Soil ecological environment damage confirmation method suitable for illegally dumping household garbage area and application of soil ecological environment damage confirmation method

Abstract

The invention discloses a soil ecological environment damage confirmation method suitable for an illegal dumping household garbage area, which comprises the following steps: screening and analyzing characteristic pollution indexes of soil samples in an evaluation area and a control area; performing high-throughput sequencing and data processing to obtain bacterial community composition structures of the soil samples in the evaluation area and the control area, and comparing and analyzing changes of bacterial communities in the evaluation area to judge whether the soil environment of the evaluation area is obviously changed or not; the method for determining and evaluating the influence of the household garbage on the soil environment is scientific and reasonable, a plurality of related data indexes can be used for quantifying the influence degree of the household garbage on the soil environment, and the method for determining and evaluating the influence degree of the household garbage on the soil environment can be used for determining the influence degree of the household garbage on the soil environment. The method can effectively assist confirmation of damage facts of damage identification and evaluation cases of illegal dumping of household garbage.

Description

Technical Field

[0001] The present invention belongs to the technical field of environmental monitoring, and in particular relates to a method for confirming soil ecological environmental damage in areas where domestic waste is illegally dumped, and an application thereof. Background Art

[0002] Although the Solid Waste Law clearly requires that solid waste must not be dumped, piled, discarded or scattered without authorization, cases of illegal dumping of solid waste still occur.

[0003] Data shows that the proportion of ecological and environmental damage compensation incidents in my country has increased, with soil and groundwater damage appraisal and assessment cases accounting for the largest proportion of such cases. Scientifically and quantifiably assessing environmental damage has become a pressing issue and challenge in the field of environmental justice. Therefore, establishing a comprehensive ecological and environmental damage appraisal and assessment technology system is not only an inevitable requirement for implementing ecological and environmental damage compensation reforms, but also a necessary guarantee for refined ecological and environmental management and maintaining environmental justice. This has far-reaching significance for promoting my country's ecological civilization construction.

[0004] For example, the patent document with application number CN202211589012.3 discloses an ecological environment damage assessment method and system applied to underground engineering water gushing events. The method includes a damage causal relationship judgment method, including the following steps: determining the source and receptor; judging homology; judging correlation; judging connectivity.

[0005] The above-mentioned existing technology combines the characteristics of the underground engineering industry, combines the legal theory of causal relationship determination with hydrogeological surveys, online monitoring, mathematical statistics and other methods, and develops a targeted method for assessing the value of ecological and environmental damage caused by underground engineering water gushing incidents, providing technical and normative guarantees for the assessment of ecological and environmental damage caused by underground engineering water gushing incidents. However, this existing technology was developed for underground engineering water gushing incidents and is not suitable for soil environmental damage identification and assessment cases.

[0006] On the other hand, the illegal dumping and landfill of domestic waste are typical types of environmental violations, and the current ecological environmental damage identification work mainly focuses on the investigation of conventional physical and chemical indicators and heavy metal indicators. However, compared with other types of environmental pollution, the pollution of soil environment by domestic waste has obvious hidden and cumulative characteristics. It often happens that when conducting environmental impact assessments, conventional physical and chemical indicators and heavy metal indicators do not exceed national standards, but the facts of illegal dumping of domestic waste and soil pollution do exist. This aggravates the difficulty of tracing the causes of pollution and defining responsibilities, and thus makes the process of confirming the facts of damage more complicated.

[0007] The present invention aims to provide an assessment method that can further quantify the damage of domestic waste to the soil ecological environment in an area where domestic waste is illegally dumped, provided that the conventional physical and chemical indicators and heavy metal indicators of the soil do not exceed the national standards. Summary of the Invention

[0008] In response to the problems in the related art, the present invention proposes a method for confirming soil ecological environment damage suitable for areas where domestic waste is illegally dumped and its application, so as to overcome the above-mentioned technical problems existing in the existing related art.

