Cat milk-derived pediococcus acidilactici M22 and application thereof

Abstract

The invention relates to the field of microorganisms, in particular to cat milk-derived pediococcus acidilactici and application thereof. The cat milk source pediococcus acidilactici provided by the invention is preserved in the China General Microbiological Culture Collection Center on July 1, 2024, the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen West Road, Chaoyang District, Beijing, China, and the preservation number is CGMCC NO.31006. The cat milk-derived pediococcus acidilactici provided by the invention is obtained by screening from healthy cat milk, has relatively strong acid resistance and bile salt resistance, is extremely strong in intestinal tract adhesion capacity, has a good inhibition effect on common pathogenic bacteria, is sensitive to various antibiotics, has good probiotic potential, and can be used for preparing a feed additive for the cat milk-derived pediococcus acidilactici. The invention provides a new candidate strain for pet food, health care and treatment products or probiotic preparations, and has wide application prospects in the aspects of preparation of pet food, health care products, medicines and the like.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a feline milk-derived Pediococcus acidilactici M22 and an application thereof. Background Art

[0002] Probiotics have received much research attention because they are safe and harmless and can be used in foods such as dairy products, cheese, functional drinks, and food protectants. The screening of animal-derived probiotics is mainly conducted from the intestines, mouth, feces, etc., because the flora in these places are easy to obtain. However, for a long time, researchers have overlooked a place rich in probiotics, namely breast milk. Breast milk is the most suitable natural food for infants because it provides unique and balanced nutrients to promote the growth and development of newborns, while also continuously providing infants with microorganisms from the mother to maintain the healthy development of their intestinal flora.

[0003] As the number of cats in households continues to increase, a survey of cat populations in Australia and New Zealand shows that there are an average of two cats in a household. Probiotics for cats are gaining more and more attention, and the development of probiotics for cats has a broad market prospect. The bacteria obtained by kittens from breast milk during lactation play a vital role in the growth and development of kittens and the establishment of early intestinal flora.

[0004] Cat milk is an essential source of nutrition for kittens in their early growth stages and plays a vital role in their growth and development. A large number of studies have confirmed the presence of lactic acid bacteria in cat milk, which has become an important source of lactic acid bacteria and has the potential to be discovered as probiotics. At present, there are relatively few probiotics from cat milk, and these strains all show certain probiotic functions. It is of great significance to continuously discover probiotics from cat milk.

[0005] Pediococcus acidilactici has the ability to produce acid, regulate gastrointestinal flora, and maintain intestinal microecological balance. It has an antagonistic effect on pathogenic microorganisms in animals, competitively inhibits pathogenic microorganisms, enhances the immune function of animals, produces beneficial metabolites, activates the activity of acid proteases, participates in the body's metabolism, and prevents the production of harmful substances.

[0006] Feline milk-derived Pediococcus acidilactici is a probiotic with excellent properties and potential benefits. It comes from cat milk and, after scientific screening, has multiple physiological functions such as regulating gastrointestinal flora, enhancing immune function, and anti-inflammatory effects. Isolating and screening out excellent feline milk-derived Pediococcus acidilactici has important scientific significance and practical application value, which can meet market demand, promote the development of the probiotics industry, promote scientific research, improve animal health levels, and promote the development of related application fields. Summary of the invention

[0007] The purpose of the present invention is to provide a cat milk-derived Pediococcus acidilactici M22 and an application thereof. The strain is a strain with excellent probiotic properties selected after separation, screening and identification from cat milk, and provides a new candidate strain for cat milk replacers, pet food and health care products.

[0008] The present invention provides a strain of feline milk-derived lactic acid Pediococcus acidilactici M22. The strain of the present invention has a preservation number of CGMCC NO.31006 and is deposited in the General Microbiology Center of China National Committee for the Preservation of Microbiological Cultures. The preservation date is June 19, 2024, and the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China.

[0009] The complete gene sequence of the feline milk-derived Pediococcus acidilactici M22 of the present invention is shown in SEQ ID NO: 01.

[0010] The feline milk-derived Pediococcus acidilactici M22 of the present invention has an inhibitory effect on Gram-positive bacteria and Gram-negative bacteria. Specifically, the bacteria of the present invention may be, but are not limited to, Staphylococcus aureus, Escherichia coli, and drug-resistant Salmonella.

