection Agency (EPA), and jointly administered by the EPA and the Department of Energy. The design of the field application involves using a flooded mine shaft as an in-situ biological reactor to which organic substrate and SRB will be added. Technical parameters required for the field application are presently being developed from laboratory testing of SRB in packed-bed reactors.

Keyword(s): sulfate-reducing bacteria, acid rock drainage.

Oral presentation in research track.


L.M. Carmichael and F.K. Pfaender, Department of Environmental Sciences and Engineering, CB #7400, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599- 7400

Creosote contaminated sites are of environmental significance due to the high concentrations of toxic and/or mutagenic PAH usually found at these sites. Microbial degradation of PAH can be seen as a novel form of contaminant detoxification. This paper describes the microbial degradation of PAH in creosote contaminated soils using (9-14C) phenanthrene as a model PAH. Microbial metabolism was assessed with a mass balance approach as well as identification of PAH metabolites by GC/MS/FTIR. The mass balance accounted for the amount portion of the added phenanthrene. To confirm the effectiveness of microbial degradation to decrease soil toxicity, the Microtox® and Mutatox® assays were used to monitor toxicity of the creosote soils throughout the experiments.

Mass balance results indicated that phenanthrene was readily mineralized in the contaminated soils, while metabolite production accounted for only a minor portion of the added phenanthrene. Toxicity of contaminated soils increased slightly early in the incubation and then decreased over longer time periods. Mutagenicity of soils, however, did not decrease appreciably over a 3-month time period. The identity of metabolic products found in the soils will be discussed.z

Keyword(s): polycyclic aromatic hydrocarbons, biodegradation, metabolite production, Microtox®/Mutatox®.

Poster presentation.


K. Chaloupka1, N. Harper1, M. Steinberg1, S. Safe1, L.V. Rodriguez2 and L.S. Goldstein3, 1Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX 77843-4466; 2Department of Molecular Pathology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030; 3Electric Power Research Institute, P.O. Box 10412, Palo Alto, CA 94303

Complex mixtures of polynuclear aromatic hydrocarbons (PAHs) are organic combustion products and are components of creosote and oily wastes which have been identified in a large number of hazardous chemical waste sites. Risk assessment of PAH mixtures must take into account the toxicity or carcinogenicity of the individual compounds and their possible additive or nonadditive interactive effects. A reconstituted PAH mixture which resembled manufactured gas plant PAH residues was prepared using 16 different compounds and the immunotoxicity and monooxygenase induction activity of the 2-ring, 3-ring and ł 4-ring PAHs were compared to that observed for the reconstituted mixtures in B6C3F1 mice. The results showed that the reconstituted mixture inhibited the splenic plaque- forming cell response to T-cell dependent and independent antigens and induced hepatic microsomal ethoxyresorufin O-deethylase activity and Cyp1a-1 mRNA levels. The relative potencies of the reconstituted mixture and its components indicated that most of the activity was associated with ł 4-ring PAHs and the interactive effects of the individual PAHs in the reconstituted PAH mixture were essentially additive. This research was supported by the Electric Power Research Institute and the National Institute for Environmental Health Sciences (P42 ES04917).

Keyword(s): polynuclear aromatic hydrocarbons, interactive effects.

Poster presentation.


P. Chen, E. Yu, W.S. Amato, Z. Zhang and L.L. Tavlarides, Department of Chemical Engineering and Materials Science, Syracuse University, 320 Hinds Hall, Syracuse, NY 13244-1190

Contamination of soils with polychlorinated biphenyls at industrial and military sites is a significant problem. A process based on supercritical carbon dioxide extraction of polychlorinated biphenyls from the soil followed by supercritical water oxidation of the extract appears to be economically attractive with the benefit of returning clean soils/sediments to the environment. Our research objective is to assess the technical feasibility of this process and provide data for economic evaluation.

Results of desorption rate studies of Aroclor 1248 from soils and sediments will be presented. Data were acquired on a continuous flow fixed- bed laboratory-scale supercritical extraction unit. The experimental range of variables include: 30 to 50 C, 75 to 400 atmospheres, soil/sediment sample size of 8 grams, carbon dioxide flow rate of 0.02 to 0.54 grams/second, modifier flows 0.0 - 5 mole %, moisture content 0.0 to 20 weight %, and initial polychlorinated biphenyl concentration of 500 to 10,000 parts per million. The soils (sand, clay, till and surfacial soil) and sediment samples are representative of a specific Superfund Hazardous Waste Site in New York State. Contact time needed for sub 5 parts per million residual polychlorinated biphenyl concentrations range from 30 to 90 minutes depending on soil type, level of contamination, and conditions of contacting. A desorption rate model is presented to model the data. Also, solubility data of specific polychlorinated biphenyl congeners were obtained and modeled using the Peng-Robinson equation of state. These results will also be presented. Future work involving bench-scale studies with a 1.0 liter system will be described.

Keyword(s): supercritical extraction, carbon dioxide, polychlorinated biphenyls.

Oral presentation in research track.


Namhyun Chung and Steven D. Aust, Biotechnology Center, Utah State University, Logan, UT 84322-4705

Biodegradation of pentachlorophenol by Phanerochaete chrysosporium was examined in nonsterile soil. The rate of mineralization of pentachlorophenol was essentially linear for at least 27 days and increased almost linearly with increasing concentration of pentachlorophenol (from 50 to 1600 parts per million). With an initial concentration of 100 parts per million, no pentachlorophenol was found at 18 days while 40% of the added pentachlorophenol was present as pentachloroanisole, and mineralization continued linearly. Both pentachlorophenol and pentachloroanisole were found after 18 days when the initial concentration of pentachlorophenol was 800 parts per million. The rate of mineralization of pentachloroanisole also increased with increasing concentration of pentachloroanisole, however the increase was not linear. Essentially no radioactivity was found in either the aqueous or volatile organic fraction during the mineralization of either pentachlorophenol or pentachloroanisole.

Keyword(s): pentachlorophenol, pentachloroanisole, Phanerochaete chrysosporium, white rot fungi.

Oral presentation in research track.


C.S. Clennan, W. Yu and M.K. Banks, Department of Civil Engineering, Kansas State University, Manhattan, KS 66506

A method for enumerating subsurface soil bacteria by direct epifluorescent microscopy was developed. The dyes DAPI (4, 6-diamidino-2 phenylindole) and CTC (5-cyano-2,3 ditolyl tetrazolium chloride) were used to assess total and viable cells, respectively. The DAPI-CTC method was compared in the laboratory to an INT-AO [(2- (p-iodopheny)-3-(p-nitropheny)-5-phenyl tetrazolium chloride]-[acridine orange] method and viable plate counts. The DAPI-CTC method was found to adequately evaluate microbial numbers.

Keyword(s): epifluorescent microscopy, subsurface, microbes, soil microflora.

Poster presentation.


B.J. Clennan, J.K. Koelliker and G.J. Kluitenberg, Kansas State University, Manhattan, KS 66506

Soil cores were obtained from the vadose zone of the Konza Prairie. Tempe cells and rigid-wall permeameters were used to determine water characteristic curves and saturated hydraulic conductivity. Runoff plots, neutron access tubes, and automated rain gauges were installed at the two sites to determine runoff, soil moisture, and rainfall amounts. A mathematical model was developed to predict the movement of water through the vadose zone using the soil hydraulic properties and surface water balance.

Keyword(s): vadose zone, soil hydraulic properties.

Poster presentation.


T.E. Clevenger1, Daryl Roberts2 and Pat Phillips2,1Water Resources Research Center, University of Missouri- Columbia, Columbia, MO 65211; 2Missouri Department of Health, Jefferson City, MO 65101

Missouri has a long history of lead mining activities. One of the most productive areas was in the southwest part of the state in what was called the "Tri-State District." It received this name because the ore deposits were in Kansas and Oklahoma, as well as Missouri. About 40% of all the ore came from Missouri. Mining in this region began in 1848, with production ending in 1957. The tailings or mining wastes were stored on the surface, where an estimated 8 million cubic yards remains today.

Because of the high levels of lead in the area, several superfund sites have now been designated in the "District." A cooperative study was done by Region VII EPA, Missouri Department of Health, and the Centers for Disease Control (CDC). Blood samples, dust, soil, water and paint samples were collected from 150 houses where young children lived in the Joplin, Missouri, area. The dust and soil samples were split and part was sent to the University of Missouri for analysis by sequential extraction in order to determine the environmental availability and possible bioavailability of lead. The dust samples were determined to be significantly more available than the soil. Other relationships including correlation with blood lead results will be presented.

Keyword(s): bioavailability, lead, soil, dust.

Oral presentation in research track.


Joan Combie and Kenneth Runnion, J. K. Research, 210 South Wallace, Bozeman, MT 59715

Manufacturing and handling of conventional munitions have produced explosives-contaminated soil at numerous military installations. These soils pose both a reactivity and toxicity hazard. Waste streams at manufacturing facilities further add to the problem. To prevent contamination of groundwater, facilitate clean up of contaminated sites, and bring current manufacturing plants into line with environmental regulations, practical remediation methods are being sought.

Ideally, biological treatment would eliminate anthropogenic substances by aerobic mineralization to carbon dioxide and water or anaerobic decomposition to carbon dioxide and methane, hydrogen sulfide or nitrogen. However, accumulation of toxic intermediates rather than complete degradation, has hindered widespread application of the biological approach. Also, biodegradation that performs well in the laboratory is not always as efficient when tested in the field.

Microorganisms from extreme environments are a rich source of unique metabolic pathways and unusually stable enzymes. Anaerobic enrichment cultures were started a year ago with TNT, nitrobenzene or nitrotoluene and samples of thermal water. Of the five cultures now running, one has developed a thriving consortium. Recently, samples were analyzed under the direction of Dr. Donald Crawford. A total of 1200 ppm TNT was added to the culture. Now only 41.4 ppm TNT and 2.5 ppm 4-amino-2,6 dinitrotoluene remains in the liquid, while the soil extract contains 50.0 ppm TNT and 1.1 ppm 4- amino-2,6 dinitrotoluene.

Keyword(s): TNT, biodegradation, nitro-substituted.

Poster presentation.


Jeffrey R. Conuel, Douglas C. Mosteller and Kenneth F. Reardon, Department of Agricultural and Chemical Engineering, Colorado State University, Fort Collins, CO 80523

Biodegradation research has focused primarily on the metabolism of single pollutants by pure strains of microorganisms despite the fact that contamination of soil and water is caused more often by chemical mixtures. When available, biodegradation data for mixtures is usually cast in terms of bulk measurements like total organic carbon, which are inadequate for many applications.

In this presentation, two aspects of our research into the biodegradation of organic chemical mixtures will be discussed. First, results of biodegradation experiments with Pseudomonas putida F1 and benzene, toluene, phenol and trichloroethylene will be presented.

Biodegradation rate data were obtained for the aromatic compounds individually and in mixtures, and cometabolism of trichloroethylene with these compounds has also been studied. Mathematical models have been proposed to describe these results.

The second part of the presentation will cover the application of these results to the development of an immobilized-cell bioreactor for the cometabolic degradation of trichloroethylene. Phenomena such as competitive inhibition, induction and energy supply have been incorporated into a mathematical model. Results from bioreactor experiments will be discussed.

Keyword(s): biodegradation, kinetics, mixture, aromatics, trichloroethylene.

Oral presentation in research track.


A. Cramer, J.E. McLean and R.C. Sims, Utah Water Research Laboratory, Utah State University, Logan, UT 84322-8200

Abiotic interactions of pentachlorophenol (PCP) on manganese oxide surfaces were investigated to determine the extent of transformation. The optimal pH and ratio of manganese oxide to PCP were determined. Sorption of PCP on manganese oxide surfaces was quantified at optimal conditions. The effectiveness of utilizing manganese oxide to remediate contaminated subsurface environments was investigated.

Keyword(s): groundwater quality, geochemistry, remediation, aquifers.

Oral presentation in research track.


K.C. Donnelly, K.W. Brown and L.Y. He, Departments of Veterinary Anatomy & Public Health and Soil & Crop Sciences, Texas A&M University, College Station, TX 77843

Microbial bioassays have been used to assess the genotoxic hazard at more than 30 different hazardous waste sites. Environmental samples were extracted with dichloromethane and methanol, and the resulting residue tested using GC/MS analysis as well as the Salmonella Microsomal and E. coli Prophage Induction assays. At a munitions wastewater contaminated site, there was no correlation between mutagenicity in bacteria, and the risk as estimated from chemical analysis data. Samples 202 and 204 from a coal gasification site contained 72 mg/kg and 9 mg/kg benzo(a)pyrene, whereas the mutagenic responses of these samples were 231 net revertants/mg and 902 revertants/mg, respectively. The data suggest that microbial bioassays provide a valuable tool for monitoring the interactions of the components of a complex mixture.

Keyword(s): bioassay, hazardous waste, benzo(a)pyrene, trinitrotoluene.

Oral presentation in research track.

This research was supported by NIH grant P42 ES04917.


W.J. Doucette1 and B. Bugbee2, 1Utah Water Research Laboratory and 2Utah Agricultural Experiment Station, Utah State University, Logan, UT 84322

Information regarding biodegradability is critical for assessing the environmental fate and impact of organic chemicals in soil/plant systems. However, quantitative information describing the biodegradation of organic chemicals in soils and the effect of vegetation is lacking, primarily due to the difficulty and expense associated with experimentally measuring biodegradation rates. Typically, biodegradation rates in soils are determined directly by measuring the disappearance of the chemical of interest over time or indirectly by measuring evolved CO2 or O2 consumption. Indirect methods are generally simpler, less expensive and less time consuming than direct methods. The focus of this study was to develop a simple microcosm approach, utilizing measurements of O2 consumption, to investigate the biodegradability of selected organic compounds in rhizosphere soil. The microcosms consisted of 100 mL glass vials fitted with Mininert® valves and typically contained 40 grams of either rhizosphere or control soil. The rhizosphere soil was obtained from a vegetated plot, while the control soil from a unvegetated plot. All compounds were added directly to the soil, without a carrier solvent, resulting in a soil concentration of approximately 200 ppm.

Headspace concentrations of O2, converted to percent of theoretical oxygen demand (%ThOD), were analyzed over time using a gas chromatograph equipped with a thermal conductivity detector. The indirect microcosm technique provided a simple, inexpensive method for comparing the aerobic biodegradation of several aromatic hydrocarbons in rhizosphere versus soil.

Keyword(s): microcosm, biodegradation rates, plants, soil, oxygen, carbon dioxide, gas chromatography.

Oral presentation in research track.


Mitchell D. Erickson1, Joseph H. Aldstadt1, Jorge S. Alvarado1, Jeffrey S. Crain2, Kent A. Orlandini1 and Lesa L. Smith2, 1Environmental Research Division and 2Chemical Technology Division, Argonne National Laboratory, 9700 S. Cass Ave., Argonne, IL 60439-4837, 708-252-7772, FAX 70X-252-9594

Our nation faces a daunting challenge to clean up and protect our environment. Before launching any cleanup, one must characterize the type, concentration and extent of the contamination. During cleanup, one must monitor the progress, and after cleanup, one must often monitor the site to ensure that the cleanup was successful. Chemical characterization and monitoring techniques and methods are at the heart of executing these efforts. R&D efforts to improve capabilities can translate into major savings and improvement in environmental cleanup by reducing the unit cost of measurements (e.g., fewer steps in an analysis), reducing the time required to provide the information to the user (e.g., field analytical), or improving the quality of information (e.g., chemical speciation).

Radiochemical analysis is of particular concern to DOE, where over $300 million is expended annually on this activity. Unlike organic and inorganic analytes, which are widely found as contaminants, there has been comparatively little effort expended on improvements of radiochemical analyses. Desirable characteristics of any new methods for characterization of DOE's radiochemical contamination are faster, to reduce horrendous turnaround times cheaper, to reduce the burden on the taxpayers better performance, to achieve desired data quality objectives reduced scale and fewer steps, to minimize secondary mixed waste Methods for chemical characterization of the environment are being developed under a multitask project for DOE's Office of Technology Development within the Office of Environmental Restoration and Waste Management. The project focuses on improvement of radioanalytical methods with an emphasis on faster and cheaper routine methods. We have developed improved methods for separation of environmental levels of technetium-99, radium and actinides from soil and water; separation of actinides from soil and water matrix interferences; and isolation of strontium. We are also developing methods for simultaneous detection of multiple isotopes (including nonradionuclides) using the new instrumental technique inductively coupled plasma/mass spectrometry (ICP/MS). These ICP/MS methods would more efficiently replace alpha and beta counting techniques. Integration and automation of the separation methods with the ICP/MS methology using flow injection analysis is underway with the objectives of achieving more reproducible results, reducing labor costs, cutting analysis time, and reducing secondary waste generation through miniaturization of the process. The final product of all activities will be methods which are available (published in DOE's analytical methods compendium) and acceptable for use in regulatory situations.

Keyword(s): analytical chemistry, flow injection analysis, inductively coupled plasma/mass spectrometry, radioanalytical, separations

Poster presentation.

This work is supported by the U.S. Department of Energy, Assistant Secretary for Environmental Remediation and Waste Management, Office of Technology Development, under contract W-310109-Eng-38.