[0009] The technical solution of the present invention is achieved as follows:

[0010] A method for confirming soil ecological environmental damage in areas where domestic waste is illegally dumped, comprising at least the following steps:

[0011] 1) Screening characteristic pollution indicators and determining characteristic pollution indicators;

[0012] 2) Determine the levels of characteristic pollution indicators in soil samples from the control area and the assessment area where domestic waste is illegally dumped, and analyze to determine whether the levels of characteristic pollution indicators in the assessment area exceed the baseline levels in the soil of the control area;

[0013] 3) Perform high-throughput sequencing on soil samples from the evaluation and control areas, perform OTU cluster analysis and α-diversity analysis based on the sequence information, and analyze whether the bacterial community in the evaluation area has changed significantly compared with the control area by at least using the Wilcoxon rank sum test;

[0014] 4) Through RDA redundancy analysis, the correlation between the changes in bacterial communities in the assessment area and characteristic pollution indicators was tested to indirectly confirm whether domestic waste has damaged the soil ecological environment, and the degree of impact of domestic waste on the soil ecological environment was quantified through changes in various indices of bacterial communities.

[0015] Preferably, since the present invention is directed to monitoring of the soil environment, i.e., solid waste, the screening of characteristic pollution indicators in step 1) adopts the following determination method:

[0016] Heavy metals were determined using a 7900 inductively coupled plasma mass spectrometer, semivolatile organic compounds were determined using a 8860-5977B gas chromatograph mass spectrometer, and volatile organic compounds were determined using an Atomx-XYZ-8890-5977B gas chromatograph mass spectrometer.

[0017] Preferably, step 2) comprises the following steps:

[0018] 2.1) Determination of characteristic pollution indicators in soil samples from the assessment area and the control area;

[0019] 2.2) Determine soil baseline level:

[0020] The specific operation is to perform statistical calculations on the characteristic pollution index content data measured in the control area to check whether it belongs to a normal distribution;

[0021] If the soil distribution is normal, the 90% upper reference limit of the control point data is used as the soil baseline, which is the arithmetic mean + 1.65 times the standard deviation; if the soil distribution is not normal, the 90th percentile of the control point data is used as the soil baseline;

[0022] 2.3) Compare with the soil baseline level to determine whether the content of characteristic pollution indicators in the assessment area exceeds the soil baseline level.

[0023] Preferably, in step 3), OTU cluster analysis is performed on the sequences obtained by high-throughput sequencing at a similarity level of 97%.

[0024] Preferably, in step 3), the α diversity analysis comprises ACE index, Chao1 index, Simpson index and Shannon-Wiener index analysis.

[0025] Preferably, in step 3), 0.25 to 0.5 g of soil sample is taken and stored in a refrigerator at ≤80°C, and high-throughput sequencing is performed within 24 hours. The sample nucleic acid is extracted using a magnetic bead method, and then PCR amplification and sequencing are performed to obtain raw sequencing data.

[0026] Preferably, in step 3), at least the following steps are included:

[0027] 3.1) High-throughput sequencing of soil samples from the evaluation and control areas;

[0028] 3.2) Analyze the number of OTUs and α-diversity index in the evaluation area and the control area to determine whether the number of OTUs and α-diversity index in the evaluation area have decreased;

[0029] 3.3) Perform taxonomic analysis of the sequence data obtained from high-throughput sequencing at least at the phylum level, and then determine the top 10 dominant bacterial phyla and their relative abundances in the evaluation area. Determine the relative abundance and magnitude of the dominant bacterial phyla in the control area.

[0030] 3.4) The Wilcoxon rank sum test was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial phylum between the evaluation area and the control area.

[0031] More preferably, in step 3), the sequence results obtained by high-throughput sequencing are further subjected to taxonomic analysis at the genus level, comprising the following steps:

[0032] 3.5) Perform taxonomic analysis of the sequence results obtained from high-throughput sequencing at the genus level to determine the top 10 dominant bacterial genera and their relative abundances in the evaluation area, and then determine the relative abundances of the corresponding dominant bacterial genera in the control area;

[0033] 3.6) The Wilcoxon rank sum test was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial genus between the evaluation area and the control area.

