Handbook of BIOREMEDIATION NORRIS • HINCHEE • BROWN • MCCARTY SEMPRM • WILSON • KAMPBELL REINHARD • BOUWER • BORDEN • VOGEL THOMAS • WARD Project Officer John E. Matthews Chief, Applications and Assistance Branch Robert S. Kerr Environmental Research Laboratory Ada, Oklahoma First published 1994 by Lewis Publishers Taylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300 Boca Raton, FL 33487-2742 Reissued 2018 by CRC Press © 1994 by Taylor & Francis CRC Press is an imprint of Taylor & Francis Group, an Informa business No claim to original U.S. Government works This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been made to publish reliable data and information, but the author and publisher cannot assume responsibility for the validity of all materials or the consequences of their use. 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ISBN 13: 978-1-138-10570-6 (hbk) ISBN 13: 978-0-203-71276-4 (ebk) Visit the Taylor & Francis Web site at http://www.taylorandfrancis.com and the CRC Press Web site at http://www.crcpress.com FOREWORD EPA is charged by Congress to protect the Nation's land, air and water systems. Under a mandate of national environmental laws focused on air and water quality, solid waste management and the control of toxic substances, pesticides, noise and radiation, the Agency strives to formulate and implement actions which lead to a compatible balance between human activities and the ability of natural systems to support and nurture life. The RobertS. Kerr Environmental Research Laboratory is the Agency's center of expertise for investigation of the soil and subsurface environment. Personnel at the laboratory are responsible for management of research programs to: (a) determine the fate, transport and transformation rates of pollutants in the soil, the unsaturated and the saturated zones of the subsurface environment; (b) define the processes to be used in characterizing the soil and subsurface environment as a receptor of pollutants; (c) develop techniques for predicting the effect of pollutants on ground water, soil, and indigenous organisms; and (d) define and demonstrate the applicability and limitations of using natural processes, indigenous to the soil and subsurface environment, for the protection of this resource. In-situ bioremediation of subsurface environments involves the use of microorganisms to convert contaminants to less harmful products and sometimes offers significant potential advantages over other remediation technologies. This report provides the most recent scientific understanding of the processes involved with soil and ground-water bioremediation and discusses the applications and limitations of the various in-situ bioremediation technologies. Clinton W. Hall Director Robert S. Kerr Environmental Research Laboratory iii Dedicated to Richard L. Raymond, Sr. Through his vision, commitment, and humanity, he created the subsurface bioremediation industry. iv CONTENTS Foreword ..........................................................................................................................................iii Figures .............................................................................................................................................xi Tables .............................................................................................................................................xiii Executive Summary...........................................................................................................................1 Section 1. Introduction ................................................................................................................13 Section 2. In-Situ Bioremediation of Soils and Ground Water Contaminated with Petroleum Hydrocarbons 2.1. Introduction .....................................................................................................17 2.2. Fundamental Principles.....................................................................................17 2.3 Historical Perspective........................................................................................19 2.4 Repositories of Expertise..................................................................................21 2.5. General Designs................................................................................................21 2.6. Laboratory Testing............................................................................................22 2.7. Contamination Limits........................................................................................22 2.8. Site Characterization.........................................................................................24 2.9. Favorable Site Conditions.................................................................................25 2.9.1. Solubility ..........................................................................................25 2.9.2. Volatility ............................................................................................25 2.9.3. Viscosity ............................................................................................25 2.9.4. Toxicity .............................................................................................26 2.9.5. Permeability of Soils and Subsurface Materials..................................26 2.9.6. Soil Type............................................................................................26 2.9.7. Depth to Water....................................................................................26 2.9.8. Mineral Content..................................................................................26 2.9.9. Oxidation/Reduction Potential............................................................26 2.9.10. pH .....................................................................................................27 2.10. Infrastructure and Institutional Issues.................................................................27 2.11. Performance ......................................................................................................28 2.12. Problems ..........................................................................................................28 2.13. Site Properties vs Cost......................................................................................30 