Paenibacillus, fungicide, preparation method and application of paenibacillus in multi-drug-resistant escherichia coli

Abstract

The invention discloses a Paenibacillus sp.HNYJ2209J, the preservation number of the Paenibacillus sp.HNYJ2209J is CGMCC (China General Microbiological Culture Collection Center) No.31256, the preservation date of the Paenibacillus sp.HNYJ2209J is July 10, 2024, the preservation unit of the Paenibacillus sp.HNYJ2209J is China General Microbiological Culture Collection Center, and the preservation number of the Paenibacillus sp.HNYJ2209J is CGMCC No.31256. The preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The preparation method of the biological agent is simple, the biological agent can achieve a good antibacterial effect on multiple drug-resistant bacteria and escherichia coli when being used in any mode of oral administration, accompanying materials or drinking water, and the biological agent is convenient to use, free of antibiotic residues, efficient and safe.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and biological products, and specifically relates to a strain of Paenibacillus, a bacterial agent, a preparation method and application thereof to multidrug-resistant Escherichia coli. Background Art

[0002] As an important zoonotic pathogen, Escherichia coli's drug resistance is a worrying issue. Currently, the treatment of Escherichia coli infection mainly relies on antibiotics. The development and utilization of anti-resistant microorganisms and their active product resources are considered to be a viable alternative to antibiotics. Numerous studies have shown that medicinal plant endophytes and their active metabolites have the potential to develop new drugs and solve the problem of bacterial multidrug resistance.

[0003] Endophytes in medicinal plants refer to those that can exist in medicinal plants and independently synthesize active products that are the same or similar to those of medicinal plants. The discovery of secondary metabolites of endophytic microorganisms in plants with antiviral, antitumor and antibacterial activities provides important research ideas for the development and production of natural medicines.

[0004] Studies have shown that endophytes in Chinese medicinal materials can produce active substances with antibacterial, anti-tumor and antioxidant effects, as well as anti-insect, anti-diabetic and other effects. Bacillus subtilis isolated from Alternantherabrasiliana can produce the same bioactive compounds as its host plant. Two isomers, pestacin and isopestacin, were isolated from the endophytic fungi of the medicinal plant iris, both of which showed antibacterial and antioxidant activities; aromatic sesquiterpenes isolated from the metabolites of endophytic fungi of Viburnum plants had significant killing activity against Plasmodium falciparum; endophytic fungi of Coptis chinensis (Phomopsis sp) can produce berberine, the main active alkaloid of Coptis chinensis; Stierle et al. discovered the anticancer compound paclitaxel from the secondary metabolites of the endophytic fungi Taxomyces andreanae of Taxus brevis; metabolites produced by the endophytic fungi Nigrospora sp. of the medicinal plant Aconitum had significant antiviral activity against influenza virus H1N1; Castillo et al. isolated four peptide compounds from the endophytic fungi Streptomyces NRRL 30562 of the medicinal plant Kennedianigriscans, which had broad-spectrum antibacterial activity; the endophytic fungi Fusarium isolated from Catharanthus roseus (Fusarium oxysporum) can produce the anticancer substance vincristine; the Streptomyces sp. microorganisms isolated from the medicinal plant Aconite can produce a 2,2′-bipyridine compound with antifungal activity; the endophytic actinomycetes from the seeds of Camptotheca acuminata can produce the secondary metabolite camptothecin.

[0005] Medicinal plants and their endophytes are important medicinal resources, which are conducive to the research and development of natural medicines. Once developed, these endophytes can be directly associated with actual production applications, generating huge economic and social value, which is of great significance to increasing the productivity of natural products and the protection of medicinal plants. Therefore, increasing the research on endophyte metabolic components and obtaining new effective ingredient pathways are the focus of future research. Further in-depth understanding of the dynamic interaction between endophytes and host plants and the origin of endophyte genes will be conducive to the huge development of the strategic industry of endophytes in traditional Chinese medicine.

