Composition of postbiotics and chlorogenic acid and its application

Abstract

The present invention discloses a combination composition of postbiotics and chlorogenic acid and its application, belonging to the technical field of pharmaceutical compositions. The postbiotic composition of the present invention is composed of the postbiotic Postbio YDFF and chlorogenic acid; the postbiotic Postbio YDFF and chlorogenic acid have a synergistic inhibitory effect on the pathogenic microorganism Escherichia coli. In addition, the present invention also provides an antibiotic combination composition, which is composed of the postbiotic Postbio YDFF, chlorogenic acid and antibiotics; the three have a synergistic inhibitory effect on Escherichia coli. The present invention lays a theoretical foundation for the application of postbiotics Postbio YDFF and chlorogenic acid in antibacterial treatment, and provides new ideas and methods for reducing the side effects of drugs and enriching the research and application in the field of antibacterial treatment. At the same time, it also provides a new direction for the screening of drugs for the prevention and treatment of diseases caused by pathogenic bacteria, which is of great significance to the promotion of human health and development.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical compositions, and particularly relates to a combination composition of postbiotics and chlorogenic acid and applications thereof. Background Art

[0002] Foodborne diseases are obviously toxic or contagious. Bacterial foodborne diseases are mainly transmitted through bacteria in food or drinking water, leading to gastrointestinal diseases and other related health problems. Their potential risks to food safety cannot be ignored. Currently, there are more than 200 types of foodborne diseases discovered, most of which are caused by microbial contamination. Common causes of bacterial foodborne diseases include infections caused by Escherichia coli, Salmonella, Shigella, etc. After infecting the human body, they mainly present symptoms such as food poisoning and can be treated with antibiotics. Antibiotics can inhibit bacterial growth, but excessive use of antibiotics will make bacteria resistant, resulting in poor treatment effectiveness. The combination of drugs can alleviate the resistance problem caused by the use of a single antibiotic to a certain extent, thereby improving the prevention and treatment of pathogenic microorganisms.

[0003] The primary benefit of postbiotics is improving intestinal health, including alleviating intestinal inflammation, improving intestinal flora, and relieving constipation. They can also act on other tissues, improving metabolic abnormalities and maintaining oral health. Furthermore, postbiotics exhibit a certain antibacterial effect. In recent years, postbiotics have also shown great potential in a wide range of fields, including as a novel food quality improver, functional food supplement, and preservative.

[0004] Chlorogenic acid is a naturally occurring active polyphenol. It exhibits broad-spectrum antibacterial activity against a variety of pathogenic bacteria, including Gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa, as well as Gram-positive bacteria such as Staphylococcus aureus and Bacillus subtilis. As a naturally occurring active substance, chlorogenic acid exhibits strong antimicrobial activity and possesses multiple biological activities, including antioxidant activity, mitigation of bacterial resistance, and anticancer effects.

[0005] However, existing research on postbiotics and chlorogenic acid mainly focuses on anti-inflammatory, antioxidant, and antiviral effects, and there are few reports on the combined use of postbiotics and chlorogenic acid with antibiotics against pathogenic microorganisms. Summary of the Invention

[0006] The present invention provides a postbiotic composition comprising the postbiotic Postbio YDFF and chlorogenic acid. The postbiotic composition exhibits a synergistic inhibitory effect against Escherichia coli. The postbiotic YDFF and chlorogenic acid ratio is 0.015625 to 0.03125 mL:1500, preferably 0.015625:1500 or 0.03125:1500.

[0007] The present invention provides an application of the above-mentioned postbiotic composition in preparing a preparation for synergistically inhibiting pathogenic microorganisms; the pathogenic microorganism is Escherichia coli.

[0008] The present invention provides an antibiotic combination composition comprising a postbiotic Postbio YDFF, chlorogenic acid, and an antibiotic selected from doxycycline, amoxicillin, levofloxacin, ampicillin, or enrofloxacin. The postbiotic Postbio YDFF, chlorogenic acid, and antibiotic exhibit a synergistic inhibitory effect against Escherichia coli. The combination ratio of Postbio YDFF, chlorogenic acid, and antibiotic can be selected from the following: 0.0009765625-0.015625:46.875-750:4-4000, in mL:μg:μg.

