A bacterial agent composition and its application in water pollution treatment

Abstract

This invention discloses a microbial agent composition, wherein the microbial agent composition contains Tropical Bacillus , Acinetobacter June , Klebsiella variicola , Enterobacter sewers , Kurthia gibsonii and Pseudomonas plecoglossicide When the above-mentioned bacterial agent composition is added to water, water pollution can be treated. Furthermore, the bacteria in the bacterial agent composition all originate from the aquatic environment, effectively avoiding the risk of invasion by exogenous species. Based on this, the present invention also provides a method for water pollution treatment, in which the above-mentioned bacterial agent composition is added to the water body to be treated at a final volume concentration of 9% to 11%, thereby achieving water pollution treatment.

Description

Technical Field

[0001] This invention relates to the field of water pollution control technology, specifically to a bacterial agent composition and its application in water pollution control. Background Technology

[0002] Aquatic products occupy an important position in the global food supply chain, and with the rapid development of aquaculture, the output of various aquatic products has increased significantly. However, the expansion of aquaculture area has not kept pace with the increase in output, which has driven the widespread application of high-density, highly intensive aquaculture models. Under this model, only about 20% to 30% of the high-protein feed fed to fish is effectively absorbed and utilized, while 70% to 80% of the nutrients are discharged into the surrounding environment. This discharge exacerbates the pollution of aquaculture water bodies, accelerates eutrophication of surrounding water bodies, and thus poses a potential threat to ecosystems and human health. At the same time, against the backdrop of the public's growing demand for healthy and green aquatic products and increasingly stringent resource and environmental constraints, the aquaculture industry is facing the dual pressures of economic and social development and ecological environmental protection. This reality requires profound transformation of the industry. Promoting green development to overcome resource and environmental constraints while meeting the demand for high-quality aquatic products is one of the key paths to achieving sustainable development in aquaculture.

[0003] The main reasons for excessive heavy metals in aquaculture water include the following: overuse of feed additives and veterinary drugs (such as disinfectants like copper sulfate and certain antibiotics containing heavy metals), industrial wastewater discharge, and agricultural runoff. Furthermore, in high-density aquaculture, overfeeding leads to increased feed residues, while insufficient water exchange weakens the self-purification capacity, exacerbating heavy metal accumulation. These factors combined result in concentrations of heavy metals such as copper and zinc in aquaculture water frequently exceeding the safety limits stipulated in the "Fishery Water Quality Standard" (Cu≤0.01mg / L, Zn≤0.1mg / L), posing a potential threat to aquatic organisms and food safety. The persistent nature of copper in wastewater stems from its non-degradable elemental nature, i.e., copper ions (Cu... 2+ Humic acid and its complexes cannot be eliminated by mineralization; they can only be removed from water bodies through speciation transformation (precipitation / adsorption) or phase transfer (separation). This process faces three core obstacles: organic ligands such as humic acid and EDTA in the effluent and Cu. 2+The formation of ultra-high stability complexes (complexation constant logK > 18) hinders the approach of precipitant / adsorbent through charge shielding effect, and imparts resistance to hydrolysis (precipitation efficiency < 50% at pH=9) and a negative shift in cathodic reduction potential (requiring -1.2 V vs. SHE), leading to a 50-80% reduction in efficiency compared to traditional technologies. Secondly, the low concentration conditions under strict emission standards (≤ 0.3 mg / L) result in insufficient mass transfer kinetics, causing the adsorption kinetic constant to decrease by 1-2 orders of magnitude, nanofiltration's Cu-citric acid rejection rate to < 50%, and electrodeposition current efficiency to < 20%. Thirdly, the coexistence of Ca... 2+ / Mg 2+ Competitive active sites, Cl - Promotes the production of byproducts and soluble organic matter, quenching free radicals (•OH quenching rate 10). 6 M -1 s -1 Complex matrix interference further inhibits the efficiency of advanced oxidation and biological treatment (microbial diversity decreases by 80%). Furthermore, studies have shown that low concentrations of ammonia and the nitrites generated during its oxidation are significantly toxic to fish, potentially leading to growth retardation, decreased immune function, and even direct threats to survival.

[0004] Therefore, in order to promote the healthy, sustainable and green development of aquaculture and protect the surrounding aquatic ecological environment, effective measures must be taken to strictly control the concentrations of ammonia nitrogen, nitrate, nitrite, phosphate, heavy metals, etc. in aquaculture water.

[0005] Meanwhile, microplastic pollution has become a global concern. Based on their source characteristics, microplastics can be categorized into three main types: (1) household microplastics, including fibers shed during the washing of synthetic textiles and plastic microbeads in personal care products; (2) transportation microplastics, mainly from wear and tear of synthetic rubber tires, brake friction, and road marking removal; and (3) industrial microplastics, represented by accidental leakage of plastic raw material particles. These microplastics mainly enter the marine environment through river transport and direct discharge. In environmental media, microplastics exhibit significant migration capabilities. Their migration pathways mainly include atmospheric transport, river transport, ocean current movement, and biological carriers, enabling microplastics to achieve widespread diffusion across media and regions. Existing research confirms that microplastic pollution has covered various ecosystems globally, from deep-sea sedimentary environments (such as the Mariana Trench) to high-altitude glaciers, and from polar ice caps to tropical soils. Due to their stable polymer structure, microplastics can persist in the environment for hundreds of years. The combined effect of this long-term environmental retention and widespread distribution means that microplastic pollution transcends the geographical boundaries of traditional pollutants, posing a persistent threat to global ecosystems. Current research indicates that no ecosystem has yet been found capable of completely avoiding the impacts of microplastic pollution.

[0006] Microplastics can adsorb and carry various toxic pollutants in water, including inorganic pollutants such as heavy metals and organic pollutants such as PAHs. ] Microplastics such as DDT and PBDEs, in large quantities, have toxic effects on aquatic organisms, posing serious ecological risks. They may also be transferred and bioaccumulated through the food chain, thus threatening human health. Therefore, with the accumulation of microplastics in the environment, microplastic pollution has become a global concern. The most commonly used plastics include PE, PP, PET, PBT, PA, PS, PVC, and PU, with PE and PP accounting for more than 40% of total production. Therefore, in the treatment of microplastics, PE-MPs, a representative pollutant, were selected for degradation experiments.