[0006] Based on this, the present invention intends to isolate, identify and screen endophytes from different tissues such as roots, stems, leaves and flowers of Forsythia suspensa, and study their antibacterial effects and antibacterial active substances, laying a foundation for the screening of new antibacterial active substances and providing a reference for the research of new anti-resistant microbial drugs. Summary of the invention

[0007] In view of the current prevention and control difficulties of Escherichia coli disease in veterinary clinics, such as the difficulty in prevention and control, strong bacterial resistance and the lack of specific drugs, the present invention provides a strain of Bacillus, a bacterial agent, a preparation method and its application on multidrug-resistant Escherichia coli, separates, identifies and screens endophytes from different tissues such as roots, stems, leaves and flowers of Forsythia suspensa, and studies their antibacterial effects and antibacterial active substances, laying a foundation for the screening of new antibacterial active substances, providing a reference for the research of new anti-resistant microbial drugs, and providing ideas for the improvement and research of antibiotic substitutes in livestock and poultry production.

[0008] The solution adopted by the present invention to solve its technical problem is: a strain of Paenibacillus, wherein the Paenibacillus is Paenibacillus sp. HNYJ2209J, with a deposit number of CGMCC No.31256, a deposit date of July 10, 2024, a deposit unit of the General Microbiology Center of China National Microbiological Culture Collection Administration, and a deposit address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0009] Furthermore, the Paenibacillus HNYJ2209J is screened from Forsythia suspensa and its plant tissues.

[0010] The present invention also provides a Paenibacillus biological agent, comprising Paenibacillus HNYJ2209J and / or a metabolite of Paenibacillus HNYJ2209J.

[0011] The present invention also provides a method for preparing a Paenibacillus biological agent, comprising the following steps:

[0012] Step 1, inoculate Paenibacillus HNYJ2209J onto a tryptic soy agar solid medium and culture at 28-30°C for 24-48 hours;

[0013] Step 2: Take the cultured bacteria in step 1 and inoculate them into tryptic soy liquid culture medium, and culture them at 28-30° C. for 24-48 hours to obtain seed liquid;

[0014] Step 3, inoculating the seed liquid into the fermentation medium at an inoculum rate of 2-3%, fermenting and culturing for 24-48 hours at 28-30°C, a rotation speed of 80-120rpm, and a pH of 6.8-7.2, and collecting the fermentation liquid to obtain a microbial agent;

[0015] Step 4: Spray the fermentation liquid obtained in step 3 into dry powder using a powder spraying device to obtain microbial powder.

[0016] Furthermore, the live bacteria concentration of the Bacillus biological agent is 10-13×10 12 cfu / mL.

[0017] The present invention also provides the use of a strain of Paenibacillus or a biological bacterial agent prepared therefrom on multi-drug resistant Escherichia coli.

[0018] Furthermore, the invention is used for preventing and treating multidrug-resistant Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5).

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention provides a strain of Paenibacillus, a bacterial agent, a preparation method and an application thereof on multidrug-resistant Escherichia coli. The Paenibacillus HNYJ2209J is separated and screened from Forsythia suspensa and its tissues, and by evaluating its antibacterial activity, it can be obtained that the diameter of the antibacterial zone of the Paenibacillus HNYJ2209J on Escherichia coli is greater than 20 mm, indicating that the Paenibacillus HNYJ2209J has a strong antibacterial effect on "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase, that is, the biological bacterial agent prepared based on the Bacillus has a strong antibacterial and bactericidal effect on multidrug-resistant Escherichia coli.

[0021] Moreover, the present invention has been verified by multiple experiments, and the biological bacterial agent prepared by the Paenibacillus subtilis HNYJ2209J of the present invention has a 100% prevention and treatment effect on Escherichia coli disease in laying hens; the biological bacterial agent prepared by the Paenibacillus subtilis HNYJ2209J of the present invention has a good prevention and treatment effect on lamb diarrhea, and can effectively solve the prevention and treatment problem of lamb diarrhea; the biological bacterial agent prepared by the Paenibacillus subtilis HNYJ2209J of the present invention has a good prevention and treatment effect on Escherichia coli disease in calves, and can effectively solve the prevention and treatment problem of Escherichia coli disease in calves.