[0011] Furthermore, the diameter of the inhibition zone of the lactic acidococcus M22 derived from cat milk of the present invention against Escherichia coli and drug-resistant Salmonella is ≥15 mm; the diameter of the inhibition zone of the lactic acidococcus M22 derived from cat milk of the present invention against Escherichia coli is ≥20 mm.

[0012] The feline milk-derived Pediococcus acidilactici M22 of the present invention has acid resistance.

[0013] Furthermore, the feline milk-derived lactic acid Pediococcus M22 of the present invention has a survival rate of ≥59% after surviving for 3 hours under pH 2.5 conditions, a survival rate of ≥94% after surviving for 3 hours under pH 3 conditions, and a survival rate of ≥95% after surviving for 3 hours under pH 4 conditions.

[0014] The feline milk-derived Pediococcus acidilactici M22 of the present invention has bile salt resistance ability.

[0015] Furthermore, the survival rate of the feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention after surviving for 3 hours under 0.3% bile salt conditions is ≥84%.

[0016] The survival rate was ≥88% after 3 h of survival under 0.1% bile salt conditions, and ≥76% after 3 h of survival under 0.2% bile salt conditions.

[0017] The feline milk-derived lactic acid Pediococcus M22 of the invention has no hemolytic property and is a safe and non-toxic strain.

[0018] The feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention has extremely strong cell adhesion ability, with an adhesion rate of 69%, and has good probiotic potential. Adhesion and colonization in the intestinal tract are important prerequisites for lactic acid bacteria to exert their ecological effects and physiological functions, and are one of the important bases for evaluating their probiotic potential. Adhesion of lactic acid bacteria to epithelial cells is the main basis for them to exert their probiotic functions. The adhered lactic acid bacteria can exist in the intestinal tract and can resist the invasion of pathogenic bacteria, produce metabolites such as antimicrobial peptides to help effectively kill pathogenic bacteria, achieve the functions of avoiding the intestinal tract from being infected by pathogenic bacteria, and enhance the immune ability of the body.

[0019] The feline milk-derived lactococcus M22 of the present invention shows certain sensitivity to a variety of antibiotics, and the antibiotics of the present invention include but are not limited to macrolide antibiotics, penicillins, aminoglycosides, lincosamides, cephalosporins, quinolones, and chloramphenicol.

[0020] Furthermore, the feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention is sensitive to gentamicin, erythromycin, ampicillin, tetracycline, penicillin, chloramphenicol, cefazolin, erythromycin, ampicillin, penicillin, chloramphenicol, cefazolin, penicillin, chloramphenicol, and cefazolin.

[0021] The feline milk-derived Pediococcus acidilactici M22 of the present invention has certain antioxidant properties.

[0022] The present invention also provides pet food and health products. The animal food and health products of the present invention contain feline milk-derived lactic acid Pediococcus acidilactici M22 and its metabolites.

[0023] Preferably, the pet food of the present invention includes, but is not limited to, yogurt, milk powder, cheese, snacks and / or freeze-dried staple food or dry pet food, etc.

[0024] The invention also provides a probiotic preparation, which contains feline milk-derived Pediococcus acidilactici M22 and metabolites thereof.

[0025] The present invention has the following beneficial effects:

[0026] (1) The present invention provides a new strain of feline milk-derived Pediococcus acidilactici M22, which has strong adhesion ability and good probiotic ability.

[0027] (2) The feline milk-derived lactic acid Pediococcus acidilactici M22 provided by the present invention is non-hemolytic and has high biological safety.

[0028] (3) The feline milk-derived lactic acid Pediococcus acidilactici M22 provided by the present invention has good acid resistance and bile salt resistance.

[0029] (4) The cat milk-derived lactic acid Pediococcus acidilactici M22 provided by the present invention is a strain with excellent probiotic properties that is isolated, screened, and identified from cat milk. It has a good inhibitory effect on common pathogenic bacteria and is sensitive to a variety of antibiotics. It provides a new candidate strain for pet food, health care products, or probiotic preparations, and has broad application prospects in the preparation of pet food, health care products, medicines, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0031] Figure 1 This is the acid resistance test result of Example 1 of the present invention at pH = 2.5.

[0032] Figure 2 This is the bile salt tolerance test result of Example 1 of the present invention at a bile salt concentration of 0.3%.