[0007] Therefore, how to effectively reduce microplastic pollution while controlling the concentrations of ammonia nitrogen, nitrate, nitrite, phosphate, and heavy metals in aquaculture water has become an urgent problem to be solved. Summary of the Invention

[0008] The purpose of this invention is to overcome the above-mentioned shortcomings of the prior art and to provide a microbial agent composition and its application in water pollution control.

[0009] The first objective of this invention is to provide a microbial agent composition.

[0010] A second objective of this invention is to provide the application of the above-mentioned bacterial agent composition in water pollution control.

[0011] A third objective of the present invention is to provide the application of the above-mentioned bacterial agent composition in the adsorption of heavy metals in water.

[0012] A fourth objective of this invention is to provide the application of the above-described microbial agent composition in the degradation of microplastics in water.

[0013] A fifth objective of this invention is to provide the application of the above-described bacterial agent composition in the removal of nitrogen and phosphorus from water.

[0014] The sixth objective of this invention is to provide a method for treating water pollution.

[0015] To achieve the above objectives, the present invention is implemented through the following solution: A microbial agent composition, said microbial agent composition containing Bacillus tropicalis ( Tropical Bacillus Acinetobacter jumbo (), Acinetobacter junii ), Klebsiella variant Klebsiella variicola Enterobacter cloacae () Enterobacter cloacae ), Curtaella gibberella ( Kurthia gibsonii ) and Pseudomonas aeruginosa ( Pseudomonas plecoglossicide ).

[0016] Preferably, in the bacterial agent composition, Bacillus tropicalis (… Tropical Bacillus The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Acinetobacter junei ( Acinetobacter junii The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Klebsiella variants ( Klebsiella variicola The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Enterobacter cloacae ( Enterobacter cloacae The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Coccidioides gibberellus ( Kurtia Gibson's The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Pseudomonas aeruginosa ( Pseudomonas plecoglossicide The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL.

[0017] More preferably, in the bacterial agent composition, Bacillus tropicalis (… Tropical Bacillus The concentration of ) is 3.0 × 10 6 cell / mL, Acinetobacter junei ( Acinetobacter junii The concentration of ) is 3.0 × 10 6 cell / mL, Klebsiella variants ( Klebsiella variicola The concentration of ) is 3.0 × 10 6 cell / mL, Enterobacter cloacae ( Enterobacter sewers The concentration of ) is 3.0 × 10 6 cell / mL, Coccidioides gibberellus ( Kurthia gibsonii The concentration of ) is 3.0 × 10 6 cell / mL, Pseudomonas aeruginosa ( Pseudomonas plecoglossicida The concentration of ) is 3.0 × 10 6 cell / mL.

[0018] More preferably, the tropical Bacillus ( Tropical Bacillus The name is KCTC 33711; the Acinetobacter juni ( Acinetobacter junii The strain is DSM 14968; the variant Klebsiella ( Klebsiella smallpox The ATCC BAA-830 is described as Enterobacter cloacae (…). Enterobacter cloacae The strain is ATCC 222; the *Cootybrio gibrini* (… Kurthia gibsonii The strain is ATCC 43195; the *Pseudomonas aeruginosa* ( Pseudomonas plecoglossicide(This is ATCC 700383.)

[0019] Preferably, the microbial agent composition further contains Scenedesmus sp. , Armed Desmodesmus and / or Poteriospumella vasocystis .

[0020] More preferably, in the microbial agent composition, Scenedesmus sp. The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL Armed Desmodesmus The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL Poteriospumella vasocystis The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL.

[0021] More preferably, in the microbial agent composition, Scenedesmus sp. The concentration is 1.0 × 10⁻⁶. 6 cell / mL Armed Desmodesmus The concentration is 1.0 × 10⁻⁶. 6 cell / mL Poteriospumella vasocystis The concentration is 1.0 × 10⁻⁶. 6 cell / mL.

[0022] More preferably, the Scenedesmus sp. for Scenedesmus sp. FACHB-489 The Armed Desmodesmus for Desmodesmus armatus FACHB-2051 The Poteriospumella vasocyst The classification number in NCBI is NCBI Taxonomy ID: 2976938.

[0023] More preferably, the Poteriospumella vasocystis Obtained from the corresponding author of the prior art DOI: 10.1016 / j.ejop.2022.125915.

[0024] When the above-mentioned bacterial agent composition is added to water bodies, water pollution can be treated. Furthermore, the bacteria and algae involved in the bacterial agent composition are all extracted from water bodies, which can effectively avoid the risk of invasion by exogenous species.

[0025] Therefore, the present invention also seeks protection for the use of any of the above-described bacterial agent compositions in water pollution control.

[0026] The present invention also seeks protection for the use of the above-described bacterial agent composition in the adsorption of heavy metals in water.

[0027] Preferably, the heavy metal is copper (Cu) and / or zinc (Zn).

[0028] The present invention also seeks protection for the use of the above-described bacterial agent composition in the degradation of microplastics in water.

[0029] Preferably, the microplastic is PE-MPS.

[0030] The present invention also claims protection for the use of the above-described bacterial agent composition in the removal of nitrogen and phosphorus from water.

[0031] The present invention also claims protection for a method for treating water pollution, characterized in that the above-described bacterial agent composition is added to the water body to be treated at a final volume concentration of 9% to 11%.

[0032] Preferably, any of the above-described bacterial agent compositions is added to the water body to be treated at a final volume concentration of 10%.

