Microorganism Characterization in the Hospital Wastewater by Epifluorescence Microscopy and Flow Cytometry with Fluorochrome
Date Issued
2004
Date
2004
Author(s)
Chiang, Wan-Chia
DOI
zh-TW
Abstract
Culture-based analysis remains the primary methods for environmental sample analysis. But, there are some limitations of this method. The results always time-consuming and it can only account for a small proportion of the total cell count .Thus, providing a rapid and accurate non-culture method is needed. This study aims to provide a non-culture method for detecting microorganisms and validating this technique to the hospital wastewater.
The nucleic-acid dyes acridine orange (AO) and 4’,6 – diamidino - 2 - phenylindole dihydrochloride (DAPI) penetrate all bacterial membranes, while YO-PRO-1 and Propidium iodide (PI) only penetrate cells with damaged membranes. After optimization, those nucleic-acid dyes were applied to estimate both total counts and viable of microorganisms in the hospital wastewater by using the epifluorescence microscopy and the flow cytometry.
The results indicated that AO, DAPI, YO-PRO-1 and PI were the suitable dyes for determining total concentration and viability of microorganisms in the hospital waste water. The fluorescent techniques in combination with FCM and EFM for assessment of total concentration and viability for microorganisms were demonstrated to successfully apply for environmental field samples. The total concentrations of microorganisms in the hospital wastewater were 1.1×107 ~ 2.4×108 cells/ml and the concentrations of bacteria were 28 ~ 449 – folds higher than those of fungi . Besides, total counts in the outgoing water from clarifier showed dramatic reduction of over one log. The standard culture method underestimated the microorganism concentration over 11 ~100 –folds when compared to non-culture method except the effluent tank. In addition, the viabilities of microorganisms were high before treatment by YO-PRO-1 (0.74) and PI (0.86). However, the low viabilities of both bacteria (YO- PRO-1：0.18、PI：0.23) and fungi (YO- PRO-1：0.38、PI：0.39) were observed after chlorination tank. Therefore, the chlorination is an effective way to induce a decrease in culturabilities and viabilities of microorganisms.
Subjects
流式細胞儀
螢光染劑
螢光顯微鏡
Epifluorescence Microscopy
Fluorochrome
Flow Cytometry
Type
thesis
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