[0033] Figure 3 This is the antibacterial ability result of feline milk-derived Pediococcus acidilactici M22 in Example 1 of the present invention.

[0034] Figure 4 It is the BLAST database annotation result in Example 2 of the present invention.

[0035] Figure 5 This is the colony morphology of the feline milk-derived Pediococcus acidilactici M22 in Example 2 of the present invention.

[0036] Figure 6 This is the growth curve of Pediococcus acidilactici M22 from feline milk in Example 3 of the present invention.

[0037] Figure 7 This is the in vitro acid resistance result of the feline milk-derived Pediococcus acidilactici M22 in Example 3 of the present invention.

[0038] Figure 8 This is the in vitro bile salt resistance result of the feline milk-derived lactic acid Pediococcus M22 in Example 3 of the present invention.

[0039] Fig. 9 This is the hemolytic ability result of lactococcus acidilactici M22 from feline milk in Example 3 of the present invention.

[0040] Fig.10 This is the adhesion test result of feline milk-derived lactic acid bacteria M22 in Example 3 of the present invention.

[0041] Fig.11This is the result of the test on scavenging DPPH free radicals by Pediococcus acidilactici from feline milk in Example 3 of the present invention. DETAILED DESCRIPTION

[0042] The following examples are provided for a better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are purchased from conventional biochemical reagent companies unless otherwise specified. The quantitative tests in the following examples are repeated three times, and the results are averaged.

[0043] Example 1: Screening and identification of feline milk-derived Pediococcus acidilactici M22

[0044] 1. Cultivation and Isolation of Strain

[0045] Fresh cat milk samples were selected, gradiently diluted with sterilized PBS buffer (purchased from Wuhan Sevier Biotechnology Co., Ltd.), and then spread on MRS solid culture medium containing 1% CaCO3, and cultured in an anaerobic incubator at 37°C for 48 hours, and single white or milky white colonies with obvious transparent circles were picked for Gram staining and microscopic examination, and rod-shaped and Gram-positive bacterial strains were selected. From the numerous strains, strains M2, M5, M6, M7, M13, and M22 with better growth potential were selected, and enriched and cultured in an anaerobic incubator at 37°C; the bacterial solution was mixed with sterilized 30% glycerol in a ratio of 1:1 and stored in a refrigerator at -80°C for use.

[0046] 2. Screening of strains

[0047] The strains of M2, M5, M6, M7, M13 and M22 were stored in glycerol and inoculated into MRS broth medium under sterile conditions, and cultured anaerobically at 37°C for 24 hours, and activated for three generations continuously; after centrifugation of the bacterial solution, the culture medium was washed 2-3 times with sterilized PBS;

[0048] The acid resistance, bile resistance and antibacterial properties of the M2, M5, M6, M7, M13 and M22 strains were tested.

[0049] The test steps for acid resistance, bile salt resistance and adhesion are as follows:

[0050] (1) Acid resistance test

[0051] The washed culture medium was resuspended in MRS broth medium with a pH of 2.5 and cultured anaerobically at 37°C. Samples were taken at 0 h and 3 h for gradient dilution, spread on MRS solid culture medium plates, and counted after anaerobically cultured at 37°C for 48 h.

[0052] Strain survival rate (%) = lg (N 1 / N 0) * 100%;

[0053] Where: N0 is the number of viable bacteria at 0h (cfu / mL); N1 is the number of viable bacteria after the strain is acid-resistant for 3h (cfu / mL).

[0054] The experimental results are as follows Figure 1 As shown, strain M22 had the highest survival rate, reaching 59.93%.

[0055] (2) Bile salt resistance test

[0056] The washed culture medium was resuspended in MRS broth medium containing 0.3% bile salts and cultured anaerobically at 37°C. Samples were taken at 0h and 3h for gradient dilution, spread on MRS solid culture medium plates, and counted after anaerobically cultured at 37°C for 48h.

[0057] Strain survival rate (%) = lg (N 1 / N 0) * 100%;

[0058] Where: N0 is the number of viable bacteria at 0h (cfu / mL); N1 is the number of viable bacteria after the strain has been resistant to bile salt for 3h (cfu / mL).

[0059] The experimental results are as follows Figure 2 As shown, the survival rate of the M22 strain was 84.38%.

