Bacterium capable of degrading pyrethroid pesticides, and preparation and application thereof
By using a mixed bacterial agent of pantrophic paracoccus BB-2 and Pseudomonas aeruginosa BS-3 to synergistically degrade pyrethroids, the high cost and low efficiency of existing technologies for treating pyrethroid pollution have been solved, achieving efficient and economical water and soil remediation.
Patent Information
- Application Number
- CN202510821901.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-19
- Publication Date
- 2026-01-23
AI Technical Summary
Existing technologies for treating pyrethroid pesticide pollution are ineffective in removing pyrethroid pollutants from water bodies. Chemical oxidation is costly and produces highly toxic intermediates, adsorption methods have low saturation adsorption capacity and require frequent replacement, and biological methods have low degradation rates.
A mixed bacterial agent of pantrophic paracoccus BB-2 and Pseudomonas aeruginosa BS-3 is used to efficiently degrade pyrethroid pesticides by secreting extracellular hydrolases and oxidoreductases through synergistic action. The preparation method includes cultivation, freeze drying and mixing in biochar and glycerol carrier.
It achieves a high degradation rate of over 90% for pyrethroids, simplifies the treatment process, reduces costs, minimizes environmental harm, and is suitable for the remediation of water and soil pollution.
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Figure CN121379865A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the field of microbial application, and particularly relates to a bacterium capable of degrading pyrethroid pesticides, a preparation thereof and application. BACKGROUND
[0002] Pyrethroids are synthetic pyrethroid insecticides widely used in agricultural production, veterinary, landscape maintenance and home. With the increase of application amount, pyrethroids have been detected in rural, urban and even animal and human tissues. Many research results show that pyrethroid pesticides have biological toxicity to fish, bees and earthworms in the environment, in addition, pyrethroid drugs also affect the reproductive system and endocrine system, and also increase the risk of 3-11 year-old children suffering from developmental and nervous system diseases. Through the research of Christopher DeMars, since 2011, the use of pyrethroids has been increased in the central coast of California, and in Monterey County alone, the agricultural use of bensultap and bifenthrin has increased by about 50%, and around 2011-2018, the positive detection rate of the two is also increasing (DeMars, Christopher, et al. Assessment of Pyrethroid Contamination and Potential Mitigation Strategies in California Central Coast Surface Waters [J]. Journal of Environmental Management, 2021, 278), which poses a potential threat to wild animals and plants and humans. At the same time, near the Pearl River Delta, Zhao's detection, 7 kinds of pyrethroid pesticides were detected, among which bensultap and bifenthrin had high content (Zhao, L. N, et al. Residue and risk assessment of 7 kinds of pyrethroids in water environment in the Pearl River Delta [D]. Shanghai Ocean University, Shanghai, China, 2014). The frequent detection of pyrethroids in water and soil has potential impact on the ecological environment and human health.
[0003] Currently, the water remediation technologies applied to pyrethroid pesticide pollution include chemical oxidation method, adsorption method, biological degradation method and membrane filtration method, etc. The chemical oxidation technology can completely degrade and remove pyrethroid pesticide pollution, but still has the disadvantages of high operation cost and poor in-situ treatment capacity, and the intermediate products generated are more toxic and more difficult to degrade. The adsorbents such as diatomite and activated carbon have low saturated adsorption capacity of pyrethroid, and need to be frequently replaced or regenerated, and the regeneration process (such as thermal desorption) has high energy consumption and may damage the structure of the adsorbent. In addition, the adsorption method only transfers the pollutants from the water body to the solid phase, and does not realize complete decomposition. The saturated adsorbent needs to be further treated as hazardous waste, increasing the disposal cost. The biological method becomes the green preferred solution for pyrethroid pollution treatment due to its environmental friendliness, low cost and in-situ remediation. However, the actual site is complex, and the indigenous microorganisms compete for nutrients, which makes the degradation rate of pyrethroid-degrading bacteria low.