[0033] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a microbial agent composition containing Bacillus tropicalis (Bt). Bacillus tropical Acinetobacter jumbo (), Acinetobacter junii ), Klebsiella variant Klebsiella smallpox Enterobacter cloacae () Enterobacter cloacae ), Curtaella gibberella ( Kurthia gibsonii ) and Pseudomonas aeruginosa ( Pseudomonas plecoglossicida When the above-mentioned bacterial agent composition is added to water, water pollution can be treated. Furthermore, the bacteria in the bacterial agent composition all originate from the aquatic environment, effectively avoiding the risk of invasion by exogenous species. Based on this, the present invention also provides a method for water pollution treatment, in which the above-mentioned bacterial agent composition is added to the water body to be treated at a final volume concentration of 9% to 11%, thereby achieving water pollution treatment. Detailed Implementation

[0034] The present invention will be further described in detail below with reference to specific embodiments. These embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0035] The tropical Bacillus used in the embodiments of the present invention ( Tropical Bacillus KCTC 33711), Acinetobacter juncus ( Acinetobacter junii DSM 14968), Klebsiella variant ( Klebsiella variicola ATCC BAA-830), Enterobacter cloacae ( Enterobacter cloacae ATCC 222), Curvatobrachium gibberi ( Kurtia Gibson's ATCC 43195) and Pseudomonas aeruginosa ( Pseudomonas plecoglossicida All (ATCC 700383) were purchased from https: / / www.biofeng.com / ; the following are examples of the embodiments of this invention. Scenedesmus sp. FACHB-489 and Desmodesmus armatus FACHB-2051 All were purchased from the Freshwater Algae Seed Bank of the Chinese Academy of Sciences; Poteriospumella vasocyst Obtained from the corresponding author of the prior art DOI: 10.1016 / j.ejop.2022.125915.

[0036] The instruments and equipment used in the embodiments of the present invention are shown in Table 1.

[0037] Table 1 Instruments and Equipment

[0038] In the embodiments of this invention, the determination of chemical oxygen demand was carried out in accordance with the standard HJ / T399-2007; the determination of ammonia nitrogen was carried out in accordance with the standard HJ535-2009; the determination of total nitrogen was carried out using the chromotropic acid method; the determination of total phosphorus was carried out in accordance with the standard GB11893-89; the determination of chemical oxygen demand was carried out in accordance with the standard HJ / T399-2007; the determination of heavy metal zinc was carried out in accordance with the standard GB / T 10656-2008; the determination of heavy metal copper was carried out using the BCA spectrophotometric method; the determination of turbidity was carried out in accordance with the standard GB13200-91; and the determination of color was carried out in accordance with the standard GB3143-1982.

[0039] Example 1: Enrichment and purification of bacterial strains and algae I. Experimental Methods The surface water and bottom sediment samples in this embodiment were all from the Dongbian Aquaculture Professional Cooperative in Sanjiao Town, Zhongshan City, Guangdong Province (113°23′26″E, 22°38′20″N). On August 8, 2024, at 4:00 PM, surface water and bottom sediment were collected from 9 ponds (pond 1 to pond 9) of the aquaculture professional cooperative, and were denoted as surface water 1 to surface water 9 and bottom sediment 1 to bottom sediment 9. Two sampling points were selected from each pond, and two samples of surface water and two samples of bottom sediment were collected and combined (the surface water was combined at a volume ratio of 1:1, and the bottom sediment was combined at a mass ratio of 1:1) as the surface water and bottom sediment of that pond.

[0040] 1. Enrichment and purification of microalgae (1) Water sample treatment In a sterile conical flask, add 10 mL of surface water 1 from pond 1, followed by 50 mL of fresh BG-11 liquid culture medium to obtain a water sample from pond 1; the components of the BG-11 liquid culture medium are shown in Table 2.

[0041] Table 2 BG-11 Liquid Culture Medium

[0042] After sealing the pond 1 water sample with sealing film, it was placed in a 27℃ incubator and cultured under continuous light for 24 hours. During the culture process, the conical flask was shaken every 6 hours. After 3 days of culture, the pond 1 water sample was transferred to a 50mL centrifuge tube and centrifuged at 3000rpm for 3min. The precipitate was collected and washed twice with PBS buffer. Then, 50mL of fresh sterile BG-11 liquid culture medium was added to the precipitate and mixed well to obtain the pretreated pond 1 water sample.

[0043] (2) Accumulation of microalgae Take six sterile 2mL centrifuge tubes and label them 1 to 6. Add 900μL of BG-11 liquid culture medium to each of the six centrifuge tubes. Then, add 100μL of pretreated pond 1 water sample to tube 1 and mix well. Take 100μL of the liquid from tube 1 and add it to tube 2 and mix well. Take 100μL of the liquid from tube 2 and add it to tube 3 and mix well. Take 100μL of the liquid from tube 3 and add it to tube 4 and mix well. Take 100μL of the liquid from tube 4 and add it to tube 5 and mix well. Take 100μL of the liquid from tube 5 and add it to tube 6 and mix well.

[0044] Next, 80 μL of liquid from tubes 4, 5 and 6 were taken and evenly spread on solid screening medium (BG-11 liquid medium with a final concentration of 20 g / L agar as shown in Table 2), and placed in a light incubator at 27°C for 24 hours of continuous light to enrich the medium until algal colonies appeared on the medium.

[0045] (3) Isolation and purification of algal strains Add agar to the BG-11 liquid medium formula shown in Table 2 to a final concentration of 20 g / L to obtain solid screening medium. Sterilize the solid screening medium at 121°C for 15 min, then cool it to 70°C~80°C and pour it out for later use.

[0046] Using a sterilized 1μL pipette tip, pick up a single algal colony that appears on the culture medium in step (3), streak it on a solid screening medium, seal it with a sealing film, and then invert it in a light incubator to be cultured under continuous light conditions of 27℃ for 24 hours. Cultivate until a single algal colony grows on the solid screening medium, pick up the single algal colony, and obtain the purified algal strain.

[0047] (4) Identification of algal species The purified algal strain obtained in step (3) was sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing analysis. Homology comparison analysis was performed using the 18S-based ID tool in the Ezbiocloud database and the Blast tool in the NCBI database. The species of microalgae in the purified algal strain was determined by using the highest homology and score of the 18S rRNA gene sequence as the classification standard.

[0048] Surface water 1 was replaced sequentially with surface water 2 to surface water 9, and the species of microalgae in the purified algae strain were identified according to the method shown above.

[0049] 2. Enrichment and purification of bacterial strains (1) Sample pretreatment Add 2g of bottom mud 1 from pond 1 into a centrifuge tube, and prepare 5 centrifuge tubes in parallel (denoted as centrifuge tube 1 to centrifuge tube 5).