Terence M. Fairbanks, Tissa Illangasekare and Dobroslav Znidarcic, Department of Civil, Environmental and Architectural Engineering, University of Colorado, Boulder, CO 80309-0428

Subsurface stratigraphy plays an important role in the migration and distribution of chemical and waste products that are in the form of nonaqueous phase liquids (NAPL). In the saturated zone, coarse layers act as preferential flow channels and as pockets for fluid entrapment. The entrapment saturations resulting from these macro-scale soil heterogeneities are much larger than the residual saturations controlled by the micro-scale pore characteristics of the soil. The phenomena of preferential flow and macro-scale entrapment have been observed in our laboratory spill simulations and in the field. It is our hypothesis that they are caused by the capillary barrier effects existing at the interfaces of the coarse and fine sand formations. Precision flow experiments were conducted in soil cells to obtain quantitative data to understand the nature of these capillary barrier effects. A complete testing program was implemented with five sands using both air and a light NAPL as the non-wetting phase. The experimental design and the results are presented. The experimental results are used to identify the characteristics of the soils in layered aquifers that produce barrier effects. The results of this study will help in developing field characterization techniques to determine soil parameters needed in modeling nonaqueous phase fluid flow and entrapment in heterogeneous aquifers. This knowledge will also be useful in the design of fine soil barriers for the containment of plumes and coarse soil traps for the recovery of waste fluids.

Keyword(s): groundwater contamination, organic wastes, nonaqueous phase fluids.

Oral presentation in research track.


R.B. Galloway, B.S. Langkopf and J.T. McCord, Environmental Restoration Department, Sandia National Laboratories, P.O. Box 5800, Albuquerque, NM 87185-0727

The Environmental Restoration Project at Sandia National Laboratories/New Mexico (SNL/NM) is tasked with performing assessments and remediation of waste sites resulting from over 40 years of weapons development and testing. Operable Unit 1295, Septic Tanks and Drainfields, includes 23 different sites at SNL/NM where hazardous and radioactive wastes may have been released into the environment. A RCRA Facility Investigation (RFI) Work Plan for the sites has been submitted to the U.S. EPA for approval. This presentation will outline the proposed technical approach for the characterization of these sites for the RFI Report and the Corrective Measures Study. The phased assessment of the sites is designed to quickly eliminate sites where contamination levels are at or below background levels and fully determine the nature and extent of contamination at the other sites. Field investigation methods and sample field-screening techniques will also be presented.

Keyword(s): septic systems, site characterization, drainfields, field screening methods.

Poster presentation.


P. Gandhi, L.E. Erickson and L.T. Fan, Department of Chemical Engineering, Durland Hall, Kansas State University, Manhattan, Kansas 66506-5102

Subsurface contaminants are frequently encountered as mixtures of nonaqueous phase liquids (NAPLs) at sites contaminated by gasoline or coal tar comprising organic mixtures. The leaching of these organic mixtures from the aquifer has been examined with and without biodegradation. The results obtained have been compared with the limiting case of a single component NAPL.

Various physical processes involved have been quantified based on the assumptions that liquid- liquid and sorption equilibria are established at the beginning of each flushing; oxygen required for biochemical oxidation is completely consumed by the end of each flushing; and the rate of biochemical oxidation obeys the Monod kinetics for a multi-substrate system, characterized by an oxygen utilization factor. This has given rise to an equilibrium model expressing the mass fraction of any component remaining in the aquifer, its aqueous concentration, and the composition of the NAPL as functions of the number of flushings. The results of the simulation with the model demonstrate that bioremediation can significantly reduce the time necessary for removing the components of intermediate solubility such as xylene. Highly soluble components of the NAPL are mainly removed by the pump-and-treat mechanism while the components of extremely low solubility are unavailable to the microbes as substrates in a multi-component system. The results also demonstrate that relatively soluble contaminants tend to persist for a longer duration in the mixture than when they exist as pure components and that this effect is magnified by the inclusion of a non-soluble component in the mixture. The proposed mass balance approach yields a useful bound of the effectiveness of biodegradation aided dissolution of a multi- component NAPL; the approach is relatively simple and computationally efficient.

Keyword(s): dissolution, biodegradation, NAPL, mixture, equilibrium.

Oral presentation in research track.


Julio Garcia1, Ranjith B. Mapa1, Tissa Illangasekare1 and Thomas Russell2, 1Department of Civil, Environmental and Architectural Engineering, University of Colorado, Boulder, CO 80309-0428; 2Department of Mathematics, University of Colorado, Denver, CO 80309

In the development and application of groundwater models, it has been recognized that the process understanding at a small scale does not necessarily lead in a straightforward manner to appropriate models at a large scale. The upscaling of models of water flow and solute transport in saturated porous media has been the topic of significant theoretical investigation. Data from a few field sites have been used to validate some of these theories. However, the field data will have limitations with respect to accuracy and the inability to fully characterize the soil conditions which define the field-scale heterogeneities. Laboratory investigations in soil columns and tanks can provide much accurate data compared to the field. Also, soils which are used to create heterogeneities can be well characterized in the laboratory. The purpose of the study reported in this paper is to develop an experimental facility to generate accurate data on water flow and solute transport in heterogeneous aquifers. The data is used to study the scale effects in solute transport (scale-dependent dispersivity). The experimental facility consists of a horizontal soil tank of dimensions 2.44 m x 1.22 m. Soil samples of approximately 5 cm thickness can be packed into the tank. Two constant head end tanks are used to create various groundwater flow velocities. Pressure measurements and solute sampling will be done over the horizontal plane using 45 sampling ports drilled through the top wall. These same ports can be used for tracer injection. The experiments will involve injection of a tracer into the flow field and measurement of water pressure and solute concentrations at prescribed locations in the flow field. Pressure measurements are done using an automated pressure scanning system. The measured values of flow, pressure and solute concentrations are used to estimate hydraulic conductivity and dispersivity using inverse problem solution techniques. Both homogeneous and heterogeneous soil packing configurations could be investigated. Results from simulations with homogeneous packing are presented.

Keyword(s): upscaling, dispersivity.

Poster presentation.


J. Golchin1 and S. Nelson2, 1Iowa Department of Natural Resources, Des Moines, IA 50319; 2Midwest Gas, Sioux City, IA

A source removal action plan was developed by Midwest Gas and the Iowa Department of Natural Resources to address the source coal tar contamination within the underground gas holder basin at Former Manufactured Gas Plant (MGP) sites. The procedure utilizes a mixture of coal, contaminated soil, and coal tar sludge to provide a material that had suitable material handling characteristic for shipment and burning in high efficiency utility boilers. Screening of the mixture was required to remove oversized debris and ferrous metal. The resulting mixture did not exhibit toxic characteristics when tested under the Toxicity Characteristics Leaching Procedure (TCLP). Test results on the coal tar sludges have indicated that the more pure coal tar materials may fail the TCLP test and be classified as a RCRA hazardous waste. The processing procedure was designed to stabilize the coal tar sludges and render those sludges less hazardous and as a result pass the TCLP test. This procedure was adopted by the Edison Electric Institute to develop a national guidance document for remediation of MGP sites. EPA Office of Solid Waste and Emergency Response recommended this strategy to the Regional Waste Management Directors as a practical tool for handling wastes that may exhibit the RCRA characteristics.

Keyword(s): stabilization, coal tar, RCRA, TCLP.

Oral presentation in technology transfer track.


M. Govindaswami, D.J. Feldhake and J.C. Loper, University of Cincinnati Medical Center, Cincinnati, OH 45262-0524

PAHs are a class of widespread pollutants, some of which have been shown to be genotoxic, hence the fate of these compounds in the environment is of considerable interest. Research on the biodegradation of 4 and 5 ring PAHs has been limited by the general lack of microbial isolates or consortia which can completely degrade these toxicants. Heitkamp and Cerniglia have described an oxidative soil Mycobacterium- strain PYR-1 that metabolizes pyrene and fluoranthene more rapidly than the 2 and 3 ring naphthalene and phenanthrene; although some metabolites of benzo-(a)-pyrene (BaP)were detected, no mineralization of BaP was observed. In 1991 Grosser et al. reported the isolation of a Mycobacterium sp. which mineralizes pyrene and also causing some mineralization of BaP. Our study describes a comparative analysis of these two strains, which show very similar colony morphology, growth rate and yellow-orange pigmentation. Genetic differences were shown by DNA amplification fingerprinting (DAF) using two arbitrary GC-rich octanucleotide primers, and by sequence comparison of PCR amplified 16S rDNA, although both strains show similarity closest to that of the genus Mycobacteria. These 16S rDNA sequences are in use for the construction of strain-specific DNA probes to monitor the presence, survival and growth of these isolates in PAH-contaminated soils in studies of biodegradation.

Keyword(s): polycyclic aromatic hydrcarbons, mycobacteria, pollution.

Poster presentation.

Supported by NIEHS Grant ES04908.


G.S. Groenewold, J.C Ingram, A.D. Appelhans, J.E. Delmore and D.A. Dahl, Idaho National Engineering Laboratory, P.O. Box 1625, Idaho Falls, ID 83415-2208

Ions derived from tributyl phosphate (TBP) are observed in the static secondary ion mass spectrum of minerals exposed to TBP; these ions are very dependent on the nature of the mineral surface. When TBP is adsorbed onto a reducing site, specifically Fe(ll), the compound will be reduced upon SIMS bombardment, and ions corresponding to tributyl phosphite will be observed. This observation can be made for TBP that is adsorbed to basalt samples having substantial reducing phases, and FeO. The tributyl phosphite that is formed from the reduction of TBP concurrently undergoes a hydride abstraction during the SIMS process, and proceeds to eliminate one and/or two C4H8 and/or one H20 molecules to form ions at m/z 193+, 137+, and 119+. These same ions can be observed in the SIM spectrum of tributyl phosphite adsorbed onto the same mineral surfaces, although ions corresponding to the protonation of tributyl phosphite are also observed. The m/z 193+ and 137+ ions are also observed in the methane chemical ionization (CI) mass spectrum of tributyl phosphite, which demonstrates that hydride abstraction is also occurring in the gas-phase (although this is a minor ion series in the CI mass spectrum).

When TBP is adsorbed onto surfaces that are not reducing, then protonation or hydride abstraction occur. Ions resulting from protonation derive from the sequential elimination of up to three C4H8 molecules (m/z 211+, 155+, 99+) from [M + H]+, and are observed from "oxidized" basalt samples. These ions are also observed from Fe2O3 although in this case substantial hydride abstraction is also observed. This latter ionization mode is followed by the elimination of one and/or two C4H8 molecules to produce ions at m/z 209+ and 153+. Both ion series (protonation and hydride abstraction) can be observed in the CI mass spectrum of TBP. Significantly, none of the ions attributed to tributyl phosphite are observed in the CI mass spectrum of TBP, which supports the idea that their formation is the result of a surface process, and not the result of gas-phase ion chemistry.

A significant ion is observed at m/z 99+ in the SIM spectrum of tributyl phosphite on oxidized surfaces: Fe2O3 and basalt This ion is interpreted in terms of tributyl phosphite being oxidized during the SIMS analysis.

Keyword(s): mass spectrometry, surface analysis, reduction.

Oral presentation in research track.


R.J. Grosser, D. Warshawsky and B. Kinkle, Department of Biological Sciences, University of Cincinnati, Cincinnati, OH 45221-0006

Degradation of many hazardous compounds in the soil environment can be affected by the different components of the soil matrix as well as the degrading microbial population present. Fulvic acids were extracted from various soils (both impacted and non-impacted) and then used to determine their effect on pyrene mineralization by an isolated pyrene-degrading Mycobacterium sp. A comparison of non-impacted to impacted soil fulvic acids was done to determine whether these fulvic acids have different binding affinities for the pyrene. To determine the effects of fulvic acid addition on mineralization, samples were prepared using sterile sand and different concentrations of fulvic acids (1, 4 and 10%) lyophilized onto the sand. Slurries were made using sterile water and added Mycobacterium cells. Serum bottle radiorespirometry was used to measure mineralization of added 14C-pyrene over a 32 day period. Results show that the addition of fulvic acid up to 5% had little overall effect on pyrene extent of mineralization by the bacteria, but the 5% fulvic acid addition resulted in a lag before mineralization began. The 10% fulvic acid addition decreased mineralization with an extended lag period. Because the addition of fulvic acids to the slurries decreases the pH, survival studies were conducted at unadjusted pH and at pH adjusted to 7, and with varying fulvic acid concentrations up to 2.5%. Activity of the Mycobacterium cells was measured as 14C-acetate incorporation into lipids and indicates that the activity in both unadjusted and pH 7 slurries was similar with up to 1% fulvic acid added. Fulvic acid addition of 2.5% resulted in six times greater activity in pH 7 slurry than in the unadjusted slurry. The decreasing mineralization of pyrene with increasing levels of fulvic acid addition appears to be due to toxicity to the microbes and possible sorption of the compound making it less bioavailable.

Keyword(s): Mycobacterium, mineralization, fulvic acid, sorption, bioavailability.

Poster presentation.


Yinghong He1, Suzette Burckhard1, A.P. Schwab2, and M.K. Banks1, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

Ion exchange, surface adsorption/desorption, complexation by organic materials, and precipitation/dissolution of discreet solid phases are typical mechanisms that control heavy metals in solution. Understanding the mechanism that is responsible for a given metal is critical to prediction of leaching behavior. Plants can affect mobility of heavy metals by releasing organic compounds into the rhizosphere. Some of the organics released by plant roots (and microorganisms) are strong complexing agents for metals and can potentially increase metal mobility. Sorption and desorption reactions (kinetics and isotherms) of a variety of heavy metals will be determined for diffusion controlled and ion exchange reactions. Organic compounds that could impact metal transport, mine tailings, and soil will be equilibrated for approximately 18 hours in batch studies. The solution phase will be separated and concentrations of target metals measured. This research will quantify heavy metal adsorption/desorption characteristics of soil influenced by plant roots.

Keyword(s): heavy metals, rhizosphere, leaching, adsorption.

Poster presentation.


B.R. Helland, J.L. Schnoor and P.J. Alvarez, Department of Civil and Environmental Engineering, University of Iowa, Iowa City, IA 52242

We are investigating the abiotic reductive dechlorination of carbon tetrachloride using elemental iron as an electron donor. Equation 1 shows the thermodynamic favorability of the reduction of carbon tetrachloride with the oxidation of iron metal at neutral pH.

Fe0 + CCl4 + H+ ® Fe2+ + CHCl3 + Cl- Eo’ = +0.99 Volts (1) Reduction of mono-, di-, and trichloromethane is also thermodynamically favorable to a lesser degree. Our work has provided preliminary feasibility data for the development of novel and inexpensive remediation techniques for carbon tetrachloride in groundwater, including an in-situ process, which may be applicable to other halogenated priority pollutants. Batch experiments were conducted in aqueous solution to determine stoichiometric and pseudo first order reaction rate coefficients for the sequential reduction of carbon tetrachloride to chloroform, methylene chloride, chloromethane and methane using powdered iron metal as the sole reductant. Aerobic reduction rates were slower than anaerobic rates, presumably due to dissolved oxygen from the ambient atmosphere competing with carbon tetrachloride as the terminal electron acceptor. Reduction rates were accelerated at higher temperatures in agreement with the Ahrennius equation. Soil column experiments which simulated reductive dechlorination of carbon tetrachloride in subsurface environments were conducted under anaerobic conditions. These experiments showed the beneficial effect of treatment with iron powder. Potential field applications include in-situ chlorinated solvent remediation and containment and treatment of off gasses.

Keyword(s): carbon tetrachloride, in-situ, iron, reductive dechlorination.

Poster presentation.


J.M. Hillary, Industrial Process Engineering Research, EG&G Idaho Inc., PO Box 1625, Idaho Falls, ID 83415-2203

Recent surveys of mixed wastes in interim storage throughout the 30-site Department of Energy complex indicate that only 12 of those sites account for 98% of such wastes by volume. Current inventories at the Idaho National Engineering Laboratory (INEL) account for 38% of total DOE wastes in interim storage, the largest of any single site. For a large percentage of these waste volumes, as well as the substantial amounts of buried and currently generated wastes, thermal treatment processes have been designated as the technologies of choice.

Current facilities and a number of proposed strategies exist for thermal treatment of wastes of this nature at the INEL. High level radioactive waste is solidified in the Waste Calciner Facility at the Idaho Central Processing plant. Low level solid wastes until recently have been processed at the Waste Experimental Reduction Facility (WERF), a compaction, size reduction, and controlled air incineration facility. WERF is currently undergoing process upgrading and RCRA Part B permitting.

Recent systems studies have defined effective strategies, in the form of thermal process sequences, for treatment of wastes of the complex and heterogeneous nature in the INEL inventory. These recommendations have resulted in the proposal of combinations of treatment, storage and disposal facilities to most effectively fill these needs.

A number of research, development and demonstration projects are also currently active in both the low temperature incineration and high temperature melting regimes, to provide the fundamental scientific understanding necessary to support design, construction and operation of the proposed facilities. These studies address the partitioning of the toxic metals and radionuclides to the treatment process products and strategies for minimizing release of such residuals to the environment.

This presentation reviews the current status of operating facilities, active studies in this area, and proposed strategies for thermal treatment of INEL wastes.

Keyword(s): thermal treatment, incineration, melting, mixed waste, heavy metals.

Oral presentation in technology transfer track.