[0009] When the antibiotic is doxycycline, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and doxycycline is 0.00390625:187.5:4, mL:μg:μg.

[0010] When the antibiotic is amoxicillin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and amoxicillin is 0.00390625:187.5:4000, mL:μg:μg.

[0011] When the antibiotic is levofloxacin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and levofloxacin is 0.0009765625:46.875:32, mL:μg:μg.

[0012] When the antibiotic is ampicillin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and ampicillin is 0.015625:750:1024, mL:μg:μg.

[0013] When the antibiotic is enrofloxacin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and enrofloxacin is 0.0009765625:46.875:64, mL:μg:μg.

[0014] The present invention provides the use of the above antibiotic combination composition in preparing a preparation for synergistically inhibiting Escherichia coli.

[0015] The present invention provides a pharmaceutical preparation, which contains the above-mentioned postbiotic composition or antibiotic combination composition.

[0016] The beneficial effects of the present invention are:

[0017] The present invention provides a postbiotic composition comprising the postbiotic Postbio YDFF and chlorogenic acid. This composition is capable of effectively inhibiting Escherichia coli, wherein the postbiotic Postbio YDFF and chlorogenic acid exhibit a synergistic antibacterial effect. Furthermore, the present invention combines the postbiotic composition with antibiotics and demonstrates a synergistic antibacterial effect between the postbiotic Postbio YDFF, chlorogenic acid, and antibiotics against Escherichia coli. This combination significantly reduces antibiotic resistance in E. coli and enhances the antibacterial activity of antibiotics against E. coli, restoring antibiotic sensitivity in E. coli that was previously highly resistant to antibiotics. This can enhance the prevention and control of pathogens and reduce antibiotic use. This invention lays a theoretical foundation for the application of the postbiotic Postbio YDFF and chlorogenic acid in antibacterial treatment, and provides new ideas and methods for reducing drug side effects and enriching the research and application of antibacterial therapy. It also provides a new direction for the screening of drugs for the prevention and treatment of diseases caused by pathogens, which is of great significance for promoting human health and development. DETAILED DESCRIPTION

[0018] The materials used in the present invention are as follows: Escherichia coli (E. coli, ATCC 8739) was purchased from the American Type Culture Collection (ATCC). LB broth was purchased from Haibo Biotechnology Co., Ltd., Qingdao High-Tech Industrial Park. Postbio YDFF (100 mL / bottle) was purchased from Qingdao Yuanda Biotechnology Co., Ltd. Chlorogenic acid, doxycycline, amoxicillin, levofloxacin, ampicillin, and enrofloxacin were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. A 3K15 medical centrifuge was purchased from SIGMA Centrifuge Co., Ltd., Germany. A DHP-9162 electric constant temperature incubator was purchased from Zhongyi Guoke (Beijing) Technology Co., Ltd. A BKQ-B7511 pressure steam sterilizer was purchased from Shandong Boco Biological Industry Co., Ltd. A Multiskan FC microplate reader and a UV spectrophotometer were purchased from Thermo Fisher Scientific (Shanghai) Instrument Co., Ltd. An SW-CJ-2FD clean bench was purchased from Suzhou Antai Air Technology Co., Ltd. The drug-resistant Escherichia coli used in the present invention was isolated from a commercial broiler farm in Jiangtun Town, Tengzhou City, Shandong Province. Drug resistance testing proved that the strain was significantly resistant to antibiotics.

[0019] The steps for bacterial activation are as follows:

[0020] Inoculate the test strain (E. coli, drug-resistant E. coli) from a 30% glycerol cryovial into LB broth and incubate in a 37°C incubator for 12 hours. Continue activation with a 2% broth inoculum until the third generation, subculturing every 12 hours to ensure viability.

[0021] Determination of bacterial concentration:

[0022] Centrifuge the culture at 4°C and 4000 rpm for 20 min, discard the supernatant, and reconstitute with 5 mL of saline. Determine the total bacterial count using a hemocytometer.

