Abstract
One of the newly developed methods for Assimilable organic carbon (AOC) determination is leveraged on the cell enumeration by flow cytometry (FC) which could provide a rapid and automated solution for AOC measurement. However, cell samples staining with fluorescence dye is indispensable to reduce background and machine noise. This step would bring additional cost and time consuming for this method. In this study, a green fluorescence protein (GFP) tagged strain derived of AOC testing strain Pseudomonas fluorescens P-17 (GFP-P17) was generated using Tn5 transposon mutagenesis. Continuous culture of this mutant GFP-P17 showed stable expression of eGFP signal detected by flow cytometry without staining step. In addition, this GFP-P17 strain displayed faster growth rate and had a wider range of carbon substrate utilization patterns as compared with P17 wild-type. With this strain, the capability of a new FC method with no dye staining was explored in standard acetate solution, which suggests linear correlation of counts with acetate carbon concentration. Furthermore, this FC method with GFP-P17 strain is applicable in monitoring GAC/BAC efficiency and condition as similar trends of AOC level in water treatment process were measured by both FC method and conventional spread plating count method. Therefore, this fast and easily applicable GFP-P17 based FC method could serve as a tool for routine microbiological drinking water monitoring.
Highlights
Growth of bacteria in drinking water treatment, distribution and storage systems can result in deterioration of water quality, violation of water quality standards and increased operating expenses
The Assimilable organic carbon (AOC) testing bacteria P. fluorescens P-17 was subjected to transposon mutagenesis for stable insertion of green fluorescence protein (GFP) operons into their genomes using electroporation method
AOC assay was of importance and relevant in determining water quality and biological stability of drinking water, it is not widely used and applicable in practice due to the time consuming and tedious nature of the conventional AOC assay
Summary
Growth of bacteria in drinking water treatment, distribution and storage systems can result in deterioration of water quality, violation of water quality standards and increased operating expenses. Bacterial growth or regrowth can result from viable bacteria surviving the disinfection process and utilizing nutrients in the water or biofilm to sustain growth[1]. Microbial activity in the distribution system largely depends on the nutrients carbon source. Controlling microbial activity in water treatment and distribution system is of great importance to prevent water quality deterioration resulting in non-compliance with regulations, consumer complaints and disease.
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