R.M. Hoffman1, V.P. Visser1, L.C. Davis2, L.E. Erickson3, N. Muralidharan3, R.M. Hammaker1 and W.G. Fateley1, Departments of Chemistry1, Biochemistry2, and Chemical Engineering3, Kansas State University, Manhattan KS, 66506

This study addresses a viable and natural solution to the elimination of volatile organic compounds (VOCs), pollutants, through the bioremediation process. Plants and associated rhizosphere bacteria have the ability for bioremediation of both volatile and non-volatile organic compounds. For volatile compounds, intersystem transfer by transpiration may be a matter for concern when plants interact with such materials. We have monitored, using FT-IR, the potential transfer of toluene from subsurface water in the presence of toluene-adapted alfalfa plants. These experiments show that the plants and/or their associated micro-organisms effectively degrade toluene so that potential intersystem transfer of VOCs by transpiration may be quite manageable with adapted-plants. Presently, we are monitoring l,l,l-trichloroethane (TCA) and trichloroethylene (TCE) from subsurface water and gas phase above plants. TCA does not show an indication of degradation whereas TCE does. Methane is produced in the groundwater but not transferred to the atmosphere, indicating the presence of a consortium of methanogens and methanotrophs in this soil. The TCE presumably is the substrate for methane production based on chloride ion accumulation. The majority of TCE must be degraded aerobically to yield CO2 in the vadose zone. The FT-IR spectrometer can quickly determine and analyze contaminants in the gas phase, groundwater and plant tissue successfully.

Keyword(s): FT-IR, bioremediation, volatile organic compounds.

Oral presentation in research track.


Andy Hong, Department of Civil Engineering, University of Utah, Salt Lake City, UT 84112

Heavy metal contamination of soil is a common problem encountered at many hazardous waste sites. Lead, chromium, cadmium, copper, zinc and mercury are among the most frequently observed metal contaminants. They are present at elevated concentrations at many National Priority List sites. Heavy metals are toxic to people and pose a great risk for safe groundwater supply. Once released into the soil matrix, most heavy metals are strongly retained and their adverse effects can last for a long time.

Chelating extraction of heavy metals from contaminated soils has recently been seen as a treatment method. However, only a few chelates, familiar ones such as EDTA and NTA, have been tried for this application, and the choice of chelates for decontamination purpose appears to be haphazard. There exists a need to assess the full potential of this technology in removing and/or recovering heavy metals from contaminated media, e.g., soils and mine tailings ponds. This paper will: present a methodology to examine a large number of chelates and to identify those chelates suitable for the selective removal or recovery of various heavy metals. Chemical equilibrium modeling has been used to examine over 100 chelates for their extraction potential; present the experimental results of heavy metal extraction from soil using a few selected chelates. The extraction of metals including cadmium, copper, lead and zinc has been studied under different pH, soil suspension, total chelate concentration, total carbonate concentration, and age conditions; demonstrate that, through a proper choice of chelate, the extraction can be made more selective toward heavy metals; demonstrate that, with the chosen chelates, the extracted metals can be readily recovered as solid precipitates and the soluble chelates reclaimed and reused.

Keyword(s): metal, soil, chelate, contamination, remediation.

Poster presentation.


M.S. Hong, K.W. Brown, B.E. Dale, K.C. Donnelly, L.Y. He and K.V. Markiewicz, Depts. of Chemical Engineering, Soil & Crop Sciences, and Veterinary Anatomy & Public Health, Texas A & M University, College Station, TX 77843

Detoxification of pentachlorophenol- containing wood preserving waste was monitored under ambient, enhanced and chemical pretreatment conditions for genotoxicity and parent compound removal. Samples were collected throughout the treatment periods and sequentially extracted with dichloromethane and methanol with the Tecator Soxtec apparatus. The organic extracts were analyzed on GC/ECD and GC/MS. The extract mutagenic and genotoxic potentials were evaluated with and without metabolic activation with the Salmonella Microsomal and E. coli Prophage Induction assays. The Salmonella mutagenic responses of extracts from Weswood soil amended with wood preserving waste and treated under ambient conditions were 2.0, 34.6 and 2.4 times greater than the solvent control on days 0, 540 and 1,200 respectively. Organic extracts of soil amended with wood preserving waste and treated under enhanced conditions in a solid-phase rotating drum bioreactor had mutagenic potentials of 3.4, 4.9 and 3.5 on days 0, 14 and 30, respectively. Extracts from wood preserving waste sludge treated with potassium polyethylene glycol were shown to have mutagenic potentials of 2.8, 6.1 and 3.8 at 0, 10 and 30 minutes. The results indicate that the initial products of the wood preserving waste detoxification under all treatment conditions appear to have greater genotoxic potentials than the starting material. The results also suggest that a more rapid detoxification occurs under enhanced and chemical pretreatment conditions.

Keyword(s): detoxification, chemical pretreatment, hazardous waste, pentachlorophenol, bioremediation.

Oral presentation in research track.

This research was supported by NIH grant P42 ES04917.


G.L. Horst1, W.L. Powers2, P.J. Shea2, D.R. Miller1 and C.L. Stuefer-Powell2, 1377 Plant Science, and 2279 Plant Science, University of Nebraska, Lincoln, NE 68583

Organic solute mobility may be better understood by simulating field conditions in the greenhouse. Turf/soil columns from a Kentucky bluegrass (Poa pratensis L.) area near Mead, NE, and Ames, IA, were excavated, concrete encased, and transported to a greenhouse. Organic solutes applied at recommended rates included the herbicides 2,4-D, dicamba, mecoprop, and pendimethalin, the insecticides isazofos and chlorpyrifos, and metalaxyl fungicide. Nitrogen (15N urea) was applied at 49 kg ha-1. Potassium bromide was applied at 100 kg ha-1 as a conservative tracer. Two irrigation water regimes (2.5 and 5.0 cm) with drainage suction (500 cm water) were imposed. Daily column and leachate weights were used to determine water balance and schedule irrigations. The distribution of organic solute residues among leachate, verdure, thatch, and soil depth to 50 cm, were determined and related to nitrogen and water tracer movement. Residual analysis pesticide in the turf/soil profile was metalaxyl > chlorpyrifos = pendimethalin > isazofos. The amount of metalaxyl residue was mainly influenced by differences in soil types. The amount of isazofos residue was affected by irrigation level in the Nebraska soil. Chlorpyrifos and pendimethalin residues were similar in both soils and were not affected by irrigation level.

Keyword(s): pesticides, irrigation, chemical properties, soil properties.

Poster presentation.


H.J. Huebner, K.W. Brown, K.C. Donnelly and L.Y. He, Department of Veterinary Anatomy & Public Health, Texas A & M University, College Station, TX 77843

The objective of this study was to evaluate the efficiencies of the automatic Soxtec and U. S. EPA SW846 Soxhlet soil extraction methods. In phases one and two of the experiment, extractions were performed on silicon dioxide matrices and silt-loam soils spiked with benz(a)pyrene, pentachlorophenol, and naphthalene at three concentration levels. Each test sample contained either an individual chemical or a 1:1:1 mixture of all three chemicals. Phase three consisted of extractions performed on a silt-loam soil spiked with a coal tar complex mixture. Soxtec samples were sequentially extracted with dichloromethane and methanol while Soxhlet samples were extracted with dichloromethane.

Gas chromatographic results obtained from sample extract analysis were used to calculate percent recoveries of the chemicals. The recoveries of benz(a)pyrene and pentachlorophenol in the Soxtec procedure ranged from 55-88% and 49-88%, respectively. For the Soxhlet method, the recoveries ranged from 46- 73% and 52-87%, respectively. Complex mixture recoveries ranged from 50-60% for both procedures. The mutagenic potentials of the solvent extracts were evaluated using Salmonella typhimurium strain TA98 with and without metabolic activation. Assay results indicated a positive correlation between mutagenic response, assay controls, and the chemical concentrations derived from GC analysis. The data indicate that the Soxtec method, which requires 2 hours, is as effective as the traditional 16 hour Soxhlet extraction procedure for recovering organic chemicals from contaminated matrices. The Soxtec method, thus, offered substantial time and cost savings.

Keyword(s): pentachlorophenol, benz(a)pyrene, extraction, bioassay, soil.

Poster presentation.

This research was supported by NIH grant P42 ES04917.


H.J. Huebner, K.W. Brown, K.C. Donnelly and L.Y. He, Department of Veterinary Anatomy & Public Health, Texas A & M University, College Station, TX 77843

Numerical models of water flow and solute transport have been extensively used in problems in groundwater quantity and quality management. The fundamental processes which govern the flow and transport behavior are fairly well understood, and the models have been adequately validated using both laboratory and field data. Field and laboratory techniques for the determination of parameters needed in these models are generally available. However, the same cannot be said about the models which have been developed to simulate multiphase flow in aquifers. The need to model multiphase flow arises in dealing with contamination problems involving organic chemicals and waste products which are in the form of nonaqueous phase liquids. Existing models use the basic equations, mathematical formulations, and numerical schemes which have been used in reservoir simulators in petroleum engineering applications. The field application of these models as prediction tools for the design of remediation schemes in hazardous waste problems have been limited for a variety of reasons. These models have not been adequately validated in the laboratory or in the field due to the scarcity of data. The models sometimes fail in attempting to simulate flow and entrapment behavior under heterogeneous soil conditions that are commonly encountered in the field. Accurate field calibration and prediction become difficult due to the limitations of the field and laboratory techniques which are used to obtain the model parameters. The assumptions which are made in modeling the mass transfer from the nonaqueous phase to aqueous phase become questionable under some conditions of ganglia formation and macro-scale entrapment. Results from laboratory spill simulations conducted in large soils tanks, soil columns, soil cells and field investigations, and model analysis are used to discuss some of these limitations. Conclusions on possible improvements to models and issues related to laboratory and field characterization are presented.

Keyword(s): remediation design, multiphase flow, numerical models, groundwater models.

Oral presentation in research track.


W.P. Inskeep, S. Sun, H. Gaber, J.M. Wraith and R. Doughten, Department of Plant, Soil and Environmental Sciences, Montana State University, Bozeman, MT 59717

An understanding of degradation rates of organic chemicals under transport conditions is critical for enhancing biodegradation rates in-situ. Currently, most solute transport models assume first-order degradation kinetics, and users generally rely on rate constants determined under batch conditions. However, the rates of solute diffusion and sorption-desorption reactions, which influence contaminant bioavailability, are dependent on pore water velocities. Consequently, batch degradation rates are probably not adequate for describing actual degradation rates occurring during transport. To test this hypothesis, we conducted transport experiments using l4C-labeled 2,4-D in disturbed soil columns at three different pore water velocities under unsaturated flow conditions. The amount of 2,4-D in effluent fractions was determined using scintillation analysis, and plotted as a function of pore volume (i.e. 2,4-D breakthrough curves, BTCs). The percent of applied 2,4-D degraded during transport was directly related to the residence time of 2,4-D in the column. For example, higher pore water velocities corresponding to lower residence times resulted in lower total amounts of 2,4-D degraded. These results can generally be predicted using the convection-dispersion equation for solute transport with a first-order model for 2,4-D degradation using batch-determined degradation rate constants. However, batch-determined degradation rates significantly underpredicted the amount of 2,4-D degraded during transport conditions. Optimized rate constants determined for the transport experiments were greater than batch-determined degradation rate constants, and increased with increasing pore water velocity. It is hypothesized that higher pore water velocities yield greater contaminant diffusion rates through interstitial pores and increase contaminant desorption rates, both resulting in enhanced bioavailability.

Keyword(s): biodegradation, transport, 2,4-D.

Poster presentation.


D.B. Jackson and D.J. Dollhopf, Reclamation Research Unit, Montana State University, Bozeman, MT 59717

Research was undertaken to assist the Bureau of Land Management and Forest Service in determining the best rehabilitation techniques for mine waste streamside deposits in the Elkhorn Mountains of Montana. These wastes have a high sulfur content, a low pH, and are contaminated with elevated levels of arsenic, lead and other trace metals.

Utilizing a randomized block experimental design, these mine wastes were amended with calcium carbonate, calcium hydroxide, manure and fertilizer. Three grass species, Agrostis alba, Deschampsia caespitosa and Elymus cinereus, were grown under greenhouse conditions to determine their performance.

Keyword(s): revegetation, trace metals, acid mine waste.

Oral presentation in research track.


S. Jennings and D.J. Dollhopf, Reclamation Research Unit, Montana State University, Bozeman, MT 59717

The oxidation of sulfide minerals in mine waste is a widespread source of resource degradation, often resulting in the generation of acidic water and mobilization of heavy metals. The quantity of acid-forming minerals present in mine waste, dominantly as pyrite (FeS2) is routinely determined by acid-base account (ABA) analytical methods. The acid-base account method specifies the use of extraction techniques to determine the total quantity of acid-forming sulfur compounds in a sample relative to the neutralizing potential. However, when common sulfide and sulfate minerals were subjected to ABA extraction methods, the ABA method failed to accurately distinguish the acid-forming from nonacid- forming minerals, resulting in errors in the determination of potential acidity. These analytical errors are subsequently reflected in inaccurate liming of acid producing waste materials resulting in either excessive cost when potential acidity is overestimated, or potential reclamation failure when potential acidity is underestimated.

Keyword(s): acid-base account, heavy metals, pyrite oxidation, acid mine drainage.

Oral presentation in research track.


Jagath J. Kaluarachchi and K.M. Mesbah-ul Islam, Utah Water Research Laboratory, Utah State University, Logan, UT 84322-8200

In-situ soil vapor extraction, commonly known as soil gas venting, has become an effective remediation technique to remove volatile organic contaminants from the subsurface. The major drawback of this scheme is that the efficiency tends to decrease with the presence of less-volatile organics. Since fluid properties are highly sensitive to ambient subsurface temperature, it is potentially feasible to perform vapor extraction under induced thermal gradients, also called thermal venting, to enhance the recovery of less- volatile fraction of the contaminant mass. The objective of this work is to demonstrate the applicability of the thermal venting technique to enhance the recovery of organics from the liquid unsaturated zone.

To investigate the applicability of the thermal venting scheme, a multicomponent, non- isothermal theoretical model was developed to predict the coupled gas flow, heat transport and mass transport in the unsaturated zone. Heat input to the system was through the incoming gas flow. As the inlet heated air passes through the contaminated zone, enhanced volatilization of liquid contaminant occurs at the air-liquid interface. Heat energy transport was considered under thermodynamic equilibrium condition between different fluid phases. Also the latent heat transfer due to moisture condensation and evaporation of the organic phase was considered. Mass transport analysis used local phase equilibrium approximation to partition component mass between the air, oil, water and adsorbed phases within the residual oil plume. Phase partitioning between the aqueous, air and solid phases were considered outside the organic plume. A number of hypothetical simulations were performed to demonstrate the applicability of the proposed technique. The results were used to demonstrate the propagation of heat through the soil due to the effects of conductive and convective heat fluxes, the latent heat absorbed during contaminant volatilization, and the latent heat released during condensation of water vapor of the incoming air. The results also showed the rate of mass recovery and oil saturation in the contaminated zone during venting. The results of the model will also demonstrate the effectiveness of various forms of heat energy input, energy balance within the system, and the efficiency of the technique with different organic compounds.

Keyword(s): thermal venting, vapor extraction, less-volatile, remediation, hydrocarbons.

Oral presentation in research track.


Aditya Khindaria, Thomas A. Grover and Steven D. Aust, Biotechnology Center, Utah State University, Logan, UT 84322-4705

Reduced radicals of highly oxidized and electron deficient halocarbons (carbon tetrachloride, chloroform, dichloromethane, trichloroethylene, 1,1,1-trichloroethane) were detected by electron paramagnetic resonance spin trapping spectroscopy in a reductive reaction system consisting of lignin peroxidase, hydrogen peroxide, veratryl alcohol, ethylenediaminetetraacetic acid or oxalate, hydrogen peroxide, phenyl-N-t-butylnitrone (as the spin trap) and 1% of the halocarbon. This is a free radical mediated process which uses the organic acid as the electron donor. To correlate the involvement of this mechanism in vivo, mineralization of trichloroethylene and carbon tetrachloride was studied by ligninolytic cultures of Phanerochaete chrysosporium. Both trichloroethylene (20.3% of 10 parts per million) and carbon tetrachloride (18.8% of 10 parts per million) were mineralized by the fungus over a period of 9 days. Mineralization of these chemicals did not take place in formaldehyde- killed control cultures or nonligninolytic cultures of the fungus, indicating that lignin peroxidases play an important role in the process. Since all the components of the reductive system are excreted entracellularly, we propose that this fungus can be used for bioremediation of these halocarbons.

Keyword(s): Phanerochaete chrysosporium, white rot fungi, aliphatic halocarbons, trichloroethylene.

Oral presentation in research track.


S.C. Kim1, M.K. Banks2 and A.P. Schwab3, 1Department of Chemical Engineering, 2Department of Civil Engineering, and 3Department of Agronomy, Kansas State University, Manhattan, KS 66506

Polycyclic aromatic hydrocarbons (PAHs) are a class of potentially hazardous chemicals that exhibit toxic, mutagenic or carcinogenic properties. Microbial degradation is the major route through which PAHs are removed from contaminated environments although other mechanisms such as volatilization, leaching and photodegradation may also be effective. The rhizosphere contains a diversity of microorganisms that contribute to plant health and soil homeostasis. Recent studies indicate that microorganisms in the rhizosphere can degrade toxicants of concern to human health and the environment. The increased density and diversity of rhizosphere microflora may be an important factor for enhanced microbial degradation of PAHs.