[0023] The calculation formula is as follows:

[0024] Bacterial concentration = N (total number of bacteria in five grids) / 80×400×10 4 × dilution factor;

[0025] Dilute the bacterial solution with physiological saline to obtain the required concentration: Escherichia coli (1×10 6 CFU / mL); drug-resistant Escherichia coli (1×10 6 CFU / mL); obtain bacterial suspension.

[0026] Preparation of Postbiotic Dilutions:

[0027] The postbiotic Postbio YDFF stock solution was placed in a centrifuge tube and diluted in equal parts with LB liquid culture medium using a gradient dilution method to obtain postbiotic Postbio YDFF solutions with concentrations of 1 / 2, 1 / 4, 1 / 8, 1 / 16, 1 / 32, 1 / 64, 1 / 128, 1 / 256, 1 / 512, 1 / 1024, and 1 / 2048, i.e., the concentrations were 0.5 mL / mL, 0.25 mL / mL, 0.125 mL / mL, 0.0625 mL / mL, 0.03125 mL / mL, 0.015625 mL / mL, 0.0078125 mL / mL, 0.00390625 mL / mL, 0.001953125 mL / mL, 0.0009765625 mL / mL, and 0.00048828125 mL / mL, respectively. The unit mL / mL refers to how many mL of postbiotic stock solution is contained in each mL of postbiotic dilution. For example, 0.5 mL / mL means that 1 mL of postbiotic dilution contains 0.5 mL of postbiotic stock solution.

[0028] Preparation of chlorogenic acid dilutions:

[0029] Chlorogenic acid was placed in a centrifuge tube containing LB liquid culture medium to obtain a chlorogenic acid stock solution with a drug concentration of 192,000 μg / mL, which was then diluted in equal multiples to obtain chlorogenic acid solutions with concentrations of 96,000 μg / mL, 48,000 μg / mL, 24,000 μg / mL, 12,000 μg / mL, 6,000 μg / mL, 3,000 μg / mL, 1,500 μg / mL, 750 μg / mL, 375 μg / mL, 187.5 μg / mL, and 93.75 μg / mL.

[0030] Preparation of doxycycline dilutions:

[0031] Take doxycycline and dissolve it in 1 mL of sterile water to prepare a doxycycline solution with a concentration of 2048 μg / mL. Then use the equal dilution method to adjust the concentration of the doxycycline solution to 1024 μg / mL, 512 μg / mL, 256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, and 1 μg / mL using LB liquid culture medium, and set the solutions of each concentration aside.

[0032] Preparation of various dilutions of amoxicillin:

[0033] Amoxicillin was dissolved in 1 mL of sterile water to prepare an amoxicillin solution with a concentration of 256000 μg / mL. The concentrations of the amoxicillin solution were adjusted to 128000 μg / mL, 64000 μg / mL, 32000 μg / mL, 16000 μg / mL, 8000 μg / mL, 4000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, 250 μg / mL, and 125 μg / mL using the equal dilution method with LB liquid culture medium. Solutions of each concentration were set aside.

[0034] Preparation of levofloxacin dilutions:

[0035] Levofloxacin was dissolved in 1 mL of sterile water to prepare a levofloxacin solution with a concentration of 1024 μg / mL. The concentrations of the levofloxacin solution were adjusted to 512 μg / mL, 256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL, and 0.5 μg / mL using the equal dilution method with LB liquid culture medium. Solutions of each concentration were set aside.

[0036] Preparation of dilutions of ampicillin:

[0037] Ampicillin was dissolved in 1 mL of sterile water to prepare an ampicillin solution with a concentration of 8192 μg / mL. The concentrations of the ciprofloxacin aqueous solution were adjusted to 4096 μg / mL, 2048 μg / mL, 1024 μg / mL, 512 μg / mL, 256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, and 4 μg / mL using the equal dilution method with LB liquid culture medium. Solutions of each concentration were set aside.

[0038] Preparation of Enrofloxacin Dilutions:

[0039] Enrofloxacin was dissolved in 1 mL of sterile water to prepare an enrofloxacin solution with a concentration of 4096 μg / mL. The concentrations of the enrofloxacin aqueous solution were adjusted to 2048 μg / mL, 1024 μg / mL, 512 μg / mL, 256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, and 2 μg / mL using the equal dilution method with LB liquid culture medium. Solutions of each concentration were set aside.