[0050] Centrifuge tube 1 culture: Inoculate centrifuge tube 1 into ammonia nitrogen utilization and phosphorus removal medium, and after culture, pick a portion and dilute the bacterial suspension to 10⁻⁶. 1 ~10 6 Then, 80 μL was inoculated into aerobic denitrification phosphorus removal medium 1, and after cultivation, the bacterial suspension was diluted proportionally to 10. 1 ~10 6 Then, 80 μL of each culture was inoculated into aerobic denitrification phosphorus removal medium 1, and after culturing, the bacterial suspension was diluted proportionally to 10⁻⁶. 1 ~10 6 Take 80 μL of each and spread it evenly on a BCIP plate. After incubation until colonies grow, pick the colonies and spread them on a bromothymol blue (BTB) plate. Incubate until colonies grow and pick the colonies to obtain the cultured bacteria in centrifuge tube 1. The culture medium and its components combined during the treatment of centrifuge tube 1 are shown in Table 3.

[0051] Table 3. Culture medium and components of centrifuge tube 1

[0052] Centrifuge tube 2: Centrifuge tube 2 was used for enrichment culture on LB medium. The cultured bacterial suspension was then diluted to 10⁻⁶. 1 ~10 6 Then, 80 μL was evenly spread on the organic matter degradation medium and cultured until a single colony grew. Iodine solution was dropped onto the organic matter degradation medium, and colonies that did not turn blue were picked. They were then separated and purified sequentially on nitrifying bacteria enrichment medium, aerobic denitrifying bacteria enrichment medium, and bromothymol blue medium to obtain the cultured bacteria in centrifuge tube 2. The culture medium and its components combined during the treatment of centrifuge tube 2 are shown in Table 4.

[0053] Table 4. Culture medium and components of centrifuge tube 2

[0054] Centrifuge tube 3: Centrifuge tube 3 was used for enrichment culture on LB medium. The cultured bacterial suspension was then diluted to 10⁻⁶. 1 ~10 6 Then, 80 μL of the culture medium was used for screening and isolation using beef extract peptone medium, heterotrophic ammonia-oxidizing bacteria medium, and heterotrophic nitrite-oxidizing bacteria medium, respectively. The isolated strains were inoculated into BTB medium and cultured until single colonies grew. Colonies in BTB medium that turned blue were selected to obtain the cultured bacteria in centrifuge tube 3. The culture medium and its components used in the treatment of centrifuge tube 3 are shown in Table 5.

[0055] Table 5. Culture media and components of centrifuge tube 3

[0056] Centrifuge tube 4 cultured bacteria: Centrifuge tube 4 was screened and separated sequentially using heterotrophic nitrification enrichment medium, DM denitrification enrichment medium, BTB medium, nitrate beef extract peptone medium and denitrification medium to obtain centrifuge tube 4 cultured bacteria; the culture media and their components combined during the centrifuge tube 4 treatment process are shown in Table 6.

[0057] Table 6. Culture medium and components of centrifuge tube 4

[0058] Centrifuge tube 5 cultured bacteria: Centrifuge tube 5 was screened and separated sequentially using enrichment medium, BTB medium, heterotrophic nitrification medium, aerobic denitrification medium I, aerobic denitrification medium II and composite medium to obtain centrifuge tube 5 cultured bacteria; the culture media and their components combined in the centrifuge tube 5 treatment process are shown in Table 7.

[0059] Table 7. Culture media and components of centrifuge tube 5

[0060] Combine the cultures from centrifuge tube 1 to centrifuge tube 5 to obtain a total of 45 strains selected from all screening.

[0061] Substrate 1 was replaced sequentially with Substrate 2 through Substrate 9, and the same treatment was performed to screen out strains 1 through 45. The screened strains 1 through 45 were then combined. For the combined strains 1 through 45, nitrate reduction tests were performed using nitrate (reduction) biochemical identification tubes (Guangdong Huankai Microbial Technology Co., Ltd., 075320) according to their instructions; nitrate (gas production) biochemical identification tubes (Guangdong Huankai Microbial Technology Co., Ltd., 075620) according to their instructions; nitrite (gas production) biochemical identification tubes (Guangdong Huankai Microbial Technology Co., Ltd., 075630) according to their instructions; and urea tests were performed using urea biochemical identification tubes (Guangdong Huankai Microbial Technology Co., Ltd., 075150) according to their instructions. The test results were recorded. The results will show any positive indicators for the strains sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing. Homology comparison analysis was performed using the 16S-based ID tool in the Ezbiocloud database and the Blast tool in the NCBI database. The species identification of the strains was based on the 16S rRNA gene sequence similarity of ≥98.62%.

[0062] II. Experimental Results 1. Among the purified algal strains, seven microalgae were identified, each belonging to a different family. Scenedesmus sp. "", Chlorella sp. "", Armed Desmodesmus "", Micropore Coelastrum "", Chlamydomonas fasciata "", Poteriospumella vasocystis "and" Chlorella sorokiniana The specific information of the 7 microalgae is shown in Table 3.

[0063] Table 3 Information on 7 microalgae strains

[0064] 2. In the bacterial enrichment and purification experiment, 45 strains were screened and merged. After species identification of the 45 strains, according to the species to which the strains belong and the number of positive biochemical indicators (nitrate gas production, nitrate reduction, nitrite gas production and urea test), 15 strains were selected that were positive for nitrate gas production, nitrate reduction, nitrite gas production and urea test, and one strain of each genus was retained. These 15 strains were named B-1 to B-15. The specific information of the 15 strains is shown in Table 4.