2.13.1. Mass of Contaminant..........................................................................30 2.13.2. Volume of Contaminated Aquifer.......................................................30 2.13.3. Aquifer Permeability/Soil Characteristics............................................30 2.13.4. Final Remediation Levels...................................................................30 2.13.5. Depth to Water....................................................................................31 2.13.6. Monitoring Requirements...................................................................31 2.13.7. Contaminant Properties.......................................................................31 2.13.8. Location of Site..................................................................................31 2.14. Previous Experience with Costs.........................................................................32 2.15. Regulatory Acceptance......................................................................................33 2.16. Knowledge Gaps...............................................................................................34 v Section 3. Bioventing of Petroleum Hydrocarbons 3.1. Fundamental Principles........................................................................................39 3.1.1. Review of the Technology....................................................................39 3.1.2. Maturity of the Technology...................................................................41 3.1.3. Repositories of Expertise.......................................................................41 3.2. Contamination that is Subject to Treatment.........................................................43 3.3. Special Requirements for Site Characterization...................................................44 3.3.1. Soil Gas Survey.....................................................................................44 3.3.2. Soil Gas Permeability and Radius of Influence.....................................44 3.3.3. In-Situ Respiration.................................................................................45 3.4. Impact of Site Characteristics on Applicability...................................................47 3.5. Process Performance............................................................................................48 3.5.1. Case Study: Hill AFB Site....................................................................50 3.5.2. Case Study: Tyndall AFB Site...............................................................52 3.5.3. Performance of Other Sites...................................................................54 3.6. Problems Encountered with the Technology........................................................55 3.7. Costs ....................................................................................................................55 3.8. Regulatory Acceptance.........................................................................................55 3.9. Knowledge Gaps and Research Opportunities.....................................................55 Section 4. Treatment of Petroleum Hydrocarbons in Ground Water by Air Sparging 4.1. Introduction .....................................................................................................61 4.2. Development of Air Sparging...........................................................................62 4.3. Principles of the Technology............................................................................63 4.4. Benefits of Air Sparging...................................................................................65 4.5. Dangers of Air Sparging...................................................................................68 4.6. Barriers to Flow...............................................................................................68 4.7. Control of Flow...............................................................................................70 4.8. Summary of Limitations...................................................................................71 4.9. System Application and Design........................................................................73 4.9.1. Nature and Extent of Site Contaminants.............................................73 4.9.2. Hydrogeologie Conditions...................................................................74 4.9.3. Potential Ground-Water and Vapor Receptors.....................................74 4.10. Field Pilot Testing.............................................................................................74 4.11. Design Data Requirements...............................................................................75 4.12. System Elements..............................................................................................77 4.13. System Examples..............................................................................................79 4.14. Cost Factors......................................................................................................82 4.15. Conclusion ......................................................................................................83 Section 5. Ground-Water Treatment for Chlorinated Solvents 5.1. Introduction .....................................................................................................................87 5.2. Biotransformation of CaHs............................................................................................89 5.2.1. Primary Substrates and Cometabolism.......................................................89 5.2.2. CAH Usage as Primary Substrates..............................................................92 5.2.3. Anaerobic Cometabolic Transformations of CAHs...................................92 5.2.4. Aerobic Microbial Transformation of Chlorinated Aliphatic Hydrocarbons................................................................................93 vi 5.3. Processes Affecting Chemical Movement and Fate............................................95 5.3.1. Effect of Sorption................................................................................95 5.4. Field Pilot Studies of CAH Transformation.......................................................97 5.4.1. Results with Methanotrophs.................................................................98 5.4.2. Results with Phenol Utilizers........................................................... 