[0034] Preferably, step 3) further comprises preparing a dilution curve to verify the comprehensiveness of the sequence data.

[0035] Preferably, the evaluation area and the control area each have at least 3 soil samples. In step 3.2), the OTU number and α diversity index of each soil sample are analyzed and determined, and then the mean plus or minus standard deviation Mean±SD is calculated. By comparing the mean plus or minus standard deviation Mean±SD of the OTU number and α diversity index of the evaluation area and the control area, it is determined whether the OTU number and α diversity index of the evaluation area have decreased.

[0036] Preferably, step 4) comprises at least the following steps:

[0037] 4.1) Using the RDA redundancy analysis method, correlation tests were conducted between the top 10 dominant bacterial phyla at the phylum level in the assessment area and characteristic pollution indicators to indirectly confirm whether domestic waste is damaging the soil ecological environment;

[0038] 4.2) Use changes in various bacterial community indices to quantitatively assess the impact of domestic waste on the soil ecological environment.

[0039] More preferably, step 4) further comprises the following steps:

[0040] Through the RDA redundancy analysis method, the correlation between the top 10 dominant bacterial genera in relative abundance at the genus level in the assessment area and characteristic pollution indicators was tested to indirectly confirm whether domestic waste has damaged the soil ecological environment, and the degree of impact of domestic waste on the soil ecological environment was quantified through changes in various indexes of bacterial communities.

[0041] As a core component of the soil ecosystem, soil microbial communities are directly involved in the decomposition of substances and nutrient cycling within the soil. Approximately 80 to 90% of soil functions are closely related to soil microorganisms, including the efficient conversion of soil nutrients, the maintenance of ecosystem stability, and resistance to external disturbances. From a biological perspective, microorganisms are considered the most sensitive indicators of soil quality. This invention uses highly sensitive microorganisms as key indicators for assessing significant changes in the soil environment, directly reflecting the toxicity and potential ecological impacts of domestic waste.

[0042] Based on the above ideas, the present invention innovatively introduces the analytical framework of the damaged soil microbial community structure and its dynamic changes into the practice of ecological environmental damage identification and assessment. With the help of high-throughput sequencing technology, OTU cluster analysis and other means, the characteristics of bacterial communities are analyzed, providing a scientific and fast method for evaluating the impact of domestic waste on microbial communities.

[0043] The method for confirming soil ecological environmental damage in domestic waste areas proposed in the present invention is comprehensive and reasonable. Specifically, the soil environment in areas where domestic waste is landfilled is measured and evaluated at multiple levels from the perspectives of characteristic heavy pollution indicators, the number of OTUs, the α-diversity index, and the increase and decrease in the relative abundance of dominant groups, and the correlation between pollution indicators and microbial community changes is analyzed to ensure that the evaluation model is scientific and reasonable. At the same time, the number of OTUs, the α-diversity index, and the increase and decrease in the relative abundance of dominant groups introduced in the present invention can be used to quantify the impact of domestic waste on the soil environment, and can effectively assist in confirming the damage facts in the identification and assessment of ecological environmental damage caused by illegal dumping of domestic waste.

[0044] The present invention also discloses the application of the above-mentioned soil ecological environment damage confirmation method to conduct preliminary source tracing in a soil area suspected of being dumped with domestic garbage. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 This is a soil sampling point layout diagram of the present invention;

[0046] Figure 2 A histogram showing the relative abundance of the dominant bacterial phyla in the evaluation area of ​​the present invention compared to the control area;

[0047] Figure 3 A histogram showing the relative abundance of the dominant bacterial genera in the evaluation area of ​​the present invention compared to the control area;

[0048] Figure 4 A redundancy analysis diagram of the dominant bacterial phyla and characteristic pollution indicators in the evaluation area of ​​the present invention;

[0049] Figure 5 This is a redundancy analysis diagram of the dominant bacterial genera and characteristic pollution indicators in the evaluation area of ​​the present invention. DETAILED DESCRIPTION

[0050] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0051] This example describes an illegal dumping of domestic waste in northern Guangdong. This area is semi-mountainous and hilly, with mountains running primarily from northeast to southwest. Mountains account for approximately 80% of the total area. The terrain slopes from northeast to southwest, with small and medium-sized basins and river terraces forming the intervals between the mountains. The climate is a mid-subtropical monsoon climate with mountainous characteristics, with an average annual temperature of 20.3°C and an average annual rainfall of 1,778 mm.