The objective of this study is to evaluate degradation of a number of different PAHs in rhizosphere and non-rhizosphere soil. It has been shown that the biodegradation rates of PAHs increase as the number of PAH rings decrease, but there is little information about the biodegradation in rhizosphere soil. This study will provide results from a microcosm experiment designed to evaluate degradation of PAHs in rhizosphere and non-rhizosphere. Also, kinetic models will be developed to represent data collected.

Keyword(s): polycyclic aromatic hydrocarbons, rhizosphere, biodegradation.

Poster presentation.


S.C. Kim1, L.E. Erickson1 and E.Y. Yu2, 1Center for Hazardous Substance Research, Kansas State University, Manhattan KS 66506; 2Department of Chemical Engineering, Chonnam National University, Kwangju 500-757, Korea,

Methane is the main component of natural gas and has been connected with global warming. The oxidative coupling of methane has been studied to enhance the C2 hydrocarbons selectivity and to reduce the formation of carbon oxides. The acid sites of supported catalysts play an important role in the formation of carbon oxides. The supported Zn-Oxide catalyst with a-Al203 shows no acidity in temperature programmed desorption by using NH3 and exhibits good C2 hydrocarbons selectivity. The optimum loading of Zn-Oxide on a-Al203 is 60wt%. The specific surface area of the catalyst appears not to influence activity. Using alkali metal salts as a promoter in the Zn- Oxide (60wt%)/a-Al203 catalyst, the activity performance for C2 hydrocarbons is LiCl>NaCl>KCl, and that performance is well correlated with the apparent molal enthalpy of formation in alkali halides. The activity performance for reducing carbon oxides is LiCl>KCl>NaCl, which is well correlated with the melting point in alkali halides.

Keyword(s): methane, oxidative coupling, C2 hydrocarbons, carbon oxides, promoter.

Poster presentation.


Olga Koper and Kenneth J. Klabunde, Department of Chemistry, Kansas State University, Manhattan, KS 66506

Conventionally prepared and autoclave prepared calcium oxides were studied to understand the decomposition of the carbon tetrachloride and trichloroethylene processes. To explain the increased ability of autoclave prepared calcium oxide for decomposition of chlorocarbons, different methods were employed. The infra-red technique for observing adsorbed species on calcium oxide after adsorbing carbon tetrachloride and spring balance studies to understand the importance of the basic and acidic sites on the oxide were done. To enlighten some other differences between these two oxides, transmission electron microscopy, thermogravimeric analysis, and instrumental analysis were used.

Keyword(s): chlorocarbon, calcium oxide, decomposition.

Oral presentation in research track.


M.M. Kori and S.K. Gupta, Centre for Environmental Science and Engineering, Indian Institute of Technology, Powai, Bombay 400 076, INDIA

Eleven polyelectrolytes were tried separately to treat the wastewater from a paint manufacturing industry. Amongst these, Zetag 66, a cationic polyelectrolyte was found to be most effective. A dosage of 5 mg/L of this polyelectrolyte was found to be adequate to achieve 65% COD removal, 97% suspended solids removal and 90% heavy metals removal. The use of this polyelectrolyte assumes significant importance as it eliminates the use of alum completely. This elimination of alum consumption results in considerable reduction of effluent treatment plant sludge which is a hazardous waste. The savings that results in the primary treatment is an added advantage.

Keyword(s): polyelectrolyte, heavy metals, alum consumption, hazardous waste.

Oral presentation in research track.


Srinivas Kothandaraman, Robert C. Ahlert and E.S. Venkataramani, Chemical and Biochemical Engineering, Rutgers University, PO Box 909, Piscataway, NJ 08855-0909, 908-932- 3399, FAX 908-932-4963

A near-infrared spectrophotometer has been interfaced with a high-pressure apparatus, utilizing a fiber optic probe. The system was designed to re-circulate carbon dioxide for studies of supercritical extraction. Near-infrared spectra, over the range of 400 to 2400 nm, have been obtained for dense carbon dioxide, at pressures to 8 MPa. Spectra for chlorinated biphenyls in supercritical carbon dioxide have been recorded also. Data is stored directly; a software package correlates solubility in dense carbon dioxide for long-term information management. New opportunities for non-invasive sampling in high- pressure environments are being explored. Solubilities in dense carbon dioxide are modeled utilizing the fugacity coefficient approach with appropriate equations of state.

Keyword(s): supercritical, sampling, solubility, extraction, fugacity.

Oral presentation in research track.


L.J. Kurimski, M.A. St. Clair, K. Kai Fan and J.L. Schnoor, Department of Civil and Environmental Engineering, University of Iowa, Iowa City, IA 52242

This study investigated the feasibility of using iron rich mine tailings in a metals removal process for leachate based on the natural iron cycle. The natural iron cycle involves the photoreductive dissolution of iron(III) (hydr)oxides resulting in soluble iron(II) during the day, and subsequent oxidation and reprecipitation to iron(III) during the night. Photolysis experiments conducted in the laboratory using mine tailings produced significant concentrations of soluble iron(II) in aqueous solution at low pH. Oxidation of soluble iron(II) to iron(III) removed soluble arsenic and cadmium from solution to concentrations less toxic, with the solid phase precipitate more concentrated for a possible metals recovery system. Factors influencing the photoproduction of iron(II) from the mine tailings included the concentration of organic matter, the presence of amorphous iron oxides, and the chemical forms of iron in the mine tailings. Solution pH and the addition of ligands also affected iron(II) photoproduction. Factors influencing the removal of arsenic and cadmium from aqueous solution included the pH of the solution, as well as the presence of organic matter and amorphous iron oxides in the mine tailings.

Keyword(s): iron, photoreduction, oxidation, adsorption, coprecipitation.

Oral presentation in research track.


G. Kyle, S. Wetzel and M.K. Banks, Department of Civil Engineering, Kansas State University, Manhattan, KS 66506

Throughout the United States, petroleum hydrocarbon contamination of soil is a severe problem. Storage tanks are quite often the source, whereby hydrocarbons are released on or near the ground surface and are free to migrate through the soil to the groundwater. Sometimes, such chemicals accumulate at interface of different soil types.

Hexadecane transport and degradation in soil reactors were studied in a series of experiments. Three reactors, consisting of soil in sand-clay- sand layers, were contaminated at single points in the center of the top sand layer just below the surface. One reactor was used as a control while the other two were equipped with influent and effluent water ports located above the sand/clay interface. The ports were utilized to add water to the soil. One reactor was supplied with water only and the other was injected with a phosphorus/nitrogen solution to enhance degradation.

The project focused on two aspects: (1) the transport of the NAPLs at an abrupt sand/clay interface and (2) the ability of the soil microbes to degrade the hexadecane under aerobic conditions. Gas chromatography analysis of the effluent water samples indicate no significant removal of the NAPL from the soil by pumping, while mass balances indicate NAPL degradation due to microbial action. In addition, gas chromatography data of soil samples trace the movement of the chemical throughout the three reactors.

Keyword(s): NAPLs, bioremediation, soil interface, transport.

Oral presentation in research track.


Euisang Lee1, M.K. Banks1, and A.P. Schwab2, 1Department of Civil Engineering and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

The objective of this research is to investigate the impact of vegetation on the fate of toxic and recalcitrant pyrene in soil as well as the influence of different plant species (alfalfa (Meticago sativa) and fescue (Festuca arundinacea)). The effect of vegetation will be evaluated by determining the distribution of 14C among soil, plant tissue, leachate, and CO2 evolved in planted and unplanted soils using highly controlled plant growth chambers during a 6-month experiment. The influence of plant species on the fate of pyrene will be estimated by comparing the dissipation rate of the 14C-target compound between alfalfa and fescue. These data will be analyzed to ascertain if there are differences between vegetated and non-vegetated soils, also between plant species with regard to leaching, degradation, plant uptake, mineralization of the 14C-labelled pyrene. The beneficial effects of vegetation planted in soil contaminated by pyrene is anticipated.

Keyword(s): vegetation, pyrene, soil, plant, rhizosphere.

Poster presentation.


Zbigniew Lewandowski, Frank Roe, Maren Twedt, Duy Nguyen and Prasad Surapanini, Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717

Low concentrations of toxic heavy metals generally encountered in wastewater treatment exclude, for economical reasons, the use of traditional removal methods such as chemical precipitation, ion exchange, filtration, membrane technology, etc. The economical feasibility of recovering heavy metals can be extended by using relatively inexpensive yet efficient natural biopolymers.

Pursuant to our goal of using these biopolymers in decontaminating natural waters, we have measured thermodynamic and kinetic parameters for combinations of metals over a series of temperatures. Copper, lead and zinc divalent metal ions were investigated, as were three different types of calcium alginate biopolymer gel beads. Water-jacketed batch reactors were used in these studies.

Keyword(s): heavy metals, toxic waste, biopolymers, water.

Oral presentation in research track.


Zhengming Li, B.L. Woodbury, J.L. Martin, S.D. Comfort and P.J. Shea, University of Nebraska, Lincoln, NE 68583-0915

Past production practices at munitions ordnance plants have resulted in contamination of terrestrial and aquatic ecosystems. Efforts to date have documented the nature of contamination and attempted to estimate potential migration routes. To predict the fate of munitions in contaminated soils, an accurate prediction of the adsorption- desorption process is critical. Soils from drainage ditches at the abandoned Nebraska Ordnance Plant were found to be highly contaminated with TNT. Equilibrium soil solution concentrations were approximately 70 mg L-1 and relatively independent of soil to solution ratios, indicating the presence of solid phase TNT. Transport experiments were conducted using solute pulses of 70 mg TNT L-1 with uncontaminated soil and with soil columns containing contaminated and uncontaminated layers. Experiments were performed by displacing 27 or 53 pore volumes of TNT-3H20 pulse through disturbed soil columns (5.1 cm diam, 7.5 cm length) at a pore water velocity of 11 cm d-1. Results using uncontaminated soils indicated that breakthrough curves of TNT effluent concentrations never reached initial solute pulse concentrations (C/Co » 0.8) . The equilibrium adsorption isotherm for TNT sorption on the uncontaminated soil was characterized as a nonlinear Freundlich type. Assuming local equilibrium and using a batch- determined distribution coefficient (Kd) in the convection-dispersion equation resulted in an over estimation of elution times. These results indicate that errors can result by assuming a linear adsorption isotherm when predicting TNT transport in highly contaminated soils.

Keyword(s): TNT, fate, transport.

Poster presentation.


B.E. Logan, K. Blue, W. Johnson and R.G. Arnold, Department of Chemical and Environmental Engineering, University of Arizona, Tucson, AZ 85721

A sticking coefficient (a) is defined as a ratio of the rate particles stick to a surface to the rate they strike a surface. A relatively non-attaching bacterium (~ 1 µm) may undergo 350 collisions while only being transported 10 m in soils (0.5 mm diameter). Bacteria do not instantaneously and completely desorb from surfaces producing two major effects. First, sticking coefficients measured from breakthrough concentrations in continuous injection tests can be overestimated since the slowly desorbing cells are incorrectly included in the "steady state" breakthrough concentration. Second, varying times to desorption and large numbers of collisions enhance longitudinal dispersion. We developed a computer program to model desorption rate as a function of time by tracking the desorption of bacteria striking soil surfaces. Model results were compared to bacterial injection experiments conducted in soil columns. We found slow desorption resulted in two factors commonly observed in bacterial breakthrough curves: a slow rise to steady state, and continuous elution of cells at 2-log concentration reductions after a pulse injection.

< b>Keyword(s): biocolloids, filtration, groundwater, particles, subsurface.

Oral presentation in research track.


A. Malathi1, M.K. Banks1 and A.P. Schwab2, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

Polycyclic aromatic hydrocarbons (PAH) are a hazardous group of compounds which are highly toxic, recalcitrant and persistent, and belong to the class of hazardous organic compounds. Petroleum refining, coal conversion and chemical manufacturing sites are most often contaminated with PAHs. The propensity for bioaccumulation and possible adverse health effects of PAH parent compounds as well as intermediates pose special problems in designing a soil treatment system that will effectively reduce the concentration of these compounds. Once incorporated into the soil, biodegradation seems to be the most significant means of PAH decomposition.

The role of vegetation to stimulate the degradation and detoxification of toxic and recalcitrant organic chemicals at low soil concentrations is brought about by several mechanisms of plant-soil interactions, including improvement of physical and chemical properties of contaminated soils, increase in soil microbial activity and increase in contact between microbes associated with the roots and toxic compounds in a contaminated soil. This represents a potential cost effective and low maintenance alternative for waste management. However, there is not enough information concerning specific application of plants, chemicals and soils either in the form of laboratory or field results.

In the research to be presented, different and diverse perennial plant species [grasses (monocot), legumes, and dicots] were collected from the native prairie grasslands and tested for their efficiency in mineralization of the target PAH contaminant—phenanthrene.

The mineralization of phenanthrene was evaluated by the measurement of 14CO2 from the radiolabeled target compound incubated in a rhizosphere soil microcosm. Results from this study will indicate the potential of using different types of plants to enhance degradation of PAHs in contaminated soils.

Keyword(s): bioremediation, phenanthrene, rhizosphere, mineralization.

Poster presentation.


J.L. Martin, Zhengming Li, T.A. Kokjohn, P.J. Shea and S.D. Comfort, University of Nebraska, Lincoln, NE 68583-0915

Many acres of soil at the former Nebraska Ordnance Plant (NOP) are contaminated with TNT and other munitions residues. In some areas, solid phase TNT is present and controls the concentration of the soil solution. Native microbial populations in uncontaminated soils similar to those at the NOP site were severely reduced when solid phase TNT was allowed to control the soil solution TNT concentration. However, examination of NOP soil revealed an active population of Pseudomonas sp. A single species that could utilize TNT as a sole C source was isolated from the contaminated soil and tentatively identified as Pseudomonas corrugata through the BIOLOG system. Subsequent growth and characterization experiments indicate that the Pseudomonad metabolizes TNT while in the exponential phase of growth in medium containing glucose as a C source. In addition, a consortium of organisms was found that could utilize TNT as a sole N source. Low TNT mineralization rates (measured by CO2 evolution) in soil and media using the various isolates suggest reduced availability due to sorption and incorporation of transformation intermediates into the organic matrix and microbial biomass. Pretreatment of TNT by acid-metal catalyzed reduction resulted in an initially higher rate of mineralization following addition to TNT- contaminated soil. Observations indicate more rapid microbial utilization of the 2,4,6- triaminotoluene (TAT) reduction product and its spontaneous decay product, methylphloroglucinol (2,4,6-trihydroxytoluene), than TNT. Abiotic pretreatment may be useful in enhancing microbial transformation and detoxification of TNT in highly contaminated soils.

Keyword(s): TNT, degradation, acid-metal catalyzed reduction.

Poster presentation.


A. Massol-Deya1, R.F. Hickey1,2, and J.M. Tiedje1, 1NSF Center for Microbial Ecology, E. Lansing, MI 48824; and 2Michigan Biotechnology Institute, Lansing, MI 48909

Scanning electron microscopy, confocal scanning laser microscopy, fatty acid methyl ester profiles and restriction fragment length polymorphism pattern analysis of total community amplified 16S ribosomal RNA genes were used to study the development, organization and structure of natural aerobic multispecies biofilm communities in laboratory and field field-scale granular activated carbon fluidized bed reactors (GAC-FBR) treating petroleum contaminated groundwater. The processes of biofilm formation were studied in a laboratory reactor, which was fed toluene-amended groundwater. The GAC was allowed to become colonized by the indigenous aquifer populations. Subsequent cell binary fission of attached cells embedded in their own polymeric matrix led to the formation of microcolonies. During the early stages of colonization, microcolonies were primarily observed in crevices and other regions sheltered from hydraulic stream stresses. Eventually, these microcolonies grew over the entire surface of the GAC. This lead to the development of highly complex multilayer biofilm structures. During this phase, spatial cell organization appeared to play key structural roles. Channel-like structures of variable sizes were observed to interconnect the surface film with the deep inner layers. Ultimately, this resulted in an increase in the overall biological surface area/volume ratio, and may facilitate transport of substrates in and waste products out of deep regions of the biofilm at rates greater than possible due to diffusion only. Common architectural features were observed among biofilms from field GAC systems that were examined. This suggests that channel formation may be a general microbial strategy to deal with diffusion limitation problems in these type of reactor systems.

Keyword(s): biofilm, bioremediation, microbial ecology.

Poster presentation.


A.L. McCloskey and T.F. McIntyre, Mine Waste Technology Pilot Program (MWTPP), MSE, Inc., Butte, MT 59701

An overview of the on-going demonstration of an Ukrainian Integrated Clay-based Grouting technology will be presented. The field application is a project under the MWTPP which is being implemented by MSE Inc., funded by the U.S. Environmental Protection Agency (EPA) and jointly administrated by the EPA and the Department of Energy. Surface water measurements taken in Mike Horse Creek indicate a “loosing reach” in an area of the creek which corresponds with a portion of underground workings of the mine and with faulting in the local geology. Tests are being conducted to determine the subsurface water courses and to characterize the project area. Results from the initial phase of the project, the Characterization Phase, being conducted at the inactive Mike Horse Mine near Lincoln, Montana, will be reported. The preliminary plans for grout placement and evaluation of the project will be included in the presentation.

Keyword(s): grouting, hydrological control, acid rock drainage.

Oral presentation in research track.