[0004] Therefore, the present application provides a kind of bacteria for pyrethroid pollutant degradation, its preparation and application, to provide a kind of fast, efficient, cheap, stable method for degrading pyrethroid in water. SUMMARY
[0005] In view of the problems of poor pyrethroid pollutant removal effect and poor economy, the present application provides a kind of bacteria capable of degrading pyrethroid pesticides and its preparation and application. The bacteria include Paracoccus pantotrophus BB-2 or Pseudomonas aeruginosa BS-3, and also include a mixed bacterial agent of Paracoccus pantotrophus BB-2 and Pseudomonas aeruginosa BS-3. By utilizing the different physiological and biochemical characteristics and metabolic pathways of Paracoccus pantotrophus BB-2 and Pseudomonas aeruginosa BS-3, the synergistic effect of different functional degrading microorganisms can be achieved, which can effectively degrade pyrethroid in water and improve water quality.
[0006] Technical scheme of the present application:
[0007] In a first aspect, the present application provides a Paracoccus pantotrophus capable of degrading pyrethroid pesticides, which is named Paracoccus pantotrophus BB-2 and was preserved in China Center for Type Culture Collection on May 26, 2025, located at No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, with the preservation number of CCTCC NO: M20251184.
[0008] In a second aspect, the present application provides a Pseudomonas aeruginosa capable of degrading pyrethroid pesticides, which is named Pseudomonas aeruginosa BS-3 and was deposited with the China Center for Type Culture Collection on May 26, 2025 at the address of No. 299, Bajiyi Road, Wuchang District, Wuhan City, Hubei Province, China, and has the accession number of CCTCC NO: M20251185.
[0009] In a third aspect, the present application provides a microbial agent capable of degrading pyrethroid pesticides, which comprises Paracoccus pantotrophus BB-2 as described herein or Pseudomonas aeruginosa BS-3 as described herein.
[0010] In some embodiments, the microbial agent comprises Paracoccus pantotrophus BB-2 and Pseudomonas aeruginosa BS-3.
[0011] In some embodiments, the microbial agent comprises Paracoccus pantotrophus BB-2 and Pseudomonas aeruginosa BS-3.
[0012] In some embodiments, the microbial agent further comprises a carrier.
[0013] In some embodiments, the mass ratio of the microorganism to the carrier in the microbial agent is 1:1.
[0014] In some embodiments, the carrier is a mixture of biochar and glycerol, and the mass ratio of biochar to glycerol in the carrier is 8:1.
[0015] In a fourth aspect, the present application provides a preparation method of a microbial agent capable of degrading pyrethroid pesticides as described herein, which comprises the following steps:
[0016] (1) culturing Paracoccus pantotrophus BB-2 or Pseudomonas aeruginosa BS-3 as described herein in a culture medium, and concentrating and freeze-drying the obtained bacterial solution to obtain Paracoccus pantotrophus BB-2 bacterial powder or Pseudomonas aeruginosa BS-3 bacterial powder;
[0017] (2) optionally, mixing the Paracoccus pantotrophus BB-2 bacterial powder and the Pseudomonas aeruginosa BS-3 bacterial powder to obtain a mixed bacterial powder;
[0018] (3) optionally, uniformly mixing the Paracoccus pantotrophus BB-2 bacterial powder, the Pseudomonas aeruginosa BS-3 bacterial powder, or the mixed bacterial powder thereof with a carrier to obtain a composite microbial agent.
[0019] In some embodiments, the culture medium comprises LB liquid medium.
[0020] In some embodiments, in step (1), Paracoccus pantotrophus BB-2 or Pseudomonas aeruginosa BS-3 is cultured to 108 ~10 9 CFU / mL.
[0021] In some embodiments, the mass ratio of the mixed Parapseudomonas BB-2 bacterial powder and Pseudomonas aeruginosa BS-3 bacterial powder in step (2) is 1:3-3:1.
