Method for researching transportation effect of microplastics on microorganisms under continuous migration

Abstract

The invention discloses a method for researching the transportation effect of microplastics on microorganisms under continuous migration, which comprises the following steps of: placing microplastics without microorganisms on the initial surface in a field environment, so that microorganisms in the original environment are planted on the surface of the microplastics, and setting two migration modes, so as to research the transportation effect of the microplastics on the microorganisms. A continuous migration mode of continuously changing the geographic position for multiple times for continuous culture or a direct migration mode of directly changing to the final geographic position for culture is adopted, and part of micro-plastics in each environment before and after migration are reserved for continuous culture to serve as a control group; the initial surface microorganisms in the migration process of the micro-plastic in the environment are known, and the migration path is controllable; after surface microorganisms are separated and extracted by recycling a micro-plastic sample, the change condition of the surface microorganisms under continuous migration of the micro-plastic is studied, and then the transport carrier effect of the micro-plastic on the surface microorganisms in the environment migration process is explained. According to the invention, transportation simulation and analysis of microplastics on microorganisms under continuous migration are realized.

Description

Technical Field

[0001] The present invention relates to the field of environmental microorganisms, and in particular to a method for the effect of microplastics on microbial transport under continuous migration. Background Art

[0002] Microplastics refer to a mixture of plastics of different shapes, colors, sizes and polymer types with a particle size not exceeding 5mm. Due to the widespread use and irrational disposal of plastic products, a large number of microplastics currently exist in the environment, causing environmental pollution. Since microplastics have a large specific surface area (the total area per unit mass of material), they can provide more attachment sites for microorganisms than large-sized plastics. Microplastics are usually light in weight and easy to float and move on the water surface. Therefore, microplastics have become a good carrier for carrying microorganisms to migrate in the environment.

[0003] Since it is difficult to fully grasp the migration path of microplastics through random sampling of microplastics, the changes in surface microorganisms when microplastics migrate in the environment are still unclear. Given the understanding of this current situation in the field, it is still practical to use microplastics as carriers of microorganisms. Summary of the invention

[0004] The present invention aims to provide a method for transporting microorganisms using microplastics to achieve continuous migration, thereby achieving the simulation and analysis of the transport of microorganisms by microplastics under continuous migration.

[0005] The present invention realizes a method for studying the effect of microplastics on microbial transport under continuous migration, comprising:

[0006] The continuous migration path is set to include m points, the first point is placed with 3m-2 groups of microplastic samples with initial surface sterility, and the remaining points are placed with m groups of microplastic samples with initial surface sterility;

[0007] Recovering part of the samples that do not change position to obtain a first control sample group; completing direct migration to obtain a first direct migration sample group; completing the first migration in the continuous migration to obtain a first continuous migration sample group;

[0008] Recover part of the samples that do not change position to obtain a second control sample group; recover part of the first continuous migration sample group, and the remaining first continuous migration sample groups complete the second migration in the continuous migration to obtain a second continuous migration sample group; complete the direct migration to obtain a second direct migration sample group;

[0009] By analogy with the above steps, recover part of the samples that do not change their positions to obtain the xth control sample group; recover part of the x-1th continuous migration sample group, and the remaining x-1th continuous migration sample group completes the xth migration in the continuous migration to obtain the xth continuous migration sample group; complete the direct migration to obtain the xth direct migration sample group, so that the continuous migration samples reach the last point, and x represents any intermediate point;

[0010] After the continuous migration is completed, the mth control sample group, the m-1th continuous migration sample group, and the first to m-1th direct migration sample groups are recovered, where m represents the number of points included in the continuous migration path;

[0011] Surface microorganisms were extracted from all samples of the recovered 1st to mth control sample groups, 1st to m-1th continuous migration sample groups, and 1st to m-1th direct migration sample groups, and the microorganisms between different groups were compared to explore the transport effect of microplastics on microorganisms during continuous migration and complete the microbial data analysis.