[0052] The land use type of the assessment area involved in this example is agricultural land. Local ecological and environmental regulatory authorities discovered that a large amount of waste, including waste fabrics and plastics, had been dumped in the area. Surveying and mapping revealed that the area covered approximately 50,000 square meters and the waste weighed approximately 100,000 tons. Environmental protection officials arrived at the site to conduct relevant work.

[0053] A. Soil sampling

[0054] A-1) Soil sampling in the assessment area is as follows: 5 sample plots of 1 m × 1 m in area are randomly selected in the assessment area. The soil 10 cm below the domestic waste is drilled in the center of the sample plot. Three soil samples are taken from this section and mixed in equal proportions. This constitutes the soil sample of one assessment area. Five soil samples are taken from the assessment area and are numbered DG1 to DG5.

[0055] A-2) The surrounding area that was obviously uncontaminated served as a control area for soil sampling.

[0056] Five soil control points at the same depth as the assessment area were set up in areas around the area to be assessed that were obviously not contaminated by domestic waste. One soil sample was collected from each point, and the soil samples were numbered NC1 to NC5.

[0057] The specific sampling point layout of the evaluation area and the control area is shown as follows: Figure 1 shown.

[0058] B. Conduct physical and chemical property analysis and heavy metal content determination on soil samples from the evaluation area and control area.

[0059] The results showed that none of them exceeded the risk screening values ​​of the "Soil Environmental Quality Agricultural Land Soil Pollution Risk Control Standards (Trial)" (GB15618-2018).

[0060] C. Using the soil ecological environment damage confirmation method applicable to the illegal dumping of domestic garbage of the present invention to assess the area, comprising the following steps:

[0061] 1) Screen soil samples from the assessment area and the control area for characteristic pollution indicators to determine the characteristic pollution indicators.

[0062] Heavy metals were determined using a 7900 inductively coupled plasma mass spectrometer, semivolatile organic compounds were determined using a 8860-5977B gas chromatograph mass spectrometer, and volatile organic compounds were determined using an Atomx-XYZ-8890-5977B gas chromatograph mass spectrometer.

[0063] According to the screening results, the characteristic pollution indicators were determined to be Cr, Ni, Zn, Ba and Pb.

[0064] 2) The content of characteristic pollution indicators in soil samples from the evaluation area and the control area were determined respectively.

[0065] 2.1) The contents of heavy metals Pb, Zn, Ni, Cr, and Ba in soil samples DG1-DG5 and NC1-NC5 were determined by 240FS AA flame atomic absorption spectrometer.

[0066] 2.2) Perform statistical calculations on the heavy metal content data of Pb, Zn, Ni, Cr, and Ba measured in the control area to check whether they are normally distributed;

[0067] If the soil distribution is normal, the 90% reference upper limit (arithmetic mean + 1.65 times the standard deviation) of the control point data is used as the soil baseline. If the soil distribution is not normal, the 90th percentile of the control point data is used as the soil baseline.

[0068] The results are shown in the following table.

[0069]

[0070] It can be seen from the above table that the Cr, Ni, Zn, Ba, and Pb test data of soil samples NC1 to NC5 in the control area and the Cr, Ni, Zn, Ba, and Pb test data of soil samples DG1 to DG5 in the evaluation area all obey the normal distribution.

[0071] 2.3) Compare with the soil baseline level to determine whether the content of characteristic pollution indicators in the assessment area exceeds the soil baseline level.

[0072] The heavy metals Pb, Zn, Ni, Cr, and Ba in soil samples DG1 to DG5 in the assessment area all exceeded the baseline values ​​to varying degrees. That is, by comparing with the baseline values, it was proved that domestic waste caused the formation of mild heavy metal complex pollution of Pb, Zn, Ni, Cr, and Ba in the soil of the assessment area.

