The primary goal of this project is to improve bioremediation efficacy in Taiwan through promoting high resolustion site characterization (HRSC) framework and proper use of molecular biology tools (MBTs) in the field of environmental remediation. To tackle these issues, the project team conducted intensive literature research and attended an international symposium to understand the current technical and policy progress as the preparatory work for developing domestic HRSC toolbox and MBTs guideline. With specific focus on chlorinated solvent contamination, this project selected 4 contaminated sites for further site visitation and the other 4 sites for HRSC/MBTs field investigations. During the project period, a bioremediation working group was formed and involved in discussions regarding Taiwan bioremediation roadmap and a workshop hosted by the project team. The contents of HRSC toolbox include technical documents review, the concept and evolution of site investigation and chacterization, the logic and workflow for choosing proper tools, and the introduction, limitation, and data interpretation of 22 tools available for HRSC in Taiwan. Three case studies are also included to give a comprehensive view on how HRSC is implemented. On the other hand, the project evaluated 4 relatively new dense nonaqueous liquid (DNAPL) investigation or assessment tools in Taiwan, including soil gas flux chamber, tree coring for environmental forensics, crosshole electrical resistivity tomography, and double packer injection, for their feasibility and potential optimization for domestic application. To demonstrate the benefits of MBTs during contamination investigation and remediation, a brief introduction to the development of environmental molecular diagnostics and literatures review including 46 MBTs-related technical papers, 25 chlorinated solvent bioremediation technical documents and 7 case studies were summarized. Evaluation of 49 domestic chlorinated contaminated sites related to bioremediation were conducted to understand the current status of MBTs application in Taiwan. MBTs guideline, in which topics cover 7 common MBTs and next generation sequence (NGS) technology, present the principle, workflow, advantages and limitations, data imterpretation, general quality assurance and quality control considerations, and case studies of each tool. A survey was conducted targeting environmental remediation professionals and researcher to understand the current status of MBTs usage in Taiwan. The survey also served as the basis for local MBTs experts database. To assess the applicability of HRSC toolbox and MBTs giudline, 49 chlorinated solvent bioremediation sites were evaluated and divided in two categories: 4 sites for on-site visitation and 4 sites for field demonstration. The purpose of on-site visitation is to understand the progress and obstacles of local ongoing bioremediation projects. With the help of bioremediation working group, issues regarding bioremediation monitoring targets, alternative substrates, remedy optimization, or subsurface environment manipulation were discussed. On the other hand, supplementary investigation following HRSC framework and MBTs field test were conducted at 4 sites (2 dormant and 2 active). HRSC tools including Multi-Level Slug Test, multiplex well logging, time-lapse electrical resistivity tomography, cross-hole electrical resistivity tomography, and two on-site screening apparatus were used. Microbiological analysis focused on targeted microbes or functional genes quatification using qPCR and community analysis through next generation sequencing were conducted. Technical advices were provided for local environmental protection bureaus with the support of updated conceptual site model, 3 water quality indicators, and 8 microbial indicators.

High resolution site characterization, Chlorinated compounds bioremediation, Molecular biology tools