[0022] The biological agent of the present invention has a simple preparation method, can achieve good antibacterial effect on multi-drug resistant bacteria: Escherichia coli when used orally, with food or in drinking water, is easy to use, has no antibiotic residue, and is highly effective and safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The antibacterial activity of Paenibacillus HNYJ2209J against "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5), wherein A: the antagonistic effect of Paenibacillus HNYJ2209J against "super-resistant" Escherichia coli carrying NDM1 / NDM5; B: the antagonistic effect of Bacillus gelatinus against "super-resistant" Escherichia coli carrying NDM4 / NDM5;

[0024] Figure 2The antibacterial activity of Paenibacillus HNYJ2209J against "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5) by the inhibition zone method; A: The antagonistic effect of Paenibacillus against "super-resistant" Escherichia coli carrying NDM1 / NDM5; B: The antagonistic effect of Paenibacillus against "super-resistant" Escherichia coli carrying NDM4 / NDM5;

[0025] Figure 3 The colony morphology of Paenibacillus sp. HNYJ2209J and the bacterial morphology under a microscope after Gram staining, including the colony morphology (left) and Gram staining (right) photos of strain HNYJ2209J;

[0026] Figure 4 This is the phylogenetic tree of Paenibacillus HNYJ2209J;

[0027] Figure 5 The results show the effect of the fermentation supernatant of Bacillus subtilis HNYJ2209J on "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5), where A: the fermentation broth of HNYJ2209J strain has an antibacterial effect on "super-resistant" Escherichia coli carrying NDM1 / NDM5; B: the fermentation broth of HNYJ2209J strain has an antibacterial effect on "super-resistant" Escherichia coli carrying NDM4 / NDM5. DETAILED DESCRIPTION

[0028] The present invention is further described below in conjunction with embodiments.

[0029] See also Figure 1-5 The present invention provides a strain of Paenibacillus, a bacterial agent, a preparation method and a technical solution for its application on multidrug-resistant Escherichia coli: the instruments and equipment involved in the following embodiments are conventional instruments and equipment unless otherwise specified; the reagents and culture media involved are conventionally commercially available unless otherwise specified; the test methods involved are conventional methods unless otherwise specified. The multidrug-resistant Escherichia coli used in the test is the multidrug-resistant Escherichia coli used in the Journal of Biotechnology Bulletin, 2022, 28(9): 106-115, "Identification and Drug Resistance Study of Escherichia coli Producing Carbapenemase NDM5 and NDM Simultaneously"; and the Escherichia coli used in the Whole Genome Sequencing Analysis of Porcine Escherichia coli Carrying New Delhi Metallo-β-lactamase and Drug Screening Study of Sensitive Plant Extracts, Chinese Journal of Veterinary Drugs. 2023, 57(6): 23-33.

[0030] Example 1: Isolation and screening of Paenibacillus sp. HNYJ2209J

[0031] (1) Isolation and purification of Paenibacillus sp. HNYJ2209J

[0032] The surface of different tissues of Forsythia suspensa, such as roots, stems, and leaves, were disinfected, and then inoculated on PDA and LB culture media, cultured at 28-30℃ for 48-72h, and colonies of different morphologies were picked and purified on TSA culture media, and then the same strains were eliminated by morphological characteristics under a microscope. The purified strains were stored at -80℃ with 10% skim milk powder.

[0033] (2) Screening of antibacterial endophytes

[0034] Endophytes stored at -80℃ were selected and inoculated into TSA culture plates. The antibacterial activity was tested by streak method. A line was drawn in the center of the TSA culture plate and inoculated with endophytes. Then, Escherichia coli was inoculated on both sides of the streaked endophytes. The plates were cultured in a 28-30℃ incubator for 24 hours. The antibacterial activity of endophytes against multi-drug resistant bacteria was observed. Figure 1 .

[0035] Depend on Figure 1 It can be seen that Bacillus sp. HNYJ2209J has a good antibacterial and bactericidal effect on "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5).

