CN113512599A - Method for quantitatively evaluating repairing capability of pollutant repairing flora - Google Patents
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Abstract
The invention relates to a method for quantitatively evaluating the repairing capacity of a pollutant repairing flora, which comprises the following steps: s01, analyzing the background characteristics of the sediment to be repaired; s02 constructing a microscopic simulation device; s03, setting the tolerance evaluation condition and the monitoring time interval of the repair flora; s04, drawing a repair curve and evaluating the repair effect; s05, synchronously drawing the gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria. The method can quantitatively evaluate the repair capability of the repair flora on the target pollutants, and is favorable for accurately guiding engineering application.
Description
Technical Field
The invention relates to the technical field of ecological environment protection, in particular to a method for quantitatively evaluating the restoration capacity of a pollutant restoration flora.
Background
Along with the rapid advance of urbanization and the rapid development of society, soil and sediments in China are generally polluted, and certain ecological risks exist. The method for repairing various pollutants in the environment mainly comprises physical, chemical and biological repairing methods, wherein the biological repairing method has the advantages of low cost and good effect. The microorganism is used as a main body for repairing pollutants in soil and sediments and is a key research object in the bioremediation process. Quantitative evaluation of the remediation capability of the pollutant remediation flora can provide an important reference for the microbial remediation application of pollutants in the environment. The stronger the repairing capability of the repairing flora on pollutants, the higher the stability of the flora, and the more beneficial to engineering application. Currently, there is no quantitative assessment method for the repair ability of flora.
Disclosure of Invention
In order to achieve the above object, the present invention discloses 1. a method for quantitatively evaluating the repairing capability of a pollutant repairing flora, comprising: analyzing the background characteristics of the sediment to be repaired; constructing a microscopic simulation device; setting tolerance evaluation conditions and monitoring time intervals of the repair flora; drawing a repairing curve and evaluating a repairing effect; and synchronously drawing a gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria. Specifically, the method comprises the following steps:
s01: and analyzing the background characteristics of the sediment to be repaired. And collecting a sediment sample to be repaired, and analyzing basic physicochemical properties and the background of target pollutants. Wherein, the basic physicochemical indexes are different according to the types of pollutants: analyzing pH, oxidation-reduction potential and organic carbon indexes of an environment sample containing heavy metal pollutants; the environmental sample containing organic pollutants is analyzed to include Total Organic Carbon (TOC), Total Nitrogen (TN), nitrate Nitrogen (NO)3) And ammoniacal Nitrogen (NH)3) And (4) indexes.
S02: and constructing a microscopic simulation device. The details of the microscopic simulation experiment device used in the present invention are as follows: adding equal amount of sediment, inorganic salt culture solution and target pollutant into a serum bottle with the same specification, plugging the bottle mouth with cotton, and culturing in a shaking table with constant temperature and humidity. And a plurality of groups of mutually independent microscopic simulation experiment devices are arranged, so that the repair capability quantitative evaluation of the multiple pollutant repair flora can be synchronously carried out.
S03: and setting the tolerance evaluation condition and the monitoring time interval of the repair flora. Based on the previous literature research or experimental research basis, tolerance evaluation conditions and monitoring time intervals suitable for different pollutants and different pollution degrees are set. Different pollution degree settings can be set in a multiple form according to the environmental concentration.
S04: and drawing a repairing curve and evaluating the repairing effect. And (3) periodically selecting a plurality of devices to detect the residual quantity or stable state of the target pollutants, dynamically monitoring the whole repairing process, and drawing a repairing curve. The time node with the highest repair efficiency, the time node approaching stability and the upper limit of the pollution concentration are mainly identified.
S05: and synchronously drawing a gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria. Periodically and synchronously analyzing the gene copy number and relative abundance change dynamics of the pollutant repairing flora; and comparing and researching the gene copy number and the relative abundance change dynamic difference of the repairing flora in the target pollutant repairing process under different evaluation conditions, and quantitatively evaluating the repairing capability of the repairing flora by combining the repairing effect.
According to the early-stage investigation and experimental research results, the quantitative evaluation of the repair flora repair capacity is realized aiming at the early-stage evaluation problem of the flora repair capacity in the environmental pollutant bioremediation, and the accurate guidance is provided for the engineering application of the repair flora.
Drawings
FIG. 1 is a flow chart of the present invention;
FIG. 2 is a graph showing the degradation repair of a sediment polycyclic aromatic hydrocarbon phenanthrene in an example of the present invention;
FIG. 3 is a graph showing the variation of gene copy number and relative abundance of sediment polycyclic aromatic hydrocarbon phenanthrene degrading bacteria in the embodiment of the invention.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
The flow chart of the invention is shown in figure 1.