[0012] In some embodiments, the step of obtaining the first control sample group further comprises: in situ culturing at each point, after several days, recovering m+1 groups of samples from the first point, recovering 1 group of samples from each of the remaining points, and recovering each group of samples recovered from all the points to the laboratory for storage as the first control sample group;

[0013] The step of obtaining the first direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point, placing them in the same culture environment, as the first direct migration sample group;

[0014] The step of obtaining the first continuous migration sample group further includes: migrating the m-1 groups of samples recovered from the first point to the second point to simulate the first migration of the continuous migration as the first continuous migration sample group.

[0015] In some embodiments, the step of obtaining a second control sample group further comprises: continuing the culture, after several days, recovering each group of samples cultured at all points to the laboratory for storage as the second control sample group;

[0016] The step of recovering part of the first continuous migration sample group further includes: recovering a group of first continuous migration sample group samples recovered from the second point to a laboratory for storage;

[0017] The step of obtaining a second direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point as a second direct migration sample group;

[0018] The step of obtaining the second continuous migration sample group further includes: migrating the m-2 groups of samples recovered from the second point and migrated thereto from the first point to the third point to simulate the second migration of the continuous migration as the second continuous migration sample group.

[0019] In some embodiments, the step of obtaining the xth control sample group further comprises: after migration and continued culture, recovering 2 groups of samples from the first point, recovering 1 group of samples that have always been cultured at the point from any intermediate point, m-x+1 groups of samples that have migrated from the first point to this point, and recovering 1 group of samples that have always been cultured at the point from each of the remaining points; recovering each group of samples that have always been cultured at the current point from all the points to the laboratory for storage as the xth control sample group;

[0020] The step of recovering part of the x-1th continuously migrated sample group further includes: recovering a group of samples that migrated from a point above any intermediate point and recovered from the current arbitrary intermediate point to the laboratory for storage;

[0021] The step of obtaining the xth continuous migration sample group further includes: migrating the mx groups of continuous migration samples recovered from any current intermediate point to any current next point to simulate the xth migration of the continuous migration as the xth continuous migration sample group;

[0022] The step of obtaining the xth direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point as the xth direct migration sample group.

[0023] In some embodiments, the step of recovering the mth control sample group, the m-1th continuous migration sample group, and the first to m-1th direct migration groups further includes: after performing the m-1th migration and continuing to culture for several days, recovering 1 group of samples that have always been cultured here from all points, recovering 1 group of samples that have continuously migrated from the first point to here from the last point, and recovering m-1 groups of samples that have directly migrated from the first point to here from the last point.

[0024] In some embodiments, the step of performing microbial data analysis further comprises:

[0025] By using samples that were always cultured at various points as controls, the microorganisms on the surface of the samples in the continuous migration group were compared to study the transport effect of microplastics on microorganisms under continuous migration.

[0026] In some embodiments, the step of performing microbial data analysis further comprises:

[0027] By comparing the microorganisms on the surface of the continuous migration group samples and the direct migration group samples, the effect of the intermediate point environment on the microorganisms on the surface of microplastic transport during the continuous migration process was studied.

[0028] In some embodiments, the step of performing microbial data analysis further comprises:

[0029] By comparing samples recovered from the same point in the control group after different culture times, the growth changes of microorganisms on the surface of microplastics at different stages can be studied.

[0030] In some embodiments, the step of performing microbial data analysis further comprises:

[0031] By comparing the microorganisms on the surface of samples that were cultured for different times at the first point and the last point but with the same total culture time, the changes in the surface microorganisms of microplastics when they migrate to the new environment when the culture time in the original environment and the current environment is different are studied.