[0036] In order to further obtain the strength of the inhibition, the size of the inhibition zone was measured by the inhibition zone method. The endophytes stored at -80°C were selected and inoculated into the center of the TSA plate, and then 1×10 9 The large intestine is spread around the endophytes, placed in a 37°C incubator for 24 hours, and the antibacterial effect of the endophytes on the pathogens is observed. If there is a transparent inhibition zone around the endophytes, it means that this strain of endophytes has an antibacterial and bactericidal effect on the pathogens. The strength of the antibacterial effect is indicated by the size of the inhibition zone. The diameter of the inhibition zone > 20mm indicates a strong antibacterial and bactericidal effect, 10mm < inhibition zone < 20mm indicates a medium antibacterial and bactericidal effect, and an inhibition zone < 10mm indicates a weak antibacterial and bactericidal effect. See details. Figure 2 , where A: The inhibition zone size of Bacillus against "super-resistant" Escherichia coli carrying NDM1 / NDM5 is 43mm; B: The inhibition zone size of Bacillus against "super-resistant" Escherichia coli carrying NDM4 / NDM5 is 47mm.

[0037] Depend on Figure 2 It can be seen that the diameter of the inhibition zone of Bacillus sp. HNYJ2209J against Escherichia coli is greater than 20 mm, indicating that Bacillus sp. HNYJ2209J has a strong antibacterial effect on "super-resistant" Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5).

[0038] (3) Observation of endophytic bacteria morphology

[0039] The strain HNYJ2209J was inoculated on a TSA culture plate and placed in a 30°C incubator for 24 h. The colony morphology of the endophytic bacteria was observed. Figure 3 Left). The colonies of strain HNYJ2209J cultured on TSA solid medium are milky white with round protrusions. After Gram staining, it is purple, a Gram-positive bacterium with rod-shaped spores and oval shape ( Figure 3 right).

[0040] Depend on Figure 3 It can be seen that strain HNYJ2209J forms round milky white colonies on the solid culture medium, the surface of the colonies is smooth and moist, and Gram staining is positive.

[0041] Depend on Figure 3 It can be seen that the endophyte is oval in shape, with blunt or truncated ends, and is a Gram-negative bacterium, 1.0-1.2μm×3-5μm. The spore is oval and located in the center of the body. The body is solitary. Combined with the morphological observation results of the colony and the body, it was preliminarily determined that the endophyte was Bacillus.

[0042] (4) Molecular identification of endophytes

[0043] 16S rRNA and 23S rRNA were used for PCR amplification and sequencing identification. The primers used are shown in Table 1 below.

[0044] Table 1 16S rRNA and 23S rRNA primer sequences

[0045]

[0046] Endophyte DNA was used as template and 50 μL amplification system was used for amplification. The amplified product was first electrophoresed using 1% agarose gel to determine the size and position of the band. The PCR product was then sent to Shanghai Shenggong Biotechnology Co., Ltd. for sequencing. The sequence is shown in SEQ ID NO.1.

[0047] TGGCGGCGTGCCTAATACATGCAAGTCGAGCGGGGTTGTTTAGAAGCTTGCTTCTAAACAACCTAGCGGCGGACGGGTGAGTAACACGTAGGCAACCTGCCCACAAGACAGGGATAACTACCGGAAACGGTAGCTAATACCCGATACATCCTTTTCCTGCATGGGAGAAGGAGGAAAGACGGAGCAATCTGTCACTTGTGGATGGGCCTGCGGCGCATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGGCGAAAGCCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGCCAGGGAAGAACGTCTTGTAGAGTAACTGCTACAAGAGTGACGGTACCTGAGAAGAAAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTCTTTAAGTCTGGTGTTTAATCCCGAGGCTCAACTTCGGGTCGCACTGGAAACTGGGGAGCTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGGCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGCTAGGTGTTAGGGGTTTCGATACCCTTGGTGCCGAAGTTAACACATTAAGCATTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCAGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCTCTGACCGGTCTAGAGATAGACCTTTCCTTCGGGACAGAGGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGT CGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGCTTAGTTGCCAGCAGGTCAAGCTGGGCACTCTAAGCAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGAC(SEQ ID NO.1)

[0048] The sequencing results were analyzed and compared in the NCBI database, and the phylogenetic tree was constructed using the Neighbor Joining method in MEGA5.0 software. Figure 4 .

