Lactobacillus plantarum with broad-spectrum antibacterial function

Abstract

The invention discloses a broad-spectrum antibacterial lactobacillus plantarum, and belongs to the technical field of microorganisms. The lactobacillus plantarum 9-6 is separated from animal feed, the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.32568, the lactobacillus plantarum 9-6 and fermentation liquor thereof have an obvious inhibition effect on common vibrio parahaemolyticus, vibrio alginolyticus, vibrio harveyi and vibrio vulnificus, and the lactobacillus plantarum 9-6 and the fermentation liquor thereof have the advantages that the lactobacillus plantarum 9-6 and the fermentation liquor thereof have the obvious inhibition effect; and in addition, the antibacterial agent has a relatively good antibacterial effect on common staphylococcus aureus, escherichia coli, listeria monocytogenes, salmonella enterica and bacillus cereus. The lactobacillus plantarum 9-6 has probiotic safety, can grow under an initial pH condition of 4-10 and a high-salt environment, and plays a role in bacteriostasis under an acidic condition. The invention lays a foundation for developing a good antibacterial probiotic preparation and application of a fermentation culture thereof.

Description

Technical Field

[0001] The present invention relates to a Lactobacillus plantarum with broad-spectrum antibacterial activity, belonging to the field of microbial technology. Background Art

[0002] Vibrio is a genus of Gram-negative pathogenic bacteria widely present in marine and freshwater environments. Among them, Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi, and Vibrio vulnificus are important pathogens in the aquaculture industry, food industry, and public health field. These vibrios can cause explosive diseases in aquatic animals (such as shrimp and fish), resulting in serious economic losses. At the same time, they can infect humans through contaminated aquatic products or wounds, potentially leading to health risks such as food poisoning and sepsis. Currently, the prevention and control of vibrios mainly rely on antibiotics and chemical disinfectants, but long-term use is likely to lead to enhanced drug resistance, environmental pollution, and food safety hazards. There is an urgent need to develop green and sustainable alternative solutions.

[0003] In recent years, probiotics have received extensive attention due to their natural antibacterial properties, safety, and environmental friendliness. Lactic acid bacteria (such as Lactobacillus plantarum) as a group of generally recognized as safe probiotics can inhibit the growth of pathogenic bacteria through mechanisms such as acid production, competitive inhibition, and secretion of antibacterial peptides (such as bacteriocins), showing application potential in animal feed, aquaculture, and food preservation. However, there are few strains with broad-spectrum anti-vibrio activity in existing studies, and many strains have insufficient adaptability to complex environments (such as high salt, extreme pH, and gastrointestinal stress), which limits their actual application effects. Therefore, there is an urgent need to find a lactic acid bacterium with broad-spectrum anti-vibrio activity and excellent tolerance. Summary of the Invention

[0004] The purpose of this part is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments.

[0005] The present invention provides a Lactobacillus plantarum 9-6, which is deposited in the China General Microbiological Culture Collection Center, deposit number: CGMCC No. 32568, deposit date: November 11, 2024, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0006] The present invention provides a microbial preparation containing the Lactobacillus plantarum L. plantarum 9-6.

[0007] In one embodiment, in the microbial preparation, the cell count of Lactobacillus plantarum 9-6 is not less than 4×10 9CFU / mL or 8×10 11 CFU / g of wet bacterial cells.

[0008] The present invention provides postbiotics prepared from the Lactobacillus plantarum 9-6, and the postbiotics contain the fermentation broth, fermentation supernatant and / or freeze-dried powder of the Lactobacillus plantarum 9-6.

[0009] The present invention provides a method for preparing the postbiotics, and the method is any one of the following:

[0010] (1) Ferment the Lactobacillus plantarum 9-6 described in claim 1, and collect the fermentation broth after fermentation;

[0011] (2) After fermenting the Lactobacillus plantarum 9-6 described in claim 1, centrifuge to remove the bacterial cells, and collect the fermentation supernatant;

[0012] (3) Freeze-dry the fermentation broth obtained after fermenting the Lactobacillus plantarum 9-6 described in claim 1, and collect the freeze-dried powder.