[0060] (3) Determination of the antibacterial ability of lactic acid bacteria M22 from feline milk

[0061] Pet food contains a lot of nutrients. In daily life, food is accompanied by corruption and deterioration, which harms the health of pets and causes economic losses. Microbial contamination and growth and reproduction are the main causes of food corruption, so the antibacterial performance of M22 is one of the safety indicators for its application in food.

[0062] Three common foodborne pathogens, Escherichia coli, Staphylococcus aureus and Salmonella typhi, were used as indicator bacteria. After the indicator bacteria were activated for three generations, the Oxford cup method was used to perform a preliminary determination of the antibacterial activity.

[0063] The Oxford cup method is used, and the specific method is as follows: take the activated strain, inoculate it in MRS broth medium with an inoculum of 2% (V / V), and culture it anaerobically at 37°C for 24h. Dilute the activated Escherichia coli, Staphylococcus aureus, and Salmonella suspensions by 10 times gradient, and select the bacterial solution with a dilution of 10-2 for standby use. Add 5mL of LB agar medium to the culture dish to seal the bottom, cool and solidify, and place the Oxford cup evenly, take 100μL of each gradient indicator bacterial solution and add it to 15mL LB agar at 50-60°C, mix well and slowly pour it into the plate with the Oxford cup, and pull out the Oxford cup with tweezers after condensation to form a circular hole of 8×6×10. Add 200L of the strain fermentation supernatant to the circular hole, diffuse it in a refrigerator at 4°C for 12h, and culture it anaerobically at 37°C for 9-12h. Observe whether there is an inhibition zone, and measure the diameter of the inhibition zone with a vernier caliper. Each group of tests is repeated three times, and the results are expressed as x±s.

[0064] The experimental results are as follows Figure 3 As shown, M22 has a strong inhibitory effect on both Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, resistant Salmonella), and the diameter of the inhibition zone is above 15 mm. Among them, the inhibitory effect on Escherichia coli is the strongest, and the diameter of the inhibition zone of the supernatant is 20 mm.

[0065] Depend on Figure 1 , Figure 2 and Figure 3 The results show that the antibacterial ability of lactic acidococcus M22 from cat milk is extremely strong, reaching 20mm, which is much higher than other lactic acidococci from cat milk; and after surviving for 3 hours under the conditions of acid resistance of pH = 2.5 and bile resistance of 0.3% bile salt concentration, the survival rate reaches 59% and 88% respectively, and the acid and bile resistance are good.

[0066] Example 2: Identification of Feline Milk-derived Pediococcus acidilactici M22

[0067]

[0068] The sequencing results were searched in the NCBI website using BLAST in the gene library for homology, and the results showed that the highest similarity with Pediococcus acidilactici was 99.81%. Figure 4 As shown, 99.81% of the genes were consistent with the genus Pediococcus acidilactici.

[0069] Dilute the strain M22 and spread it to observe the colony status. Figure 5 As shown, combined with the morphological characteristics of M22 colonies, strain M22 was identified as Pediococcus acidilactici.

[0070] Example 3: Determination of the probiotic properties of feline milk-derived Pediococcus acidilactici M22

[0071] 1. Growth curve determination of Pediococcus acidilactici M22 from feline milk

[0072] The cultured M22 bacterial liquid was activated for three generations, inoculated into MRS broth medium, placed in a 37°C incubator, and cultured anaerobically. The viable count of each bacterium was determined at 0, 3, 6, 9, 15, 18, 21, and 24 hours.

[0073] The growth curve of feline lactic acid Pediococcus M22 was drawn. The results are shown in Figure 6 shown.

[0074] 2. Determination of in vitro acid and bile resistance of Pediococcus acidilactici M22 from feline milk

[0075] The cultured M22 bacterial liquid was activated for three generations, inoculated into MRS broth medium with pH of 3 and 4 and bile salt concentration of 0.1% and 0.2%, respectively, placed in a 37°C incubator, cultured anaerobically, and samples were taken at 0h and 3h for plate counting;

[0076] The experimental results are shown in Figure 7 and Figure 8 ;Depend on Figure 7 It can be seen that when the bacteria are cultured in an environment of pH 3 and pH 4 for 0-3h, the bacterial growth is stable, the number of viable bacteria is above 108 cfu / mL, and the acid resistance is strong.

