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Most photographs reproduce acceptably on positive microfilm or microfiche but lack the clarity on xerographic copies made from the microfilm. For an additional charge, 35mm slides of 6”x 9” black and white photographic prints are available for any photographs or illustrations that cannot be reproduced satisfactorily by xerography. Order Number 8726585 Soils containing 2,3,7,8-Tetrachlorodibenzo-p-dioxin: Aspects of their microbial activity and the potential for their microbially-mediated decontamination Arthur, Mickey Francis, Ph.D. The Ohio State University, 1987 UMI 300 N. Zeeb Rd. Ann Arbor, MI 48106 PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a checkV . mark 1. Glossy photographs or _____pages 2. Colored illustrations, paper or______ print 3. Photographs with dark background____ 4. Illustrations are poor copy______ 5. 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Other_____________________________________________________________________ University Microfilms International SOILS CONTAINING 2,3,7,8-TETRACHLORODIBENZO-P-DIOXIN: ASPECTS OF THEIR MICROBIAL ACTIVITY AND THE POTENTIAL FOR THEIR MICROBIALLY-MEDIATED DECONTAMINATION DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of the Ohio State University By Mickey Francis Arthur, B.S., M.S. The Ohio State University 1987 Dissertation Committee: Approved by J .I . Frea R.M. Pfister W .R. Strohl 0. H . Tuovinen Department of Microbiology ACKNOWLEDGEMENTS I express sincere appreciation to the many people who helped make the completion of this work possible. My thanks go to Dr. James I. Frea for his guidance, insight, and patience throughout this research. I also thank Drs. Robert M. Pfister, William R. Strohl, and Olli H. Tuovinen for their advice and suggestions during the General and Final Examinations. I express gratitude to Drs. Barney W. Cornaby, James A. Fava, L. Barry Goss, and Danny R. Jackson at Battelle-Columbus Division for their constant support and encouragement. The technical assistance of Thomas C. Zwick, G. Kelly O ’Brien, Thomas deJolsvay, John Steichen, Dan Aichele, and Lisa M. Hendrickson is gratefully acknowledged. Finally, I am indebted to my wife, Vicki, and my children, Melissa, Thomas, and Natalie, for their patience and love during this long effort. ii VITA November 18, 1950 ....... Born - Columbus, Ohio 1976 ..................... B.S., The Ohio State University, Columbus, Ohio 1978 ..................... M.S., The Ohio State University, Columbus, Ohio 1978-1983 ................Researcher, Bi©environmental Sciences, Battelle-Columbus Division, Columbus, Ohio 1983-Present .......... .Research Scientist, Environmental Technology and Assessment, Battelle-Columbus Division, Columbus, Ohio PUBLICATIONS Litchfield, J.H. and M.F. Arthur. 1982. Growth of selected ectomycorrhizal fungi in aerated liquid culture. Dev. Indust. Microbiol. 24:289-293. Arthur, M.F., C.K. Wagner, and P. Van Voris (eds.). 1983. Response of agricultural soils to acid deposition. Environ. Experiment. Botany 23:197-284. Arthur, M.F. and C.K. Wagner. 1983. Response of agricultural soils to acid deposition: supplemental literature review. Environ. Experiment. Botany 23:259-279. Tolle, D.A., M.F. Arthur, and P. Van Voris. 1983. Microcosm/field comparison of trace element uptake in crops grown in fly ash-amended soil. Sci. Total Environ. 31:243- 261. Arthur, M.F., T.C. Zwick, D.A. Tolle, and P. Van Voris. 1984. Effects of fly ash on microbial C02 evolution from an agricultural soil. Water Air Soil Pollution 22:209-216. iii Zwick, T.C., M.F. Arthur, D.A. Tolle, and P. Van Voris. 1984. A unique laboratory method for evaluating agro­ ecosystem effects of an industrial waste product. Plant Soil 77:395-399. Tolle, D.A., M.F. Arthur, J. Chesson, and P. Van Voris. 1985. Comparison of pots versus microcosms for predicting agroecosystem effects due to waste amendment. Environ. Toxicol. Chem. 4:501-509. Van Voris, P., D.A. Tolle, M.F. Arthur, and J. Chesson. 