Abstract
We have utilized a most-probable-number polymerase chain reaction (MPN-PCR) procedure to estimate gene numbers and biodegradative potential at a jet fuel (JP-5) contaminated site undergoing the first phase of bioremediation. Nucleic acid analysis was used to determine whether a lack of genetic potential for bioremediation was responsible for low levels of oxygen utilization at the site. Total community DNA was extracted and analyzed by PCR for genes (nahAc,alkB, andxylE) known to be involved in the degradation of certain JP-5 constituents. Results indicate that significant aromatic biodegradative potential exists at the site and outlying areas not subjected to engineered remediation, suggesting that physical and/or chemical factors are inhibiting oxygen delivery.xylE andnahAc were often present in significant portions of the microbial community, whereasalkB was rarely detected. This study illustrates the utility of molecular techniques in evaluating biodegradative potential in the field during active bioremediation.
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Atlas, R. M., Sayler, G., Burlage, R. S., and Bej, A. K. (1992),BioTechniques 12, 706–717.
Sayler, G. S. and Layton, A. C. (1990),Annu. Rev. Microbiol. 44, 625–648.
Steffan, R. J. and Atlas, R. M. (1991),Annu. Rev. Microbiol. 45, 137–161.
Liesack, W. and Stackebrandt, E. (1992),Biodiversity and Conservation 1, 250–262.
Ward, D. M., Bateson, M. M., Weiler, R., and Ruff-Roberts, A. L. (1992),Adv. Microb. Ecology 12, 219–286.
Brockman, F. J., Li, S. W., Fredrickson, J. K., Ringelberg, D. B., Kieft, T. L., Spadoni, C. M., McKinley, J. P., and White, D. C. (1995),Appl. Environ. Microbiol. Submitted.
Brockman, F. J., Kieft, T. L., Fredrickson, J. K., Bjornstad, B. N., Li, S. W., Spangenburg, W., and Long, P. E. (1992),Microb. Ecology 23, 279–301.
Haldeman, D. L., Amy, P. S., White, D. C., and Ringelberg, D. B. (1994),Appl. Environ. Microbiol. 60, 2697–2703.
Walia, S., Khan, A., and Rosenthal, N. (1990),Appl. Environ. Microbiol. 56, 254–259.
Erb, R. W. and Wagner-Döbler, I. (1993),Appl. Environ. Microbiol. 59, 4065–4073.
Fleming, J. T., Sanseverino, J., and Sayler, G. S. (1993),Environ. Sci. Technol. 27, 1068–1074.
Ka, J. O., Holben, W. E., and Tiedje, J. M. (1994),Appl. Environ. Microbiol. 60, 1116–1120.
Ogunseitan, O. A. (1994), inProcess Engineering for Pollution and Control and Waste Minimization, Wise, D. L. and Trantolo, D. A., eds., Marcel Dekker, NY, pp. 171–192.
National Research Council (1993), InSitu Bioremediation, When Does It Work? National Academy Press, Washington, DC.
Kittel, J. A. (1995), Personal communication.
Kittel, J. A., Hinchee, R. E., Hoeppel, R., and Miller, R. (1994), inProceedings of the 1994 Petroleum Hydrocarbons and Organic Chemicals in Groundwater: Prevention, Detection, and Remediation, Groundwater Publishing, Dublin.
Miller, R. N., Vogel, C. C., and Hinchee, R. E. (1991), in InSitu Bioreclamation: Applications and Investigations for Hydrocarbon and Contaminated Site Remediation, Hinchee, R. E. and Olfenbuttel, R. F., eds., Butterworth-Heinemann, Boston, pp. 283–302.
US Environmental Protection Agency (1986), Vol. v. 1B, method 5030, Office of Solid Waste and Emergency Response, Washington, DC.
Ogram, A., Sun, W., Brockman, F. J., and Fredrickson, J. K. (1995),Appl. Environ. Microbiol. 61, 763–768.
More’, M. I., Herrick, J. B., Silva, M. C., Ghiorse, W. C., and Madsen, E. L. (1994),Appl. Environ. Microbiol. 60, 1572–1580.
Xia, X., Bollinger, J., and Ogram, A. (1995),Mol. Ecology 4, 17–28.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989),Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Brock, T. D. and Madigan, M. T. (1991),Biology of Microorganisms, Prentice-Hall, Englewood Cliffs, NJ.
Kok, M., Oldenhuis, R., van der Linden, M. P. G., Raatjes, P., Kingma, J., van Lelyveld, P. H., and Witholt, B. (1989),J. Biol. Chem. 264, 5435–5441.
Eggink, G., Lageveen, R. G., Altenburg, B., and Witholt, B. (1987),J. Biol. Chem. 262, 17,712–17,718.
Winstanley, C., Morgan, J. A. W., Pickup, R. W., Jones, J. G., and Saunders, J. R. (1989),Appl. Envrion. Microbiol. 55, 771–777.
Herrick, J. B., Madsen, E. L., Batt, C. A., and Ghiorse, W. C. (1993),Appl. Environ. Microbiol. 59, 687–694.
Denome, S. A., Stanley, D. C., Olson, E. S., and Young, K. D. (1993),J. Bacteriology 175, 6890–6901.
Takizawa, N., Kaida, N., Torigoe, S., Moritani, T., Sawada, T., Satoh, S., and Kiyohara, H. (1994),J. Bacteriology 176, 2444–2449.
Recorbet, G., Picard, C., Normand, P., and Simonet, P. (1993),Appl. Environ. Microbiol. 59, 4289–4294.
Picard, C., Ponsonnet, C., Paget, E., Nesme, X., and Simonet, P. (1992),Appl. Environ. Microbiol. 58, 2717–2722.
Tebbe, C. C., and Vahjen, W. (1993),Appl. Environ. Microbiol. 59, 2657–2665.
Brockman, F. J. (1995),Mol. Ecology 4, 567–578.
Steffan, R. J., Goksoyr, J., Bej, A. K., and Atlas, R. M. (1988),Appl. Environ. Microbiol. 54, 2908–2915.
Kieft, T. L., Amy, P. S., Brockman, F. J., Fredrickson, J. K., Bjornstad, B. N., and Rosacker, L. L. (1993),Microb. Ecology 26, 59–78.
Colwell, F. S. (1989),Appl. Environ. Microbiol. 55, 2420–2423.
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Chandler, D.P., Brockman, F.J. Estimating biodegradative gene numbers at a JP-5 contaminated site using PCR. Appl Biochem Biotechnol 57, 971–982 (1996). https://doi.org/10.1007/BF02941777
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DOI: https://doi.org/10.1007/BF02941777