[0065] Table 4. Detailed information on the 15 strains

[0066] Example 2: Wastewater Degradation Test I. Preparation of Synthetic Wastewater 1 (Effective Wastewater) Formula 1 for artificially synthesized wastewater contains 150.0 mg / L inositol, 80.0 mg / L sodium acetate, 180.0 mg / L magnesium sulfate heptahydrate, 10.6 mg / L calcium chloride, 80.0 mg / L sodium bicarbonate, 3.0 mg / L ethylenediaminetetraacetic acid, 0.45 mg / L ferric chloride hexahydrate, 0.036 mg / L manganese chloride hexahydrate, 0.045 mg / L boric acid, 0.036 mg / L zinc sulfate heptahydrate, 0.054 mg / L copper sulfate pentahydrate, 0.054 mg / L potassium iodide, 150.0 mg / L peptone, 80.0 mg / L ammonium chloride, 26.0 mg / L potassium dihydrogen phosphate, and 150.0 mg / L carbon source (glucose), with water as the solvent.

[0067] SBR reactor treatment method: Intelligent artificial climate chamber (500L, RGC-500, Zhengzhou Shengyuan), operated in aerobic mode with external aerator, temperature controlled at 27℃, using intensity 2 and intensity 5 of the intelligent artificial climate chamber (500L, RGC-500, Zhengzhou Shengyuan) for 12h each, treatment time is 7 / 10 days.

[0068] II. Degradation rate test of single bacteria and single algae 1. Experimental Methods The 7 algal strains (shown in Table 3) and 15 bacterial strains (shown in Table 4) obtained from enrichment and purification in Example 1 were cultured to the logarithmic growth phase, respectively. Then, single bacteria and single algae were inoculated into conical flask SBR reactors containing 500 mL of synthetic wastewater 1 (where the final inoculation concentration of bacteria was 1.0 × 10⁻⁶). 6 The final inoculum concentration was 1.0 × 10⁻⁶ cells / mL. 6 The wastewater was treated for 7 days using an SBR reactor (ccells / mL). On days 0, 2, 5, and 7, the chemical oxygen demand (COD), total nitrogen (TNO), and total phosphorus (TP) in the synthetic wastewater 1 inoculated with each single bacterium and algae were measured using a portable multi-parameter water quality analyzer. The COD, TNO, and TPO degradation rates were calculated on day 7. Then, the degradation rates of each single bacterium and algae in the synthetic wastewater 1 were calculated using TNO degradation rate (30% weight), TPO degradation rate (30% weight), and COD degradation rate (40% weight). The synthetic wastewater 1 without inoculation with any single bacterium or algae served as a blank control group. Each single bacterium and algae was measured in triplicate.

[0069] 2. Experimental Results The chemical oxygen demand (COD) of each single bacterium and single algae treating synthetic wastewater 1 is shown in Table 5; the total phosphorus (TP) of each single bacterium and single algae treating synthetic wastewater 1 is shown in Table 6; the total nitrogen (N) of each single bacterium and single algae treating synthetic wastewater 1 is shown in Table 7; and the total degradation rate of each single bacterium and single algae treating synthetic wastewater 1 is shown in Table 8.

[0070] Table 5 Results of Chemical Oxygen Demand Measurement

[0071] Table 6 Total Phosphorus Determination Results

[0072] Table 7 Results of Total Nitrogen Determination

[0073] Table 8. Results of total degradation rate

[0074] The results showed that in the degradation experiments of artificially synthesized wastewater using single bacteria and single algae, 6 out of 15 bacteria and 3 out of 7 algae were able to effectively degrade total nitrogen, total phosphorus, and chemical oxygen demand in the water; the 6 bacteria were respectively Bacillus tropical (B-7) Acinetobacter junii (B-8) Klebsiella variicola (B-9) Enterobacter sewers (B-11) Kurthia gibsonii (B-12) and Pseudomonas plecoglossicida (B-13); the three algae are respectively Scenedesmus sp. (A-2) Armed Desmodesmus (A-5) and Poteriospumella vasocyst (A-9).

[0075] III. Test of Combined Degradation Rate of Bacteria and Algae 1. Experimental Methods Based on the 6 types of bacteria and 3 types of algae identified in step two, purchase the corresponding species of bacteria and algae, specifically as follows: Bacillus tropical (B-7) is KCTC 33711. Acinetobacter junii (B-8) is DSM 14968. Klebsiella smallpox (B-9) is ATCC BAA-830, Enterobacter cloacae (B-11) is ATCC 222. Kurtia Gibson's (B-12) is ATCC 43195. Pseudomonas plecoglossicida(B-13) is ATCC 700383; Scenedesmus sp. (A-2) is Scenedesmus sp. FACHB-489 , Armed Desmodesmus (A-5) is Desmodesmus armatus FACHB-2051 , Poteriospumella vasocystis (A-9) Obtained from the corresponding author of prior art DOI: 10.1016 / j.ejop.2022.125915 (its NCBI classification number is NCBITaxonomy ID: 2976938).

[0076] Next, 23 experimental groups (G-1 to G-23) were set up. Among them, G-1 to G-18 and G-23 used glucose as the carbon source, and G-19 to G-22 used starch as the carbon source. The details of each experimental group are as follows: G-1: A-2×B-7 (indicating inoculation with A-2 algae and B-7 bacteria), with a bacteria-to-algae ratio of 3:1 and a final bacterial concentration of 3.0×10⁻⁶. 6 The final inoculum concentration was 1.0 × 10⁻⁶ cells / mL. 6 cell / mL.

[0077] G-2: A-2×B-8, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those in group G-1.

[0078] G-3: A-2×B-9, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are consistent with those of group G-1.

[0079] G-4: A-2×B-11, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0080] G-5: A-2×B-12, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those of group G-1.

[0081] G-6: A-2×B-13, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0082] G-7: A-5×B-7, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those in group G-1.

[0083] G-8: A-5×B-8, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those of group G-1.

[0084] G-9: A-5×B-9, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those of group G-1.

[0085] G-10: A-5×B-11, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those of group G-1.

[0086] G-11: A-5×B-12, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0087] G-12: A-5×B-13, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0088] G-13: A-9×B-7, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are consistent with those of group G-1.

[0089] G-14: A-9×B-8, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are consistent with those of group G-1.

[0090] G-15: A-9×B-9, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are consistent with those of group G-1.

[0091] G-16: A-9×B-11, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0092] G-17: A-9×B-12, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0093] G-18: A-9×B-13, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0094] G-19: A-5×B-7, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0095] G-20: A-5×B-8, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are the same as those of group G-1.

