Pediococcus acidilactici aui2505 and uses thereof

Abstract

This invention provides a *Pediococcus lactis* strain AUI2505 and its applications. The *Pediococcus lactis* strain AUI2505 of this invention is a probiotic that grows rapidly, requires simple fermentation conditions, is biosafe, and exhibits good acid and bile salt tolerance, auto-aggregation ability, and surface hydrophobicity. It survives for a long time and maintains high activity in the gastrointestinal tract, contributing to positive health effects. Furthermore, this strain possesses antioxidant, anti-inflammatory, weight-loss, blood sugar-lowering, and adjunctive blood sugar-lowering effects, filling a gap in existing technologies and providing a new and effective means to improve metabolic health.

Description

Technical Field

[0001] This invention relates to biotechnology, and more particularly to a type of lactic acid cocci, AUI2505, and its applications. Background Technology

[0002] With global economic development and changing lifestyles, metabolic diseases such as obesity and type 2 diabetes have become major health challenges threatening human public health. Meanwhile, oxidative stress, as a common pathological basis for many chronic diseases, is increasingly attracting attention due to the resulting damage to the body. These metabolic diseases are often accompanied by multiple pathological states, including insulin resistance, chronic low-grade inflammation, gut microbiota dysbiosis, and oxidative stress imbalance. This not only severely impacts patients' quality of life but also easily induces various complications such as cardiovascular disease, retinopathy, and nephropathy, placing a heavy medical burden on individuals, families, and society.

[0003] Currently, interventions for weight loss and blood sugar control mainly include lifestyle modifications, drug therapy, and metabolic surgery. While lifestyle interventions are the basic approach, they suffer from poor adherence and difficulty in long-term maintenance. Chemotherapy (such as GLP-1 receptor agonists) can control weight and blood sugar to some extent, but may be accompanied by side effects such as gastrointestinal discomfort and increased burden on liver and kidney function, and cannot fundamentally improve the core pathological mechanisms of metabolic disorders. Metabolic surgery is only suitable for severely obese individuals and has limitations such as surgical risks and postoperative complications. Therefore, developing safe, effective, and highly adherent natural products or functional ingredients has become a research hotspot for the prevention and adjunctive treatment of obesity and diabetes, as well as the alleviation of oxidative stress. Summary of the Invention

[0004] The purpose of this invention is to provide a novel strain of Pediococcus lactis with significant weight loss, antioxidant, anti-inflammatory, hypoglycemic or adjuvant hypoglycemic activities and its applications, so as to fill the gap in the existing technology and provide a new and effective means to improve metabolic health.

[0005] This invention provides a *Pediococcus acidilactici* strain AUI2505, which was deposited on March 16, 2026, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCC NO.67956 and classification name Pediococcus acidilactici.

[0006] The present invention provides a culture, fermentation product, or fermentation supernatant of the above-mentioned Pediococcus lactis AUI2505.

[0007] The present invention provides a microbial inoculant, comprising the above-mentioned Pediococcus lactis AUI2505 and / or the above-mentioned culture or fermentation product or fermentation supernatant.

[0008] This invention provides the application of the above-mentioned *Pediococcus lactis* AUI2505, the above-mentioned culture or fermentation product or fermentation supernatant, and the above-mentioned microbial agent in the preparation of products for weight loss, anti-oxidation, anti-inflammation, blood sugar reduction, or adjuvant blood sugar reduction.

[0009] In the application described above, the viable count of *Pediococcus lactis* AUI2505 in the product is 1×10⁻⁶. 6 -10 10 CFU / mL or 1×10 6 -10 10 CFU / g.

[0010] In the applications described above, the product also includes a carrier and / or physiologically acceptable excipients;

[0011] The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes;

[0012] The excipients include at least one of the following: fillers, flavoring agents, diluents, wetting agents, binders, disintegrants, lubricants, color, flavor and aroma modifiers, solvents, solubilizers, co-solvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

[0013] As described above, the product includes at least one of food, functional food, health food, and pharmaceutical.

[0014] As described above, functional foods include at least one of the following: foods for special dietary purposes, foods for special medical purposes, infant formula, children's formula, adolescent formula, adult formula, functional powders, functional granules, functional capsules, functional beverages, and sports nutrition foods.

[0015] As described above, the health food includes at least one of the following: health foods for weight loss, anti-oxidation, anti-inflammation, and blood sugar reduction.

[0016] As described above, the drug includes at least one of the following: drugs for the prevention or treatment of hyperglycemia, drugs for the prevention or treatment of diabetes, drugs for the prevention or treatment of obesity, and drugs for the prevention or treatment of diseases caused by overweight.