[0049] from Figure 4 It can be seen that there are 19 strains with high homology to HNYJ2209J, with a homology of more than 99%, of which the homology to Paenibacillus is 100%. According to the sequencing results, HNYJ2209J strain is determined to be Paenibacillus.

[0050] (5) Antibacterial activity test of Paenibacillus HNYJ2209J fermentation broth

[0051] In order to further verify the antibacterial and bactericidal effect of the endophytic fermentation broth, the HNYJ2209J strain was fermented and the activity of the fermentation broth was detected. 100 μL of endophytic fermentation broth was added to each plate, and 4 replicates were added. After 24 hours of culture, the size of the inhibition zone was measured. Figure 5 Among them, A: The inhibition zone of Paenibacillus against Escherichia coli carrying NDM1 / NDM5 "super-resistant" is 29±0.20mm; B: The inhibition zone of Paenibacillus against Escherichia coli carrying NDM4 / NDM5 "super-resistant" is 27±0.22mm.

[0052] Example 2: Preparation of Bacillus sp. HNYJ2209J bacterial agent

[0053] (1) Strain culture and preparation

[0054] Paenibacillus HNYJ2209J was inoculated onto tryptone soy agar plates and cultured at 30°C for 24 h;

[0055] (2) Preparation of seed solution

[0056] The bacteria cultured in step (1) were inoculated into a tryptone soy broth medium and cultured at 30° C. for 24 h to obtain a seed solution;

[0057] (3) Liquid fermentation production

[0058] The seed liquid was inoculated into the fermentation medium at a rate of 3%, and fermented at 30°C, 120 rpm, and pH 7.0 for 48 hours. The fermentation liquid was collected to obtain a microbial agent, and the live bacterial concentration of the agent was 10-13×10 12 cfu / mL.

[0059] (4) Preparation of powder

[0060] The fermentation liquid obtained in step (3) is sprayed into dry powder using a powder spraying device to obtain a microbial powder.

[0061] Test Example 1: Experimental verification of the Bacillus HNYJ2209J bacterial agent (bacterial powder prepared in Example 2) in the prevention and treatment of Escherichia coli disease in laying hens

[0062] This experimental verification was conducted in a laying hen farm in Shangqiu in April 2023. Amoxicillin and blank were used as controls.

[0063] The specific steps are as follows:

[0064] (1) A total of 500 laying hens with Escherichia coli infection were selected and divided into experimental groups with different bacterial agents, amoxicillin control group and blank control group.

[0065] (2) Paenibacillus HNYJ2209J and amoxicillin were used for treatment, and the treatment effect was observed and statistically analyzed. The results of the treatment experiment are shown in Table 2:

[0066] Table 2 The control effect of Paenibacillus HNYJ2209J on Escherichia coli in laying hens

[0067]

[0068]

[0069] It can be seen from Table 2 that both the 5× microbial agent experimental group and the 10× microbial agent experimental group had 100% control effect on Escherichia coli disease in laying hens.

[0070] Test Example 2: Experimental verification of the effect of Paenibacillus HNYJ2209J bacterial agent (bacterial powder prepared in Example 2) on the prevention and treatment of Escherichia coli diarrhea in lambs

[0071] The test verification was carried out in a sheep farm in Xinxiang in February 2023. The lambs on the sheep farm had diarrhea, excreting milky white or grayish white, pasty feces, smelly and sticky. After clinical symptom observation, autopsy changes and laboratory diagnosis, it was confirmed that the lambs had diarrhea.

[0072] An oral therapeutic trial was conducted on sick lambs using 5× bacterial agent. During the treatment, the clinical symptoms of the sick lambs gradually alleviated and returned to normal; all sick lambs were cured after 3 days of treatment.

[0073] In summary, the experimental results show that the biological bacterial agent prepared by the Bacillus sp. HNYJ2209J of the present invention has a good preventive and therapeutic effect on lamb diarrhea, and can effectively solve the problem of prevention and treatment of lamb diarrhea.