[0040] The other materials used in the present invention, unless otherwise stated, can be obtained through commercial channels. Unless otherwise specified, other terms used in the present invention generally have the meanings commonly understood by those of ordinary skill in the art. The present invention will be further described in detail below with reference to specific examples and data. The following examples are merely for illustration of the present invention and are not intended to limit the scope of the present invention in any way.

[0041] Example 1

[0042] The steps for verifying the synergistic antibacterial effect of the postbiotic combination are as follows:

[0043] This example verifies the synergistic inhibitory effect of the postbiotic Postbio YDFF and chlorogenic acid on Escherichia coli.

[0044] 1. Postbiotic YDFF and chlorogenic acid minimum inhibitory concentration determination

[0045] In a 96-well ELISA plate, after adding 100 μL of bacterial suspension, add 100 μL of postbiotic PostbioYDFF or chlorogenic acid solution to each well, add 200 μL of liquid culture medium as a blank control well, and add 100 μL of bacterial suspension and 100 μL of LLB liquid culture medium to mix as a growth control well. Repeat three times for each concentration for parallel experiments, and culture in a 37°C constant temperature incubator for 12 hours. Measure the OD value of each well at 600 nm using a microplate reader.

[0046] The calculation formula is as follows:

[0047] Bacterial growth percentage = (OD value of each well - OD value of blank control well) / (OD value of growth control well - OD value of blank control well) × 100%;

[0048] Bacterial growth inhibition percentage = 1-bacterial growth percentage;

[0049] The final growth inhibition percentage of the obtained bacteria was measured, and the minimum inhibitory concentration (MIC) at which the inhibition rate reached more than 80% compared with the growth control well was taken as the individual MIC value of the postbiotic Postbio YDFF and chlorogenic acid against Escherichia coli.

[0050] 2. Determination of the minimum inhibitory concentration when Postbio YDFF is used in combination with chlorogenic acid

[0051] The postbiotic Postbio YDFF with a concentration of 1 / 2 and chlorogenic acid with a concentration of 12000 μg / mL were diluted in equal parts using the gradient dilution method to obtain postbiotic solutions with concentrations of 1 / 4, 1 / 8, 1 / 16, 1 / 32, 1 / 64, 1 / 128, 1 / 256, and 1 / 512, as well as chlorogenic acid solutions with concentrations of 6000 μg / mL, 3000 μg / mL, 1500 μg / mL, 750 μg / mL, 375 μg / mL, 187.5 μg / mL, 93.75 μg / mL, and 46.875 μg / mL.

[0052] After adding 100 μL of bacterial suspension to each well of a 96-well microtiter plate, 50 μL of each postbiotic solution and 50 μL of each chlorogenic acid solution were added to each well. Within the selected range, different concentrations of postbiotic YDFF and chlorogenic acid were randomly matched in different wells at any concentration and all possible combinations were exhausted. Each concentration was repeated three times for parallel experiments. After incubation in a 37°C incubator for 12 hours, the OD value of each well was measured at 600 nm using a microtiter plate reader.

[0053] The percentage of bacterial growth inhibition was calculated using the two formulas above. The combined MIC value for Postbio YDFF and chlorogenic acid against E. coli was determined based on the obtained percentage of bacterial growth inhibition, achieving an 80% inhibition rate compared to the growth control wells. The drug interaction effect was assessed by calculating the combined inhibitory index (FICI).

[0054] The calculation formula of combined antibacterial index is:

[0055] FICI=MIC 后生元联合1 / MIC 后生元单独1 +MIC 绿原酸联合1 / MIC 绿原酸单独1 ;

[0056] Where, MIC后生元单独1 MIC 绿原酸单独1 Represent the MIC values ​​of postbiotics and chlorogenic acid when used alone, MIC 后生元联合1 MIC 绿原酸联合1 Respectively represent the MIC values ​​of postbiotics and chlorogenic acid when used in combination.