100 5.4.3. Comparison Between the Methane and Phenol Studies..................... 102 5.4.4. Anaerobic Transformation of Carbon Tetrachloride.......................... 102 5.5. Procedures for Introducing Chemicals into Ground Water.............................. 103 5.6. The Effect of Site Conditions on Remediation Potential................................. 107 5.6.1. Microorganism Presence.................................................................. 109 5.7. Summary ....................................................................................................... 110 Section 6. Bioventing of Chlorinated Solvents for Ground-Water Cleanup through Bioremediation 6.1. Fundamental Principles.................................................................................. 117 6.2. Maturity of the Technology........................................................................... 118 6.3. Primary Repositories of Expertise................................................................... 118 6.4. Contamination Subject to Treatment............................................................... 119 6.5. Special Requirements for Site Characterization............................................... 120 6.6. Site Characteristics that are Particularly Favorable......................................... 121 6.7. Site Characteristics that are Particularly Unfavorable..................................... 121 6.8. Performance under Optimal Conditions.......................................................... 122 6.8.1. Importance of the Rate Law............................................................ 122 6.8.2. Importance of Partitioning............................................................... 122 6.9. Problems Encountered with the Technology................................................... 125 6.10. Relevant Experience with System Design....................................................... 125 Section 7. In-Situ Bioremediation Technologies for Petroleum Derived Hydrocarbons Based on Alternate Electron Acceptors (other than molecular oxygen) 7.1. Fundamental Principles.................................................................................. 131 7.1.1. Comparison of Oxygen and Alternate Electron Acceptor Based In-Situ Bioremediation Technologies................................................ 131 7.1.2. Hydrocarbon Transformation Based on Alternate Electron Acceptors........................................................................... 133 7.1.2.1. Laboratory Studies........................................................... 133 7.1.2.2. Large Scale Bioremediation Studies Using Nitrate........... 136 7.1.3. Maturity of the Technology.............................................................. 136 7.1.4. Primary Respository of Expertise...................................................... 137 7.2. Contamination that is Subject to Treatment.................................................... 138 7.2.1. Chemical Nature.............................................................................. 138 7.2.2. Range of Concentration................................................................... 138 7.3. Requirements for Site Characterization and Implementation of the Technology ............................................................................................. 138 7.4. Favorable Site Characteristics......................................................................... 139 7.5. Unfavorable Site Characteristics..................................................................... 139 7.5.1. Chemical and Physical Nature of the Contamination....................... 139 7.5.2. Site Hydrogeology and Source Characteristics................................. 139 7.5.3. Infrastructure and Institutional Issues............................................... 139 7.6. Optimal Site Conditions................................................................................. 139 vii 7.7. Problems Encountered with the Technology................................................... 140 7.8. Properties of Site and Contaminants Determining the Cost of Remediation.... 140 7.9. Previous Experience with Cost of Implementing the Technology................... 141 7.10. Factors Determining Regulatory Acceptance of the Technology..................... 141 7.11. Primary Knowledge Gaps and Research Opportunities.................................... 141 7.11.1. Degradation Under Ideal Conditions................................................ 142 7.11.2. Degradation Under Ground-Water and Soil Conditions..................... 142 7.11.3. Degradation at Sites........................................................................ 142 7.11.4. Degradation at the Hydrocarbon/Water Interface and Within the Nonaqueous Phase........................................................................... 143 7.11.5. Methods for Monitoring Performance of Bioremediation Process .... 143 7.11.6. Design of Optimal Nutrient and Electron Acceptor Systems............ 143 Section 8. Bioremediation of Chlorinated Solvents Using Alternate Electron Acceptors 8.1. Introduction .................................................................................................. 149 8.2. Metabolism and Alternate Electron Acceptors................................................ 150 8.3. Biotransformation of Chlorinated Solvents in the Presence of Alternate Electron Acceptors........................................................................................ 151 8.3.1. Carbon Tetrachloride Biotransformation.......................................... 155 8.3.2. Tetrachloroethene and Trichloroethene Biotransformation................. 155 8.4. Approaches for Treatment.............................................................................. 156 8.5. Field Experience............................................................................................. 158 8.6. Sequential Anaerobic/Aerobic Transformations of Chlorinated Solvents........ 158 8.7. Performance .................................................................................................. 