[0032] Compared with the prior art, the advantages and positive technical effects achieved by the present invention are as follows:

[0033] 1. Based on setting multiple points and multiple sampling time points in the environment, the continuous migration process of microplastics can be simulated in a controllable manner, specifically including recovering some samples that do not change their positions to obtain the xth control sample group; recovering some of the x-1th continuous migration sample groups, and the remaining x-1th continuous migration sample groups complete the xth migration in the continuous migration to obtain the xth continuous migration sample group; completing direct migration to obtain the xth direct migration sample group; making the continuous migration samples reach the last point; after completing the continuous migration and continuing to culture for several days, all samples are recovered; surface microorganisms are extracted from all recovered samples, and microorganisms between different groups are compared to explore the transport effect of microplastics on microorganisms during migration, and complete microbial data analysis. By recovering microplastic samples and extracting the microbial community on their surface, the effect of microplastics on microbial transport under continuous migration is studied, thereby studying the use of microplastics to achieve microbial transport under continuous migration, which is suitable for the simulation of all microplastic migration paths and different microplastic residence times;

[0034] 2. The differences between the microbial communities on the surface of microplastics and those in the surrounding matrix were fully considered, and all comparisons were made between the microorganisms on the surface of microplastic samples;

[0035] 3. It can simultaneously accomplish multiple research objectives, including the transport effect of microplastics on microorganisms under continuous migration, the control and simulation of the growth of microorganisms on the surface of microplastic samples under in situ cultivation corresponding to different cultivation times, and the transport effect of microplastics on microorganisms after migration at different residence times. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a schematic diagram of the overall process of a method for studying the effect of microplastics on microbial transport under continuous migration of the present invention;

[0037] Figure 2 It is a flowchart of the implementation process of a method for studying the effect of microplastics on microbial transport under continuous migration of the present invention;

[0038] Figure 3 It is a schematic diagram of placing microplastic samples at various points of in situ culture;

[0039] Figure 4 This is a schematic diagram of the first migration of microplastic samples;

[0040] Figure 5 This is a schematic diagram of the second migration of microplastic samples;

[0041] Figure 6 It is a schematic diagram of completing continuous migration. DETAILED DESCRIPTION

[0042] The technical solution will be clearly described below in conjunction with the accompanying drawings and embodiments.

[0043] The present invention changes the culture position to control the migration process of microplastics in the environment, sets different culture times and recovery times to fully represent the actual situation of different residence time of microplastics in the environment, and uses microplastics with the same culture time as a control group to avoid the difference between free growth and attached growth of microorganisms, so as to fully analyze the changes in the microbial community on the surface of microplastics before and after migration.

[0044] like Figure 1 As shown, a method of the present invention for studying the effect of microplastics on microbial transport under continuous migration includes the following steps:

[0045] Step 1: In situ culture: Set a continuous migration path including m points, and place multiple groups of microplastic samples with sterile surfaces at different points;

[0046] Step 1-1: To ensure that the number of microorganisms on the surface of the microplastic samples is sufficient for analysis after recovery, each group of microplastic samples includes multiple microplastics; before placing the microplastics, rinse the surface with 75% ethanol and pure water three times each to ensure that the surface of the microplastics is initially sterile;

[0047] Step 1-2: Select m points to simulate the points that microplastics pass through when migrating in the environment, and set the continuous migration path to start from the first point S1, pass through the second point S2 to the m-1th point S m-1 , and finally arrive at the mth point S m , then place 3m-2 groups of microplastic samples at the first point, and place m groups of microplastic samples at the remaining points, where m represents the number of points included in the continuous migration path, such as Figure 3 As shown;

[0048] Step 2: Recover part of the sample and change the position of part of the microplastics to complete the first migration in the continuous migration, such as Figure 4 As shown;

[0049] Step 2-1: After culturing at each point for several days, recover m+1 groups of samples from S1, and recover 1 group of samples from each of the remaining points;

[0050] Step 2-2: After the sample is recovered, gently wash it to remove the influence of microbial attachment in the surrounding environment; if cultured in water, use 0.45μm filter membrane to gently wash it three times in situ water; if cultured in soil, use ultrapure water to gently wash it three times;