[0096] G-21: A-5×B-9, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae are consistent with those of group G-1.

[0097] G-22: A-5×B-11, with a bacteria-to-algae ratio of 3:1, and the final inoculation concentrations of bacteria and algae were the same as those in group G-1.

[0098] G-23 (Blank Group): The blank group differs from groups G-1 to G-22 in that no bacteria or algae are added; otherwise, they are completely identical.

[0099] Next, G-1 to G-23 were inoculated into conical flask SBR reactors containing 500 mL of synthetic wastewater 1 at a final volume concentration of 10% (the final concentration of bacteria in 500 mL of synthetic wastewater 1 was 3.0 × 10⁻⁶). 5 The final concentration of algae in 500 mL of synthetic wastewater 1 was 1.0 × 10⁻⁶ cells / mL. 5(cell / mL) was treated in an SBR reactor for 7 days. On days 0, 3 and 7 of treatment, the chemical oxygen demand (COD), ammonia nitrogen, total nitrogen and total phosphorus in each group of synthetic wastewater 1 were measured using a portable multi-parameter water quality analyzer. The COD degradation rate, ammonia nitrogen degradation rate, total nitrogen degradation rate and total phosphorus degradation rate on day 7 of treatment were calculated. Then, the total degradation rate was calculated according to the COD degradation rate, ammonia nitrogen degradation rate, total nitrogen degradation rate and total phosphorus degradation rate with a weight of 30%, 20% and 20% respectively.

[0100] 2. Experimental Results The chemical oxygen demand (COD) of G-1 to G-23 in the combined bacterial and algal degradation rate test is shown in Table 9, the ammonia nitrogen is shown in Table 10, the total nitrogen is shown in Table 11, the total phosphorus is shown in Table 12, and the degradation rate and total degradation rate are shown in Table 13.

[0101] Table 9 Results of Chemical Oxygen Demand Measurement for G-1 to G-23

[0102] Table 10 Ammonia nitrogen determination results for G-1 to G-23

[0103] Table 11 Total Nitrogen Determination Results for G-1 to G-23

[0104] Table 12 Total phosphorus determination results for G-1 to G-23

[0105] Table 13 Degradation rate and total degradation rate results for G-1 to G-23

[0106] The results showed that experimental groups G-6, G-11, G-15, G-16 and G-17 had excellent degradation effects on chemical oxygen demand, ammonia nitrogen, total nitrogen and total phosphorus in synthetic wastewater 1, and showed a better total degradation rate than other experimental groups (ranking among the top 5 in total degradation rate among 24 groups). Therefore, the above 5 experimental groups were used to conduct in-situ tailwater degradation experiments.

[0107] IV. In-situ tailwater degradation rate test 1. Experimental Methods Wastewater from a fish farm belonging to the Dongbian Aquaculture Professional Cooperative in Sanjiao Town, Zhongshan City, Guangdong Province, was filtered through a 0.22μm filter membrane and used as experimental wastewater.

[0108] G-6, G-11, G-15, G-16, and G-17 were inoculated into conical flask SBR reactors containing 500 mL of experimental wastewater at a final volume concentration of 10% (the final concentration of bacteria in 500 mL of synthetic wastewater 1 was 3.0 × 10⁻⁶). 5 The final concentration of algae in 500 mL of synthetic wastewater 1 was 1.0 × 10⁻⁶ cells / mL. 5 The wastewater was treated using an SBR reactor for 7 days (cell / mL). On days 0, 3, 7, and 10 of the treatment, the chemical oxygen demand (COD), ammonia nitrogen (ANO), total nitrogen (TNO), and total phosphorus (TP) in each group of experimental wastewater were measured using a portable multi-parameter water quality analyzer. The COD degradation rate, ANOD degradation rate, TNO degradation rate, and TNO degradation rate were calculated on day 7 of the treatment. Then, the total degradation rate was calculated using COD degradation rate (30% weight), ANOD degradation rate (30% weight), TNO degradation rate (20% weight), and TNO degradation rate (20% weight).

[0109] 2. Experimental Results The results of the in-situ tailwater degradation rate test are shown in Table 14, Table 15, Table 16, Table 17, and Table 18.

[0110] Table 14 Results of Chemical Oxygen Demand Measurement in In-situ Tailwater Degradation Rate Test

[0111] Table 15 Results of ammonia nitrogen determination in in-situ tailwater degradation rate test experiment

[0112] Table 16 Results of Total Nitrogen Determination in In-situ Tailwater Degradation Rate Test

[0113] Table 17 Results of Total Phosphorus Determination in In-situ Tailwater Degradation Rate Test

[0114] Table 18 Degradation rate and total degradation rate results in the in-situ tailwater degradation rate test experiment.

[0115] The results showed that the bacterial and algal combinations shown in G-6, G-11, G-15, G-16 and G-17 could effectively degrade chemical oxygen demand, ammonia nitrogen, total phosphorus and total nitrogen in actual effluent, thus effectively degrading the actual effluent.

[0116] V. Degradation Test of the Mixture in Wastewater 1. Experimental Methods Set up group 6B, group 3A6B, and blank group respectively, as follows: Group 6B: Contains KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195, and ATCC700383, with a concentration of 3.0 × 10⁻⁶ for each of the six bacteria. 6 cell / mL.

[0117] Group 3A6B: Contains KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195, ATCC700383. Scenedesmus sp. FACHB-489 , Desmodesmus armatus FACHB-2051 and Poteriospumella vasocystis The concentration of bacteria in each sample was 3.0 × 10⁻⁶. 6 The concentration of algae was 1.0 × 10⁻⁶ cells / mL. 6 cell / mL.

[0118] Blank group 1: Replace the bacteria and algae with an equal amount of distilled water.