[0013] The present invention also provides a product containing the Lactobacillus plantarum 9-6, the microbial preparation or the postbiotics.

[0014] In one embodiment, the product includes food, medicine, health product or feed additive.

[0015] The present invention also provides the application of the Lactobacillus plantarum 9-6, the microbial preparation or the postbiotics in the preparation of a bacterial inhibitor.

[0016] In one embodiment, the bacteria include one or more of Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella enterica, Bacillus cereus.

[0017] In one embodiment, the bacterial inhibitor includes an oral preparation and / or a topical preparation.

[0018] Beneficial effects:

[0019] The present invention isolated a strain of Lactobacillus plantarum 9-6 from a feed sample, and the preservation number is CGMCC No. 32568. The Lactobacillus plantarum 9-6 has good antibacterial effects against Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella enterica, Bacillus cereus.

[0020] The Lactobacillus plantarum 9-6 provided by the present invention has probiotic safety, can grow under the initial pH conditions of pH = 4 - 10 and in a high-salt environment of ≤100 g / L, and this bacterium is sensitive to a variety of antibiotics. The antibacterial component of this strain acts on Vibrio by secreting it extracellularly, and this antibacterial component has pH stability and temperature stability and functions under acidic conditions.

[0021] Biological material preservation

[0022] A strain of Lactobacillus plantarum 9-6, taxonomically named Lactobacillus plantarum, was deposited at the General Microbiological Center of the China Committee for Culture Collection of Microorganisms on November 11, 2024, with the deposit number CGMCC No. 32568 and the deposit address at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Description of the drawings

[0023] Figure 1 Colony characteristics and Gram staining of strain 9-6;

[0024] Figure 2 Antibacterial spectrum of strain 9-6;

[0025] Figure 3 Growth curve of strain 9-6 under different NaCl concentrations;

[0026] Figure 4 Growth curve of strain 9-6 under different initial pH conditions;

[0027] Figure 5 Antibacterial effect after reconstitution of the freeze-dried sample of the fermentation supernatant of strain 9-6;

[0028] Figure 6 Experiment on excluding hydrogen peroxide from the antibacterial component;

[0029] Figure 7 Optimal pH of the antibacterial active component;

[0030] Figure 8 pH stability of the antibacterial component in the fermentation broth supernatant;

[0031] Figure 9 Temperature stability of the antibacterial component in the fermentation broth supernatant;

[0032] Figure 10 Comparison of the vibrio-inhibiting abilities of the fermentation supernatants of strain 9-6 and commercial strains. Detailed implementation manners

[0033] It should be noted that the following detailed description is illustrative and aims to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention pertains.

[0034] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application.

[0035] The preparation method of the Lactobacillus plantarum 9-6 bacterial suspension involved in the following examples is as follows: Inoculate the single colony on the plate into MRS liquid medium and culture it in a shaker at 37°C and 220 rpm for 5-6 h until OD 600nm = 1.0 to obtain the seed liquid, inoculate it into MRS liquid medium at an inoculation amount of 5% (v / v), and culture it in a shaker at 37°C and 220 rpm for 18-20 h until the stationary phase is reached, and OD 600nm ≈ 3.4 - 3.8.

[0036] The methods for obtaining the fermentation supernatant and freeze-dried powder samples of Lactobacillus plantarum 9-6 involved in the following examples are as follows: Place the bacterial suspension in the late logarithmic growth phase or stationary phase in a low-temperature high-speed centrifuge at 4°C, centrifuge at 10,000 rpm for 10 minutes, and collect the supernatant, which is the bacterial fermentation broth. Transfer the obtained fermentation broth to a 50 mL sterile centrifuge tube and perform pre-freezing treatment in a -80°C ultra-low temperature freezer. Subsequently, perform freeze-drying treatment using freeze-vacuum drying technology: Place the pre-frozen sample in a pre-cooled drying chamber, start the vacuum pump to maintain a low-pressure environment, and achieve water sublimation through gradient heating to finally obtain the freeze-dried powder sample.