Raj Mirpuri, Warren Jones, Eva Krieger and Gordon McFeters, Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717

Biodegradation of volatile organic compounds such as petroleum hydrocarbons and xenobiotic agents in the vapor phase is a promising new concept in well-head and end-of-pipe treatment which may have wide application where in-situ approaches are not feasible . The microbial degradation of the volatile organics can be carried out in vapor phase bioreactors which contain inert packing materials. Scale-up of these reactors from a bench scale to a pilot plant can best be achieved by the use of a predictive model, the success of which depends on accurate estimates of parameters defined in the model such as biodegradation kinetic and stoichiometric coefficients. The phenomena of hydrocarbon stress and injury may also affect performance of a vapor phase bioreactor. Batch kinetic studies on the biodegradation of toluene by P. putida 54G will be compared to those obtained from continuous culture studies for both suspended and biofilm cultures of the same microorganism. These results will be compared to the activity of the P. putida 54G biofilm in a vapor phase bioreactor to evaluate the impact of hydrocarbon stress and injury on biodegradative processes.

Keyword(s): biodegradation, vapor phase bioreactor, toluene.

Oral presentation in research track.


Cathy Mohs and Kenneth J. Klabunde, Department of Chemistry, Kansas State University, Manhattan, KS 66506

Photocatalytic oxidation of organic compounds in aqueous solution has gained interest as a possibility for purifying waste water and drinking water. Aqueous suspensions of titanium dioxide when irradiated with ultraviolet light are capable of decomposing some organic water contaminants. In this regard, different size titanium dioxide particles were prepared by an aerogel technique to investigate the effect of particle size on the photocatalytic activity of titanium dioxide. Different sized titanium dioxide particles were made, characterized and compared with a common industrial source of titanium dioxide, Degussa P-25, in the photocatalytic oxidation of chloroform.

Keyword(s): photocatalytic oxidation, aerogel, chloroform, titanium dioxide.

Oral presentation in research track.


R.J. Montgomery, Atlantic Richfield Company, 307 E. Park Ave., Suite 400, Anaconda, MT

The Anaconda Revegetation Treatability Study (ARTS) is being conducted to demonstrate in- place treatments of mill tailings, smelter wastes, and soils contaminated by smelter emissions at the Anaconda Smelter National Priorities List Site in Anaconda, Montana. A study area of approximately 11,000 acres has been divided into designated subareas: Opportunity Tailings Ponds, Anaconda Tailings Ponds, Smelter Hill, Old Works, and Adjacent Areas. Acidic mill tailings which contain elevated metal levels are contained in the Opportunity and Anaconda Tailings Ponds systems. The other three areas have been affected by smelting processes and stack emissions. The materials also contain elevated metal concentrations and have pH values ranging from acidic to neutral.

ARTS is being conducted in four phases. The first, completed in 1993, included reviews of reclamation literature and data searches to determine factors controlling the revegetation of acid metalliferous materials at the Anaconda site and at other locations with problems similar to Anaconda. Other Phase I activities included physical and chemical data collection and the selection of demonstration sites within the designated subareas. The second Phase (currently ongoing) includes laboratory and greenhouse tests which are being used to develop effective amendment/vegetation treatments. Large demonstrations have been implemented in three of the subareas as Part of Phase III. On Smelter Hill, a 5 acre site was treated with selected amendments using specialized equipment and seeded with selected vegetation in the fall of 1993. A one acre demonstration site in the Old Works area was treated with various combinations of amendments and two different pieces of heavy equipment were used to mix the amendments into these wastes. This site is to be seeded with selected plant species in the spring of 1994. Another one acre site located on contaminated soils will test the effectiveness of different sources of organic matter (wood wastes, commercial compost, and composted manure) in combination with different lime materials in providing a suitable rootzone. Field demonstration on the tailings ponds will begin in the summer of 1994. In Phase IV the effectiveness of the treatments at each site will be monitored in terms of reducing contaminant movement in the different pathways. Changes in waste chemistry, rootzone hydrology, and vegetation response will be measured.

Keyword(s): revegetation, smelter, tailings, soils.

Oral presentation in research track.

ARTS is funded by Atlantic Richfield Company (ARCO) and is under the direction of a Technical Committee comprised of representatives of the Environmental Protection Agency, the Montana Department of Health & Environmental Sciences, and ARCO. The ARTS study is being conducted by the Reclamation Research Unit at Montana State University.


Wesley K. Moore1, Deborah J. Mossman1, A. Paul Schwab2 and Thomas L. Feldbush3, 1Department of Civil Engineering, Coordinated Engineering Program, University of Missouri- Columbia, 5100 Rockhill Road, Kansas City, MO 64110-2499; 2Department of Agronomy, Kansas State University; 3Associate Dean of Medicine, Northwestern University

Current methods of detecting sorbed soil pollutants require that the contaminant be extracted from the soil. In an effort to make detection simpler and safer, standard fluorescent immunoassay techniques are being modified to allow fluorescent tags on the pollutant to be viewed and photographed with epifluorescent microscopy. Initial research focuses on detecting chlorinated benzenes on various soil types and developing a technique for tagging these pollutants with appropriate antibodies. This should lead to detection in actual soil cores and a better understanding of how contaminants progress through different soils.

The following flowchart illustrates the antibody linkage mechanism used. The compounds listed were used for the initial feasibility tests of this method.

dinitrobenzene (DNB) (pollutant) ß rabbit anti-DNB (primary antibody) ß goat anti-rabbit (secondary gamma globulin/ antibody/ fluorescein fluorescent tag) The figures following are fluorescent microscopy photos acquired using this method. The light areas indicate brick chips that were contaminated with DNB, while the uncontaminated chips remain dark.

DNB-contaminated brick Uncontaminated brick Keyword(s): fluorescence microscopy, immunoassay, sediments, soils.

Oral presentation in research track.


F. Munshower, Reclamation Research Unit, Montana State University, Bozeman, MT 59717

Revegetation is normally proposed as the ultimate fate of ash disposal ponds to alleviate the potential for contamination of water systems and to improve the appearance of the landscapes that contain these structures. This ash is usually phytotoxic due to large quantities of salts. Revegetation is most often accomplished by capping followed by seeding. The question becomes one of the depth of soil necessary to support adequate vegetation, to protect the surface from erosion by surface water flows, and to protect groundwater from salts leaching from the ash pond. Western fly ash ponds are especially sensitive to these problems because of the scarcity of water in this region and the presence of high salt loads in most groundwater systems.

A coarse textured diffusion barrier and variable soil depths were evaluated for their ability to stimulate vegetation development, prohibit salt movement into surface soil horizons, and permit plant roots to harvest water that infiltrates into the soil. After three growing seasons the vegetation appears to be flourishing. It has stabilized the surface of the test plots and plant roots appear to be harvesting water before it can penetrate below the root zone.

Keyword(s): revegetation, coal ash, salinity.

Oral presentation in technology transfer track.


N. Muralidharan1, Lawrence C. Davis2, John C. Tracy3, Larry E. Erickson1 and Ryan Green1, 1Department of Chemical Engineering, Kansas State University, Manhattan, KS 66506; 2Department of Biochemistry, Kansas State University, Manhattan, KS 66506; and 3Department of Civil Engineering, South Dakota State University, Brookings, SD 57007

Experimental investigations were carried out in the laboratory to study the impact of vegetation in bioremediating soil and groundwater contaminated with hazardous organic substances. A 90 cm long chamber with 2 U-shaped channels, each 10 cm in width and 35 cm in depth, was set up. Alfalfa plants were chosen and they have been growing in the channels under laboratory conditions for nearly 2 years. The channels were packed with fine sandy soil collected from near a landfill. The "groundwater" fed to one channel was water contaminated with toluene solution at saturated concentrations @ 25 C, whereas, the other channel was fed with water contaminated with phenol solution @ 500 ppm (v/v). The contaminant concentrations in the groundwater of the channels were monitored at the sampling wells located along each of the channels. The influent and effluent flow rates from each channel were measured every day and recorded.

"Evapotranspiration" significantly influenced the fate of the pollutants. Dispersion and adsorption processes in the channel were studied by introducing bromide tracer, as a broad pulse, into the toluene fed channel and by observing toluene and phenol concentrations, following a feed step change to pure water. Tracer studies indicated that short-circuiting in the end of the channel was quite significant. Models which were previously developed to describe the fate of the contaminants in variably-saturated soils in the presence of vegetation are being employed to simulate the fate of these hazardous organic substances in the laboratory chamber.

Keyword(s): bioremediation, evapotranspiration, adsorption, rhizosphere.

Oral presentation in research track.


M. Narodoslawsky and C. Krotscheck, Institute of Chemical Engineering, Graz University of Technology, Inffeldgasse 25, A-8010 Graz, Austria

Evaluation of processes according to their environmental compatibility is necessary at a relatively early stage of planing, since this becomes a more and more decisive feature of competitiveness.

However, at an early stage in the development, data are scarce and the effort to investigate various process alternatives using full blown ecobalances or comparable methods is prohibitive. The SPI offers a chance to evaluate the ecologic compatibility of a process (or process revamping) in a quick and reliable way. It is based on the concept of sustainable development and uses only information about mass flows, energy flows and investment needs besides readily available environmental data. It allows clear distinction between environmentally compatible and problematic processes and indicates those steps within the process which pose the largest obstacles on a way to environmental compatibility.

Keyword(s): environmental compatibility, process evaluation.

Oral presentation in research track.


S. Nedunuri and Rao S. Govindaraju, Department of Civil Engineering, Seaton Hall, Kansas State University, Manhattan, KS 66506

Spatial variability exhibited by many field soils necessitates the use of stochastic methods for prediction of average solute movement. Data from a field experiment were analyzed to characterize the random nature of the velocity and dispersion of solute (Potassium Bromide) in field scale vertical transport experiments. Solute concentrations were measured at over fifty spatial locations and at six depths within the soil. The analysis indicates that solute velocities at deeper soil layers exhibit a statistically homogeneous behavior. Dispersion was determined from breakthrough curves using a standard nonlinear regression model. These results will be presented, and the implications of modeling average solute behavior will be discussed.

Keyword(s): spatial variability, stochastic methods, transport.

Oral presentation in research track.


D. Neuman, F. Munshower, D. Dollhopf, S. Jennings and J. Goering, Reclamation Research Unit, Montana State University, Bozeman, MT 59715

Assessment of in-place chemical immobilization of metal contaminants in mine wastes/soils followed by revegetation as a potential remedial alternative for cleanup of a Superfund site is described. This six year treatability study evaluated the use of different amendments, various incorporation techniques, and selected plant species in both laboratory/greenhouse tests and at five waste/contaminated soil locations along Silver Bow Creek between Butte and Opportunity, Montana.

Amendments (mainly calcium hydroxide and calcium carbonate) found in the laboratory to be most effective in controlling pH of these acid producing metalliferous wastes and reducing their soluble metal levels were mixed with the wastes/soils in replicated greenhouse trials. Amendments and greenhouse selected plant species were combined into replicated field trials using different amendment incorporation techniques. Vegetation response and components of the soil chemistry and soil hydrology were measured, and differences among treatments were assessed.

Data support coversoil as most effective, but coversoils are not available in quantities adequate for the rehabilitation of all the disturbances along Silver Bow Creek. Pressure injection of lime slurry into the wastes did not provide an adequate rootzone for long term plant growth. Agricultural tilling of amendments provided only 15 centimeters of adequate rootzone materials. A plow capable of mixing waste to a depth of 122 centimeters provided an ameliorated rootzone that varied from 30 to 60 centimeters deep. Plant cover and production were greater on deep plowed plots than on plots prepared by the two other incorporation techniques. Deeper root penetration was also found in the deep plowed plots than in plots treated by agricultural tillage or the injection technique. Surface runoff was markedly reduced and surface water quality improved by all treatments.

Keyword(s): revegetation, tailings, metal, immobilization.

Oral presentation in research track.


M.R. Norland and D.L. Veith, Reclamation Technology, U.S. Bureau of Mines, Twin Cities Research Center, 5629 Minnehaha Ave. South, Minneapolis, MN 55417- 3099

On Minnesota’s Mesabi Iron Range, coarse taconite iron ore tailing is often used as the principal material in the construction of dams for large tailing impoundments. Mineland reclamation rules in Minnesota require that tailing dams be vegetated to control erosion for dam stability and safety. Vegetation helps to maintain the structural integrity of the dam by resisting erosion resulting from the forces of wind and water. Coarse taconite iron ore tailing is characterized chemically by its alkaline reaction, low water-holding capacity, and dark color; and biologically by its lack of microorganisms. To ameliorate these conditions and make the material more amenable to plant establishment and growth, the U.S. Bureau of Mines implemented a series of factorial experiments at two active taconite mine sites in northeastern Minnesota. At each experimental site vegetative cover has improved depending on the type of municipal solid waste compost used and rate of application. At site I, overall plant cover across all treatments had improved from 0% prior to experimental manipulation to 72% after 4 years, with 7 treatment combinations exceeding 90% cover. At site II, overall plant cover has improved from 0% prior to experimental manipulation to 83% after 4 years, with 23 treatment combinations exceeding 90% cover. At both sites, total cover has progressively increased over 4 years and has not reached steady state conditions. These results suggest a possible new strategy for reclaiming difficult sites through the use of municipal solid waste compost.

Keyword(s): taconite, tailing, revegetation, compost, cover.

Oral presentation in research track.


M.H. Ochoa and J.B. Hughes, Department of Environmental Science and Engineering, Rice University, PO Box 1892, Houston, TX 77251

Studies have been conducted assessing the influence of competitive substrate interactions on the rate/extent of trichloroethene degradation by Nitrifying mixed cultures. Specifically, the effect of ammonia, hydroxylamine, tetrachloroethene and cis-dichloroethene have been examined. Results demonstrate that rates of degradation and finite transformation capacity are significantly influenced by these interactions. Adding hydroxylamine as an exogenous source of energy in the absence of the primary substrate produced an increase in the degradation capacity of these cultures up to threefold. Models are being tested to predict these phenomena.

Keyword(s): biodegradation, nitrifiers, trichloroethene, substrate interactions.

Oral presentation in research track.


R.A. Petrie, J.E. McLean and R.C. Sims, Utah Water Research Laboratory, Utah State University, Logan, UT 84322-8200

Molecular oxygen is one of the strongest naturally occurring redox species. In subsurface environments, oxygen is quickly consumed by microorganisms and is slow to recharge. When oxygen becomes limiting, microorganisms must utilize alternative terminal electron acceptors to mediate growth. Iron and Manganese have high redox potentials and are commonly found in the subsurface. Abiotic and microbially mediated reactions are potentially energetically favored by electron transport systems that utilize these metal species.

Calculated free energy changes are used to show the potential use of iron and manganese as terminal electron acceptors in the transformation of pentachlorophenol in anaerobic environments. Microbial yields based on free energy liberated from pentachlorophenol transformation are estimated under iron and manganese reducing conditions using a simple bioenergetic growth model.

Iron and manganese was extracted from aquifer solids obtained at a superfund site and mixed with pentachlorophenol under anaerobic conditions. The rate of appearance of reduced metal species was correlated with the disappearance of pentachlorophenol. Sequential utilization of iron and manganese based on calculated free energy changes was investigated.

Keyword(s): redox, electron acceptors, remediation, aquifers.

Oral presentation in research track.


B.M. Peyton, R.S. Skeen and B.S. Hooker, Engineering Technology Center (P7-41), Pacific Northwest Laboratory, Richland, WA 99332

Nutrient cycling has been shown to enhance the degradation rate of carbon tetrachloride in anaerobic batch cultures. In addition, nutrient pulsing has been suggested as a method for controlling near-bore biofouling in application of in-situ bioremediation. To determine the effects of nutrient pulsing on biomass accumulation, soil columns were fed with nitrate and acetate to develop a denitrifying biofilm within a porous sand matrix to compare two nutrient delivery strategies. The strategies that were compared were continuous nutrient feeding and pulsed nutrient feeding.

Acetate, as the sole carbon and energy source, was fed to the columns continuously at 83 mg/L or pulsed at 2222 mg/L for 30 minutes every 12 hours. This resulted in the same time-averaged substrate loading to the soil columns. Final biomass profiles indicate a much more uniform biomass accumulation profile using a pulsed nutrient strategy. Also, notable differences in the effluent suspended cell concentrations were measured. This data was used to help calibrate a detailed model for bacterial transport in porous media, which will be used to aid in the design and implementation of in-situ bioremediation at the Hanford site.

Keyword(s): in-situ bioremediation, biofilm, biofouling, denitrification, nutrient delivery.

Oral presentation in research track.

This work was supported by the U.S. Department of Energy as part of Pacific Northwest Laboratory’s Laboratory Directed Research and Development. Pacific Northwest Laboratory is operated for the U.S. Department of Energy by Battelle Memorial Institute under contract DE-AC06-76RLO 1830.


J. Powers1, R. Luyendijk1, J. Mathur2, V. Ceci3 and D. Eyman4, 1Waste Policy Institute, 1872 Pratt Drive, Suite 1600, Blacksburg, VA 24060, (703) 231-3324; 2U.S. Department of Energy, Office of Environmental Restoration and Waste Management, Trevion II, EM-551, Room 426, 12800 Middlebrook Road, Germantown, MD 20874, (301) 990-7922; 3BDM/GER Quince Diamond, 555 Quince Orchard Road, Suite 400, Gaithersburg, MD 20878-1437, (301) 212-6208; and 4Waste Policy Institute, Quince Diamond Executive Center, 555 Quince Orchard Road, Suite 600, Gaithersburg, MD 20878-1437, (301) 990-7200.