[0074] Test Example 3: Experimental verification of the Bacillus HNYJ2209J bacterial agent (bacterial powder prepared in Example 2) in the prevention and treatment of calf Escherichia coli disease

[0075] This experimental verification was conducted in a cattle farm in Shangqiu in December 2022. Enrofloxacin and blank were used as controls.

[0076] The specific steps are as follows:

[0077] (1) Fifty calves with diarrhea were selected and divided into five groups, with 10 calves in each group, including different bacterial agent experimental groups, enrofloxacin control group and blank control group.

[0078] (2) Paenibacillus HNYJ2209J and enrofloxacin were used for treatment, and the treatment effects were observed and statistically analyzed. The results of the treatment experiment are shown in Table 3:

[0079] Table 3 Effect of Paenibacillus HNYJ2209J inoculation on the prevention and treatment of Escherichia coli disease in calves

[0080]

[0081] It can be seen from Table 3 that both the 5× microbial agent experimental group and the 10× microbial agent experimental group had 100% control effect on calf Escherichia coli disease.

[0082] Test Example 4: Experimental verification of the Bacillus HNYJ2209J bacterial agent (bacterial powder cultured in Example 2) in the prevention and treatment of calf Escherichia coli disease

[0083] The test verification was conducted in a beef cattle farm in Nanzhao County in March 2023. Some calves in the beef cattle farm had diarrhea, blood clots in their feces, foul odor, cellulose flakes, and mucous membranes. After clinical symptom observation and laboratory diagnosis, it was confirmed to be calf Escherichia coli disease.

[0084] A therapeutic trial was conducted on sick calves using 10× bacterial agent. During the treatment, the clinical symptoms of the sick calves gradually alleviated, the body temperature gradually returned to normal, the appetite increased, and the cough and diarrhea symptoms alleviated and returned to normal; all sick calves were cured after 3 days of treatment.

[0085] In summary, the experimental results prove that the biological bacterial agent prepared by the Paenibacillus sp. HNYJ2209J of the present invention has a good prevention and treatment effect on calf Escherichia coli disease, and can effectively solve the problem of prevention and treatment of calf Escherichia coli disease.

[0086] The above description is only a preferred embodiment of the present invention and does not limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A strain of Paenibacillus, characterized in that: The bacillus is a bacillus Paenibacillus sp. )HNYJ2209J, deposit number: CGMCC No.31256, deposit date: July 10, 2024, deposit unit: General Microbiology Center of China Culture Collection Administration, deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

2. The Paenibacillus according to claim 1, characterized in that: The Paenibacillus HNYJ2209J is screened from Forsythia suspensa and its plant tissues.

3. A bacillus biological agent, characterized in that: It includes Paenibacillus sp. HNYJ2209J and / or metabolites of Paenibacillus sp. HNYJ2209J.

4. A method for preparing a Paenibacillus biological agent, characterized in that: The following steps are involved: Step 1, inoculate Paenibacillus HNYJ2209J onto a tryptic soy agar solid medium and culture at 28-30°C for 24-48 hours; Step 2: Take the cultured bacteria in step 1 and inoculate them into tryptic soy liquid culture medium, and culture them at 28-30°C for 24-48h to obtain seed liquid; Step 3, inoculating the seed liquid into the fermentation medium at an inoculation rate of 2-3%, fermenting and culturing for 24-48 hours at 28-30°C, a rotation speed of 80-120rpm, and a pH of 6.8-7.2, and collecting the fermentation liquid to obtain a microbial agent; Step 4: Spray the fermentation liquid obtained in step 3 into dry powder using a powder spraying device to obtain microbial powder.

5. The method for preparing the Paenibacillus biological agent according to claim 4, characterized in that: The live bacteria concentration of the Bacillus biological agent is 10-13×10 12 cfu / mL.

6. Application of a strain of Bacillus or a biological agent prepared based on it on multidrug-resistant Escherichia coli.

7. The use according to claim 6, characterized in that: Its application in preventing and treating multidrug-resistant Escherichia coli carrying New Delhi metallo-β-lactamase (NDM1 / NDM4 / NDM5).

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