[0037] The percentages involved in the following examples, unless otherwise specified, all refer to mass / volume fraction (m / V).

[0038] The Vibrio parahaemolyticus involved in the following examples is Vibrio parahaemolyticus ATCC17802, the Vibrio alginolyticus is Vibrio alginolyticus ATCC33787, the Vibrio harveyi is Vibrio harveyi BB170, the Vibrio vulnificus is Vibrio vulnificus ATCC27562, the Staphylococcus aureus includes Staphylococcus aureus ATCC25923 and Staphylococcus aureus ATCC6538, the Escherichia coli includes Escherichia coli ATCC25922 and Escherichia coli ATCC8739, the Listeria monocytogenes is Listeria monocytogenes ATCC19115, the Salmonella enterica is Salmonella enterica ATCC14028, the Bacillus cereus is Bacillus cereus ATCC 11077, and the Bacillus subtilis is Bacillus subtilis ATCC6688.

[0039] Example 1: Screening and identification of strains

[0040] Strain screening:

[0041] Accurately weigh 1.0 g of the animal feed sample, add 10 mL of sterile 0.9% NaCl solution, and mix well by shaking for 30 min (rotation speed ≥ 200 rpm).

[0042] Use the gradient dilution coating method for strain isolation, select the appropriate dilution (10 -3 ~10 -6 ) of the gradient dilution solution and coat it on the MRS plate, and culture it at 37 °C for 24 - 48 h. Purify and culture it by the streak plate method (repeat the operation 2 - 3 times) to obtain a pure culture with uniform morphology.

[0043] Transfer the purified strain to the MRS liquid medium, and culture it in a shaker at 37 °C for 20 h (220 rpm, liquid loading volume ≤ 1 / 3). Centrifuge (4 °C, 10000 rpm, 10 min) to collect the supernatant for the Oxford cup antibacterial experiment. Use Vibrio parahaemolyticus as the indicator bacterium, and measure the diameter of the clear zone to screen out 5 strains with the strongest vibrio-inhibiting ability. The rescreening experiment sets four vibrios as the indicator bacteria, and selects strain 9 - 6 with inhibitory effects on all four vibrios through the antibacterial experiment.

[0044] Further conduct colony morphological characteristics, Gram staining and molecular identification on this bacterium.

[0045] Strain characteristics: Streak the glycerol bacteria onto the MRS solid medium, then place it in an incubator at 37 °C and continuously culture for 20 h, then take it out to observe the colony morphology and size and take pictures for record.

[0046] Gram staining: Use a Gram staining kit to perform Gram staining on the single colonies on the plate or the strains on the MRS medium plate, and observe and take pictures with an optical microscope at 100× oil immersion. The results show that this bacterium is a Gram-positive bacterium, and the cells are short rod-shaped.

[0047] 16S rRNA gene molecular identification:

[0048] Use the DNA extraction kit (product number: DP304) produced by Tiangen Biotech Co., Ltd. to extract genomic DNA, and the specific operation is carried out according to the product instruction manual. Use the genomic DNA of the target strain as the template, and use the universal primers 27F and 1492R to amplify the 16S rRNA gene. PCR reaction system (total 25 μL): genomic DNA 0.5 μL, 2×Taq 12.5 μL, upstream and downstream primers each 0.5 μL, ddH2O 11.0 μL.

[0049] PCR reaction conditions: Pre-denaturation at 95 °C for 4 min; denaturation at 94 °C for 30 s, annealing at 55 °C for 20 s, extension at 72 °C for 40 s, 35 cycles; repair extension at 72 °C for 10 min.

[0050] Electrophoresis was performed using 1% agarose gel (containing ethidium bromide) in 1×TAE buffer. The target band was observed to be near 1500 bp by a gel imager (reference Marker: DL2000). The PCR products with the correct molecular weight of the band were entrusted to Ascenta Biotechnology Co., Ltd. for Sanger sequencing.

[0051] The 16S rDNA sequence of strain 9-6 is shown in SEQ ID No.1. After BLAST alignment in the NCBI database, the strain 9-6 was identified as Lactobacillus plantarum and named Lactobacillus plantarum 9-6. The strain has been deposited in the China General Microbiological Culture Collection Center with the deposit number CGMCC No.32568.