S01: and analyzing the background characteristics of the sediment to be repaired. MiningAnd collecting sediment samples to be repaired, and analyzing basic physicochemical properties and the background of target pollutants. Wherein, the basic physicochemical indexes are different according to the types of pollutants: analyzing pH, oxidation-reduction potential and organic carbon indexes of an environment sample containing heavy metal pollutants; the environmental sample containing organic pollutants is analyzed to include Total Organic Carbon (TOC), Total Nitrogen (TN), nitrate Nitrogen (NO)3) And ammoniacal Nitrogen (NH)3) And (4) indexes.
S02: and constructing a microscopic simulation device. The details of the microscopic simulation experiment device used in the present invention are as follows: adding equal amount of sediment, inorganic salt culture solution and target pollutant into a serum bottle with the same specification, plugging the bottle mouth with cotton, and culturing in a shaking table with constant temperature and humidity. And a plurality of groups of mutually independent microscopic simulation experiment devices are arranged, so that the repair capability quantitative evaluation of the multiple pollutant repair flora can be synchronously carried out.
S03: and setting the tolerance evaluation condition and the monitoring time interval of the repair flora. Based on the previous literature research or experimental research basis, tolerance evaluation conditions and monitoring time intervals suitable for different pollutants and different pollution degrees are set. Different pollution degree settings can be set in a multiple form according to the environmental concentration.
S04: and drawing a repairing curve and evaluating the repairing effect. And (3) periodically selecting a plurality of devices to detect the residual quantity or stable state of the target pollutants, dynamically monitoring the whole repairing process, and drawing a repairing curve. The time node with the highest repair efficiency, the time node approaching stability and the upper limit of the pollution concentration are mainly identified.
S05: and synchronously drawing a gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria. Periodically and synchronously analyzing the gene copy number and relative abundance change dynamics of the pollutant repairing flora; and comparing and researching the gene copy number and the relative abundance change dynamic difference of the repairing flora in the target pollutant repairing process under different evaluation conditions, and quantitatively evaluating the repairing capability of the repairing flora by combining the repairing effect.
Example (b):
the present invention is described in detail below with reference to an example of a mycobacterial bacteria degradation sediment tolerance simulation experiment for polycyclic aromatic hydrocarbon phenanthrene contamination. The method comprises the following steps:
s01: and analyzing the background characteristics of the sediment to be repaired. And collecting a sediment sample to be repaired, and analyzing basic physicochemical properties and the background of target pollutants. The basic physicochemical properties of the coastal mangrove forest sediments studied in this example are: TOC 11.91% +/-0.35%, TN 0.23% +/-0.01%, NO311.91 +/-0.35 mg-kg-1，NH3224.47 + -21.83 mg-kg-1. The background concentration of the target organic pollutant phenanthrene in the sediment to be repaired is 416.00 +/-82.76 mu g/kg-1。
S02: and constructing a microscopic simulation device. The details of the microscopic simulation experiment device used in the embodiment of the present invention are as follows: adding equal amount of sediment, inorganic salt culture solution and polycyclic aromatic hydrocarbon phenanthrene into a serum bottle with the same specification, plugging the bottle mouth with cotton, and placing into a shaking table with constant temperature of 28 ℃ and constant humidity of 80% for culture.
S03: and setting the tolerance evaluation condition and the monitoring time interval of the repair flora. Based on the previous literature research or experimental research basis, tolerance evaluation conditions and monitoring time intervals suitable for different pollutants and different pollution degrees are set. The examples of the present invention set the conditions for evaluating the resistance of the comparative analysis of the control group, the low concentration group and the high concentration group. Examples the background values of the polycyclic aromatic hydrocarbon phenanthrene in the sediment are known; phe added to the low concentration group was 1500. mu.g.kg-1Background Phe values in the low-concentration test sediment were 1868.45. + -. 244.95. mu.g.kg-1(ii) a Phe added to the high concentration test group was 5000. mu.g.kg-1The background value of Phe in the soil of the high-concentration experimental group is 5538.75 +/-419.25 mu g/kg-1. The time interval was set to 14 days on a preliminary study basis.
S04: and drawing a repairing curve and evaluating the repairing effect. In the embodiment of the invention, 3 devices are selected for detection of phenanthrene residue in each batch at intervals of 14 days to form a phenanthrene degradation and remediation curve (as shown in fig. 2). The degradation efficiency is highest in the first 28 days, especially in the low-concentration pollution group; the degradation of phenanthrene is not obvious in 28-42 days; the degradation rate of phenanthrene is slightly increased in 42-56 days; the first 28 days are important time nodes for efficient degradation of the polycyclic aromatic hydrocarbon phenanthrene, and more than 60% of degradation (except for a high-concentration group) is completed; meanwhile, the degradation capability of the related degradation flora to organic pollutant phenanthrene with 10 times of environmental concentration is limited, care needs to be taken during engineering application, and the repair period is prolonged.