[0051] Step 2-3: Collect each group of samples from all points and store them in the laboratory as a control for the microorganisms on the surface of microplastics at each point. The naming rule is "point + pair + total number of recovery times including all samples", that is, the sample recovered from S1 at this time is named "S1 pair 1", and the naming method is the same thereafter;

[0052] Step 2-4: Migrate a set of samples recovered from S1 directly to S m , to simulate direct migration, ice packs are used during transportation to protect the microorganisms on the surface of microplastics. After being transported to the target location, the microplastic samples are placed in the same culture environment as the microplastic samples in step 1-2. The naming rule is "straight + total number of recycling times including all samples", that is, the sample at this time is named "straight 1", and the naming method is the same thereafter;

[0053] Step 2-5: Migrate the m-1 group of samples recovered from S1 to S2 to simulate the first migration of continuous migration. Use ice packs to preserve the microorganisms on the surface of microplastics during transportation. After being transported to the target location, place the microplastic samples in the same culture environment as the microplastic samples in step 1-2. The naming rule is "the total number of recovery times including all samples", that is, the sample at this time is named "Lian 1", and the naming method is the same thereafter;

[0054] Step 3: Recover part of the sample and change the position of some microplastics to complete the second migration in the continuous migration, such as Figure 5 As shown;

[0055] Step 3-1: After continuing to culture for several days, according to the sample recovery method in step 2-2, 2 groups of samples are recovered from S1, 1 group of samples that have always been cultured at this point are recovered from S2, and m-1 groups of samples that have migrated from S1 to this point are recovered, and 1 group of samples that have always been cultured at this point are recovered from each of the remaining points;

[0056] Step 3-2: Collect the samples from all the sites and keep them in the laboratory. Name them according to the method of step 2-3. That is, the samples collected from S1 are named "S1 to 2".

[0057] Step 3-3: Migrate a group of samples recovered from S1 directly to S m , to simulate direct migration, the samples were named according to steps 2-4, i.e., the samples at this time were named “Direct 2”;

[0058] Step 3-4: Migrate the m-2 group of samples recovered from S2 and migrated from S1 to S3 to simulate the second migration of continuous migration, migrate and continue to culture according to the sample migration method in step 2-5, and name them according to the method in step 2-5, that is, the samples at this time are named "Lian 2";

[0059] Step 3-5: A group of samples "Link 1" that migrated from S1 to S2 was recovered from S2 and stored in the laboratory to represent part of the process of continuous migration;

[0060] Step 4: Continue to culture and migrate the sample for the xth migration, until the continuously migrated sample reaches the last point, that is, the m-1th migration is completed;

[0061] Step 4-1: After migration and continued cultivation, recover 2 sets of samples from S1 and 2 sets of samples from S according to the sample recovery method in step 2-2. x Recover 1 group of samples that have always been cultured at this point, m-x+1 group of samples that migrated from S1 to this point, and recover 1 group of samples that have always been cultured at this point from each of the remaining points, where x represents any intermediate point;

[0062] Step 4-2: Collect one group of samples collected from all the points and keep them in the laboratory, and name them as "S1,...,S2" in the same way as in step 2-3. m for x";

[0063] Step 4-3: Migrate a group of samples recovered from S1 directly to S m , to simulate direct migration, named "straight x" in the same way as steps 2-4;

[0064] Step 4-4: From S x The recovered mx group continuously migrated samples to S x+1 , to simulate the xth migration of continuous migration, migrate and continue to culture according to the sample migration method in steps 2-5, and name it "Continuous x" according to the method in steps 2-5;

[0065] Step 4-5: From S x The recovered group 1 was composed of S x-1Migrate to S x The sample "Lian x-1" is recovered and stored in the laboratory to represent the part of the process that has undergone continuous migration;

[0066] Step 5: After completing the continuous migration and continuing the cultivation for several days, recover all the samples, as shown in Figure 6 shown;

