Citation: LI Ke-Xin, WU Man-Li, GAO Huan, LIU Heng. Determination of Active Polycyclic Aromatic Hydrocarbons Degrading Bacteria Using Flow Cytometry[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(8): 1357-1365. doi: 10.19756/j.issn.0253-3820.201598 shu

Determination of Active Polycyclic Aromatic Hydrocarbons Degrading Bacteria Using Flow Cytometry

Shaanxi Key Laboratory of Environmental Engineering, Key Laboratory of Northwest Water Resource, Environment and Ecology, School of Environmental and Municipal Engineering, Xi'an University of Architecture and Technology, Xi'an 710055, China

Corresponding author: WU Man-Li, 447005853@qq.com
Received Date: 24 December 2020
Revised Date: 27 April 2021

Fund Project: Supported by the National Natural Science Foundation of China (Nos.52070154, 21577109).

A method for determination of the number of active polycyclic aromatic hydrocarbons (PAHs)-degrading bacteria using flow cytometry was developed. PAHs degrading bacteria suspension (1×10³-1×10⁵ cells/mL) was prepared by using ultra-pure water as dilution. After stained using SG/PI and incubated at 35 ℃ for 10 min in the dark, the number of PAHs-degrading bacteria was determined using flow cytometry. The parameters of flow cytometry were set as follows: the green (FL1) and red (FL3) fluorescent acted as detector and the threshold value was 600; the flow rate of bacteria suspension was 33 mL/min and the counting rate kept below 1000 cells/s. Experimental results showed that there was a good correlation relationship between the results of flow cytometry and the plate counting determination, and the correlation coefficients were 0.9742-0.9962. To determine PAH-active bacteria, the bacteria during the growth phase were sterilized using NaClO that acted as negative control, and the activated bacteria were used as positive control. Both negative and positive bacteria were determined using flow cytometry after stained using SYBR Green I and propidium iodide, and the active and inactive cells regions were respectively classified. Compared to the plate counting method, the flow cytometry for counting bacteria number could greatly reduce determination time and improve the accuracy. This study provided a fast and efficiency method for determination of active bacteria in pollutant degrading systems.
- Flow cytometry,
- Polycyclic aromatic hydrocarbons degrading bacteria,
- Active bateria,
- Liquid phase degradation,
- Fast detection
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