[0057] The results of drug combination effects were as follows: when FICI was less than or equal to 0.5, it was a synergistic effect; when FICI was greater than 0.5 and less than or equal to 4, it was an unrelated effect; and when FICI was greater than 4, it was an antagonistic effect.

[0058] The test results are shown in Table 1:

[0059] Table 1 Antibacterial effect of Postbio YDFF combined with chlorogenic acid on Escherichia coli

[0060]

[0061] As shown in Table 1, the MIC value of Postbio YDFF against E. coli when used alone is 1 / 8, while the MIC value of chlorogenic acid against E. coli when used alone is 6000 μg / mL. The MIC value of Postbio YDFF against drug-resistant E. coli when used alone is 1 / 4, while the MIC value of chlorogenic acid against drug-resistant E. coli when used alone is still 6000 μg / mL.

[0062] When the postbiotic Postbio YDFF and chlorogenic acid were used together to treat E. coli, the minimum inhibitory concentrations were 1 / 64 and 1500 μg / mL. When the postbiotic Postbio YDFF and chlorogenic acid were used together to treat drug-resistant E. coli, the minimum inhibitory concentrations were 1 / 32 and 1500 μg / mL.

[0063] According to the formula, the FICI values ​​of the combined action of Postbio YDFF and chlorogenic acid on Escherichia coli and drug-resistant Escherichia coli were both 0.375, indicating that the combined use of Postbio YDFF and chlorogenic acid has a synergistic antibacterial effect on Escherichia coli and drug-resistant Escherichia coli.

[0064] Example 2

[0065] The steps for verifying the synergistic antibacterial effect of the antibiotic combination are as follows:

[0066] This example demonstrates the synergistic inhibitory effect of a postbiotic composition (Postbio YDFF and chlorogenic acid) in combination with antibiotics against a strain of drug-resistant Escherichia coli (isolated from a commercial broiler farm in Jiangtun Town, Tengzhou City, Shandong Province. Drug resistance testing showed that the strain was significantly resistant to antibiotics such as doxycycline, amoxicillin, levofloxacin, and ampicillin).

[0067] 1. Determination of minimum inhibitory concentration of antibiotics

[0068] Prepare the desired concentrations of doxycycline, amoxicillin, levofloxacin, ampicillin, and enrofloxacin antibiotic solutions. Add 100 μL of bacterial suspension to each well of a 96-well microtiter plate. Add 100 μL of each antibiotic solution to each well. Add 200 μL of LB liquid medium as a blank control well. Add a mixture of 100 μL of bacterial suspension and 100 μL of LB liquid medium as a growth control well. Perform the experiment in triplicate for each concentration. After incubation at 37°C for 12 hours, measure the OD value of each well at 600 nm using a microplate reader.

[0069] The growth inhibition percentage of E. coli was calculated according to the above formula. Finally, the minimum inhibitory concentration (MIC) at which the inhibition rate reached 80% or more compared with the growth control well was taken as the MIC value of the antibiotic against the above E. coli.

[0070] 2. Determination of the minimum inhibitory concentration when postbiotic compositions are used in combination with antibiotics

[0071] The postbiotic Postbio YDFF and chlorogenic acid were used together as a whole (postbiotic composition) and then combined with doxycycline, amoxicillin, levofloxacin, ampicillin, and enrofloxacin. The minimum inhibitory concentrations of the postbiotic Postbio YDFF and chlorogenic acid combined obtained in the above experiment and the minimum inhibitory concentrations of the five antibiotics when used alone were applied.