Surface and subsurface sources of groundwater contamination is a major problem at Department of Energy (DOE) facilities and is found at more than seventy percent of the sites currently on the Environmental Protection Agency (EPA) National Priority List (NPL). These water supplies are vital national resources to be utilized as sources for drinking, industrial, commercial, municipal, governmental, recreational and agricultural water uses. Once contaminated, these resources are very expensive to recover due to the large volumes and relatively low contaminant concentrations. This paper discusses a systematic approach to recover contaminated waters and to produce marketable products that is amenable to market drivers and optimization. This systems configuration is utilized to identify potential treatment processes, define process constraints, assemble treatment technologies for insertion into a logical and feasible order, and define technology deficiencies. The systems configuration displays conventional and novel technologies to be considered in combinations as well as individually, with respect to their technical feasibility, effectiveness, applicability to DOE needs, implementability, cost, regulatory acceptability, and readiness for demonstration. The strategy addresses the innovative technologies being developed by DOE and integrating novel DOE technologies with the conventional and innovative technologies of industries to develop the Berkeley Pit resource recovery plan. To offset the cost of resource recovery, the scope of this research includes the vision toward the recovery and marketability of the valuable mineral and water resources contained in the Berkeley Pit.

Keyword(s): heavy metals, systems configuration, flowchart, resource recovery, contaminated water.

Oral presentation in research track.


R.N Prabhushankar1, Lakshmi N. Reddi1 and A. Paul Schwab2, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

In addition to advective and diffusive transport of contaminants in the subsurface, clay colloid- associated transport in the shallow subsurface regions was recently found to provide a significant component. This study dealt with an assessment of clay colloid migration and the associated transport of contaminants from surface run-off into the subsurface. The study consisted of two phases of experimental investigations. In the first phase, a series of tests were conducted in permeameters on sandy soils of various densities. Suspensions of montmorillonite particles were introduced into the samples to obtain deposition profiles of montmorillonite particles. In the second phase, zinc was used as the sorptive contaminant in the influent. Breakthrough curves and zinc profiles in the soil sample were obtained. Results from these experiments were used to assess the impact of clay colloid-associated transport of contaminants on soil and groundwater quality under a range of in-situ conditions. Finally, an approach was outlined to model this transport mechanism. A sensitivity analysis was conducted to identify the effects of key parameters such as soil density, soil-contaminant interactive properties such as the distribution coefficient, and residence time of influent.

Keyword(s): clay colloids, colloid-associated transport.

Oral presentation in research track.


S. Rao1, L.L. Barton2 and T.J. Ross1, 1Environmental Scanning Electron Microscope Laboratory, Department of Civil Engineering; and 2Laboratory of Microbial Chemistry, Department of Biology, the University of New Mexico, Albuquerque, NM 87131

The metals iron, stainless steel, copper, nichrome, titanium, aluminum and aluminum alloys were exposed to different cultures of bacteria. The microorganisms included pure strains of sulfate-reducing bacteria, cocultures and mixed cultures. The results of the biological activity such as biocorrosion, scaling and precipitation of metals was studied by using Environmental Scanning Electron Microscopy, gravimetric analysis, and by Energy Dispersive X- ray analysis. The effect of different nutrients, cultures, and the presence of oxygen were studied. The observation of bacteria, colloids and plaque of iron sulfide, crystals and tubercules in their natural and wet state was facilitated by the environmental microscopy.

Keyword(s): environmental microscopy, biocorrosion, sulfate-reducing bacteria.

Oral and/or poster presentation in research track.


Lakshmi N. Reddi and Hui Wu, Department of Civil Engineering, Kansas State University, Manhattan, KS 66506

The effects of vibrations on soils containing immiscible liquids were described in the context of soil remediation. Vibrations when augmented with flow gradients have the beneficial effect of dislodging immiscible liquid blobs and mobilizing them toward the point of intended collection. Several bench-scale experiments were conducted using a laboratory vibrator on sandy soils contaminated with Soltrol. The experiments were conducted under controlled conditions using a 4-ft diameter tank. The effect of vibrations on the soil- contaminant medium was measured via pore pressure transducers installed at various radial distances from the vibrator. Soil samples from the tank were analyzed for contaminant concentrations before and after inducing vibrations. The effectiveness of vibrations was assessed in terms of reduction of residual saturation of the liquids. The results were analyzed to assess the effects of various test parameters on the process such as frequency of vibrations, flow velocities in the tank, duration of vibrations, and density of soil. The lateral zone of influence of vibratory mobilization was also assessed. In order to predict the in-situ effectiveness of the process, mobilization criteria were developed as functions of soil, contaminant and vibration parameters. The zone of influence obtained using these mobilization criteria agreed well with the observed values for bench-scale experiments. Finally, a prototype design integrating vibrator and pumping mechanism was recommended for in-situ use at hazardous waste sites.

Keyword(s): immiscible (nonaqueous phase) liquids, vibrations, in-situ remediation.

Oral presentation in research track.


Karl M. Regan and Ronald L. Crawford, Institute for Molecular and Agricultural Genetic Engineering (IMAGE), Food Research Center 103, University of Idaho, Moscow, ID 83844-1052

A strain of Clostridium bifermentans isolated from a munitions-supplemented enrichment was able to remove both TNT (2,4,6-trinitrotoluene) and RDX (1,3,5-triaza 1,3,5-trinitrocyclohexane) from its growth media. Biotransformations of TNT and RDX by cometabolism in a nutrient rich medium reduced the removal time from several days to a few hours, as compared to a nutrient limited medium. Redox potential (Eh) of the media had important effects on the biological and abiological transformations of the munition compounds.

Keyword(s): biotransformation, cometabolism, 2,4,6-trinitrotoluene, 1,3,5-triaza 1,3,5-trinitrocyclohexane, clostridium.

Poster presentation.


G. Rieck and Lakshmi N. Reddi, Department of Civil Engineering, Kansas State University, Manhattan, KS 66506

A survey of industrial waste generated in the state of Kansas revealed that foundry sand is produced in significant quantities and that its disposal costs are high. A new research project was initiated in 1993 to identify ways to utilize the foundry sand such that the waste components in the sand are stabilized and rendered harmless. Chemical analyses of the waste material showed significant quantities of phenolic compounds. A series of experiments were conducted on mixtures of foundry sand and cementitious materials to identify useful mechanical properties and to assess leachability of chemicals. Three types of cementitious materials were used as binders: i) naturally available clay minerals such as kaolinite and montmorillonite, ii) waste by-products such as fly ash which are known to possess cementitious nature, and iii) portland cement. The mechanical properties (primarily compressive strength) and the leachability of the mixes were analyzed as functions of various test parameters such as mix proportions and setting time. The results show that waste materials from foundries can adequately be stabilized using cementitious binders such that the leachates do not contain detectable levels of harmful chemicals. Finally, some engineering uses of the stabilized mixes were identified.

Keyword(s): waste stabilization, waste utilization, leachability.

Oral presentation in research track.


Jerome Rigot and Fumio Matsumura, Department of Environmental Toxicology, University of California, Davis

Recently, contamination levels of vegetated soil in polluted areas were found to be lower than contamination levels of non-vegetated soil in the same area. The greater mineralization of pollutants observed in vegetated soils may be a result of the higher microbial and metabolic activities observed in the plant's rhizosphere environment which may lead to enhanced biodegradation of xenobiotics in contaminated vegetated soils.

After determining the different factors affecting the fate and the metabolic activity of microorganisms in the rhizosphere, we plan to study the possibility of introducing specialized organisms in the rhizosphere in order to enhance the biodegradation of target compounds. Our purpose is to develop a soil bioremediation technology based on stimulating synergistic interactions existing between the rhizosphere environment and selected microorganisms to enhance the biodegradation of organic pollutants in contaminated soil. If effective, this technology could be used worldwide over large areas to effectively and inexpensively treat contaminated soil.

Keyword(s): soil bioremediation, rhizosphere, metabolic activity, synergistic interactions, enhanced biodegradation of xenobiotics.

Oral presentation in research track.


T.M Roane and S.T. Kellogg, Microbiology, Molecular Biology & Biochemistry Department, University of Idaho, Moscow, ID 83843

As a result of 100 years of mining, the largest lead-contaminated Superfund site, located in Silver Valley, Idaho, is highly contaminated with lead, mercury, and arsenic, among other metals. Despite attempts to lessen the impact of these metals upon the environment, the problem of heavy metal contamination still exists. Microbial communities are known to possess resistance to a variety of metals raising the possibility of using such microbes to detoxify heavy metal contaminated soils and waters. However, little is known about the microbial communities in these soils. Microbial soil communities from soils collected from the Silver Valley region were analyzed for total and viable numbers, metabolic activity, and resistance to lead. Different microbial communities were found to respond differently to varying lead concentrations resulting in the isolation of members of a variety of genera, including Bacillus spp., Pseudomonas spp., and Corynebacterium spp. representing common lead-resistant isolates from these soils. An Enterobacter spp., isolated from a pristine soil, also exhibited lead resistance. The basis of lead resistance was explored using plasmid analyses, microscopy, and scanning electron microscopy equipped with electron diffraction X- ray analysis. Early results indicate that lead was excluded rather than sequestered intracellularly.

Keyword(s): lead, bacteria, resistance.

Oral presentation in research track.


S. Santharam, L.E. Erickson and L.T. Fan, Department of Chemical Engineering, Kansas State University, Manhattan, KS 66506

Polynuclear aromatic hydrocarbons (PAHs) are a major source of contaminants at coal gas manufacturing sites, wood treating operations, and petroleum land farms. They are potentially carcinogenic and mutagenic. It has been known that vegetation can enhance the rate and extent of degradation of PAHs in contaminated soil. Plant roots release root exudates capable of supplying carbon and energy to microflora for degrading PAHs. It is a well established fact that the population of microorganisms in the rhizosphere is significantly greater than that in non-vegetated soil. The microbial population in the rhizosphere appears to be responsible for the enhanced biodegradation of PAHs. Since their solubility in water is low, significant quantities of PAHs often form a hydrocarbon phase; moreover, some quantities of them are adsorbed on soil. A model has been derived for describing the rates of disappearance of anthracene and pyrene in the rhizosphere. This model takes into account dissolution, adsorption, desorption and biodegradation of these compounds, without neglecting the size distribution of the organic phase droplets; the rate of biodegradation is expressed in terms of the Monod kinetics. The model is in good accord with the available experimental data

Keyword(s): polynuclear aromatic hydrocarbons, rhizosphere, soil, biodegradation, model.

Poster presentation.


J.R. Schneider, R.J. Grosser, K. Jayasimhulu and D. Warshawsky, Department of Environmental Health, University of Cincinnati, Cincinnati, OH 45267-0056

Utilization of microbial processes for bioremediation requires measuring compound degradability as well as characterization of the intermediates formed. A Mycobacterium sp. and Xanthomonas sp. were isolated from coal gasification site soils using standard enrichment techniques. The Mycobacterium sp. was isolated for its ability to mineralize pyrene, but was also found to be capable of degrading other polycyclic aromatic compounds. The Xanthomonas sp. was isolated for its ability to mineralize carbazole and showed very high substrate specificity. Through analysis, metabolites formed during pyrene degradation were found to include 4,5-pyrene- dihydrodiol, 4-phenanthroic acid, and 4,5- diphenanthroic acid. This Mycobacterium sp. was also found to biodegrade benz(a)anthracene and benzo(a)pyrene producing ring cleavage products. For benz(a)anthracene, analyses of organic extracts of the growth medium show a dihydrodiol intermediate formed as well as a ring cleavage product. A dihydrodiol intermediate and ring cleavage product were also found with benzo(a)pyrene. Fluorescence spectra of the benzo(a)pyrene dihydrodiol intermediate was characteristic of that for benz(a)anthracene. The Xanthomonas sp. was incubated with carbazole as sole added carbon source for approximately two weeks. Analyses of organic extracts show the presence of at least three cleavage products of carbazole. These have been tentatively identified as indoleacetic acid, cis-indoleacrylic acid, and indolepropionaldehyde. A pathway for carbazole degradation by this species is proposed in which the indole moiety is maintained. These results show that microbes isolated for their ability to degrade a specific compound can show some diversity in their substrate specificity.

Keyword(s): Mycobacterium, Xanthomonas, biodegradation, metabolites, polycyclic aromatic hydrocarbons.

Poster presentation.


C. Shapiro, K. Garcia, C. Barnes and J. Beller, Idaho National Engineering Laboratory, EG&G Idaho Inc., PO Box 1625, Idaho Falls, ID 83415-3765

Supercritical Water Oxidation is being developed by the Department of Energy as a promising technology for treatment of mixed waste—that is waste that contains hazardous and radioactive constituents. The aim of this program has been to identify and resolve key technical challenges that have been barriers to commercialization of supercritical water oxidation. Initial testing with benchscale systems indicated that the overriding technical challenge for this technology was prevention of corrosion and deposition inside the reactor. A number of approaches are being pursued to resolve the issue of corrosion and deposition including alternate reactor designs, advanced material development and testing, salt separation processes, and analytical modeling of supercritical water oxidation systems. Ongoing projects in these areas are discussed along with plans for pilot plant testing with promising configurations.

Keyword(s): oxidation, mixed waste.

Oral presentation in research and technology transfer tracks.


Robert Sharp1, Malcolm Shields2, Tracy Moody2 and Warren Jones1, 1Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717; and 2Center for Diagnostics and Bioremediation, University of West Florida, Pensacola, FL

The pTOM31C plasmid was derived from the large plasmid of Pseudomonas cepacia G4. Strain G4 is an environmental isolate that is capable of cometabolic mineralization of trichloroethylene (TCE). The degradation of TCE by strain G4 is a result of the induction of enzymes in a non- constitutive aromatic degradative pathway that is present on the large plasmid of G4. A Tn5 insertion into the large plasmid of G4 resulted in a TCE constitutive plasmid designated as pTOM31C. A number of laboratory and field studies have been performed to determine the feasibility of applying pTOM31C, via a host microbe designated Ps. cepacia PR1-(pTOM31C), into vapor phase TCE bioreactors. Results from degradation studies show a linear degradation rate of TCE by PR1- (pTOM31C). The growth kinetics of PR1- (pTOM31C) on phthalate followed the Monod kinetics model. However, bench scale studies showed that PR1-(pTOM31C) growing on phthalate was incapable of developing a substantial biofilm. The results from a number of bench scale and field scale reactor studies showed that PR1-(pTOM31C) could not compete with invading microorganisms, even when selective pressures for PR1 were applied to the systems. In the field scale reactors, PR1-(pTOM31C) was able to colonize only the top fraction of the oyster shell support media. During the progression of the field study, the activity in the top of-the column also diminished due to the washout of PR1- (pTOM31C). Field studies showed that the combination of the crushed oyster shell support media and the physiology of PR1-(pTOM31C) is an inadequate system for developing an effective TCE degrading biofilm. Additional studies are focused on conjugal transfer of pTOM31C into superattaching and competitive microorganisms. We have had successful transfers of pTOM31C to various Pseudomonad strains. Currently we are determining the stability and TCE degradation activity of pTOM31C in its new host strains. It is the longterm goal of this research to exploit the capability of pTOM31C to cometabolically and constitutively mineralize TCE in TCE biofilm reactors.

Keyword(s): trichloroethylene (TCE), bioreactor, pTOM31C, constitutive, conjugal transfer.

Oral presentation in research track.


Roger C. Sokol, Charlotte M. Bethoney and G-Yull Rhee, School of Public Health, State University of New York at Albany and Wadsworth Laboratories, NYS Department of Health, Albany, NY 12201-0509

Reductive dechlorination of polychlorinated biphenyls primarily removes chlorines (Cls) from the meta and para positions although ortho dechlorination has also been reported. This preferential removal is determined by the pattern of Cl substitution. Thus, the final products may not only be orth-substituted congeners. In fact, certain congeners such as 2,4'-, 2,5,2', 2,4,4'-, 2,4,2',5'- and 2,4,2',4'-chlorobiphenyls accumulated at high concentrations in both the laboratory and natural sediments in the Hudson and St. Lawrence Rivers. Many of these accumulating congeners are known to be neurotoxic by reducing cellular dopamine content. Therefore, although in-situ dechlorination may reduce the overall degree of chlorination, it is important to consider the health implications of accumulating products.

Dechlorination competence varies with sediment microorganisms from different sites, resulting in the accumulation of dissimilar products. Microorganisms from the General Motors site in the St. Lawrence River removed a higher proportion of para Cls relative to meta whereas those organisms at the Reynolds and ALCOA sites removed a higher proportion of meta Cls. Sediment amendments with H2 or biphenyl also resulted in the accumulation of different products. These results suggest that it may be possible to alter the final products through artificial manipulations.

We have identified several conditions which are critical for dechlorination. Dechlorination is most effective under methanogenic conditions; sulfidogenic or nitrate-reducing conditions inhibited it partly or completely. H2 promoted dechlorination, probably acting as an electron donor to the sediment microorganisms. Heat- killed cyanobacterial cells could serve as a carbon and energy source. A periodic addition of a nonionic surfactant at a level below the critical micelle concentration also accelerated dechlorination. Therefore, it appears possible to remediate contaminated sediments and soils with further optimization of dechlorinating activities.

Keyword(s): polychlorinated biphenyls (PCBs), reductive dechlorination, bioremediation, electron acceptors, surfactant.

Poster presentation.