[0022] In some embodiments, the mass ratio of the mixed microorganism and the carrier in step (3) is 1:1.
[0023] In some embodiments, the carrier is a mixture of biochar and glycerol, and the mass ratio of biochar to glycerol in the carrier is 8:1.
[0024] In a fifth aspect, the present application provides a culture, fermentation, metabolite or intracellular substance of Parapseudomonas BB-2, Pseudomonas aeruginosa BS-3 or the microbial agent as described herein.
[0025] In a sixth aspect, the present application provides the use of Parapseudomonas BB-2, Pseudomonas aeruginosa BS-3, the microbial agent or the culture, fermentation, metabolite or intracellular substance thereof as described herein in degrading pyrethroid pesticides.
[0026] In some embodiments, the use comprises degrading pyrethroid pesticides in a natural environment.
[0027] In some embodiments, the natural environment comprises a water body or soil.
[0028] In some embodiments, the pyrethroid pesticides comprise lambda-cyhalothrin, deltamethrin, cypermethrin, bifenthrin.
[0029] Advantages of the present application
[0030] The present application screens Parapseudomonas BB-2 and Pseudomonas aeruginosa BS-3 by domestication and passage culture, and each of the two bacteria has good pyrethroid degradation effect. More particularly, the present application finds that Parapseudomonas BB-2 and Pseudomonas aeruginosa BS-3 can synergistically coexist, secrete extracellular hydrolases and various oxidoreductases, and efficiently degrade pyrethroids, with removal efficiency of more than 90% for various pyrethroids. By adding the mixed microbial agent of the present application, the harm of pyrethroids to the ecological environment can be effectively controlled, which is simple, efficient and reliable, and has a broad application prospect. BRIEF DESCRIPTION OF DRAWINGS
[0031] Figure 1 The degradation effect of various single bacteria screened in Example 1 on bifenthrin.
[0032] Figure 2The degradation effect of the bacteria screened in Example 1 on bifenthrin was determined.
[0033] Figure 3 The degradation effect of the composite microbial agent of Example 2 on various pyrethroids was determined. DETAILED DESCRIPTION
[0034] The present application will be further described in conjunction with specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and devices used in the present application are conventional reagents, methods and devices in the technical field.
[0035] Example 1: Screening of Bifenthrin-degrading Bacteria
[0036] (1) 1 g of soil sample was taken from various bifenthrin pesticide pollution areas in the country and added to MSM medium containing 50 mg / L of bifenthrin. After incubation at 30°C and 140 r / min for 7 days, the culture was transferred to MSM medium containing 100 mg / L of bifenthrin at a 5% (v / v) inoculation ratio and incubated for another 7 days. The concentration of bifenthrin in the medium was gradually increased to reach a final concentration of 800 mg / L.
[0037] (2) After domestication, the final culture was diluted and plated on LB plates containing 100 mg / L of bifenthrin and incubated at 30°C and 140 r / min for 3 days. Single colonies were picked and successively streaked until strains with consistent colony morphology and rapid growth were obtained. Four strains were obtained, which were identified as Pseudomonas aeruginosa, Paracoccus pantotrophus, Micrococcus luteus and Pseudomonas putida by 16S rDNA. The four strains were stored at -50°C in 60% glycerol.
[0038] To determine the final immobilized microorganisms, we conducted bifenthrin degradation experiments with the four strains Figure 1 ). According to the physiological and biochemical characteristics of different bacteria, four groups of bacteria were selected for synergistic degradation experiments, including Paracoccus pantotrophus + Pseudomonas aeruginosa, Paracoccus pantotrophus + Micrococcus luteus, Pseudomonas putida + Pseudomonas aeruginosa, and Pseudomonas putida + Micrococcus luteus, numbered as groups 1, 2, 3 and 4 Figure 2 The following is a detailed description of the degradation experiment.