[0052] Example 2: Antibacterial spectrum of Lactobacillus plantarum 9-6

[0053] The indicator bacteria used in this example are: Vibrio parahaemolyticus ATCC17802, Vibrio alginolyticus ATCC33787, Vibrio harveyi BB170, Vibrio vulnificus ATCC27562, Staphylococcus aureus ATCC25923, Staphylococcus aureus ATCC6538, Escherichia coli ATCC25922, Escherichia coli ATCC8739, Listeria monocytogenes ATCC19115, Salmonella enterica ATCC14028, Bacillus cereus ATCC 11077, Bacillus subtilis ATCC6688.

[0054] The vibrio inhibition test was carried out by the Oxford cup double-layer agar plate method: 2% agar was prepared, sterilized and poured onto the bottom layer of the plate. The Oxford cup was placed on it, and the indicator bacteria (1×10 5 -1×10 6 CFU / mL) were added to the medium containing 0.8% agar for double-layer plating. The Oxford cup was taken out, and 200 μL of the fermentation supernatant of Lactobacillus plantarum 9-6 (the added supernatant was obtained from the bacterial liquid with a viable count of 4×10 9 CFU / mL) was added. Static culture was carried out according to the culture conditions of the indicator bacteria, and the situation of the transparent circle was observed. The results are as Figure 2 shown.

[0055] Due to the differences in the culture conditions and growth rates of different indicator bacteria, the culture media, culture times and diameters of the antibacterial transparent circles used for each indicator bacteria are listed in Table 1.

[0056] Table 1 Culture conditions of indicator bacteria and diameters of antibacterial circles

[0057]

[0058] Example 3: Safety and Tolerability of Lactobacillus plantarum 9-6

[0059] Safety and tolerability of Lactobacillus plantarum 9-6, including antibiotic sensitivity, salt ion tolerance, and initial pH growth stability.

[0060] 1) Antibiotic sensitivity

[0061] After the MRS solid medium containing 0.8% agar was sterilized and cooled to about 50°C, OD 600nm =1.0, so that the final concentration of Lactobacillus plantarum 9-6 in the solid medium is 1×10 6 CFU / mL, shake well and pour the plate. After the plate solidifies, gently place the antibiotic-containing drug sensitivity sheet on the surface. Place the plate in a 37°C incubator for 24 hours to observe the size of the inhibition zone, measure and record the diameter. Refer to YY / T 1191-2011 and the interpretation standard of the diameter of the drug sensitivity inhibition zone commonly used in clinical microbiology testing): sensitive (S): inhibition zone diameter ≥15mm; intermediate (I): inhibition zone diameter between 10-14mm; resistant (R): inhibition zone diameter <10mm. Table 2 records the types of strains that are sensitive to antibiotics (S).

[0062] Table 2: Antibiotic susceptibility tablets inhibition zone diameter

[0063]

[0064] 2) High salt tolerance

[0065] The growth kinetics of the strains under different NaCl concentration gradients were determined using the Xinzhi Biotechnology MGC-500 fully automatic microbial growth curve analyzer. The experiment used MRS liquid medium as the basal medium, and a series of mediums with final concentrations of 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100 g / L were prepared by gradient addition of NaCl. After sterilization at 121°C with high-pressure steam for 20 minutes, the medium was cooled and used for later use. The activated seed solution (OD 600nm =1.0) was transferred to the above culture medium at a 5% (v / v) inoculum volume, and 200 μL of the initial bacterial suspension was taken into a sterile 96-well plate, with 4 replicates for each sample. The 96-well plate was placed in a 37°C constant temperature chamber for shaking culture for 48 hours. The growth curve was obtained by real-time monitoring of the instrument ( Figure 3 ), the data showed that when the NaCl concentration reached 100g / L, the bacterial growth was significantly inhibited, but it could still maintain slow growth; under 60g / L NaCl conditions, OD 600nm It reached a plateau phase, which was equivalent to the maximum biomass under natural culture conditions; and no significant inhibitory effect was observed on strain growth at salt ion concentrations of 50 g / L and below.