J.D. Stahl and S.D. Aust, Biotechnology Center, Utah State University, Logan, UT 84322-4705

Biodegradation of trinitrotoluene by the white rot fungus Phanerochaete chrysosporium was studied in both liquid reactors and soil microcosms. It has been previously shown that trinitrotoluene mineralization by P. chrysosporium only occurs under lignin-degrading conditions. In this study trinitrotoluene biotransformation rates were dependent on initial trinitrotoluene concentration and fungal mycelia present. Nonligninolytic reactors containing 50 g/L mycelia were able to degrade up to 100 parts per million trinitrotoluene within four hours but the reduced aminonitrotoluenes were not degraded, even after 70 days incubation. On the other hand, ligninolytic reactors containing 8 g/L mycelia required 6 days to degrade 100 parts per million trinitrotoluene, and aminonitrotoluenes were completely degraded within 10 days.

Ligninolytic conditions could be maintained for 18 days by addition of 3.0 g/L glucose and 0.25 mM ammonia to the reactors every three days. In soil, P. chrysosporium was able to metabolize 200 mg/kg trinitrotoluene to less than 5 mg/kg within 21 days. The trinitrotoluene metabolites, aminonitrotoluenes reached a maximum concentration of 35 mg/kg on day 14 and 10 mg/kg on day 28. During the same time, trinitrotoluene was decreased to 150 mg/kg and aminonitrotoluenes increased to 40 mg/kg in uninoculated soil.

Keyword(s): Phanerochaete chrysosporium, white rot fungi, trinitrotoluene, bioremediation.

Oral presentation in research track.


P.J. Sturman1, A.B. Cunningham1, J. Wolfram2 and S. Niehaus3, 1Center for Biofilm Engineering, Montana State University, Bozeman, MT; 2Idaho National Engineering Laboratory, Idaho Falls, ID; and 3Gosling-Czubak Associates, Traverse City, MI

A sandy aquifer contaminated with dissolved phase BTEX compounds is currently being remediated through a combination of physical processes (pump and treat) and enhanced in-situ biodegradation. Extensive data collected from site monitoring and pump-back wells, along with knowledge of site sorptive and hydrodynamic properties, allowed the calculation of a contaminant mass balance, from which in-situ biotransformation was estimated. Data indicates that pump and treat removals of dissolved BTEX accounts for approximately half of the total reduction in BTEX mass on the site. Oxygen utilization data implicates biodegradation as responsible for the majority of remaining contaminant mass removal. Contaminant reductions in-situ appear to be associated with zones of increased contaminant-oxygen mixing located immediately downgradient from site injection wells.

Keyword(s): BTEX, in-situ, biodegradation, pump and treat.

Oral presentation in research track.


S. Sun, W.P. Inskeep and S.A. Boyd, Department of Plant, Soil and Environmental Science, Montana State University, Bozeman, MT; and Department of Crop and Soil Science, Michigan State University, East Lansing, MI

The solubility enhancement of nonionic organic compounds (NOCs) by surfactants may represent an important tool in chemical and biological remediation of contaminated soils. In aqueous systems, the presence of dissolved surfactant emulsions or micelles may enhance the solubility of NOCs by acting as a hydrophobic partitioning phase for the NOCs. However, most environmental remediation efforts involve soil- water or sediment-water systems, where surfactant molecules may also interact with the solid phase. An understanding of the effect of surfactants on the sorption and distribution of NOCs in soil or sediment environments will provide an essential basis for utilizing surfactants in environmental remediation. In this study, we examined the effect of a micelle-forming surfactant (Triton X-100) on the sorption of 2,2',4,4',5,5'-PCB, 1,1-bis(p- chlorophenyl)-2,2,2-trichloroethane (p,p'-DDT) and 1,2,4-trichlorobenzene (1,2,4-TCB). Our results show that the apparent distribution coefficient (K*) describing the partitioning of the NOC by the soil solid phase exhibited a similar pattern as a function of aqueous phase Triton X- 100 concentrations for all three solutes studied. At aqueous concentrations of Triton X-100 below 200 mg L-1, K* values actually increased with increasing surfactant concentration. At Triton X- 100 concentrations above 200 mg L-l (approximately the critical micelle concentration, CMC, for Triton X-100), K* values decreased with increasing surfactant concentration. At surfactant concentrations below the CMC, surfactant monomers in the aqueous phase are relatively ineffective as a partitioning phase for the NOCs. However, bound surfactant molecules increase the sorptive capacity of the solid phase. At surfactant concentrations above the CMC, surfactant micelles in the aqueous phase begin to compete with the solid phase as an effective partitioning phase. A conceptual model, which accurately describes the functional dependence of K* on Triton X-100 concentration, was developed based on the partition coefficients of these NOCs by soil, soil-surfactant, surfactant monomer and surfactant micelle phases. This model can be further modified to provide quantitative prediction of K* of a given NOC at different surfactant concentrations.

Keyword(s): nonionic organic compounds, surfactant, sorption, remediation.

Poster presentation.


W. Svee and F. Diebold, Chemistry and Geochemistry Department, Montana College of Mineral Science and Technology, Butte, MT 59701-8997

A Savannah River Laboratory nuclear waste glass (SRL) and a plasma furnace nuclear waste slag were subjected to vapor hydration as small suspended rectangular samples within a sealed reaction bomb at 200 C for time periods of up to 26 days. The samples were then removed from the reaction bomb and the residual water analyzed by ICP and ICP-MS for trace elements and bulk glass constituents. The sample hydration rind thickness was measured and photographed with an optical microscope and the data plotted as thickness vs. time profiles which are presented as an indicator of the relative durability of the waste medium. The chemical transport of various dissolved glass constituents from the bulk medium to the sample surface developed residual diffusion textures within the hydration rind which have been investigated relative to the residual bomb water chemistry and SEM-EDX analysis of the hydration rind. These textures reveal that the water vapor diffuses into the surface of the nuclear waste slag along the path of least resistance and selectively dissolves the more soluble constituents within the slag. The development of secondary silicate phases and selective dissolution of soluble elements contribute to extensive fracturing of the hydrated region. This enhances material transport as insoluble crystals are entrained within a surface debris flow composed of gelatinous silica, which slowly flows down the sample surface. In contrast, a nuclear waste glass hydrates more uniformly and, initially, the solvation appears to be evenly distributed within the hydration layer. Eventually diffusion gradients are established within the hydration layer. These diffusion gradients eventually result in the formation of double to triple layers within the hydration rind which have differing chemical composition. The development of these variable composition layers can impede further vapor hydration of the glass.

Keyword(s): slag, glass, vapor hydration.

Oral presentation in research track.


David C. Szlag and Tissa Illangasekare, Department of Civil, Environmental and Architectural Engineering, University of Colorado, Boulder, CO 80309-0428, 303-492- 6644

Spills of nonaqueous phase liquids (NAPLs) in the environment pose a serious long-term threat to groundwater resources through slow dissolution of entrapped NAPL in the groundwater saturated zone. Based on various spill simulation experiments conducted in large soil flumes, we conclude that the NAPL can exist as a “residual” saturation held by capillary forces, or at higher saturations through macroscopic entrapment. The “micro-scale” residual entrapment is controlled by the pore characteristics of the soil, whereas the macroscopic entrapment is caused by much larger scale macroscopic soil heterogeneities in the aquifer. A study to evaluate mass transfer under both residual and macro-scale entrapment is currently underway. This paper focuses on dissolution of NAPLs entrapped at residual saturation. Current practice usually assumes local equilibrium at the NAPL source. We have found that this approximation may underestimate the length of time needed for clean-up in some cases. An existing flow model was modified to account for the reduced relative permeability in the NAPL source zone. This model was in turn coupled to a method of characteristics (MOC) based advection- dispersion model that accounted for rate limited mass transport between the NAPL and aqueous phases. Using this hybrid model, we have estimated overall mass transfer rate coefficients (time-1) for NAPL sources over a range of saturations and groundwater velocities. This information will be useful in remediation design in aquifers contaminated with organic waste chemicals.

Keyword(s): groundwater contamination, NAPL, dissolution.

Oral presentation in research track.


B.M. Thomson1, L.L. Barton2, K. Steenhoudt1 and M.D. Tucker1, 1Department of Civil Engineering and 2Department of Biology, University of New Mexico, Albuquerque NM 87131

Sulfate-reducing bacteria, nitrate-reducing bacteria and bacteria present in sewage sludge were examined for their ability to reduce the level of soluble U(VI) in enriched media. Cultures of Desulfovibrio desulfuricans, D. gigas, and D. vulgaris were grown in sulfate-containing media while Pseudomonas putida and P. denitrificans were cultivated in nitrate media. The amount of U(VI) removed from solution was dependent on metabolism because greater levels of uranium were removed when U(VI) was added to a growing culture than when added to a culture in stationary phase. The presence of vanadate, arsenate, selenate or molybdate at 0.1 and 0.01 M levels in sulfate-reducing cultures, nitrate- respiring cultures or in sludge cultures did not have an effect on the amount of uranium removed. In all cultures the amount of uranium in solution was markedly reduced after 10 to 20 days and reduced uranium, as U(IV), was detected in several cultures. Present in the cultures of D. desulfuricans were crystals of uranium. Examination of these cultures by electron microscopy indicates that the uranium (IV) is deposited outside of the cell and these needle-like crystals are associated with cellular material. X- ray probe analysis with the electron microscope gave an image that was in close agreement with U(IV). With D. desulfuricans in a continuous stirred tank reactor, kinetic parameters have been calculated for uranium reduction. Over a period of 20 to 60 hours, the amount of soluble uranium removed from the bioreactor was proportional to residence time over a period of 20 to 60 hours.

Keyword(s): uranium, denitrification, sulfate reduction, anaerobisis.

Poster presentation.


E. Tiffany-Castiglioni, W.H. Hanneman, M.E. Legare, S.J. Hong, R. Barhoumi, R.C. Burghardt and S. Safe, Department of Veterinary Anatomy and Public Health, Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX 77843

Recent attention has focused on the neurotoxicity of polychlorinated biphenyls, dibenzofurans, and quaterphenyls and related compounds. The hippocampus may be an important target for neurotoxic compounds because of its role in short-term memory and learning. We report preliminary experiments on the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as a model for the neurotoxicity of halogenated aromatic hydrocarbons. Exposure of cultured rat hippocampal neurons and glia to TCDD (10-100 nM) invoked a rapid (within 3 min), concentration dependent increase in intracellular calcium ion concentration as determined by microscopic image analysis of cells noninvasively labeled with fluo-3. This rapid increase was blocked by the addition of EDTA (2 mM) or nifedipine (100 µM) to the external medium. In contrast, the nontoxic congener 1,2,3,4-TCDD was inactive at concentrations up to 10 µM. Other effects of TCDD on cultured neurons and astroglia were measured, including cell-cell communication via gap junctions, which was down-regulated, and cytosolic glutathione content, which was depressed in astroglia. Astroglial cells serve vital roles in regulating the neuronal environment. We have also conducted pilot experiments on TCDD effects on synaptic function in hippocampal slices. In these experiments, the excitatory postsynaptic potential was decreased. A cytosolic aryl hydrocarbon receptor was identified in the astroglia, but the neuronal cultures have not been studied. However, nuclear extracts from the astroglia treated with TCDD did not bind to a dioxin responsive element in a gel shift assay. These findings suggest that TCDD has effects on neural cells that are not due to the classical aryl hydrocarbon receptor- mediated signal transduction pathway. Calcium ions may play a role in TCDD-induced cell dysfunction and death, as has been previously shown for thymocytes.

Keyword(s): dioxin, neurotoxicity, hippocampus, neuron, astroglia.

Oral presentation in research track.


J.C. Tracy and H. Ramireddy, Northern Great Plains Water Resources Research Center and Department of Civil Engineering, South Dakota State University, Brookings, SD 57007

Recently, much work has been performed on the use of vegetative systems for remediating soil and groundwater contaminated with hazardous organic substances. A model to describe the performance of these vegetative systems under varying hydrogeochmeical conditions has been previously developed by several of the authors. Studies of the variability of difference plant characteristics on the performance of vegetative remediation systems have been reported.

However, a greater factor in the variability of vegetative remediation system performance could be caused by yearly changes in climatological conditions at a site. This paper will present the results of model analyses that use historical climatological data on the performance of vegetative remediation systems at a site. The results of these analyses will produce an estimate of the performance of the vegetative remediation system as well as a measure of the anticipated variability of the system for a particular site. The results could then be used to provide guidelines for design of a vegetative remediation system at a site.

Keyword(s): groundwater, phytoremediation, hazardous organic contaminants.

Oral presentation in research track.


M. Truex, B.M. Peyton and Y. Gorby, Pacific Northwest Laboratory, Richland, WA 99352

Some bacteria can destabilize the soluble uranyl carbonate complex prevalent in mine drainage by enzymatically reducing U(VI) to U(IV). Reduced uranium is highly insoluble and precipitates from solution as the U(IV) oxide, uraninite. The advantage of this technology is that the uranium is easily separated from the aqueous phase resulting in a small volume of relatively pure uraninite waste. Although a wide range of bacteria can reduce U(VI) to U(IV), many of the bacteria exhibit undesirable characteristics for large-scale treatment of contaminant waste streams. In this technology, dissimilatory iron reducers are used to reduce uranium under anaerobic conditions. The process is also applicable to other metal species such as chrome and technecium.

In initial experiments, cells from a 100-ml culture were introduced into a 1.5-L bioreactor containing 10-mM uranyl carbonate and reduced 100% of the U(VI) to U(IV). Pure uraninite precipitated from solution and was recovered from the vessel in a concentrated 20-ml suspension resulting in a 75-fold reduction in uranium waste volume. Additional experiments were conducted to describe the process kinetics using Monod expressions. These process kinetics and the pre- and post-treatment requirements of this technology will be presented.

Keyword(s): uranium, bioprecipitation, heavy metals, mine drainage.

Oral presentation in research track.

This work was supported by the U.S. Department of Energy as part of Pacific Northwest Laboratory’s Laboratory Directed Research and Development. Pacific Northwest Laboratory is operated for the U.S. Department of Energy by Battelle Memorial Institute under contract DE-AC06-76RLO 1830.


B. Tucker, C. Radtke, K. Sun-Il and A.J. Anderson, Department of Biology, Program in Toxicology, Utah State University, Logan, UT 84322-5305

The timing of onset of mineralization of pyrene by P. chrysosporium correlates with the production of ligninase activity. Ligninase activity is monitored by the decolorization of the polymeric dye Poly R-478. A layer of native soil decreased the growth of P. chrysosporium by visual assessment of hyphal mass and measurement of ergosterol, a fungal membrane sterol. The native soil layer slowed the production of ligninase as determined by Poly R-478 decolorization. The rate of mineralization of pyrene also was decreased. Mineralization, fungal growth, and rate of decolorization of the dye was improved when soil sterilized by autoclaving was used. Extracts from the native soil contained a consortium of bacteria and fungi. Several of the fungi and the bacteria were antagonistic to the growth of P. chrysosporium on solid medium. The bacteria caused antagonism as well as hyphal lysis. The onset of dye decolorization by P. chrysosporium on a nitrogen-limited medium was delayed by certain of the bacteria. These findings suggest that in certain soils, bioremediation by P. chrysosporium could be suppressed by indigenous soil microbes.

Keyword(s): P. chrysosporium, soil microbes, suppression.

Oral presentation in research track.


J.B. Varga2, F. Friedler l,2, and L.T. Fan1,1Department of Chemical Engineering, Kansas State University, Manhattan, KS 66506; and 2Department of Systems Engineering, Chemical Engineering Institute, Hungarian Academy of Sciences, Veszprem, Pf. 125, H-8201, Hungary

A simplistic approach to designing a waste minimizing chemical process is to synthesize the structure of the process first, followed by the design of the treatment system for the waste generated. In general, such an approach does not lead to a truly optimal process with integrated in- plant waste treatment because of the unavoidable interaction between the product generation and waste treatment. Thus, the waste generation and treatment should be taken into account from the outset in designing or synthesizing the process. This, however, will inevitably overburden the already complex task of process synthesis or design. Avoiding this difficulty requires a drastically improved algorithmic approach. The often adopted mixed-integer nonlinear programming (MINLP) model of a total flowsheet synthesis problem usually contains a large number of discrete or binary variables. This makes the model difficult to solve by methods that do not exploit the unique combinatorial feature of feasible process structures. The combinatorially accelerated branch and bound algorithm developed by us previously for the general total flowsheet synthesis minimizes the complexity involved in applying the conventional branch and bound algorithm by reducing the number of subproblems generated and by decreasing the size of each subproblem. This accelerated branch and bound algorithm has been extended to the optimal integrated synthesis of a total process flowsheet without neglecting waste treatment from the outset of design. The efficiency of the algorithm is demonstrated by synthesizing a relatively simple, commercial-scale process together with its waste treatment system; the stepwise procedure is given in detail.

Keyword(s): process design, waste minimization, algorithm.

Oral presentation in research track.


