Final Report: Remediation of TNT-Contaminated Soil by Cyanobacterial Mat.

EPA Grant Number: R825513C030
Subproject: this is subproject number 030 , established and managed by the Center Director under grant R825513
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: HSRC (1989) - South and Southwest HSRC
Center Director: Reible, Danny D.
Title: Remediation of TNT-Contaminated Soil by Cyanobacterial Mat.
Investigators: Mondecar, M. C. , Saunders, F. Michael
Institution: Clark Atlanta University , Georgia Institute of Technology
EPA Project Officer: Hahn, Intaek
Project Period: January 1, 1995 through January 1, 1997
Project Amount: Refer to main center abstract for funding details.
RFA: Hazardous Substance Research Centers - HSRC (1989) RFA Text |  Recipients Lists
Research Category: Hazardous Substance Research Centers , Land and Waste Management


The objectives for this study are:
1. To develop bacterial consortia and Oscillatoria spp. for tolerance to TNT, to give "TNT-Oscillatoria".
2. To determine rates of degradation and mineralization in TNT-spiked soil, using TNT-mat, i.e. mat containing TNT-tolerant bacteria.
3. To test mat for TNT degradation in actual field samples of soil contaminated with TNT.

Summary/Accomplishments (Outputs/Outcomes):

Bacteria were isolated from TNT-contaminated soil collected from Alabama Army Ammunition Plant (AAAP), Childersburg, AL. Enrichment cultures were started by placing 1 g soil in a flask of 50 ml nutrient broth (NB), then incubating on a rotary shaker. Subenrichment cultures were established with a 2% inoculum of the previous culture, along with TNT. Oscillatoria was cultured by placing mat onto Allen-Arnon (AA) agar (Allen and Arnon, 1955), then cutting out filaments of the algae growing from the mat, and adding the filaments to AA liquid medium. It was also grown from Oscillatoria which had previously been isolated from other mat. New "control" mat was prepared by blending previous mat with AA and Silage Medium (Judith Bender, personal communication), then adding silage. Both Oscillatoria and mat were maintained in am incubator set to a 12:12 LD cycle. This light regime was also used in all degradation and mineralization experiments.

To develop tolerance for TNT, bacterial consortia were incubated in increasing concentrations of TNT in NB. Consortia were cultured in mineral salts medium (MSM) to determine if TNT could be used as a sole carbon source and in a modified MSM supplemented with glucose and without any nitrogen compounds to determine if it could be used as a sole nitrogen source. Increases in growth were measured by increases in optical density at 600 nm on a Milton Roy Spectronic 21D Spectrophotometer. Mat which had not previously been exposed to TNT was incubated in 50 ppm and 100 ppm TNT. TNT-bacteria were integrated into mat. TNT-bacteria, TNT-mat and control mat were exposed to 100 ppm TNT in a NaOH plate assay to screen for degradative ability of TNT-bacteria and TNT-mat (Osmon and Klausmeier, 1972).

Although these bacteria can not use TNT as a sole carbon source or nitrogen source, they do degrade it. Both TNT-mat and mar lacking TNT-bacteria are tolerant of TNT. Mineralization was low. Using spiked soil, most degradation by mat occurred in the first four days. When "naturally" contaminated soil was used, soil treated with mat had a greater decrease in TNT concentrating than did untreated soil. However, the mat did not appear to take up any TNT. This may be due to either the very high concentration of TNT in soil or due to TNT being bound to the humic fraction of the soil to the TNT: the degree of binding would increase over time. This binding may make it more difficult for the components of the mat to get to the TNT in order to degrade it. It may be desirable in future field studies using mat to periodically till the soil to the mat.