[0066] 3) Acid and alkali tolerance

[0067] Adjust the pH of MRS liquid medium to 1 - 11 gradients with 1 mol / L NaOH and 1 mol / L HCl respectively. After autoclaving at 121 °C for 20 minutes and cooling, it is ready for use. The activated strain (OD 600nm = 1.0) was transferred to the above - treated medium at an inoculation amount of 5% (v / v). Take 200 μL of the bacterial suspension and inoculate it into a 96 - well plate, and incubate it in a constant - temperature incubator (temperature not marked) for a specific time. The growth curve was plotted by measuring the OD 600nm value ( Figure 4 ). The experimental results showed that the strain could not proliferate at all in an extremely acidic environment (pH = 1, 2, 3); it could survive within the pH range of 4 - 11, and there was no significant difference in the growth rate within the pH range of 5 - 10, indicating that it had a relatively broad pH adaptation range.

[0068] Example 4: Vibrio - inhibiting ability after re - dissolving the freeze - dried powder of the fermentation supernatant

[0069] The methods for obtaining the fermentation supernatant and freeze - dried powder of Lactobacillus plantarum 9 - 6 were as follows: The bacterial suspension in the late logarithmic growth phase or stationary phase was placed in a low - temperature high - speed centrifuge at 4 °C and centrifuged at 10,000 rpm for 10 minutes. The supernatant collected was the bacterial fermentation broth. The obtained fermentation broth was transferred to a 50 mL sterile centrifuge tube and pre - frozen in a - 80 °C ultra - low - temperature refrigerator. Subsequently, freeze - drying treatment was carried out using freeze - drying vacuum technology: The pre - frozen sample was placed in a pre - cooled drying chamber, the vacuum pump was started to maintain a low - pressure environment, and water sublimation was achieved by gradient heating to finally obtain the freeze - dried powder sample.

[0070] The freeze - dried sample was configured into a 50 mg / mL solution with ddH2O. Using Vibrio parahaemolyticus ATCC17802, Vibrio alginolyticus ATCC33787, Vibrio harveyi BB170, and Vibrio vulnificus ATCC27562 as indicator bacteria, the Oxford cup antibacterial experiment was carried out with reference to the method of Example 2 ( Figure 5 ). The results showed that the fermentation broth supernatant still had antibacterial activity against the four Vibrio strains after freeze - drying and re - dissolving.

[0071] Example 5: Characterization of Lactobacillus plantarum 9 - 6

[0072] Hydrogen peroxide elimination experiment of the fermentation supernatant of strain 9 - 6, the optimal antibacterial pH, pH stability of antibacterial activity, and temperature stability. The indicator bacteria used in this example were Vibrio parahaemolyticus and Vibrio alginolyticus.

[0073] 1) Hydrogen peroxide elimination experiment

[0074] Hydrogen peroxide has broad-spectrum antibacterial activity and can inhibit the growth of pathogenic bacteria by causing oxidative damage and destroying cell membranes. It has a particularly strong inhibitory effect on Vibrio parahaemolyticus. To determine whether the antibacterial component is hydrogen peroxide, after adjusting the pH of the fermentation supernatant to 7.0, catalase (5 mg / mL) was added, and after standing in a water bath at 37 °C for 2 hours, it was adjusted to the initial pH. Referring to the method of Example 2, the Oxford cup experiment was carried out respectively and the diameter of the antibacterial zone ( Figure 6 ) was measured. The results showed that adding catalase did not affect the size of the antibacterial zone, indicating that the effective anti-vibrio component in the fermentation supernatant was not hydrogen peroxide.

[0075] 2) Optimal antibacterial activity pH

[0076] The pH of the fermentation supernatant was adjusted to the gradient range of 2 - 9 with 1M NaOH and 1M HCl respectively. Referring to the method of Example 2, the antibacterial effect was verified by the Oxford cup method and the diameter of the antibacterial zone was measured. The results showed that the antibacterial component exhibited antibacterial activity in the acidic range of pH 2.0 - 5.0, especially the antibacterial activity was significantly enhanced at pH = 2.0 - 4.0. Among them, the average diameter of the antibacterial zone of the fermentation supernatant (natural pH = 3.67) was about 26.5 mm, suggesting that its antibacterial active component played a role in an acidic environment ( Figure 7 ).