159 8.7.1. Physical/Chemical Properties........................................................... 159 8.7.2 Concentration Range........................................................................ 160 8.7.3. Favorable Redox Conditions............................................................ 161 8.8. Biotransformation Stoichiometry.................................................................... 163 8.9. Biotransformation Rates................................................................................ 166 8.10. Limitations ................................................................................................... 167 8.11. Research Needs.............................................................................................. 169 8.12. Concluding Remarks...................................................................................... 169 Section 9. Natural Bioremediation of Hydrocarbon-Contaminated Ground Water 9.1. General Concept of Natural Bioremediation.................................................. 177 9.2. Hydrocarbon Distribution, Transport and Biodégradation in the Subsurface ... 178 9.2.1. Petroleum Hydrocarbon Biodégradation.......................................... 179 9.2.2. Subsurface Microorganisms............................................................. 179 9.2.3. Use of Different Electron Acceptors for Biodégradation.................. 180 9.2.3.1. Aerobic Biodégradation.................................................. 180 9.2.3.2. Biodégradation via Nitrate Reduction.............................. 181 9.2.3.3. Biodégradation using Ferric Iron..................................... 181 9.2.3.4. Biodégradation via Sulfate Reduction and Methanogenesis ............................................................. 182 9.2.4. Effect of Environmental Conditions on Biodégradation................... 182 9.3. Natural Bioremediation of a Hydrocarbon Plume............................................ 183 9.4. Case Studies of Natural Bioremediation......................................................... 185 9.5. Site Characterization for Natural Bioremediation............................................ 186 9.5.1. Is the Contaminant Biodegradable?.................................................. 187 viii 9.5.2. Is Biodégradation Occurring in the Aquifer?..................................... 187 9.5.3. Are Environmental Conditions Appropriate for Biodégradation?...... 188 9.5.4. If the Waste Doesn’t Completely Biodegrade, Where Will It Go? ... 188 9.6. Monitoring Natural Bioremediation Systems..................................................... 189 9.7. Performance of Natural Bioremediation Systems.............................................. 190 9.8. Predicting the Extent of Natural Bioremediation............................................... 190 9.9. Issues That May Affect the Costs of This Technology..................................... 192 9.10. Knowledge Gaps and Research Opportunities.................................................. 192 Section 10. Natural Bioremediation of Chlorinated Solvents 10.1. Summary .......................................................................................................201 10.2. Fundamental Principles..................................................................................201 10.3. Chemical Reactions........................................................................................203 10.4. Microbiological Reactions..............................................................................204 10.4.1. Aerobic ..........................................................................................204 10.4.2. Anaerobic ........................................................................................206 10.5. Predictions of Product Distribution................................................................208 10.6. Rationale for Technology...............................................................................211 10.7. Practical Implications.....................................................................................215 10.8. Special Requirements for Site Characterization...............................................216 10.9. Favorable Site Characteristics.........................................................................216 10.10. Unfavorable Site Characteristics.....................................................................218 10.11. Cost Evaluations.............................................................................................220 10.12. Knowledge Gaps............................................................................................220 10.13. Conclusion ....................................................................................................220 Section 11. Introduced Organisms for Subsurface Bioremediation 11.1. Fundamental Principles of the Technology..............................................................227 11.1.1. Review of the Development of the Technology.....................................227 11.1.2. Matrix Properties that Affect Transport...................................................228 11.1.3. Properties of Organisms that Affect Transport......................................230 11.1.4. Operational Factors that Affect Transport..............................................231 11.1.5. Environmental Factors that Affect Survivability of Added Organisms.........................................................................................233 11.1.6. Field Demonstrations of Microbial Transport.........................................233 11.1.7. Inoculation to Enhance Biodégradation of Hydrocarbons.....................235 11.1.8. Inoculation to Enhance Biodégradation of Chlorinated Compounds ..................................................................................................236 11.2. Maturity of the Technology.......................................................................................237 11.3. Primary Repositories of Expertise..............................................................................238 11.4. Other Factors Concerning Application.....................................................................238 11.5. State of the Art of Transport of Microorganisms with Specialized Metabolic Capabilities and Research Opportunities..............................................239 ix