A simple and effective plating method to screen polycyclic aromatic hydrocarbon-degrading bacteria under various redox conditions |
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Authors: | Youngsoon Um Matthew Wook Chang Tracey Pulliam Holoman |
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Institution: | (1) Clean Energy Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seongbuk-gu, Seoul, 136-791, South Korea;(2) School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, 637459, Singapore;(3) Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD 20742, USA |
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Abstract: | Agar plates with a polycyclic aromatic hydrocarbon (PAH) layer have been used to screen for microorganisms that degrade PAHs,
leaving clear zones around colonies; however, there are several problems with previous methods such as undesired contamination
in the fume hood and difficulty in controlling the amount of PAH on the plates. In this study, we developed a modified screening
method to address the drawbacks encountered with previous screening methods. A uniform white layer of PAHs was generated by
spreading PAHs dissolved in volatile solvents over a surface of solidified agar medium, followed by the evaporation of the
solvents. An inoculation was then performed by spreading a molten agar medium containing microbial samples over the solidified
agar medium with a PAH layer. Subsequently, the white PAH layer migrated to the surface of the molten agar medium. This essential
modification enabled us not only to solve problems of the previous screening methods but also to prepare an agar plate with
a PAH layer without a complicated experimental scheme in the anaerobic chamber. After solidification of the molten agar medium
and incubation of the plates, clear zones were successfully detected around colonies with aerobic and anaerobic PAH-degrading
microbial cultures. |
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Keywords: | |
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