Method for measuring active oxygen level in phanerochaete chrysosporium in treated wastewater

Abstract

The invention discloses a method for measuring the active oxygen level in phanerochaete chrysosporium in treated wastewater. The method comprises the following steps: washing a phanerochaete chrysosporium bacterium ball in the treated wastewater, adding a liquid culture medium for further culture, further adding 2',7'-dichloro dihydro fluorescein sodium diacetate into the liquid culture medium after culture to obtain a mixture solution, carrying out incubation on the mixture solution, filtering, then performing ultrasonic crushing and centrifuging on the bacterium ball obtained through filtration, extracting a supernate, and finally measuring the fluorescence intensity of the oxidized dichlorofluorescein in the supernate. The method has the advantages that the operation condition is simple, the application is easy and the active oxygen level in the phanerochaete chrysosporium in the treated wastewater can be accurately and visibly measured.

Description

technical field

[0001] The invention relates to the field of microorganism application and waste water treatment, in particular to a method for measuring the active oxygen level in Phanerochaete chrysosporium cells after waste water is treated. Background technique

[0002] At present, the harm of heavy metals and organic pollutants to the environment has become a global problem. In the current field of wastewater treatment, the existing technology has been able to use Phanerochaete chrysosporium to remove heavy metals and organic matter in wastewater. Receptivity also deserves our attention.

[0003] Existing detection methods for reactive oxygen species are mainly used in the detection of reactive oxygen species in higher organisms such as animals and plants. The detection method used is basically flow cytometry, but the price of flow cytometry is high, the detection cost is high, and the operation of the instrument is complicated. , and it is not suitable for batch test...

Examples

Embodiment 1

[0028] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0029] (1) Growth stage: Scrape the Phanerochaete chrysosporium spore powder from the slant medium, make a spore suspension in sterile water, put the spore suspension into a measuring bottle, and adjust the spore suspension with a turbidimeter. The turbidity value of the liquid is 60%, and each milliliter of spore suspension contains 2.5×10 6 spores; then the spore suspension was inoculated into the Kirk liquid medium in the Erlenmeyer flask, 200mL of Kirk liquid medium was housed in the 500mL Erlenmeyer flask, and cultivated in a constant temperature shaking box, the cultivation temperature was 37°C, and the shaking table speed was 150r / min, the cultivation time is 60h, and the Phanerochaete chrysosporium suspension is obtained.

[0030] (2) Wastewater treatment stage: Add the bacterial balls in the above-mentioned Phan...

Embodiment 2

[0034] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0035] (1) Growth stage: this step is the same as step (1) of Example 1;

[0036](2) Adsorption degradation stage: add the bacterial balls in the Phanerochaete chrysosporium bacterial suspension prepared in step (1) of this example to 20 mg / L self-prepared dichlorophenol wastewater, and the chrysosporium in each liter of wastewater The quality of Phanerochaete bacteria balls is 0.4g by dry weight, adjust the acidity and alkalinity of waste water to pH value to 6.5, and carry out oscillation reaction under 37 ℃, 150r / min conditions for 12h, complete the degradation of dichlorophenol in waste water, After the reaction, the waste water is filtered and the bacteria balls are recovered.

[0037] (3) Fluorescence analysis: this step is the same as step (3) in Example 1.

[0038] The measurement results are shown in Table 1. ...

Embodiment 3

[0044] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0045] (1) Growth stage: this step is the same as step (1) of Example 1;

[0046] (2) Adsorption degradation stage: Add the bacterial balls in the suspension of Phanerochaete chrysosporium prepared in step (1) of this example to 50 mg / L industrial cadmium wastewater, and the level of Phanerochaete chrysosporium in each liter of wastewater The quality of the chrysalis balls is 0.4g by dry weight, adjust the pH value of the wastewater to 6.5, and carry out the shaking reaction at 37°C and 150r / min for 12h to complete the adsorption of cadmium in the wastewater. After the reaction, filter the wastewater and recover Bacteria.

[0047] (3) Fluorescence analysis: this step is the same as step (3) in Example 1;

[0048] The results are shown in Table 2.

technical field

[0001] The invention relates to the field of microorganism application and waste water treatment, in particular to a method for measuring the active oxygen level in Phanerochaete chrysosporium cells after waste water is treated. Background technique

[0002] At present, the harm of heavy metals and organic pollutants to the environment has become a global problem. In the current field of wastewater treatment, the existing technology has been able to use Phanerochaete chrysosporium to remove heavy metals and organic matter in wastewater. Receptivity also deserves our attention.

[0003] Existing detection methods for reactive oxygen species are mainly used in the detection of reactive oxygen species in higher organisms such as animals and plants. The detection method used is basically flow cytometry, but the price of flow cytometry is high, the detection cost is high, and the operation of the instrument is complicated. , and it is not suitable for batch test...

Examples

Embodiment 1

[0028] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0029] (1) Growth stage: Scrape the Phanerochaete chrysosporium spore powder from the slant medium, make a spore suspension in sterile water, put the spore suspension into a measuring bottle, and adjust the spore suspension with a turbidimeter. The turbidity value of the liquid is 60%, and each milliliter of spore suspension contains 2.5×10 6 spores; then the spore suspension was inoculated into the Kirk liquid medium in the Erlenmeyer flask, 200mL of Kirk liquid medium was housed in the 500mL Erlenmeyer flask, and cultivated in a constant temperature shaking box, the cultivation temperature was 37°C, and the shaking table speed was 150r / min, the cultivation time is 60h, and the Phanerochaete chrysosporium suspension is obtained.

[0030] (2) Wastewater treatment stage: Add the bacterial balls in the above-mentioned Phan...

Embodiment 2

[0034] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0035] (1) Growth stage: this step is the same as step (1) of Example 1;

[0036](2) Adsorption degradation stage: add the bacterial balls in the Phanerochaete chrysosporium bacterial suspension prepared in step (1) of this example to 20 mg / L self-prepared dichlorophenol wastewater, and the chrysosporium in each liter of wastewater The quality of Phanerochaete bacteria balls is 0.4g by dry weight, adjust the acidity and alkalinity of waste water to pH value to 6.5, and carry out oscillation reaction under 37 ℃, 150r / min conditions for 12h, complete the degradation of dichlorophenol in waste water, After the reaction, the waste water is filtered and the bacteria balls are recovered.

[0037] (3) Fluorescence analysis: this step is the same as step (3) in Example 1.

[0038] The measurement results are shown in Table 1. ...

Embodiment 3

[0044] The method for determining the level of active oxygen in Phanerochaete chrysosporium cells after wastewater treatment comprises the following steps:

[0045] (1) Growth stage: this step is the same as step (1) of Example 1;

[0046] (2) Adsorption degradation stage: Add the bacterial balls in the suspension of Phanerochaete chrysosporium prepared in step (1) of this example to 50 mg / L industrial cadmium wastewater, and the level of Phanerochaete chrysosporium in each liter of wastewater The quality of the chrysalis balls is 0.4g by dry weight, adjust the pH value of the wastewater to 6.5, and carry out the shaking reaction at 37°C and 150r / min for 12h to complete the adsorption of cadmium in the wastewater. After the reaction, filter the wastewater and recover Bacteria.

[0047] (3) Fluorescence analysis: this step is the same as step (3) in Example 1;

[0048] The results are shown in Table 2.

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