S05: and synchronously drawing a gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria. In the embodiment of the invention, based on the analysis of the functional gene sequencing result, the gene copy number and abundance of the phenanthrene degrading bacterium Mycobacterium are continuously changed along with the increase of the degradation time (as shown in figure 3); the copy number of the gene of the Mycobacterium in the sediment shows a trend of decreasing firstly and then increasing in both a low-concentration group and a high-concentration group, and reaches the lowest value in 28 days; the control group showed a decreasing trend overall. The early-stage rapid degradation of the phenanthrene is demonstrated, and the capacity of degrading bacteria reaches saturation; after the release (28-42 days), the activity starts again, the copy number is gradually recovered, and the degradation rate of phenanthrene is increased again. Meanwhile, the relative abundance of the Mycobacterium bacteria has consistent trend of change under different pollution degree conditions, but the size difference is obvious: the higher the concentration of the contaminant, the lower the relative abundance; at the same time, the degradation efficiency of the high concentration group decreased, which indicates that maintaining a certain abundance helps to maintain an efficient degradation effect. From the change of degrading bacteria, the degradation repair of phenanthrene with 10 times of environmental concentration is feasible, but the repair time needs to be prolonged.
Claims (6)
1. A method for quantitatively evaluating the repairing capability of a pollutant repairing flora is characterized by comprising the following steps: s01, analyzing the background characteristics of the sediment to be repaired; s02 constructing a microscopic simulation device; s03, setting the tolerance evaluation condition and the monitoring time interval of the repair flora; s04, drawing a repair curve and evaluating the repair effect; s05, synchronously drawing the gene copy number and relative abundance change curve of the repairing bacteria, and quantitatively evaluating the repairing capability of the repairing bacteria.
2. The method for quantitatively evaluating the repairing capability of a contaminant repairing flora according to claim 1, wherein the method comprises the following steps: the analysis of the background characteristics of the sediment to be repaired comprises the following steps: collecting deposit sample to be repaired, analyzing basic physicochemical property and target pollutant costAnd (4) bottom. Wherein, the basic physicochemical indexes are different according to the types of pollutants: analyzing pH, oxidation-reduction potential and organic carbon indexes of an environment sample containing heavy metal pollutants; the environmental sample containing organic pollutants is analyzed to include Total Organic Carbon (TOC), Total Nitrogen (TN), nitrate Nitrogen (NO)3) And ammoniacal Nitrogen (NH)3) And (4) indexes.
3. The method for quantitatively evaluating the repairing capability of a contaminant repairing flora according to claim 1, wherein the method comprises the following steps: the construction of the microscopic simulation device is as follows: adding equal amount of sediment, inorganic salt culture solution and target pollutant into a serum bottle with the same specification, plugging the bottle mouth with cotton, and culturing in a shaking table with constant temperature and humidity. And a plurality of groups of mutually independent microscopic simulation experiment devices are arranged, so that the repair capability quantitative evaluation of the multiple pollutant repair flora can be synchronously carried out.
4. The method for quantitatively evaluating the repairing capability of a contaminant repairing flora according to claim 1, wherein the method comprises the following steps: the setting of the tolerance evaluation condition and the monitoring time interval of the repair flora comprises the following steps: based on the previous literature research or experimental research basis, tolerance evaluation conditions and monitoring time intervals suitable for different pollutants and different pollution degrees are set. Different pollution degree settings can be set in a multiple form according to the environmental concentration.
5. The method for quantitatively evaluating the repairing capability of a contaminant repairing flora according to claim 1, wherein the method comprises the following steps: the drawing of the repair curve and the evaluation of the repair effect comprise the following steps: and (3) periodically selecting a plurality of devices to detect the residual quantity or stable state of the target pollutants, dynamically monitoring the whole repairing process, and drawing a repairing curve. The time node with the highest repair efficiency, the time node approaching stability and the upper limit of the pollution concentration are mainly identified.
6. The method for quantitatively evaluating the repairing capability of a contaminant repairing flora according to claim 1, wherein the method comprises the following steps: the method is characterized in that the gene copy number and relative abundance change curve of the repairing bacteria is synchronously drawn, and the repairing capability of the repairing bacteria is quantitatively evaluated, and the method mainly comprises the following steps: periodically and synchronously analyzing the gene copy number and relative abundance change dynamics of the pollutant repairing flora; and comparing and researching the gene copy number and the relative abundance change dynamic difference of the repairing flora in the target pollutant repairing process under different evaluation conditions, and quantitatively evaluating the repairing capability of the repairing flora by combining the repairing effect.
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| CN114410444A (en) * | 2022-01-11 | 2022-04-29 | 中国地质大学（武汉） | Method for acquiring microbial community change information in contaminated site remediation process |
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| CN114410444A (en) * | 2022-01-11 | 2022-04-29 | 中国地质大学（武汉） | Method for acquiring microbial community change information in contaminated site remediation process |
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