[0039] Degradation experiment: Bifenthrin standard solution was added to 50 mL of inorganic salt liquid culture medium to adjust its concentration to 100 mg / L. The medium was then shaken for 30 min at 30℃ and 140 rpm in the dark. 5% bacterial suspension was inoculated into the above inorganic salt culture medium; the medium was then incubated at 30℃ and 140 rpm in the dark for 7 consecutive days. A blank control was set up with no inoculated culture medium. Each treatment was performed in triplicate. Samples were taken and extracted at day 7, and the degradation efficiency of the strain was calculated after high-performance liquid chromatography (HPLC) detection.
[0040]
[0041] A1 – Bifenthrin pesticide residue concentration (mg / L) after treatment with degrading bacteria
[0042] A0 – Bifenthrin pesticide residue concentration (mg / L) after control treatment
[0043] Degradation experiment results are as follows Figure 1 and Figure 2 As shown, only the combination of *Paracoccus pantotrophus* and *Pseudomonas aeruginosa* achieved a synergistic effect. The *Paracoccus pantotrophus* strain screened in this embodiment is named *Paracoccus pantotrophus* BB-2, and it has been deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: M20251184, deposited on May 26, 2025. The *Pseudomonas aeruginosa* strain screened in this embodiment is named *Pseudomonas aeruginosa* BS-3, and it has been deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: M 20251185, deposited on May 26, 2025.
[0044] Example 2: Preparation of Mixed Microbial Agent
[0045] (1) Paracoccus BB-2 and Pseudomonas aeruginosa BS-3 were cultured separately in LB liquid medium. When the colony-forming units of the two bacteria reached 102, the colony-forming units were determined to be 102. 8 ~10 9 At CFU / mL, the two bacterial solutions were centrifuged and concentrated, and then freeze-dried to obtain the original bacterial powders of *Paragonimococcus pantrophicans* BB-2 and *Pseudomonas aeruginosa* BS-3, which were then frozen and stored at -20°C.
[0046] (2) Mix the original bacterial powder of Paracoccus BB-2 and the original bacterial powder of Pseudomonas aeruginosa BS-3 at a mass ratio of 1:3 to obtain a bacterial powder mixture. Mix the mixture with the carrier (biochar:glycerol=8:1) at a mass ratio of 1:1 to prepare a mixed bacterial agent.
[0047] Example 3: Degradation of various pyrethroids by the mixed bacterial agent
[0048] In 50 mL of inorganic salt liquid medium, high-efficiency chlorofluoro-cyhalothrin, deltamethrin, cypermethrin, and bifenthrin standard solutions were added respectively to adjust the concentration to 100 mg / L, and then shaken at 30℃ and 140 rpm in the dark for 30 min. The bacterial agent with a mass ratio of 1% was inoculated into the above inorganic salt medium, and then continuously cultured at 30℃ and 140 rpm in the dark for 7 days. A blank control without inoculation of the bacterial agent was set, and each treatment was set in triplicate. Sampling and extraction were performed at 7 d, and the degradation efficiency of the bacterial agent was calculated after high-performance liquid chromatography detection:
[0049]
[0050] A1 - Pyrethroid pesticide residue concentration (mg / L) after degradation of the bacterial agent
[0051] A0 - Pyrethroid pesticide residue concentration (mg / L) after control treatment
[0052] The degradation experiment results are shown in Table 2. Figure 3 It can be seen that the mixed bacterial agent prepared in Example 2 achieves good degradation effect on various pyrethroid pesticides.
[0053] It should be noted that the specification and drawings of the present application give the preferred embodiments of the present application, but the present application can be implemented in many different forms and is not limited to the embodiments described in the specification. These embodiments are not additional limitations on the content of the present application, and the purpose of providing these embodiments is to make the disclosure of the present application more thorough and comprehensive. Furthermore, the above technical features continue to be combined with each other to form various embodiments not listed above, which are all considered to be within the scope of the present application. Furthermore, for those skilled in the art, the above description can be improved or changed, and all these improvements and changes should be within the scope of the appended claims of the present application.
