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Estimating biodegradative gene numbers at a JP-5 contaminated site using PCR

  • Session 5 Environmental Biotechnology
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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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References

  1. Atlas, R. M., Sayler, G., Burlage, R. S., and Bej, A. K. (1992),BioTechniques 12, 706–717.

    CAS  Google Scholar 

  2. Sayler, G. S. and Layton, A. C. (1990),Annu. Rev. Microbiol. 44, 625–648.

    Article  CAS  Google Scholar 

  3. Steffan, R. J. and Atlas, R. M. (1991),Annu. Rev. Microbiol. 45, 137–161.

    Article  CAS  Google Scholar 

  4. Liesack, W. and Stackebrandt, E. (1992),Biodiversity and Conservation 1, 250–262.

    Article  Google Scholar 

  5. Ward, D. M., Bateson, M. M., Weiler, R., and Ruff-Roberts, A. L. (1992),Adv. Microb. Ecology 12, 219–286.

    CAS  Google Scholar 

  6. 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.

  7. 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.

    Article  Google Scholar 

  8. Haldeman, D. L., Amy, P. S., White, D. C., and Ringelberg, D. B. (1994),Appl. Environ. Microbiol. 60, 2697–2703.

    CAS  Google Scholar 

  9. Walia, S., Khan, A., and Rosenthal, N. (1990),Appl. Environ. Microbiol. 56, 254–259.

    CAS  Google Scholar 

  10. Erb, R. W. and Wagner-Döbler, I. (1993),Appl. Environ. Microbiol. 59, 4065–4073.

    CAS  Google Scholar 

  11. Fleming, J. T., Sanseverino, J., and Sayler, G. S. (1993),Environ. Sci. Technol. 27, 1068–1074.

    Article  CAS  Google Scholar 

  12. Ka, J. O., Holben, W. E., and Tiedje, J. M. (1994),Appl. Environ. Microbiol. 60, 1116–1120.

    CAS  Google Scholar 

  13. 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.

    Google Scholar 

  14. National Research Council (1993), InSitu Bioremediation, When Does It Work? National Academy Press, Washington, DC.

    Google Scholar 

  15. Kittel, J. A. (1995), Personal communication.

  16. 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.

    Google Scholar 

  17. 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.

    Google Scholar 

  18. US Environmental Protection Agency (1986), Vol. v. 1B, method 5030, Office of Solid Waste and Emergency Response, Washington, DC.

  19. Ogram, A., Sun, W., Brockman, F. J., and Fredrickson, J. K. (1995),Appl. Environ. Microbiol. 61, 763–768.

    CAS  Google Scholar 

  20. More’, M. I., Herrick, J. B., Silva, M. C., Ghiorse, W. C., and Madsen, E. L. (1994),Appl. Environ. Microbiol. 60, 1572–1580.

    CAS  Google Scholar 

  21. Xia, X., Bollinger, J., and Ogram, A. (1995),Mol. Ecology 4, 17–28.

    Article  CAS  Google Scholar 

  22. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989),Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  23. Brock, T. D. and Madigan, M. T. (1991),Biology of Microorganisms, Prentice-Hall, Englewood Cliffs, NJ.

    Google Scholar 

  24. 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.

    CAS  Google Scholar 

  25. Eggink, G., Lageveen, R. G., Altenburg, B., and Witholt, B. (1987),J. Biol. Chem. 262, 17,712–17,718.

    CAS  Google Scholar 

  26. Winstanley, C., Morgan, J. A. W., Pickup, R. W., Jones, J. G., and Saunders, J. R. (1989),Appl. Envrion. Microbiol. 55, 771–777.

    CAS  Google Scholar 

  27. Herrick, J. B., Madsen, E. L., Batt, C. A., and Ghiorse, W. C. (1993),Appl. Environ. Microbiol. 59, 687–694.

    CAS  Google Scholar 

  28. Denome, S. A., Stanley, D. C., Olson, E. S., and Young, K. D. (1993),J. Bacteriology 175, 6890–6901.

    CAS  Google Scholar 

  29. Takizawa, N., Kaida, N., Torigoe, S., Moritani, T., Sawada, T., Satoh, S., and Kiyohara, H. (1994),J. Bacteriology 176, 2444–2449.

    CAS  Google Scholar 

  30. Recorbet, G., Picard, C., Normand, P., and Simonet, P. (1993),Appl. Environ. Microbiol. 59, 4289–4294.

    CAS  Google Scholar 

  31. Picard, C., Ponsonnet, C., Paget, E., Nesme, X., and Simonet, P. (1992),Appl. Environ. Microbiol. 58, 2717–2722.

    CAS  Google Scholar 

  32. Tebbe, C. C., and Vahjen, W. (1993),Appl. Environ. Microbiol. 59, 2657–2665.

    CAS  Google Scholar 

  33. Brockman, F. J. (1995),Mol. Ecology 4, 567–578.

    Article  CAS  Google Scholar 

  34. Steffan, R. J., Goksoyr, J., Bej, A. K., and Atlas, R. M. (1988),Appl. Environ. Microbiol. 54, 2908–2915.

    CAS  Google Scholar 

  35. 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.

    Article  Google Scholar 

  36. Colwell, F. S. (1989),Appl. Environ. Microbiol. 55, 2420–2423.

    Google Scholar 

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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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