[0072] Postbio YDFF, chlorogenic acid, doxycycline, amoxicillin, levofloxacin, ampicillin, and enrofloxacin were dissolved in 1 mL of sterile water to prepare 1 / 32 postbiotic solution, 3000 μg / mL chlorogenic acid solution, 32 μg / mL doxycycline solution, 32000 μg / mL amoxicillin solution, 128 μg / mL levofloxacin solution, 4096 μg / mL ampicillin solution, and 256 μg / mL enrofloxacin solution. The concentrations of the postbiotic solutions were adjusted to 1 / 32 using LB liquid culture medium by equal dilution. 1 / 64, 1 / 128, 1 / 256, 1 / 512, 1 / 1024, 1 / 2048, 1 / 4096, 1 / 8192; the concentration of chlorogenic acid solution was adjusted to 1500μg / mL, 750μg / mL, 375μg / mL, 187.5μg / mL, 93.75μg / mL, 46.875μg / mL, 23.4375μg / mL, 11.71875μg / mL; the concentration of doxycycline solution was adjusted to 16μg / mL, 8μg / mL, 4μg / mL, 2 μg / mL, 1μg / mL, 0.5μg / mL, 0.25μg / mL, 0.125μg / mL, and each concentration solution was prepared for use; the concentration of amoxicillin solution was adjusted to 16000μg / mL, 8000μg / mL, 4000μg / mL, 2000μg / mL, 1000μg / mL, 500μg / mL, 250μg / mL, 125μg / mL; the concentration of levofloxacin solution was adjusted to 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 4μg / mL, 2μg / mL, 1μg / mL, 0.5μg / mL; adjust the concentration of ampicillin solution to 2048μg / mL, 1024μg / mL, 512μg / mL, 256μg / mL, 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL; adjust the concentration of enrofloxacin solution to 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 4μg / mL, 2μg / mL, 1μg / mL.

[0073] After adding 100 μL of bacterial suspension to each well of a 96-well ELISA plate, 25 μL of each concentration of postbiotic solution and 25 μL of each concentration of chlorogenic acid solution were added to each well. Finally, 50 μL of each concentration of five antibiotics were added. Within the selected range, different concentrations of postbiotics Postbio YDFF, chlorogenic acid, and different concentrations of various antibiotics were mixed in different wells at any concentration of three and exhausted all possible combinations. 200 μL of LB liquid medium was added as a blank control well, and 100 μL of bacterial suspension and 100 μL of LB liquid medium were added as a growth control well. Each concentration was repeated three times for parallel experiments. After incubation in a 37°C constant temperature incubator for 12 hours, the OD value of each well was measured at 600 nm using a microplate reader.

[0074] The calculation formula of combined inhibition index (FICI) is as follows:

[0075] FICI=MIC 药物1联合 / MIC 药物1单独 +MIC 抗生素联合 / MIC 抗生素单独 ;

[0076] Where, MIC 药物1单独 The MIC value of Postbio YDFF and chlorogenic acid when used as a drug combination is shown. 药物1联合 Indicates the MIC value when Postbio YDFF and chlorogenic acid are used together as one drug. 抗生素单独 Indicates the MIC value when the antibiotic is used alone, MIC 抗生素联合 It indicates the MIC value of antibiotics when the three substances PostbioYDFF, chlorogenic acid and antibiotics are used in combination.

[0077] The results of drug combination effects were as follows: when FICI was less than or equal to 0.5, it was a synergistic effect; when FICI was greater than 0.5 and less than or equal to 4, it was an unrelated effect; and when FICI was greater than 4, it was an antagonistic effect.

[0078] The test results are shown in Table 2:

[0079] Table 2 Inhibitory effects of postbiotics, chlorogenic acid and antibiotics on Escherichia coli

[0080]

[0081] According to CLSIM100 33ed, E. coli is considered resistant to antibiotics when the MICs for doxycycline, amoxicillin, levofloxacin, and ampicillin are ≥16 μg / mL, ≥32 μg / mL, ≥2 μg / mL, and ≥32 μg / mL, respectively. This experiment verified that the MICs for these four antibiotics against the E. coli strain isolated from the chicken farm were 16 μg / mL, 16,000 μg / mL, 64 μg / mL, and 2,048 μg / mL, respectively. These values ​​are all higher than the standard MICs, and therefore the E. coli strain is considered resistant.