[0073] Based on the above situation, the present invention aims to prove that the soil environment has changed through changes in the microbial community, and at the same time analyze whether the changes in the microbial community are caused by the above-mentioned mild metal complex pollution.

[0074] 3) High-throughput sequencing

[0075] 3.1) 16S rRNA high-throughput sequencing was performed on soil samples from the evaluation area and the control area.

[0076] All soil samples collected this time were stored in a refrigerator ≤80℃ and sent to the laboratory for relevant sequencing work within 24 hours after sampling.

[0077] 3.1a) 0.35 g of soil samples were collected from each of DG1-DG3 and NC1-NC3. Nucleic acid was extracted using the TGuide S96 magnetic bead method and the nucleic acid concentration was measured using a Synergy HTX microplate reader.

[0078] 3.1b) PCR amplification was performed using a Veriti 96-well 9902 gradient amplification instrument, and the integrity of the PCR product was checked by agarose gel electrophoresis (1.8% mass concentration);

[0079] 3.1c) Perform library testing using the Qsep-400 method. If the test is qualified, proceed to the next step;

[0080] 3.1d) High-throughput sequencing was performed using a Novasek 6000 sequencer to obtain raw sequencing data.

[0081] 3.1e) High-throughput sequencing data were processed. The raw sequencing data were quality-filtered using Trimmomatic 0.33. Primer sequences were identified and removed using Cutadapt 1.9.1. Paired-end reads were then spliced ​​and chimeras were removed using USEARCH 10.0 to obtain high-quality sample sequence data.

[0082] 3.1f) OTU clustering analysis was performed on the sequences at the 97% similarity level using USEARCH 10.0, and the quality-controlled data were denoised using the DADA2 method in QIIME2.

[0083] 3.2) Verify the comprehensiveness of the test data volume.

[0084] Random sampling of test sequences was used for verification. When the number of test sequences reached 60,000, the coverage was ≥99%, and the growth of the number of OTUs slowed significantly, approaching saturation. In other words, the amount of sequencing data used in this example was sufficient and reasonable, the sequencing depth was sufficient to cover most bacteria, and the evaluation data was reliable.

[0085] 3.3) OTU number records and α diversity index analysis

[0086] Alpha diversity analysis includes ACE index, Chao1 index, Simpson index and Shannon-Wiener index analysis.

[0087] The results are shown in the following table.

[0088]

[0089]

[0090] The tabular data show that the number of OTUs in the soil samples of the evaluation area is significantly lower than that in the control area. At the same time, the α diversity indices (ACE, Chao1, Shannon, and Shannon-Wiener) of the soil samples in the evaluation area are all lower than those in the evaluation area, indicating that the bacterial diversity and richness of the soil environment in the evaluation area are significantly lower than those in the control area.

[0091] 3.4) Perform taxonomic analysis of the sequence results obtained from high-throughput sequencing at the phylum level, and then determine the top 10 dominant bacterial phyla and their relative abundance in the evaluation area. Determine the relative abundance and increase or decrease of the dominant bacterial phyla in the control area.

[0092] 3.5) The Wilcoxon rank sum test (P < 0.05) was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial phylum between the evaluation area and the control area.

[0093] The results are shown in the following table.

[0094]

[0095] 3.6) Perform taxonomic analysis at the genus level on the sequence data obtained from high-throughput sequencing to determine the top 10 dominant bacterial genera and their relative abundances in the evaluation area. Furthermore, determine the relative abundance and magnitude of the corresponding dominant bacterial genera in the control area.

[0096] 3.7) The Wilcoxon rank sum test (P < 0.05) was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial genus between the evaluation area and the control area.

[0097] The results are shown in the following table.

[0098]

[0099] 3.8) Create a distribution histogram of the top 10 dominant taxa in relative abundance in the assessment area and compare it with the control area to visually demonstrate the changes in relative abundance of dominant taxa at the phylum and genus levels.