[0067] Step 5-1: After the (m-1)th migration and continuing the cultivation, according to the sample recovery method in Step 2-2, recover one set of samples that have been cultivated here all the time from all positions to the laboratory for storage, and name them "S1,..., S m to x" in the way of Step 2-3;

[0068] Step 5-2: Recover one set of samples "Lian m-1" that have migrated continuously from S1 to here from S m and recover them to the laboratory for storage;

[0069] Step 5-3: Recover (m-1) sets of samples "Zhi 1" to "Zhi m-1" that have migrated directly from S1 to here from S m and recover them to the laboratory for storage;

[0070] Step 6: Data analysis: Extract the microorganisms on the surface of the microplastic samples, compare the microorganisms among different groups to explore the transportation effect of microplastics on microorganisms during continuous migration;

[0071] Step 6-1: Immediately after all the microplastic samples are recovered, clean them in situ according to the method in Step 2-2, transport them back to the laboratory as soon as possible under the preservation of ice bags, and perform subsequent microbial analysis as soon as possible after returning to the laboratory;

[0072] Step 6-2: By using the samples that have been cultivated at each position all the time as a control and comparing the microorganisms on the surface of the samples in the continuous migration group, the transportation effect of microplastics on microorganisms under continuous migration can be studied;

[0073] Step 6-3: By comparing the microorganisms on the surface of the samples in the continuous migration group and the direct migration group, the effect of the intermediate position environment on the microorganisms transported on the surface of microplastics during the continuous migration process can be studied;

[0074] Step 6-4: By comparing the samples recovered at the same position in the control group after different cultivation times, the growth changes of the microorganisms on the surface of microplastics at different stages can be studied;

[0075] Step 6-5: By comparing the microorganisms on the surface of the samples cultivated at different times but with the same total cultivation time in S1 and S m respectively, the changes of the microorganisms on the surface of microplastics after migration to the new environment can be studied when the cultivation times in the original environment and the current environment are different.

[0076] Furthermore, the point selection in the present invention can select different numbers of points and continuous migration paths passing through different points according to research requirements.

[0077] Furthermore, the culture time of each point in the present invention may be equal or different.

[0078] In summary, the present invention has the following advantages:

[0079] 1. The present invention is applicable to the simulation of all microplastic migration paths and all microplastic residence times;

[0080] 2. The present invention fully considers the differences between the microbial communities on the surface of microplastics and the microbial communities in the surrounding matrix, and all comparisons are made between the microorganisms on the surface of microplastics;

[0081] 3. The present invention can simultaneously achieve multiple research objectives, including the transport effect of microplastics on microorganisms under continuous migration, the growth of microorganisms on the surface of microplastics at different culture times under in situ culture, and the transport effect of microplastics on microorganisms under migration after different residence times.

[0082] Therefore, the present invention places microplastics with no microorganisms attached to the surface in a wild environment, so that the surface is colonized with the microbial community in the original environment, and then sets up two migration modes, a continuous migration mode in which the geographical location is changed repeatedly for continued cultivation, or a direct migration mode in which the final geographical location is changed directly for cultivation, and retains some microplastics in each environment before and after migration for continued cultivation as a control group, so that the initial surface microorganisms of the microplastics in the environment are known and the migration path is controllable; after recovering the microplastic samples and separating and extracting the surface microorganisms, the changes in the surface microorganisms of the microplastics under continuous migration are studied, thereby explaining the role of microplastics as transport carriers for surface microorganisms during environmental migration.

[0083] The embodiments of the present invention are described in detail as follows.

[0084] 1. In situ culture: Place multiple groups of microplastic samples with sterile surfaces in S A , S B , S C Three points were used for cultivation;

[0085] 1-1. To simulate the process of plastic bottles drifting and migrating with water in the environment, polyethylene terephthalate (PET) plastic bottles, which are the most widely used beverage packaging today, were selected as the research object. PET plastics were cut into microplastics of 3mm×3mm in size. The surface of the microplastics was rinsed 3 times with 75% ethanol and then rinsed 3 times with ultrapure water to make the surface of the PET microplastics sterile at the beginning. 20 pieces of PET microplastics were placed in a nylon mesh bag of 10cm×10cm in size, and the PET microplastics were prevented from contacting the edge of the mesh bag as much as possible. One mesh bag was regarded as one group of microplastic samples.