[0017] This invention provides a novel strain of *Pediococcus lactis*, AUI2505. This strain is a probiotic, exhibiting rapid growth, simple fermentation conditions, and practicality, facilitating large-scale industrial production. It also possesses strong acid and bile salt resistance, spontaneous aggregation ability, and hydrophobic surface properties, allowing for long survival and high activity in the gastrointestinal tract, thus enhancing its positive health effects. Furthermore, it demonstrates good biocompatibility, exhibiting good antibiotic resistance without causing hemolysis. In addition, this strain possesses antioxidant, anti-inflammatory, weight-loss, hypoglycemic, and adjunctive hypoglycemic effects. Therefore, this new strain fills a technological gap and provides a new and effective means to improve metabolic health. Attached Figure Description

[0018] Figure 1 This is a colony morphology diagram of AUI2505 in one embodiment of the present invention;

[0019] Figure 2 This is a morphological diagram of strain AUI2505 in one embodiment of the present invention;

[0020] Figure 3 This is a growth curve of AUI2505 in one embodiment of the present invention;

[0021] Figure 4 This is a graph showing the carbohydrate metabolism results of AUI2505 in one embodiment of the present invention;

[0022] Figure 5 This is a hemolysis result diagram of AUI2505 in one embodiment of the present invention;

[0023] Figure 6 This is a statistical chart showing the mortality rate of zebrafish juveniles in one embodiment of the present invention;

[0024] Figure 7 This is a weight chart of zebrafish juveniles in one embodiment of the present invention;

[0025] Figure 8 This is a statistical chart of blood glucose concentration in juvenile zebrafish according to one embodiment of the present invention;

[0026] Figure 9 This is a statistical chart of insulin content in juvenile zebrafish in one embodiment of the present invention;

[0027] Figure 10 This is a statistical chart of glucagon content in juvenile zebrafish in one embodiment of the present invention;

[0028] Figure 11 This is a statistical chart of the inflammatory factor IL-6 in juvenile zebrafish in one embodiment of the present invention;

[0029] Figure 12 This is a statistical chart of the inflammatory factor TNF-α in juvenile zebrafish according to one embodiment of the present invention;

[0030] Figure 13 This is a statistical chart of glucose tolerance in juvenile zebrafish according to one embodiment of the present invention;

[0031] Figure 14 This is a statistical graph showing the area under the blood glucose concentration-time curve of zebrafish juveniles in one embodiment of the present invention. Detailed Implementation

[0032] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below. The specific embodiments listed below are merely descriptions of the principles and features of the present invention, and the examples are only for explaining the present invention and are not intended to limit the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] Probiotics, as a class of live microorganisms beneficial to the host's health, are widely used in the development of functional foods and health products due to their unique advantages in regulating gut microbiota balance, improving metabolic disorders, and inhibiting inflammatory responses. Studies have shown that gut microbiota dysbiosis is closely related to obesity and the development of type 2 diabetes. Obese individuals have an increase in harmful bacteria and a decrease in beneficial bacteria in their gut, leading to insufficient synthesis of short-chain fatty acids, which in turn exacerbates insulin resistance and metabolic disorders. Supplementing with specific probiotics can improve insulin sensitivity, regulate blood glucose homeostasis, and reduce fat deposition through mechanisms such as reshaping the gut microbiota structure, producing short-chain fatty acids, and inhibiting the release of pro-inflammatory factors. Furthermore, probiotics can alleviate oxidative stress and reduce oxidative damage to body tissues by enhancing the activity of antioxidant enzymes and scavenging reactive oxygen free radicals, thereby reducing the risk of metabolic diseases.

[0034] Pediococcus acidilactici is a Gram-positive probiotic with good gastrointestinal tolerance and colonization ability. In recent years, research on the probiotic functions of Pediococcus acidilactici has deepened, confirming its significant effects in regulating gut health, enhancing immunity, and improving neurodegenerative diseases. Although existing research has preliminarily revealed some of the probiotic properties of Pediococcus acidilactici, its specific mechanisms of action in weight loss, anti-oxidation, anti-inflammation, and hypoglycemia-lowering or adjuvant hypoglycemia-lowering still need further clarification. Furthermore, the probiotic functions of different strains exhibit significant strain specificity; not all Pediococcus acidilactici possess the aforementioned activities. Currently, most commercially available probiotic products targeting weight loss and hypoglycemia use common strains such as Bifidobacterium and Lactobacillus. Products specifically targeting Pediococcus acidilactici in this area are scarce, lacking clear strain identification and sufficient efficacy verification. Therefore, identifying Pediococcus acidilactici strains with clear weight loss, anti-oxidation, anti-inflammation, and hypoglycemic activities and applying them to the preparation of related products is of great significance for enriching the variety of functional probiotic products and meeting consumers' demand for natural health interventions. Based on this, the present invention aims to provide a type of Pyrococcus lactis with significant weight loss, antioxidant, anti-inflammatory, hypoglycemic or adjuvant hypoglycemic activities and its applications, in order to fill the gap in the prior art and provide a new and effective means to improve metabolic health.