[0077] Depend on Figure 8 It can be seen that the bacteria grow relatively stably in a culture medium with a bile salt concentration of 0.1%; after being cultured in a culture medium with a bile salt content of 0.2% for 3 hours, the survival rate decreased by 3%, and the bacteria have a strong bile salt tolerance.

[0078] 3. Antibiotic sensitivity test of lactic acid bacteria M22 from feline milk

[0079] Excessive use of antibiotics in the pharmaceutical industry can lead to increased drug resistance in pathogenic strains, and the flora in the human intestine is prone to disorder and imbalance. Therefore, obtaining lactic acid bacteria that are sensitive to antibiotics has become one of the important indicators of whether they are safe for use in food.

[0080] The concentration of the fermentation liquid of each strain cultured for 24 hours was adjusted to 108 CFU / mL. 100uL of the bacterial liquid was added to the MRS solid plate and evenly spread and dried. An average of 5 to 6 drug-susceptibility sheets were placed on each plate. After diffusion in a 4°C refrigerator for 12 hours, the plates were anaerobically cultured at 37°C for 9 to 12 hours. The sensitivity of the strains to drugs was judged based on the results according to the instructions for use of the drug-susceptibility sheets.

[0081] The results are shown in Table 1. It can be seen from Table 1 that Pediococcus acidilactici M22 is extremely sensitive to gentamicin, erythromycin, ampicillin, penicillin, chloramphenicol, cefazolin (cephalosporin), and tetracycline, and is resistant to vancomycin, ciprofloxacin, co-trimoxazole, amikacin, and norfloxacin.

[0082] Table 1 Antibiotic sensitivity test

[0083]

[0084]

[0085] S: susceptible; I: intermediate; R: resistance

[0086] 4. Determination of hemolytic ability of lactic acid bacteria M22 from feline milk

[0087] Use an inoculation loop to dip a small amount of bacterial liquid and streak it on Columbia blood agar plate, incubate anaerobically at 37℃ for 24-36h, and observe whether there is a hemolysis circle on the plate. According to the hemolysis phenomenon, it can be divided into a hemolysis (grass green hemolysis), β hemolysis (completely transparent hemolysis circle) and y hemolysis (no hemolysis).

[0088] Hemolysis test can test the safety of strains and whether they are hemolytic. Fig. 9 As shown, this strain did not undergo hemolysis on the blood plate, which was y hemolysis. Staphylococcus aureus was selected as a positive control, and a completely transparent hemolytic ring was found around Staphylococcus aureus, which was β hemolysis. The above results show that this lactic acid Pediococcus strain is non-hemolytic and is a safe and non-toxic strain.

[0089] 5. Adhesion test

[0090] After Caco-2 cells (human colon cancer cells) were revived, they were inoculated into cell culture bottles, and fresh DMEM complete medium (DMEM high-glucose medium containing double antibodies purchased from Beijing Solebow Technology Co., Ltd., 20% fetal bovine serum) was added, and cultured in an incubator at 37°C and 5% CO2. The cell culture medium was changed every two days. When the cell adherence growth and fusion state reached about 70%-80%, 0.25% trypsin-EDTA (purchased from Beijing Solebow Technology Co., Ltd.) mixed digestion solution was used for subculture, and the last culture medium used DMEM incomplete medium.

[0091] Preparation of FITC solution: FITC (fluorescein isothiocyanate) labeling solution (purchased from Beijing Solebow Technology Co., Ltd.): Place FITC stored at 4°C at room temperature for 30 min before preparation, accurately weigh 0.5 g and dissolve it in 1 mL of dimethyl sulfoxide (DMSO) to prepare a 500 mg / mL solution, dilute 1000 times (take 10 ul and add it to 9.99 ml of DMSO in a brown 15 ml EP tube), and obtain a labeling solution with a working concentration of 500 μg / mL, and store it at 4°C.

[0092] Fluorescence labeling: Take 1ml of freshly cultured probiotic suspension and centrifuge it in a brown 1.5ml EP tube (4℃, 4000rpm, 5min), wash it with sterile PBS for 3 times in a row, and resuspend the bacteria with 1ml FITC solution that has been placed at room temperature for 30min after the last wash, and incubate it in the dark at 37℃ for 2h. After the dark incubation, centrifuge (4℃, 4000rpm, 5min) and wash the strain 3 times with sterile PBS to wash away the unbound FITC labeling solution, and resuspend it with 1ml DMEM cell culture medium. Take 200ul and add it to a black 96-well plate to measure the relative fluorescence intensity value (RFU) of the strain at a wavelength of 485nm (absorption wavelength) and a wavelength of 530nm (emission wavelength), which is recorded as the relative fluorescence intensity value R0 of the strain before adhesion.