[0086] 1-2. In order to study the important pathway for microplastics to enter the ocean, namely, from rivers to the sea, S A , S B , S C Three points, continuous migration path from S A To S B To S C , in S A Place 7 groups of microplastic samples in S B Place 3 groups of microplastic samples in S C Place 3 groups of microplastic samples; fix the samples to the shore with nylon ropes and let them float freely on the water surface, simulating the state of PET plastic bottles floating on the water surface;

[0087] 2. First migration: After 15 days of cultivation, some samples were recovered and the positions of some microplastic samples were changed to complete the first migration in the continuous migration;

[0088] 2-1. After 15 days of culture, A Four groups of microplastic samples were recovered from S B , S C One group of microplastic samples were recovered respectively;

[0089] 2-2. After the microplastic samples were dragged to the shore by nylon ropes, the PET microplastics were taken out of the mesh bag and gently washed three times with in-situ water filtered with a 0.45 μm filter membrane to remove the influence of microbial attachment in the surrounding environment. The PET microplastics taken out from the same mesh bag were then placed back into a new mesh bag.

[0090] 2-3. From S A , S B , S C The recovered samples were stored in the laboratory and used as controls to represent the microorganisms on the surface of microplastics at each location, and were recorded as “S A For 1", "S B For 1", "S C 1”;

[0091] 2-4. From SA The recovered samples migrated directly to S C To simulate direct migration, ice packs were used during transportation to protect the microorganisms on the surface of microplastics. C After that, the microplastic sample was placed in the same culture environment as the microplastic sample in step 1-2, recorded as "direct 1";

[0092] 2-5. From S A The two recovered samples migrated to S B To simulate the first migration of continuous migration, ice packs were used during transportation to protect the microorganisms on the surface of microplastics. B Afterwards, the microplastic sample was placed in the same culture environment as the microplastic sample in step 1-2, recorded as "Link 1";

[0093] 3. Second migration: After 20 days of continuous culture, some samples were recovered and the positions of some microplastic samples were changed to complete the second migration in the continuous migration;

[0094] 3-1. After 20 days of continuous cultivation, the S A Recover 2 groups of samples from S B One group of samples were collected from the same spot and the other two groups were collected from S A The samples migrated here from S C Recover a group of samples that have always been cultured at this point;

[0095] 3-2. From S A , S B , S C The samples of each group that had been cultured here were recovered and stored in the laboratory and recorded as "S A For 2, "S B For 2, "S C For 2”;

[0096] 3-3. From S A The recovered samples migrated directly to S C Continue to culture to simulate direct migration, recorded as “direct 2”;

[0097] 3-4. From S B 1 group of recovered A Samples that migrate here migrate to S C The culture was continued to simulate the second migration of the serial migration, which was recorded as “serial 2”;

[0098] 3-5. S B 1 group from S AThe sample "Lian 1" transferred here is recycled to the laboratory for storage to represent the part of the process that has undergone continuous migration;

[0099] 4. After continued cultivation, all samples are recycled;

[0100] 4-1. After 25 days of continued cultivation, according to the recycling method in step 2-2, from S A 、S B 、S C One group of samples that have always been cultivated here are recycled respectively, and are denoted as "S A Dui 3", "S B Dui 3", "S C Dui 3";

[0101] 4-2. Two groups of samples "Zhi 1" and "Zhi 2" that are directly transferred here from S C are recycled from S A ;

[0102] 4-3. One group of samples "Lian 2" that have migrated continuously here are recycled from S C ;

[0103] 5. Data analysis: Extract the bacterial communities on the surface of microplastics, compare the bacterial communities among different groups to explore the transportation effect of PET microplastics on bacterial communities during the process from river to sea;