[0077] 3) Acid-base stability of the antibacterial component

[0078] The pH of the fermentation supernatant was adjusted to the gradient range of 2 - 10 with 1M NaOH and 1M HCl respectively, and it was left standing at room temperature (25 ± 2 °C) for 4 h, and then the solution was adjusted to the initial pH with 1M NaOH and 1M HCl. Referring to the method of Example 2, the antibacterial effect was verified by the Oxford cup method and the diameter of the antibacterial zone was measured ( Figure 8 ). The results showed that the antibacterial active component remained stable in the pH range of 2 - 10, and no significant activity decay was observed, indicating that it had excellent acid-base tolerance and stability.

[0079] 4) Thermal stability of the antibacterial component

[0080] The fermentation broth was heated at 60 °C, 70 °C, 80 °C, 90 °C, 100 °C and 121 °C for 15 min respectively and then restored to room temperature. The fermentation supernatant was collected, and the Oxford cup experiment was carried out referring to the method of Example 2, with the fermentation broth without high-temperature treatment as the negative control ( Figure 9 ). The results showed that the fermentation supernatant or the antibacterial agent component had good thermal stability.

[0081] Comparative Example 1:

[0082] The specific implementation is as in Example 2, with the difference that the fermentation supernatant of the commercial strain Lactococcus lactis subsp. lactis CICC 6242 that produces nisin was used for the antibacterial control experiment, and the antibacterial effects were compared. The results are as follows: Under the same culture conditions, the fermentation supernatant of the CICC 6242 strain also has an inhibitory effect on the four vibrios, but the diameters of the inhibition zones are all smaller than those of Lactobacillus plantarum 9-6( Figure 10 ), and Lactobacillus plantarum and its fermentation broth in the present invention have obvious advantages.

[0083] Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any person familiar with this technology can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims

1. A Lactobacillus plantarum 9-6, which has been deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number CGMCC No. 32568.

2. A microbial preparation containing the Lactobacillus plantarum 9-6 as claimed in claim 1.

3. The microbial preparation according to claim 2, wherein In the microbial preparation, the cell count of Lactobacillus plantarum 9-6 is not less than 4×10 9 CFU / mL or 8×10 11 CFU / g of wet cells.

4. Postbiotics prepared from Lactobacillus plantarum 9-6 according to claim 1, characterized in that, The postbiotic contains the fermentation broth, fermentation supernatant and / or freeze-dried powder of the Lactobacillus plantarum 9-6.

5. A method for preparing the postbiotic according to claim 4, characterized in that, It is any one of the following: (1) Collect the fermentation broth after fermenting the Lactobacillus plantarum 9-6 as claimed in claim 1; (2) After fermenting the Lactobacillus plantarum 9-6 as claimed in claim 1, centrifuge to remove the bacterial cells and collect the fermentation supernatant; (3) Freeze-dry the fermentation broth obtained after fermenting the Lactobacillus plantarum 9-6 as claimed in claim 1 and collect the freeze-dried powder.

6. A product containing the Lactobacillus plantarum 9-6 as claimed in claim 1, or the microbial preparation as claimed in claim 2 or 3, or the postbiotic as claimed in claim 4.

7. The product according to claim 6, wherein The product includes food, medicine, health products or feed additives.

8. Use of the Lactobacillus plantarum 9-6 as claimed in claim 1, or the microbial preparation as claimed in claim 2 or 3, or the postbiotic as claimed in claim 4 in the preparation of a bacterial inhibitor.

9. The application according to claim 8, characterized in that The bacteria include one or more of Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi, Vibrio vulnificus, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella enterica, Bacillus cereus.

10. The application according to claim 8, characterized in that The bacterial inhibitor includes an oral preparation and / or an external preparation.

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