[0100] Figure 2 The relative abundance histogram of the top 10 dominant bacterial phyla in soil samples from the evaluation area and the control area is shown in the figure. Figure 3 The relative abundance bar graphs of the top 10 dominant bacterial genera in soil samples from the evaluation area and the control area.

[0101] In summary, compared with the control area, the relative abundance of dominant groups in the assessment area at the phylum and genus levels changed significantly, indicating that the soil ecological environment in the assessment area has changed.

[0102] 4) The correlation between the changes in bacterial communities in the assessment area and characteristic pollution indicators was tested through RDA redundancy analysis.

[0103] 4.1) Using the RDA redundancy analysis method, correlation tests were performed between the top 10 dominant bacterial phyla with the highest relative abundance at the phylum level in the assessment area and characteristic pollution indicators.

[0104] See the results Figure 4 .

[0105] Figure 4 The analysis results showed that Cr, Ni, Zn, Ba, and Pb were positively correlated with Thermodesulfobacteriota, Gemmatimonadota, Bacillota, Chloroflexota, and Pseudomonadota, and negatively correlated with Bacteroidota and Actinobacteriota. That is, changes in the content of Cr, Ni, Zn, Ba, and Pb had a significant impact on as many as 7 of the top 10 dominant bacterial phyla in relative abundance.

[0106] 4.2) Using the RDA redundancy analysis method, correlation tests were conducted between the top 10 dominant bacterial genera with the highest relative abundance at the genus level in the assessment area and characteristic pollution indicators.

[0107] See the results Figure 5 .

[0108] Figure 5 The analysis results showed that Pb, Zn, Ni, Cr, and Ba were negatively correlated with the top 10 dominant bacterial genera in relative abundance, that is, changes in the content of Cr, Ni, Zn, Ba, and Pb had a significant impact on the top 10 dominant bacterial phyla in relative abundance; at the same time, the bacterial community structure between the two groups of soil samples was quite different, specifically, the three soil samples NC in the control area (i.e., the green dots in the figure) showed a more compact distribution pattern, showing a higher intra-group similarity, while the three soil samples DG in the evaluation area (i.e., the yellow dots in the figure) showed a more dispersed distribution pattern, indicating the complexity of its bacterial community structure.

[0109] 4.3) There was a significant correlation between the changes in the relative abundance of the top 10 dominant bacterial groups at the phylum and genus levels in the assessment area relative to the control area and the characteristic heavy metal indicators, indicating that the changes in the soil ecological environment in the assessment area were caused by domestic waste. The number of OTUs, α-diversity index, and the difference in the relative abundance of dominant groups in the assessment area relative to the control area can be used as quantitative indicators to assess the impact of domestic waste on the soil environment, which can assist in the identification and confirmation of damage facts in cases of illegal dumping of domestic waste.

[0110] Based on the disclosure and teachings of the above description, those skilled in the art may also make changes and modifications to the above embodiments. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and some modifications and variations of the present invention should also fall within the scope of protection of the claims of the present invention. In addition, although some specific terms are used in this description, these terms are only for convenience of description and do not constitute any limitation to the present invention.

Claims

1. A method for confirming soil ecological environment damage in areas where domestic waste is illegally dumped, characterized in that: At least the following steps are included: 1) Screening characteristic pollution indicators and determining characteristic pollution indicators; 2) Determine the levels of characteristic pollution indicators in soil samples from the control area and the assessment area where domestic waste is illegally dumped, and analyze to determine whether the levels of characteristic pollution indicators in the assessment area exceed the baseline levels in the soil of the control area; 3) Perform high-throughput sequencing on soil samples from the evaluation and control areas, perform OTU cluster analysis and α-diversity analysis based on the sequence information, and analyze whether the bacterial community in the evaluation area has changed significantly compared with the control area by at least using the Wilcoxon rank sum test; 4) Through RDA redundancy analysis, the correlation between the changes in bacterial communities in the assessment area and characteristic pollution indicators was tested to indirectly confirm whether domestic waste has damaged the soil ecological environment, and the degree of impact of domestic waste on the soil ecological environment was quantified through changes in various indices of bacterial communities.

2. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 1, characterized in that: In step 3), the following steps are included: 3.1) High-throughput sequencing of soil samples from the evaluation and control areas; 3.2) Analyze the number of OTUs and α-diversity index in the evaluation area and the control area to determine whether the number of OTUs and α-diversity index in the evaluation area have decreased; 3.3) Perform taxonomic analysis of the sequence data obtained from high-throughput sequencing at least at the phylum level, and then determine the top 10 dominant bacterial phyla and their relative abundances in the evaluation area. Determine the relative abundance and magnitude of the dominant bacterial phyla in the control area. 3.4) The Wilcoxon rank sum test was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial phylum between the evaluation area and the control area.

3. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 2, characterized in that: In step 3), the sequence results obtained by high-throughput sequencing are also subjected to taxonomic analysis at the genus level, including the following steps: 3.5) Perform taxonomic analysis of the sequence results obtained from high-throughput sequencing at the genus level to determine the top 10 dominant bacterial genera and their relative abundances in the evaluation area, and then determine the relative abundances of the corresponding dominant bacterial genera in the control area; 3.6) The Wilcoxon rank sum test was used to determine whether there was a significant difference in the relative abundance of the same dominant bacterial genus between the evaluation area and the control area.

4. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 3, characterized in that: Step 4) includes at least the following steps: 4.1) Using the RDA redundancy analysis method, correlation tests were conducted between the top 10 dominant bacterial phyla at the phylum level in the assessment area and characteristic pollution indicators to indirectly confirm whether domestic waste is damaging the soil ecological environment; 4.2) Use changes in various bacterial community indices to quantitatively assess the impact of domestic waste on the soil ecological environment.

5. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 4, characterized in that: Step 4) also includes: Through the RDA redundancy analysis method, the correlation between the top 10 dominant bacterial genera in relative abundance at the genus level in the assessment area and characteristic pollution indicators was tested to indirectly confirm whether domestic waste has damaged the soil ecological environment, and the degree of impact of domestic waste on the soil ecological environment was quantified through changes in various indexes of bacterial communities.

6. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 2, characterized in that: Step 3) includes making a dilution curve to verify the comprehensiveness of the sequence data; The α diversity analysis includes ACE index, Chao1 index, Simpson index and Shannon-Wiener index analysis.

7. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 6, characterized in that: The evaluation area and the control area each have at least 3 soil samples. In step 3.2), the OTU number and α diversity index of each soil sample are analyzed and determined, and then the mean plus or minus standard deviation Mean±SD is calculated. By comparing the mean plus or minus standard deviation Mean±SD of the OTU number and α diversity index in the evaluation area and the control area, it is determined whether the OTU number and α diversity index in the evaluation area have decreased.

8. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 1, characterized in that: The screening of characteristic pollution indicators in step 1) uses the following measurement methods: Heavy metals were determined using a 7900 inductively coupled plasma mass spectrometer, semivolatile organic compounds were determined using a 8860-5977B gas chromatograph mass spectrometer, and volatile organic compounds were determined using an Atomx-XYZ-8890-5977B gas chromatograph mass spectrometer.

9. The method for confirming soil ecological environment damage applicable to areas where domestic waste is illegally dumped according to claim 8, characterized in that: Step 2) includes the following steps: 2.1) Determination of characteristic pollution indicators in soil samples from the assessment area and the control area; 2.2) Determine soil baseline level: The specific operation is to perform statistical calculations on the characteristic pollution index content data measured in the control area to check whether it belongs to a normal distribution; If the soil distribution is normal, the 90% upper reference limit of the control point data is used as the soil baseline, which is the arithmetic mean + 1.65 times the standard deviation; if the soil distribution is not normal, the 90th percentile of the control point data is used as the soil baseline; 2.3) Compare with the soil baseline level to determine whether the content of characteristic pollution indicators in the assessment area exceeds the soil baseline level.

10. Application of the soil ecological environment damage confirmation method applicable to areas where domestic waste is illegally dumped as described in any one of claims 1 to 9 to conduct preliminary tracing of the soil in areas suspected of having dumped domestic waste.

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