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Bioaugmentation of microbial communities in laboratory and pilot scale sequencing batch biofilm reactors using the TOL plasmid

Authors:	S Venkata Mohan Christina Falkentoft Y Venkata Nancharaiah Belinda S McSwain Sturm Pierre Wattiau Peter A Wilderer Stefan Wuertz Martina Hausner

Institution:	1. Institute of Water Quality Control and Waste Management, Technical University of Munich, Am Coulombwall, D-85748 Garching, Germany;2. Bioengineering and Environmental Centre, Indian Institute of Chemical Technology, Hyderabad 500 007, India;3. European Patent Office, Erhardtstrasse 27, 80331 Munich, Germany;4. Water and Steam Chemistry Laboratory, BARC Facilities, Kalpakkam 603102, India;5. Department of Civil, Environmental and Architectural Engineering, The University of Kansas, Learned Hall, 1530 W 15th Street, Lawrence, KS, USA;6. Veterinary and Agrochemical Research Institute, Department of Bacteriology, Groeselenberg 99, B-1180 Brussels, Belgium;7. Institute for Advanced Studies on Sustainability, Amalienstr. 75, 80799 Munich, Germany;8. Department of Civil and Environmental Engineering, University of California, One Shields Avenue, Davis, CA 95616, USA;9. Department of Chemistry and Biology, Ryerson University, 350 Victoria Street, Toronto, Ontario, Canada M5B 2K3;1. Laboratory of Environmental Microbiology & Ecology (LEnME), Department of Life Science, National Institute of Technology, Rourkela 769008, Odisha, India;2. Biofouling & Biofilm Processes Section, Water & Steam Chemistry Division, BARC Facilities, Kalpakkam 603102, Tamil Nadu, India;1. Singapore Centre on Environmental Life Sciences Engineering (SCELSE), School of Biological Sciences SBS-B1N-27, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore;2. Department of Civil and Environmental Engineering, 2001 Ghausi Hall, One Shields Avenue, University of California, Davis, CA 95616, USA;3. Department of Civil Engineering, Indian Institute of Technology, Guwahati, 781039, India;4. HDR Engineering, Inc., 2365 Iron Point Road, Suite 300, Folsom, CA 95630-8709, USA;5. School of Civil and Environmental Engineering, 50 Nanyang Ave, Nanyang Technological University, Singapore 639798, Singapore;1. Center for Microbial Communities, Department of Chemistry and Bioscience, Aalborg University, Fredrik Bajers Vej 7H, 9220 Aalborg East, Denmark;2. Novozymes Biologicals, Inc., 5400 Corporate Circle, Salem, VA 24153, United States

Abstract:	The aim of this study was to investigate the effectiveness of bioaugmentation and transfer of plasmid pWWO (TOL plasmid) to mixed microbial populations in pilot and laboratory scale sequencing batch biofilm reactors (SBBRs) treating synthetic wastewater containing benzyl alcohol (BA) as a model xenobiotic. The plasmid donor was a Pseudomonas putida strain chromosomally tagged with the gene for the red fluorescent protein carrying a green fluorescent protein labeled TOL plasmid, which confers degradation capacity for several compounds including toluene and BA. In the pilot scale SBBR donor cells were disappeared 84 h after inoculation while transconjugants were not detected at all. In contrast, both donor and transconjugant cells were detected in the laboratory scale reactor where the ratio of transconjugants to donors fluctuated between 1.9 × 10^?1 and 8.9 × 10^?1 during an experimental period of 32 days. BA degradation rate was enhanced after donor inoculation from 0.98 mg BA/min prior to inoculation to 1.9 mg BA/min on the seventeenth day of operation. Survival of a bioaugmented strain, conjugative plasmid transfer and enhanced BA degradation was demonstrated in the laboratory scale SBBR but not in the pilot scale SBBR.

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