[0093] Adhesion assay: Caco-2 cells were passaged for more than 3 generations and prepared for the assay.

[0094] Before the test, the cell concentration was adjusted to 5×105cell / mL with cell culture medium, and 1mL of cell solution was spread in each well of a 24-well cell culture plate for further culture. After the cells in the 24-well plate were cultured to a monolayer (about 24 hours of culture), the cell culture medium was discarded, washed three times with sterile PBS, and 0.6mL of the labeled probiotic suspension was added. Three replicate wells were set for each strain of bacteria, and the cells were incubated in the dark at 37°C and 5% CO2 for 2 hours. The culture medium was discarded after being taken out, and the strains that did not adhere to the cells were washed three times with sterile PBS; 0.3mL of trypsin was added to each well to digest the cells for 5 minutes, and the digestion was terminated with DMEM cell culture medium. The relative fluorescence intensity value (RFU) of the cell suspension was measured under the same wavelength conditions and recorded as the relative fluorescence intensity value R of the strain after adhesion.

[0095] Adhesion rate (%) calculation formula: R / R0×100%

[0096] CFU (colony forming unit): the total number of bacterial colonies in a unit volume; when counting live bacteria in culture, the colony formed by a single bacterium or multiple bacteria clustered together growing and multiplying on a solid culture medium is called a colony forming unit, which is used to express the number of live bacteria.

[0097] The experimental results are shown in Fig.10 As shown, the adhesion rate of strain M22 was as high as 69.23%.

[0098] 6.DPPH free radical scavenging test

[0099] The bacterial cells were collected and the concentration of the bacterial solution was adjusted to 10-8. A DPPH free radical scavenging ability kit (purchased from Nanjing Jiancheng Bioengineering Institute) was used, and the specific operations were carried out according to the steps in the instruction manual.

[0100] Sample DPPH free radical scavenging rate (%) = (1-(A determination-A control) / A blank) × 100%

[0101] The experimental results are shown in Fig.11 The DPPH free radical scavenging rate of strain M22 was 23.53%, which showed certain antioxidant activity.

[0102] Finally, it should be noted that the above invention is only a preferred embodiment of the invention and is not intended to limit the invention. Although the invention is described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions described in the above embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the invention shall be included in the protection scope of the invention.

Claims

1. A strain of Pediococcus acidilactici M22, characterized in that: The preservation number of this feline milk-derived lactic acid bacteria is CGMCC NO.31006. It is deposited in the General Microbiology Center of China National Culture Collection Administration. The preservation date is June 19, 2024. The preservation address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China.

2. The feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The complete gene sequence of the feline milk-derived Pediococcus acidilactici is shown in SEQ ID NO:

01.

3. The feline milk-derived Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived lactic acid Pediococcus has an inhibitory effect on Gram-positive bacteria and Gram-negative bacteria, including Staphylococcus aureus, Escherichia coli, and drug-resistant Salmonella.

4. The feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived Pediococcus acidilactici has acid resistance.

5. The feline milk-derived Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived Pediococcus acidilactici has bile salt resistance.

6. The feline milk-derived lactic acid Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived lactic acid Pediococcus is non-hemolytic and is a safe and non-toxic strain.

7. The feline milk-derived Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived lactic acid Pediococcus has adhesion ability, and the adhesion rate reaches 69%.

8. The feline milk-derived Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived lactic acid Pediococcus is sensitive to macrolide antibiotics, penicillins, aminoglycosides, lincosamides, cephalosporins, quinolones, and chloramphenicol.

9. The feline milk-derived Pediococcus acidilactici M22 of the present invention according to claim 1, characterized in that: The feline milk-derived Pediococcus acidilactici has antioxidant properties.

10. A pet food, health product, or probiotic preparation, characterized in that: The animal food, health product and probiotic preparation contain the feline milk-derived lactic acid Pediococcus acidilactici M22 and its metabolites according to claim 1 of the present invention.

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