[0082] As shown in Table 2, when the postbiotic Postbio YDFF is used in combination with chlorogenic acid and doxycycline, amoxicillin, levofloxacin, ampicillin, and enrofloxacin alone, the minimum inhibitory concentrations are 1 / 32 + 1500 μg / mL, 16 μg / mL, 16000 μg / mL, 64 μg / mL, 2048 μg / mL, and 128 μg / mL, respectively. When the postbiotic Postbio YDFF, chlorogenic acid, and doxycycline are used in combination, the minimum inhibitory concentrations against drug-resistant Escherichia coli are 1 / 256, 187.5 μg / mL, and 4 μg / mL, respectively. When the postbiotic Postbio YDFF, chlorogenic acid, and amoxicillin are used in combination, the minimum inhibitory concentrations against Escherichia coli are 1 / 256, 187.5 μg / mL, and 4000 μg / mL, respectively. When the postbiotic Postbio YDFF, chlorogenic acid, and levofloxacin were used in combination, the minimum inhibitory concentrations against Escherichia coli were 1 / 1024, 46.875 μg / mL, and 32 μg / mL, respectively. When the postbiotic Postbio YDFF, chlorogenic acid, and ampicillin were used in combination, the minimum inhibitory concentrations against Escherichia coli were 1 / 64, 750 μg / mL, and 1024 μg / mL, respectively. When the postbiotic Postbio YDFF chlorogenic acid and enrofloxacin were used in combination, the minimum inhibitory concentrations against Escherichia coli were 1 / 1024, 46.857 μg / mL, and 64 μg / mL, respectively. According to the formula, the FICI of the postbiotic Postbio YDFF combined with chlorogenic acid on Escherichia coli were 0.5, 0.5, 0.5625, 1.5, and 0.5625, respectively. This shows that the combination of postbiotic PostbioYDFF, chlorogenic acid, doxycycline, and amoxicillin has a synergistic antibacterial effect on Escherichia coli, and the postbiotic Postbio YDFF, chlorogenic acid, levofloxacin, ampicillin, and enrofloxacin have no synergistic antibacterial effect.

[0083] The above results also show that the combined use of Postbio YDFF and chlorogenic acid not only has in vitro antibacterial activity against Escherichia coli, but also can increase the sensitivity of Escherichia coli to doxycycline and amoxicillin, and reduce the resistance of Escherichia coli to these two antibiotics.

[0084] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any other manner. Any person skilled in the art may utilize the above-disclosed technical content to modify or modify the present invention into equivalent embodiments. However, any simple modifications, equivalent variations, and modifications to the above embodiments that do not depart from the technical content of the present invention and are based on the technical essence of the present invention remain within the scope of protection of the present invention.

Claims

1. A postbiotic composition, characterized in that The postbiotic composition consists of the postbiotic Postbio YDFF and chlorogenic acid.

2. The postbiotic composition according to claim 1, characterized in that The combination ratio of the postbiotic Postbio YDFF and chlorogenic acid is 0.015625~0.03125:1500, mL:μg.

3. Use of the postbiotic composition according to claim 1 in the preparation of a preparation for synergistic inhibition of Escherichia coli.

4. An antibiotic combination composition, characterized in that: The antibiotic combination composition consists of postbiotic PostbioYDFF, chlorogenic acid and antibiotics.

5. The antibiotic combination composition according to claim 4, characterized in that The antibiotic is selected from doxycycline, amoxicillin, levofloxacin, ampicillin or enrofloxacin.

6. The antibiotic combination composition according to claim 4, characterized in that The combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and antibiotic is selected from: 0.0009765625~0.015625: 46.875~750: 4~4000, mL: μg: μg.

7. The antibiotic combination composition according to claim 4, characterized in that: When the antibiotic is doxycycline, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid, and doxycycline is 0.00390625: 187.5: 4, mL: μg: μg; When the antibiotic is amoxicillin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and amoxicillin is 0.00390625: 187.5: 4000, mL: μg: μg; When the antibiotic is levofloxacin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and levofloxacin is 0.0009765625: 46.875: 32, mL: μg: μg; When the antibiotic is ampicillin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and ampicillin is 0.015625:750:1024, mL:μg:μg; When the antibiotic is enrofloxacin, the combination ratio of the postbiotic Postbio YDFF, chlorogenic acid and enrofloxacin is 0.0009765625:46.875:64, mL:μg:μg.

8. Use of the antibiotic combination composition according to claim 4 in the preparation of a preparation for synergistic inhibition of Escherichia coli.

9. A pharmaceutical preparation, characterized in that The preparation contains the postbiotic composition according to claim 1 or the antibiotic combination composition according to claim 4.

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