Gabriele Walser and Tissa Illangasekare, Dept. of Civil Engineering, University of Colorado, Boulder, CO 80309-0428

Spills of organic chemicals and waste products in the form of nonaqueous phase fluids (NAPL) has the potential to contaminate groundwater. Accidental spills at the soil surface or chemicals applied to the soil from leaking underground tanks result in the chemical to first travel through the unsaturated zone before reaching the groundwater in the saturated zone. During the migration through the vadose zone, a significant fraction of the contaminant, however, stays behind in the unsaturated soil as an entrapped phase. These entrapped chemicals act as a continuous source of contamination, as small amounts of the NAPL will continue to move within the soil or dissolve into the aqueous phase. The rising water table which results from the transient behavior of the regional groundwater system may also mobilize large amounts of the contaminant entrapped in the vadose zone. Thus, for cleanup measures it is very important to evaluate the process of entrapment and quantify the entrapped saturations based on the characteristics of the fluids and soils. In spill simulations which have been conducted in large tanks with various soil packing configurations, we have observed that entrapment is especially large in heterogeneous and layered soils. As only qualitative and semi-quantitative data can be obtained in large two-dimensional tanks, an experimental program was developed to investigate and obtain accurate data on entrapment in layered sand in vertical soil columns. In the descriptions of entrapment of nonaqueous phase contaminants in layered soils, it is assumed that the residual saturation can be determined using static pressure. The results from the column experiments are used to test this assumption. A test NAPL was spilled on top of a sand-filled layered column. The saturation and pressure distribution are monitored over a longer period of time. A dual-gamma system is used to determine the phase saturations. Specially designed ring-tensiometers are used to measure the average pressures at selected sections along the column. The transient pressure values are used to determine whether the entrapment is controlled by the static pressure distribution or is a steady state flow process which is reflected as a permanent entrapment. A simple steady state model will be evaluated to determine whether it can accurately predict this entrapment behavior. Then a non-steady state finite difference model developed by us will be used to see whether it can predict the steady state behavior.

Keyword(s): nonaqueous phase liquids, entrapment.

Oral presentation in research track.


G.A. Walter1, H.D. Stensel1 and S.E. Strand2, 1Department of Civil Engineering, FX-10; and 2College of Forest Resources, AR-10, University of Washington, Seattle, WA 98195

A mixed, suspended-growth, methanotrophic bioreactor was operated with continuous aqueous feed and increasing trichloroethylene loading until failure due to toxicity. Biomass and TCE degradation were continuously maintained as TCE loading increased from 4 to 10 µg TCE/mg protein/day. The maximum sustainable ratio of TCE degraded to methane consumed was 6 µg TCE/mg methane. Biomass, methane consumption, and TCE degradation fell precipitously when TCE loading exceeded 10 µg TCE/mg protein/day. Reactor failure was preceded by a fall in soluble methane monooxygenase activity.

Keyword(s): trichloroethylene, methanotrophic, bioreactor, toxicity, monooxygenase.

Poster presentation.


K.S. Washburn and T.D. Phillips, Toxicology, Department of Veterinary Anatomy and Public Health, Texas A&M University, College Station, TX 77843

Water-solvated chlorophenols (CPs) are environmental toxins associated with wood preservation and pesticide synthesis and usage. Their toxicity and association with dioxin- contaminated wastes are well-documented, as is their stability in most environmental settings. Several analytical procedures, mainly HPLC and GC/MS, are currently used to detect and quantify CPs, but these procedures are based on expensive equipment and technical expertise in a laboratory setting. We have developed an inexpensive, field- practical method for CPs, utilizing a small, packed glass minicolumn and derivatization of target CP molecules with dansyl chloride (5- dimethylaminonaphthalene-1sulfonyl chloride), or DsCl. A nonfluorescent borosilicate glass tube was used to house an array of inorganic sorbent materials, including preparative layers and a reactive neutral alumina interface separated by sand. DsCl is a substituted naphthalene with a conjugated X system that is responsible for its fluorescent complexation. Amines that reacted with DsCl were removed with a small amount of phyllosilicate clay to avoid interference. A neutral alumina/sand interface was used to strongly bind and immobilize the dansylated CPs. Activities greater than 3.0 for the alumina were avoided to prevent loss of selectivity, intensity and color of the fluorescence at the reactive interface. Our results indicated that this assay was capable of rapidly screening potable water samples and detecting CP contamination at very low concentrations (i.e., 1.0 ppb of pentachlorophenol in drinking water).

Keyword(s): detoxification, chlorophenols, rapid analysis.

Poster presentation.


C. Wend and W. Jones, Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717

Vapor-phase bioreactors (VPBRs) have been considered for treating gas streams contaminated by volatile organic compounds (VOCs). A VPBR is a gas absorption column that employs a biofilm as a heterogeneous catalyst growing on an artificial porous medium.

To aid in the design, scale-up and operation of a VPBR, a phenomenologically-based mathematical model has been developed to describe the steady-state operation of a VPBR. The present version tracks the electron-donor (VOC) and the electron-acceptor (oxygen) in the gas/liquid/biofilm phases within the bioreactor. The model uses a single species in the biofilm with reaction rates modeled by Monod kinetics for the electron acceptor and Haldane kinetics for the electron donor. Bench-scale VPBRs and the model show evidence of electron-acceptor limitation under certain operating conditions. Due to the effects of the inorganic salts and biomass on the apparent Henry's law coefficients, these coefficients were experimentally determined using reactor effluent. The coefficients were found to be approximately 50% of the published values for the VOCs.

Mass-transfer coefficients for the gas-liquid interface are predicted by Onda correlations. Literature shows these correlations to be within 20% of actual coefficients for abiotic systems. Bench-scale experiments using a non-reactive tracer showed that the Onda correlations can overestimate mass-transfer coefficients by as much as three times.

Keyword(s): modeling, mass-transfer coefficients, Onda, biofilm, Henry's law.

Poster presentation.


S. Wetzel, G. Kyle and M.K. Banks, Department of Civil Engineering, Kansas State University, Manhattan, KS 66506

Contamination of soil by petroleum hydrocarbons is a significant problem in the United States. Release of nonaqueous phase liquids (NAPLs) petroleum contaminants at or near the ground surface can migrate through the vadose zone to the groundwater. In contaminated coarse-textured soils containing abrupt interfaces with clay layers, petroleum hydrocarbons can accumulate at the clay interface due to restricted water flow through the clay layer. In addition, the soil matrix can act as a sink to which the petroleum contaminant becomes adsorbed, resulting in a difficult remediation scenario. Soil columns were erected using six inch PVC pipe. Three types of columns were constructed: one comprised only of sand, one of only clay, and one with both clay and sand layers. Each column was 24 inches long and duplicate columns were prepared, as well. Dodecane was introduced into the columns at a single mid-point at the top of each column as a one-time event. Four sets of each type of column were constructed to permit sampling of the column soil at four subsequent time intervals. Each column was disassembled at specified times and the contaminant concentration levels were quantified as a function of distance into the column. Results of this study contribute to a more comprehensive understanding of the behavior of NAPLs in contaminated heterogeneous soils with high clay content.

Keyword(s): petroleum hydrocarbons, NAPLs, vadose zone, heterogeneous soil.

Poster presentation.


D.R. Wildcat1, G.L. Godfrey1, D.L. Tillison1 and S.C. Grant2, 1Haskell Indian Nations University, Dept. of Natural and Social Sciences, 155 Indian Ave., Box 1227, Lawrence, KS 66046; and 2Great Plains-Rocky Mountain Hazardous Substance Research Center, Kansas State University, 101 Ward Hall, Manhattan, KS 66506- 2502

In 1993 Congress appropriated funds to the Environmental Protection Agency for the five national Hazardous Substance Research Centers to begin programs to serve minority educational institutions. The Great Plains-Rocky Mountain Hazardous Substance Research Center (GP-RM HSRC) serves EPA Regions VII and VIII. The states that the GP-RM HSRC serves contain 17 predominantly Native American colleges and universities, two predominantly black universities, and two predominantly Hispanic colleges. Due to the prominence of Native American educational institutions in its regions, the GP-RM HSRC has decided to make the emphasis of its program toward American Indian and Alaska Native institutions, but the program will also serve other minority institutions in EPA Regions VII and VIII. The GP-RM HSRC has joined with Haskell Indian Nations University to begin the Native American and Other Minority Institutions (NAOMI) Program. The NAOMI Program will involve minority educational institutions in research, training and technology transfer. This will be accomplished primarily through the establishment of an outreach office of the GP-RM HSRC on the campus of Haskell Indian Nations University. This office will be called the Haskell Environmental Research Studies Center (HERS). The NAOMI Program will include a seminar program, a summer research participation program at GP-RM HSRC consortium universities, and projects at NAOMI consortium schools.

Keyword(s): Native American, minority, research, technology transfer, hazardous substances.

Poster presentation.


P. Williamson and V. Prabhakar, Roux Associates, Inc., 1401 17th Street, Suite 400, Denver, CO 80202

Characterization and delineation of the source of volatile organic compounds (VOCs) detected in a groundwater plume was performed using a cone penetrometer testing (CPT) rig equipped with a drive point groundwater sampling system. Attempts to delineate the source area using a soil gas survey, borings and soil analyses were not successful. The VOCs were apparently released at the surface of a 45-foot-thick interlayered sand and clay alluvium overlying a limestone aquifer. Depth to groundwater was approximately 25 feet below ground surface. Lithologic data and groundwater samples were collected with the CPT rig along four sampling lines located perpendicular to the direction of groundwater flow. The high resolution lithologic data obtained at each sampling point delineated transition zones from coarse-grained sand to clay soils. Groundwater samples were collected from the saturated transition zones. Expedited analyses of groundwater samples and on-site development of geologic cross sections determined the locations of the additional sampling lines. A total of 24 CPT soundings and 31 groundwater samples were required to delineate the lithology and distribution of VOCs in four distinct source areas. The CPT sounding and sampling technique produced high resolution hydrogeologic and groundwater chemical data of the source area for approximately one-third of the cost of a traditional boring and monitoring well program. In addition, soil cuttings, well development water, or well purge water were not produced.

Keyword(s): site characterization, volatile organic compounds, cone penetrometer testing.

Oral presentation in technology transfer track.


W.E. Woldt, M.F. Dahab and Jan Hygnstrom, Department of Biological Systems Engineering, 253 L.W. Chase Hall, University of Nebraska, Lincoln, NE 68583-0771

Pollution prevention is considered as the principal means for true waste reduction in rural areas since traditional waste minimization activities such as reuse, recycling and recovery are not easily implemented in such areas. This presentation will focus on the final products resulting from the development of a pollution prevention educational program for small quantity hazardous waste generators within the EPA Regions VII and VIII. The principal goal of the educational program is to provide additional incentives for true hazardous waste prevention through source reduction in commercial businesses (small quantity generators) such as vehicle maintenance, dry cleaning operations, agricultural cooperatives, pesticide applicators, and others. The presentation will center on results of development of waste prevention "tool kits" which will be targeted for use by small quantity generators. These "tool kits" will be made available to small quantity generators which in turn can be used in the implementation of waste prevention programs within their facilities. The "tool kits" consist of self assessment and self help "how to" documents and checklists that can be used by individual small quantity generators. Five different types of "tool kits" will be presented, each for a different type of hazardous waste small quantity generator.

Keyword(s): pollution, prevention, hazardous, waste, reduction.

Oral presentation in technology transfer track.


J.H. Wolfram and R.D. Rogers, EG&G Idaho Inc., Idaho Falls, ID 83415

The potential of biological processing of mixed hazardous waste has not been determined. However, the use of selected microorganisms for the degradation and/or detoxification of hazardous organic compounds is gaining wide acceptance as an alternative waste treatment technology. The isolation of a unique strain of Pseudomonas Putida Idaho seems well adapted to withstand the demands of the input stream comprised of liquid scintillation waste. This paper describes the results from the continuous processing of a mixture comprised of p-xylene and surfactant as well as commercial liquid scintillation formulations. The two formulations tested contained xylene and pseudocumene as the solvent base. The process is now at the demonstration phase at one of DOE’s facilities which has a substantial amount of stored waste of this type. The system at the DOE facility is comprised of two CSTR units in series.

Keyword(s): bioremediation, mixed waste, organic solvents.

Oral presentation.


Xiaoqing Yang, Larry E. Erickson and L.T. Fan, Department of Chemical Engineering, Durland Hall, Kansas State University, Manhattan, KS 66506-5102

Since many petroleum components are barely soluble in water, they are frequently present as nonaqueous phase liquids (NAPLs) in subsurface soils. Bioremediation of soils contaminated by such residual hydrocarbons appears to be cost- effective under various situations. More often than not, however, bioremediation can be rate-limited by dissolution, solute transport or biotransformation in a four-phase system comprising the NAPL, aqueous, solid and microbial phases. This paper proposes a model elucidating the microbial assimilation of hydrocarbon contaminants on the solid surface and in the aqueous phase. It focuses on those situations in which dissolution is the main rate- limiting factor. An investigation has been carried out on the rates of biodegradation of some common petroleum components such as benzene, ethyl-benzene, toluene and xylene in the four- phase system. The effects considered are those of the mass-transfer area, specific growth rate of biomass, and velocity of pore-water flow.

Keyword(s): NAPL, bioremediation, soil, BETX, batch reactor.

Poster presentation in research track.


W. Yu1, C.S. Clennan1, D. Sotomayor2, M.K. Banks1 and C.W. Rice2, 1Department of Civil Engineering, and 2Department of Agronomy, Kansas State University, Manhattan, KS 66506

The number and activity of microorganisms in the unsaturated zone of both pristine and agricultural soils have been examined in depth profiles. Numbers of total and actively respiring microbes were enumerated by DAPI ( 4’,6- diamidino-2-phenylindole ) and CTC (5-cyano- 2,3-ditolyl tetrazolium chloride) epifluorescent microscopic direct count methods, respectively. Viable aerobes were also assessed by plate counts. Denitrifiers were enumerated by most probable number (MPN) method. Results show that there is little variability in total numbers of microbes among the depths for both sites and between the two sites. The numbers of actively respiring bacteria in the pristine site decreased with depth above groundwater table and increased thereafter. The numbers of actively respiring bacteria in agricultural site, however, showed little variability with depth. The results of plate counts show a similar pattern throughout the depth profile to the CTC counts. Numbers of denitrifiers in the agricultural site increased with depth up to two meters from the surface, but stayed constant between a depth of two meters and water table. The numbers of denitrifiers in pristine site were lowest at water table and became consistently high in the saturated zone. The results suggest that there was low microbial activity around the groundwater table at pristine soils. The results also show the numbers of microbes and denitrifiers are higher in the agricultural site than in the pristine soils. The number and activity of microbes in the unsaturated soil appear to vary with the land use and methods of assessment.

Keyword(s): unsaturated, soil, microorganism, direct count.

Oral presentation in research track.


T.C. Zhang, Y.C. Fu and P.L. Bishop, Department of Civil and Environmental Engineering, University of Cincinnati, Cincinnati, OH 45221

Microelectrodes and a microslicing technique have been used to elucidate changes in the structure of biofilms with depth and to determine the influence of these changes on mass transport in biofilms treating wastewaters containing azo dyes. Biofilm physical properties such as density, porosity and mean pore size were measured at various depths within a biofilm, as were several biological properties such as total plate count, viable cell populations and microbial activity. In addition, microelectrodes were used to measure profiles of dissolved oxygen, pH, ammonium nitrogen and nitrate nitrogen. These data were coupled to determine variations in diffusivity with depth in the biofilm and to show where various reactions were occurring. The results show that biofilms are non-homogeneous and highly stratified. They are characterized by an increase of biofilm density, a decrease of metabolically active biomass, and a decrease of porosity with depth. The ratio of effective diffusivity for dissolved oxygen inside the biofilm to the diffusivity in the bulk solution, as well as the effectiveness factor, decrease with biofilm depth. This stratified structure is created by the competition for substrate and space by microorganisms in the biofilm. Commonly used biofilm models do not account for this stratification and need to be modified.

Keyword(s): biofilm, mass transport, microelectrode, micro-slicing, diffusivity.

Oral presentation in research track.


W. Zhang1, E. Bouwer1 and Al Cunningham2, 1Department of Geography and Environmental Engineering, the Johns Hopkins University, Baltimore, MD 21218; 2Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717

In-situ bioremediation has been applied successfully at a few sites. Several restrictions presently exist which could greatly limit the effectiveness of this promising technology. Hydrophobic organic contaminants tend to sorb onto soil. However, microorganisms are most effective in utilizing substrates from the aqueous phase. Sorption tends to separate the direct contact between microorganisms and contaminants necessary for biodegradation to occur. Effects of sorption on biodegradation in the subsurface can be classified into two categories: (1) concentration effect. Sorption reduces concentrations of organic contaminants in bulk water. This can increase microbial growth if the organic compound is toxic to microorganisms or it can decrease or even stop the microbial growth by lowering the bulk water concentration for a non-toxic biodegradable compound. (2) Rate limitation by mass transfer processes. The movement of organic compounds from solid phases to the aqueous phase is subject to mass transfer resistance before reaching microorganisms. Recent evidence from both laboratory and field work indicates that the mass transfer rate could be slow.

A series of experiments, which represented scenarios with fast sorption/desorption, slow sorption/desorption, mass transfer across boundary layer and mass transfer within attached microorganisms (biofilm), was conducted to demonstrate the concentration effect and the mass transfer effect. A method has been developed to quantify bioavailability of organic contaminants in aquatic environments. Bioavailability Factor (Bf), a dimensionless parameter derived from mathematical models and verified by experimental results, has been formulated to describe the impact of equilibrium sorption, nonequilibrium sorption, and mass transfer processes on the rate and extent of biodegradation. This method can be used to improve design and operation, such as scale-up of laboratory results for bioremediation. Shortcomings in the reported studies on sorption/biodegradation interactions have been identified by this method.

Keyword(s): sorption, biodegradation, bioavailability, in-situ bioremediation, petroleum hydrocarbons.

Oral presentation in research track.