Development of an Improved Sulfur-Oxidizing Bacteria-Based Ecotoxicity Test for Simple and Rapid On-Site Application

Microbial toxicity tests are considered efficient screening tools for the assessment of water contamination. The objective of this study was to develop a sulfur-oxidizing bacteria (SOB)-based ecotoxicity test with high sensitivity and reproducibility for simple and rapid on-site application. To attain this goal, we developed a 25 mL vial-based toxicity kit and improved our earlier SOB toxicity test technique. The current study applied a suspended form of SOB and shortened the processing time to 30 min. Moreover, we optimized the test conditions of the SOB toxicity kit in terms of initial cell density, incubating temperature, and mixing intensity during incubation. We determined that 2 × 10⁵ cells/mL initial cell density, 32 °C incubating temperature, and 120 rpm mixing intensity are the optimal test conditions. Using these test conditions, we performed SOB toxicity tests for heavy metals and petrochemicals, and obtained better detection sensitivity and test reproducibility, compared to earlier SOB tests. Our SOB toxicity kit tests have numerous advantages, including a straightforward test protocol, no requirement of sophisticated laboratory equipment, and no distortion of test results from false readings of end-points and properties of test samples, making it suitable for simple and rapid on-site application.