[0119] Next, groups 6B, 3A6B, and control group 1 were inoculated into conical flask SBR reactors containing 500 mL of synthetic wastewater 1 at a final volume concentration of 10% (the final concentration of bacteria in 500 mL of synthetic wastewater 1 was 3.0 × 10⁻⁶). 5 The final concentration of algae in 500 mL of synthetic wastewater 1 was 1.0 × 10⁻⁶ cells / mL. 5 (cell / mL) was treated in an SBR reactor for 10 days. On days 0, 3, 7 and 10 of treatment, the chemical oxygen demand (COD), ammonia nitrogen, total nitrogen and total phosphorus in the synthetic wastewater 1 of each group were measured using a portable multi-parameter water quality analyzer. The degradation rates of COD, ammonia nitrogen, total nitrogen and total phosphorus were calculated. Then, the total degradation rate was calculated according to the COD degradation rate, ammonia nitrogen degradation rate, total nitrogen degradation rate and total phosphorus degradation rate with a weight of 30%, 20% and 20% respectively.

[0120] 2. Experimental Results The chemical oxygen demand (COD) determination results in the tailwater degradation test of the mixture are shown in Table 19, the ammonia nitrogen determination results are shown in Table 20, the total nitrogen determination results are shown in Table 21, the total phosphorus determination results are shown in Table 22, and the degradation rate and total degradation rate determination results are shown in Table 23.

[0121] Table 19 Results of Chemical Oxygen Demand (COD) Measurement in the Tailwater Degradation Test of the Mixture

[0122] Table 20 Ammonia nitrogen determination results in the tailwater degradation test of the mixture

[0123] Table 21 Total nitrogen determination results in the tailwater degradation test of the mixture

[0124] Table 22 Total phosphorus determination results in the tailwater degradation test of the mixture

[0125] Table 23 Degradation rate and total degradation rate results of the mixture degradation rate test experiment

[0126] The results showed that in the effluent degradation experiments of groups 6B and 3A6B, the chemical oxygen demand (COD), ammonia nitrogen content, total nitrogen content, and total phosphorus content in the synthetic wastewater 1 decreased significantly with increasing treatment time. This indicates that the mixture of six bacteria (KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195, and ATCC 700383) and the bacteria themselves significantly reduced the chemical oxygen demand (COD), ammonia nitrogen content, total nitrogen content, and total phosphorus content in the synthetic wastewater 1. Scenedesmus sp. FACHB-489 , Desmodesmus armatus FACHB-2051 and Poteriospumella vasocystis The mixture of 6 bacteria and 3 algae can effectively degrade the wastewater and achieve water pollution control.

[0127] Example 3 Heavy metal degradation test I. Experimental Methods 1. Preparation of wastewater samples Artificially synthesized wastewater 2 contains 75.0 mg / L inositol, 40.0 mg / L sodium acetate, 90.0 mg / L magnesium sulfate heptahydrate, 5.3 mg / L calcium chloride, 40.0 mg / L sodium bicarbonate, 1.5 mg / L ethylenediaminetetraacetic acid, 0.225 mg / L ferric chloride hexahydrate, 0.018 mg / L manganese chloride hexahydrate, 0.0225 mg / L boric acid, 0.018 mg / L zinc sulfate heptahydrate, 0.027 mg / L copper sulfate pentahydrate, 0.027 mg / L potassium iodide, 75.0 mg / L peptone, 40.0 mg / L ammonium chloride, and 18.0 mg / L potassium dihydrogen phosphate, with water as the solvent.

[0128] Using Cu 2+ The stock solution will contain Cu from artificially synthesized wastewater 2 2+The concentration was adjusted to 2 mg / L, using Zn 2+ The stock solution will contain Zn from artificially synthesized wastewater 2 2+ The concentration was adjusted to 1 mg / L; of which Cu 2+ Preparation method for 50 mL of stock solution: The solution was prepared using copper chloride dihydrate CuCl2·2H2O. 0.1355 g of CuCl2·2H2O powder (purity: 99%) was weighed quantitatively using an analytical balance, dissolved in pure water, and diluted to a volumetric flask of 50 mL. The solution was then filtered to remove bacteria.

[0129] Zn 2+ Preparation method for 50 mL of stock solution: ZnCl2 was used for preparation. 0.1053 g of ZnCl2 (purity: 99%) was weighed quantitatively using an analytical balance, dissolved in a small amount of hydrochloric acid solution, and diluted to a volumetric flask with pure water. The solution was then filtered to remove bacteria.

[0130] Group 3B: Contains DSM 14968, ATCC BAA-830, and ATCC 43195, with a concentration of 3.0 × 10⁻⁶ for each of the three bacteria. 6 cell / mL.

[0131] Group 6B: Contains KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195, and ATCC700383, with a concentration of 3.0 × 10⁻⁶ for each of the six bacteria. 6 cell / mL.

[0132] Group 3A6B: Contains KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195, ATCC700383. Scenedesmus sp. FACHB-489 , Desmodesmus armatus FACHB-2051 and Poteriospumella vasocystis The concentration of bacteria in each sample was 3.0 × 10⁻⁶. 6 The concentration of algae was 1.0 × 10⁻⁶ cells / mL. 6 cell / mL.

[0133] Groups 3B, 6B, and 3A6B were inoculated into conical flask SBR reactors containing 500 mL of synthetic wastewater 2 at a final volume concentration of 10% (the final concentration of bacteria in 500 mL of synthetic wastewater 1 was 3.0 × 10⁻⁶). 5 The final concentration of algae in 500 mL of synthetic wastewater 1 was 1.0 × 10⁻⁶ cells / mL. 5The samples were treated using an SBR reactor for 10 days (cell / mL). The concentrations of heavy metals copper and zinc in each group of synthetic wastewater 2 were measured daily during each treatment period using a portable multi-parameter water quality analyzer. The degradation rates of heavy metals copper and zinc were calculated on the 8th and 10th days of treatment.

[0134] II. Experimental Results The concentration results of heavy metal zinc are shown in Table 24, and the concentration results of heavy metal copper are shown in Table 25.

[0135] Table 24 Results of Concentration Determination of Heavy Metal Zinc

[0136] Table 25 Results of concentration determination of heavy metal copper

[0137] The results showed that when treating synthetic wastewater 2, groups 6B and 3A6B exhibited significantly higher degradation rates of zinc than group 3B on day 8, and were also effective in degrading zinc on day 10. For copper degradation, groups 6B and 3A6B showed significantly better degradation effects, achieving rates exceeding 80% when treating the wastewater.