Journal Articles:

No journal articles submitted with this report: View all 1 publications for this subproject

Supplemental Keywords:

bacterial consortia, degradation, and mineralization., RFA, Scientific Discipline, Geographic Area, Waste, Water, Ecology, Chemical Engineering, Contaminated Sediments, Remediation, Environmental Chemistry, Chemistry, State, Biochemistry, Bioremediation, Ecology and Ecosystems, Biology, Environmental Engineering, plant-based remediation, in situ remediation, Alabama (AL), contaminated soil, soils, bioremediation of soils, contaminants in soil, biotransformation, phytoremediation, plant mediated contaminants, TNT, cyanobacteria, munitions, zooplankton

Progress and Final Reports:

Original Abstract
  • 1995

  • Main Center Abstract and Reports:

    R825513    HSRC (1989) - South and Southwest HSRC

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R825513C001 Sediment Resuspension and Contaminant Transport in an Estuary.
    R825513C002 Contaminant Transport Across Cohesive Sediment Interfaces.
    R825513C003 Mobilization and Fate of Inorganic Contaminant due to Resuspension of Cohesive Sediment.
    R825513C004 Source Identification, Transformation, and Transport Processes of N-, O- and S- Containing Organic Chemicals in Wetland and Upland Sediments.
    R825513C005 Mobility and Transport of Radium from Sediment and Waste Pits.
    R825513C006 Anaerobic Biodegradation of 2,4,6-Trinitrotoluene and Other Nitroaromatic Compounds by Clostridium Acetobutylicum.
    R825513C007 Investigation on the Fate and Biotransformation of Hexachlorobutadiene and Chlorobenzenes in a Sediment-Water Estuarine System
    R825513C008 An Investigation of Chemical Transport from Contaminated Sediments through Porous Containment Structures
    R825513C009 Evaluation of Placement and Effectiveness of Sediment Caps
    R825513C010 Coupled Biological and Physicochemical Bed-Sediment Processes
    R825513C011 Pollutant Fluxes to Aquatic Systems via Coupled Biological and Physicochemical Bed-Sediment Processes
    R825513C012 Controls on Metals Partitioning in Contaminated Sediments
    R825513C013 Phytoremediation of TNT Contaminated Soil and Groundwaters
    R825513C014 Sediment-Based Remediation of Hazardous Substances at a Contaminated Military Base
    R825513C015 Effect of Natural Dynamic Changes on Pollutant-Sediment Interaction
    R825513C016 Desorption of Nonpolar Organic Pollutants from Historically Contaminated Sediments and Dredged Materials
    R825513C017 Modeling Air Emissions of Organic Compounds from Contaminated Sediments and Dredged Materials title change in last year to "Long-term Release of Pollutants from Contaminated Sediment Dredged Material"
    R825513C018 Development of an Integrated Optic Interferometer for In-Situ Monitoring of Volatile Hydrocarbons
    R825513C019 Bioremediation of Contaminated Sediments and Dredged Material
    R825513C020 Bioremediation of Sediments Contaminated with Polyaromatic Hydrocarbons
    R825513C021 Role of Particles in Mobilizing Hazardous Chemicals in Urban Runoff
    R825513C022 Particle Transport and Deposit Morphology at the Sediment/Water Interface
    R825513C023 Uptake of Metal Ions from Aqueous Solutions by Sediments
    R825513C024 Bioavailability of Desorption Resistant Hydrocarbons in Sediment-Water Systems.
    R825513C025 Interactive Roles of Microbial and Spartina Populations in Mercury Methylation Processes in Bioremediation of Contaminated Sediments in Salt-Marsh Systems
    R825513C026 Evaluation of Physical-Chemical Methods for Rapid Assessment of the Bioavailability of Moderately Polar Compounds in Sediments
    R825513C027 Freshwater Bioturbators in Riverine Sediments as Enhancers of Contaminant Release
    R825513C028 Characterization of Laguna Madre Contaminated Sediments.
    R825513C029 The Role of Competitive Adsorption of Suspended Sediments in Determining Partitioning and Colloidal Stability.
    R825513C030 Remediation of TNT-Contaminated Soil by Cyanobacterial Mat.
    R825513C031 Experimental and Detailed Mathematical Modeling of Diffusion of Contaminants in Fluids
    R825513C033 Application of Biotechnology in Bioremediation of Contaminated Sediments
    R825513C034 Characterization of PAH's Degrading Bacteria in Coastal Sediments
    R825513C035 Dynamic Aspects of Metal Speciation in the Miami River Sediments in Relation to Particle Size Distribution of Chemical Heterogeneity