Molecular biology techniques used in wastewater treatment: An overview |
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Institution: | 1. Astani Department of Civil and Environmental Engineering, University of Southern California, 3620 South Vermont Avenue, Los Angeles, CA 90089, USA;2. mBioInform ApS, Ole Maaloes Vej 3, 2200 Copenhagen N, Denmark;3. Department of Environmental Sciences, Aarhus University, Frederiksborgvej, 399, 4000 Roskilde, Denmark;4. Department of Civil and Environmental Engineering, Rice University, 6100 Main Street, TX 77005, USA;5. Soil and Crop Sciences Section, Cornell University, 306 Tower Road, Ithaca, NY 14853, USA;1. Laboratory for Process Microbial Ecology and Bioinspirational Management (PME&BIM), Department of Microbial and Molecular Systems, KU Leuven, Campus De Nayer, Fortsesteenweg 30A, B-2860 Sint-Katelijne-Waver, Belgium;2. Process and Environmental Technology Lab (PETLab), Department of Chemical Engineering, KU Leuven, Campus De Nayer, B-2860 Sint-Katelijne-Waver, Belgium;3. Chemical and Biochemical Process Technology and Control (BioTeC), Department of Chemical Engineering, KU Leuven, Technology Campus Ghent, B-9000 Ghent, Belgium |
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Abstract: | Identification of microorganisms by conventional methods requires the isolation of pure cultures followed by laborious characterization experiments. These procedures are therefore inadequate for study of the biodiversity of a natural or engineered ecosystem. A new set of molecular techniques developed during the 1990s revolutionized microbial ecology research. Among these techniques, cloning and the creation of a gene library, denaturant gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization with DNA probes (FISH) stand out. Cloning provides very precise taxonomical information, but is time consuming and requires specialized personnel and so its introduction in wastewater treatment has been slow. DGGE is a rapid and simple method that provides characteristic band patterns for different samples, allowing quick sample profiling, while retaining the possibility of a more thorough genetic analysis by sequencing of particular bands. FISH makes possible to identify microorganisms at any desired taxonomical level, depending on the specificity of the probe used. It is the only quantitative molecular biology technique, although quantification is either complex or tedious and subjective. Combination with a confocal laser-scanning microscope allows the visualization of three-dimensional microbial structures (granules, biofilms). The methods discussed have deepened our understanding of the microbiology of biological wastewater treatment. PCR-based methods (cloning and DGGE) have proved suitable for identifying the microorganisms that form the sludge. Both DGGE and FISH have been extensively employed. FISH is currently being used for elucidation of the composition, quantification and distribution of different bacterial groups in granules and biofilms, as well as their structure and architecture. |
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