[0138] Example 4 Microplastic Degradation Test I. Experimental Methods 1. Microplastic pretreatment Cleaning and drying: Remove impurities and contaminants from the surface of the microplastics (PE-MPS) to obtain cleaned and dried microplastics; Particle size control: After cleaning and drying, the microplastics are pulverized or screened to a specific particle size of 500µm to obtain pulverized microplastics; Sterilization treatment: Sterilize the pulverized microplastics. Weigh 2g of PE-MPs and pour it into a 150ml Erlenmeyer flask. Immerse the microplastics in 2% (w / w) SDS, 75% (w / w) ethanol and 95% (w / w) ethanol respectively and wash them thoroughly for 3 hours. After immersion, rinse three times with sterile water. Dry in a 55℃ oven to constant weight. Pour the completely dried PE-MPs into a wide-mouth blue-cap reagent bottle for storage to obtain pretreated microplastics.

[0139] 2. Degradation test KCTC 33711, DSM 14968, ATCC BAA-830, ATCC 222, ATCC 43195 and ATCC 700383 were cultured to the logarithmic phase using LB medium to obtain bacterial suspensions for each strain.

[0140] PEB group: 1 mL of bacterial culture (OD 1.0) of each bacterium was inoculated into 400 mL of mineral salt culture medium containing 2 g (initial mass) of pretreated microplastics (PE-MPS, sole carbon source). The culture was incubated at 37℃ and 200 rpm for 60 days. The OD and pH values ​​of the culture medium were recorded on day 0 and day 60 of incubation. The final mass of the microplastics on day 60 was also recorded, and the degradation rate of the microplastics was calculated. The degradation rate was calculated as (initial mass - final mass) / initial mass. The determination was performed in quadruplicate. The mineral salt culture medium contains 1.0 g of MgSO4·7H2O, 1.0 g of NH4NO3, 1.0 g of KH2PO4, 1.0 g of K2HPO4, 1 mL of 1000× trace element culture medium and 1 L of pure water, with a pH of 7.0 to 7.2. The 1000× trace element culture medium contains 5g FeCl3, 2g CaCl2, 0.5g NaCl, 0.2g FeSO4·7H2O, 0.2g ZnSO4·7H2O, 0.1g MnSO4·H2O, and 100mL of pure water, with concentrated sulfuric acid added dropwise until completely dissolved.

[0141] The difference between the PE group and the PEB group is that no bacterial solution is added to the PE group, but the other treatments are exactly the same.

[0142] The difference between Group B and Group PEB is that 1 mL of bacterial suspension (OD1.0) of each bacterium was inoculated into mineral salt culture medium. No microplastics were added to Group B, and the other treatments were exactly the same.

[0143] II. Experimental Results The test results for the PEB group, PE group, and B group are shown in Table 25.

[0144] Table 25 Test results for PEB group, PE group and B group

[0145] In the three replicates of the PEB group, the final mass of microplastics after 60 days of culture was 1.5106 g, 1.444 g, and 1.5005 g, respectively, and the degradation rates of microplastics were 24.47%, 27.80%, and 24.98%, respectively, indicating that the combination of the six strains can effectively degrade microplastics.

[0146] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description and ideas, and it is neither necessary nor possible to exhaustively describe all implementation methods here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A microbial agent composition, characterized in that, The bacterial agent composition contains Bacillus tropicalis (… Bacillus tropicus Acinetobacter jumbo (), Acinetobacter junii ), Klebsiella variant Klebsiella variicola Enterobacter cloacae () Enterobacter cloacae ), Curtaella gibberella ( Kurthia gibsonii ) and Pseudomonas aeruginosa ( Pseudomonas plecoglossicida ).

2. The microbial agent composition according to claim 1, characterized in that, In the bacterial agent composition, Bacillus tropicalis (… Bacillus tropicus The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Acinetobacter junei ( Acinetobacter junii The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Klebsiella variants ( Klebsiella variicola The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Enterobacter cloacae ( Enterobacter cloacae The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Coccidioides gibberellus ( Kurthia gibsonii The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL, Pseudomonas aeruginosa ( Pseudomonas plecoglossicida The concentration of ) is 2.7–3.3 × 10⁻⁶. 6 cell / mL.

3. The microbial agent composition according to claim 2, characterized in that, The tropical spores ( Bacillus tropicus The name is KCTC 33711; the Acinetobacter juni ( Acinetobacter junii The strain is DSM 14968; the variant Klebsiella ( Klebsiella variicola The ATCC BAA-830 is described as Enterobacter cloacae (…). Enterobacter cloacae The strain is ATCC 222; the *Cootybrio gibrini* (… Kurthia gibsonii The strain is ATCC 43195; the *Pseudomonas aeruginosa* ( Pseudomonas plecoglossicida (This is ATCC 700383.) 4. The microbial agent composition according to claim 1, characterized in that, The microbial agent composition also contains Scenedesmus sp. , Desmodesmus armatus and / or Poteriospumella vasocystis .

5. The microbial agent composition according to claim 4, characterized in that, In the bacterial agent composition Scenedesmus sp. The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL Desmodesmus armatus The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL Poteriospumella vasocystis The concentration is 0.9–1.1 × 10⁻⁶. 6 cell / mL.

6. The microbial agent composition according to claim 5, characterized in that, The Scenedesmus sp. for Scenedesmus sp. FACHB-489 The Desmodesmus armatus for Desmodesmus armatus FACHB- 2051 The Poteriospumella vasocystis The classification number in NCBI is NCBI Taxonomy ID: 2976938.

7. The use of the bacterial agent composition according to any one of claims 1 to 6 in the adsorption of heavy metals in water.

8. The use of the bacterial agent composition according to any one of claims 1 to 6 in the degradation of microplastics in water.

9. The use of the bacterial agent composition according to any one of claims 1 to 6 in the removal of nitrogen and phosphorus from water.

10. A method for treating water pollution, characterized in that, The bacterial agent composition according to any one of claims 1 to 6 is added to the water body to be treated at a final volume concentration of 9% to 11%.

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