[0104] 5-1. After all microplastic samples are recycled and cleaned, they are transported back to the laboratory as soon as possible under the preservation of ice packs. After returning to the laboratory, the DNA of the surface bacterial communities is extracted and analyzed as soon as possible; if the extraction cannot be carried out immediately, the samples are stored at -80 °C; the extracted DNA is sequenced to analyze the community composition;

[0105] 5-2. Take the bacterial communities on the surfaces of "S A Dui 1", "S B Dui 1", "S C Dui 1", "S A Dui 2", "S B Dui 2", "S C Dui 2", "S A Dui 3", "S B Dui 3", "S C Dui 3" as controls, and compare the bacterial communities on the surfaces of "Lian 1" and "Lian 2", so as to study the transportation effect of PET microplastics on bacterial communities under continuous migration during the process from river to sea;

[0106] 5-3. By comparing the bacterial communities on the surfaces of "Lian 1", "Lian 2" and "Zhi 1", "Zhi 2", the effect of the intermediate point environment on the transportation of bacterial communities by PET microplastics during the process from river to sea can be studied;

[0107] 5-4. By comparing "SA "Pair 1", "S" A "Pair 2", "S" A For the bacterial communities on the surfaces of "Pair 3", it is possible to study S A In the environment, the growth changes of the bacterial communities on the surfaces of PET microplastics at different stages; by comparing "S" B "Pair 1", "S" B "Pair 2", "S" B For the bacterial communities on the surfaces of "Pair 3", it is possible to study S B In the environment, the growth changes of the bacterial communities on the surfaces of PET microplastics at different stages; by comparing "S" C "Pair 1", "S" C "Pair 2", "S" C For the bacterial communities on the surfaces of "Pair 3", it is possible to study S C In the environment, the growth changes of the bacterial communities on the surfaces of PET microplastics at different stages;

[0108] 5-5. By comparing the bacterial communities on the surfaces of "Straight 1" and "Straight 2", it is possible to study in S A and S C In these two different environments, when the respective cultivation times are different but the total cultivation time is the same, the changes in the bacterial communities on the surfaces of PET microplastics after migration in S C The changes in the surface bacterial communities in the new environment.

[0109] Based on the embodiments of the present invention, all other embodiments and technical substitutions and deformations of the embodiments obtained by those of ordinary skill in the art without departing from the spirit of the present invention and without making creative efforts shall fall within the protection scope of the present invention.

[0110] The above is only the preferred embodiment of the present invention. The embodiments described below with reference to the drawings are exemplary and are only used to explain the present invention, and cannot be construed as a limitation of the present invention. It should be noted that for those of ordinary skill in the art in this technical field, several improvements and refinements made without departing from the principle of the present invention should also be regarded as falling within the protection scope of this application of the present invention.

Claims

1. A method for studying the effect of microplastics on microbial transport under continuous migration, characterized in that: include: The continuous migration path is set to include m points, the first point is placed with 3m-2 groups of microplastic samples with initial surface sterility, and the remaining points are placed with m groups of microplastic samples with initial surface sterility; Recovering a portion of the samples that do not change position to obtain a first control sample group; completing direct migration to obtain a first direct migration sample group; completing the first migration in the continuous migration to obtain a first continuous migration sample group; Recovering part of the samples that do not change position to obtain a second control sample group; recovering part of the first continuous migration sample group, and allowing the rest of the first continuous migration sample group to complete the second migration in the continuous migration to obtain a second continuous migration sample group; completing direct migration to obtain a second direct migration sample group; By analogy with the above steps, recover part of the samples that do not change their positions to obtain the xth control sample group; recover part of the x-1th continuous migration sample group, and the remaining x-1th continuous migration sample group completes the xth migration in the continuous migration to obtain the xth continuous migration sample group; complete the direct migration to obtain the xth direct migration sample group, so that the continuous migration samples reach the last point, and x represents any intermediate point; After the continuous migration is completed, the mth control sample group, the m-1th continuous migration sample group, and the first to m-1th direct migration sample groups are recovered; Surface microorganisms were extracted from all samples of the recovered 1st to mth control sample groups, 1st to m-1th continuous migration sample groups, and 1st to m-1th direct migration sample groups, and the microorganisms between different groups were compared to explore the transport effect of microplastics on microorganisms during continuous migration and complete the microbial data analysis.

2. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The step of obtaining the first control sample group further includes: in situ culturing at each point, after a number of days, recovering m+1 groups of samples from the first point, recovering 1 group of samples from each of the remaining points, and recovering each group of samples recovered from all the points to the laboratory for storage as the first control sample group; The step of obtaining the first direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point, placing them in the same culture environment, as the first direct migration sample group; The step of obtaining the first continuous migration sample group further includes: migrating the m-1 groups of samples recovered from the first point to the second point to simulate the first migration of the continuous migration as the first continuous migration sample group.

3. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The step of obtaining a second control sample group further includes: continuing to culture, after several days, recovering each group of samples cultured at all points to the laboratory for storage as the second control sample group; The step of recovering part of the first continuous migration sample group further includes: recovering a group of first continuous migration sample group samples recovered from the second point to a laboratory for storage; The step of obtaining a second direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point as a second direct migration sample group; The step of obtaining the second continuous migration sample group further includes: migrating the m-2 groups of samples recovered from the second point and migrated thereto from the first point to the third point to simulate the second migration of the continuous migration as the second continuous migration sample group.

4. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The step of obtaining the xth control sample group further includes: after migration and continued cultivation, recovering 2 groups of samples from the first point, recovering 1 group of samples that have always been cultivated at the point from any middle point, m-x+1 groups of samples that have migrated from the first point to this point, and recovering 1 group of samples that have always been cultivated at the point from each of the remaining points; recovering each group of samples that have always been cultivated at the current point from all the points to the laboratory for storage as the xth control sample group; The step of recovering part of the x-1th continuously migrated sample group further includes: recovering a group of samples that migrated from a point above any intermediate point and recovered from the current arbitrary intermediate point to the laboratory for storage; The step of obtaining the xth continuous migration sample group further includes: migrating the mx groups of continuous migration samples recovered from any current intermediate point to any current next point to simulate the xth migration of the continuous migration as the xth continuous migration sample group; The step of obtaining the xth direct migration sample group further includes: directly migrating a group of samples recovered from the first point to the last point as the xth direct migration sample group.

5. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The step of recovering the mth control sample group, the m-1th continuous migration sample group, and the first to m-1th direct migration groups further includes: after the m-1th migration and continued cultivation for several days, recovering one group of samples that have always been cultivated here from all points, recovering one group of samples that have continuously migrated from the first point to here from the last point, and recovering m-1 groups of samples that have directly migrated from the first point to here from the last point.

6. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The steps to complete the microbial data analysis further include: By using samples that were always cultured at various points as controls, the microorganisms on the surface of the samples in the continuous migration group were compared to study the transport effect of microplastics on microorganisms under continuous migration.

7. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The steps to complete the microbial data analysis further include: By comparing the microorganisms on the surface of the continuous migration group samples and the direct migration group samples, the effect of the intermediate point environment on the microorganisms on the surface of microplastic transport during the continuous migration process was studied.

8. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The steps to complete the microbial data analysis further include: By comparing samples recovered from the same point in the control group after different culture times, the growth changes of microorganisms on the surface of microplastics at different stages can be studied.

9. A method for studying the effect of microplastics on microbial transport under continuous migration as claimed in claim 1, characterized in that: The steps to complete the microbial data analysis further include: By comparing the microorganisms on the surface of samples that were cultured for different times at the first point and the last point but with the same total culture time, the changes in the surface microorganisms of microplastics when they migrate to the new environment when the culture time in the original environment and the current environment is different are studied.

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