Claims
1. Paracoccus pantotrophus capable of degrading pyrethroid pesticides, characterized in that, The Paraprevotella sp. is named Paraprevotella sp. BB-2, which was preserved in China Center for Type Culture Collection on May 26, 2025, and the preservation number is CCTCC NO: M 20251184.
2. A Pseudomonas aeruginosa capable of degrading pyrethroid pesticides, characterized in that, The Pseudomonas aeruginosa is named Pseudomonas aeruginosa BS-3, which was preserved in China Center for Type Culture Collection on May 26, 2025, and the preservation number is CCTCC NO: M 20251185.
3. A bacterial agent capable of degrading pyrethroid pesticides, characterized in that, The microbial agent comprises the Paraprevotella sp. BB-2 according to claim 1 or the Pseudomonas aeruginosa BS-3 according to claim 2.
4. The bacterial agent of claim 3, characterized in that, The microbial agent comprises the Paraprevotella sp. BB-2 and the Pseudomonas aeruginosa BS-3.
5. The bacterial agent of claim 4, characterized in that, The microbial agent comprises the Paraprevotella sp. BB-2 and the Pseudomonas aeruginosa BS-3 in a mass ratio of 1:3 to 3:
1.
6. The inoculant of any one of claims 3-5, wherein, The microbial agent further comprises a carrier; Further, the mass ratio of the microorganism to the carrier in the microbial agent is 1:
1. Further, the carrier is a mixture of biochar and glycerol, and the mass ratio of biochar to glycerol in the carrier is 8:
1.
7. A method for preparing a microbial agent capable of degrading pyrethroid pesticides according to any one of claims 3 to 6, characterized in that, The method comprises the following steps: (1) culturing the Paraprevotella sp. BB-2 according to claim 1 or the Pseudomonas aeruginosa BS-3 according to claim 2 in a culture medium, and centrifuging and freeze-drying the obtained bacterial solution to obtain Paraprevotella sp. BB-2 bacterial powder or Pseudomonas aeruginosa BS-3 bacterial powder; (2) optionally, mixing the Paraprevotella sp. BB-2 bacterial powder and the Pseudomonas aeruginosa BS-3 bacterial powder to obtain mixed bacterial powder; (3) optionally, uniformly mixing the Paraprevotella sp. BB-2 bacterial powder, the Pseudomonas aeruginosa BS-3 bacterial powder, or the mixed bacterial powder with a carrier to obtain a composite microbial agent.
8. The preparation method according to claim 7, characterized in that, In step (1), Parvularubrum curvum BB-2 or Pseudomonas aeruginosa BS-3 was cultured to 10 8 ~10 9 CFU / mL; Further, the mass ratio of the Paraprevotella sp. BB-2 bacterial powder to the Pseudomonas aeruginosa BS-3 bacterial powder mixed in step (2) is 1:3 to 3:
1. Further, the mass ratio of the microorganism to the carrier mixed in step (3) is 1:
1. Further, the carrier is a mixture of biochar and glycerol, and the mass ratio of biochar to glycerol in the carrier is 8:
1.
9. A culture, fermentation, metabolite, or intracellular substance of the Paraprevotella sp. BB-2 of claim 1, the Pseudomonas aeruginosa BS-3 of claim 2, or the microbial agent of any one of claims 3-6.
10. Use of the Paraprevotella sp. BB-2 of claim 1, the Pseudomonas aeruginosa BS-3 of claim 2, the microbial agent of any one of claims 3-6, or the culture, fermentation, metabolite, or intracellular substance of claim 9 in degrading pyrethroid pesticides; Further, the use comprises degrading pyrethroid pesticides in a natural environment. Further, the natural environment comprises water or soil. Further, the pyrethroid pesticides comprise lambda-cyhalothrin, deltamethrin, cypermethrin, and bifenthrin.