Development of Comprehensive Detection Method of Pathogenic Bacteria from River Water Combining Foam Concentration and Bacterial Flora Analysis

摘要

Waterborne diseases due to pathogenic bacteria in the water environment are a serious problem all over the world. Therefore, it is extremely important to grasp the actual condition of their existences. In this study, comprehensive detection of pathogenic bacteria from river water using a combined process of foam concentration and Next Generation Sequencer (NGS) was developed. As a result of the foam concentration, the DNA in the extract of concentrate from each river water sample was detected in the range of 88-215 ng/μL. By the foam concentration, it was possible to obtain a sufficient amount of DNA extract from a river water sample for NGS analysis. When the DNA extracts from all the five rivers pass through the large cities were analyzed by the NGS, 42 genera of pathogenic bacteria were detected, and the relative abundance ratio accounted for 0.79 to 3.0% of the total bacteria. The major pathogenic bacteria were Acinetobacter, Mycobacterium, and Pseudomonas. In addition, 7 genera of pathogenic bacteria were detected at the relative abundance ratio more than 0.01%. From the above, it is possible to detect the pathogenic bacteria comprehensively from river water by combining foam concentration and bacterial flora analysis using NGS.