1985. Terrestrial microcosms: applications, validation and cost-benefit analysis, pp. 117-142. In: J. Cairns (ed.). Multispecies Toxicity Testing. Pergamon Press, New York. DeGraeve, G.M., W.H. Clement, M.F. Arthur, R.B. Gillespie, and G.K. O ’Brien. 1987. Environmental persistence/ degradation of toxicity in complex effluents: laboratory simulations of field conditions. In: Tenth Symposium on Aquatic Toxicology, American Society for Testing and Materials (in press). Arthur, M.F. and J.I. Frea. 1987. Microbial activity in soils contaminated with 2,3,7,8-tetrachlorodibenzo-p- dioxin. Environ. Toxicol. Chem. (in press). FIELDS OF STUDY Major Field: Environmental Microbiology Studies in Biochemistry: L. Johnson; G. Means Plant Physiology: C. Swanson Soil Chemistry: E.O. McClean Soil Fertility: T.A. Arscott Soil Microbiology: R.H. Miller iv TABLE OF CONTENTS ACKNOWLEDGEMENTS .................................... ii VITA ................................................ ill LIST OF TABLES ...................................... vii LIST OF F I G U R E S .................................... xii INTRODUCTION ........................................ 1 SCOPE AND OBJECTIVES............................... 5 APPROACH ......................... 5 Y CHAPTER PAGE I. LITERATURE REVIEW .......................... 9 Physicochemical Properties of TCDD .... 9 Sources of TCDD........................... 12 Biological Properties of TCDD............ 13 Environmental Properties of TCDD ..... 30 Potential Microbial Metabolism: TCDD and Other Chlorinated Hydrocarbons ........................... 34 Summary.................................... 53 II. MATERIALS AND METHODS........................ 55 Soil Characterization..................... 55 Microbial Activity in TCDD Soils ........ 68 Microbial Mineralization of TCDD ........ 74 Surfactant Experiments ................... 76 HCB Degradation Experiments............... 84 Surfactant-Mediated TCDD Degradation Experiments................. 88 III. RESULTS AND DISCUSSION..................... 90 Soil Characterization..................... 90 Microbial Activity in TCDD Soils ........ 94 v Surfactant Experiments ................... 116 HCB Biodegradation Experiments .......... 138 TCDD Biodegradation Experiments.......... 153 IV. GENERAL DISCUSSION .................. 164 Soil Characterization..................... 164 Microbial Activity in TCDD Soils ........ 166 Surfactant Experiments ................... 179 HCB Biodegradation Experiments .......... 184 TCDD Biodegradation Experiments.......... 186 V. SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS............................. 191 Summary.................................... 191 Conclusions............................... 194 Recommendat ions........................... 195 APPENDIX ............................................ 199 BIBLIOGRAPHY ........................................ 210 vi LIST OF TABLES TABLE PAGE 1. Possible chlorinated compounds of dibenzo-p-dioxin................ ......... 10 2. Physicochemical properties of 2,3,7,8- tetrachlorodibenzo-p-dioxin ................. 11 3. Acute toxicity of TCDD compared to other toxic materials, and the relative toxicity of TCDD among species ....................... 16 4. TCDD Bioconcentration Factors (BCF) in aquatic organisms grown in model ecosystems in the laboratory........................... 27 5. Physicochemical parameters of experimental soils........................... 91 6. Mean microbial numbers in TCDD soils and a non-TCDD control soil (n=5). Within columns, means followed by a common letter are not significantly different at the 99% confidence level..................... 96 7. Mean activity of selected enzymes in TCDD soils and a non-TCDD control soil (n=3). Within columns, means followed by a common letter are not significantly different at the 99% confidence level .... 100 8. Mean cumulative