[0035] The first aspect of this invention provides a *Pediococcus acidilactici* strain AUI2505, which was deposited on March 16, 2026, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCCNO.67956 and classified as *Pediococcus acidilactici*.

[0036] Experiments have shown that this new strain has the following advantages:

[0037] (1) Rapid growth and simple fermentation conditions:

[0038] Pediococcus lactis AUI2505 has high metabolic activity and strong growth ability. After being inoculated into MRS broth medium, it can quickly enter the fermentation state at 37°C, and its metabolites can accumulate in large quantities in the fermentation system. Moreover, the carbohydrates required by this strain are inexpensive and readily available, and can be D-glucose, D-lactose, D-maltose or D-sucrose.

[0039] (2) Strong resistance to acids and bile salts:

[0040] Pediococcus lactis AUI2505 can effectively tolerate acids, bile salts, or proteases in gastric or intestinal juices and still has good activity in the human gastrointestinal tract. Therefore, it may have good gastrointestinal survival ability, which helps it exert its probiotic effects.

[0041] (3) High security:

[0042] On the one hand, Pediococcus acidilactici has good antibiotic resistance, thus exhibiting high biosafety; on the other hand, its resistance to some clinically important antibiotics allows it to be used as an effective strain in combination with antibiotics to treat bacterial infections.

[0043] On the other hand, Pediococcus lactis AUI2505 is not hemolytic, meaning it does not produce hemolysin to cause red blood cell rupture and thus hemolysis.

[0044] (4) Excellent self-aggregation ability and surface hydrophobicity:

[0045] Pediococcus lactis AUI2505 has a high autoaggregation ability and a hydrophobic surface, which can enhance its ability to adhere to the intestinal epithelial mucus layer, increase its survival rate and residence time in the gastrointestinal tract, thereby exerting a positive health effect.

[0046] (5) Good antioxidant capacity:

[0047] Pediococcus lactis AUI2505 can effectively scavenge 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) and hydroxyl radicals, thereby alleviating chronic oxidative damage or oxidative stress caused by excessive free radicals.

[0048] (6) Good ability to lower blood sugar or assist in lowering blood sugar:

[0049] Pediococcus lactis AUI2505 can effectively inhibit α-glucosidase, reduce blood glucose concentration, and alleviate metabolic disorders and glucose load in hyperglycemic bodies.

[0050] (7) Good weight loss ability:

[0051] Lactococcus lactis AUI2505 can effectively reduce weight.

[0052] (8) Good anti-inflammatory effect:

[0053] Pediococcus lactis AUI2505 can reduce the levels of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) in hyperglycemic organisms, thereby exerting a good anti-inflammatory effect.

[0054] In summary, the *Pediococcus lactis* AUI2505 strain provided in the first aspect of this invention is a probiotic. Firstly, it grows rapidly and requires simple fermentation conditions, making it practical and suitable for large-scale industrial production. Secondly, it exhibits strong acid and bile salt resistance, excellent auto-aggregation ability, and superior surface hydrophobicity, allowing it to survive for a long time and maintain high activity in the gastrointestinal tract, thus enhancing its positive health effects. Thirdly, it demonstrates high safety, exhibiting good antibiotic resistance without hemolysis. Finally, it possesses good antioxidant, anti-inflammatory, weight-loss, hypoglycemic, and adjunctive hypoglycemic effects. Therefore, this novel strain of *Pediococcus lactis* can fill a gap in existing technologies and provide a new and effective means to improve metabolic health.

[0055] A second aspect of the present invention provides a culture, fermentation product, or fermentation supernatant of the above-mentioned Pediococcus lactis AUI2505.

[0056] Among them, the fermentation product of Pediococcus lactis AUI2505 can be a fermentation system (e.g., fermentation broth) obtained by fermenting Pediococcus lactis AUI2505, which includes Pediococcus lactis AUI2505 cells, fermentation medium and metabolites produced during fermentation.

[0057] The fermentation supernatant of Pediococcus lactis AUI2505 can be the supernatant liquid phase obtained after removing bacterial cells from the above fermentation material by separation methods such as centrifugation and filtration, and contains metabolic products secreted by bacterial cells.

[0058] The culture of Pediococcus lactis AUI2505 may include at least one of the following: fermentation product of Pediococcus lactis AUI2505 and its concentrated product, extract or dried product; fermentation supernatant of Pediococcus lactis AUI2505 and its concentrated product, extract or dried product; spore suspension of Pediococcus lactis AUI2505; fermentation precipitate of Pediococcus lactis AUI2505; or metabolites of Pediococcus lactis AUI2505.

[0059] In one embodiment, the method for preparing the ferment of Pediococcus lactis AUI2505 includes: inoculating Pediococcus lactis AUI2505 into MRS broth medium at an inoculum of 1-5% (v / v) and culturing at 30-45℃ for 12-36 h to obtain the ferment of Pediococcus lactis AUI2505.

[0060] In one embodiment, the method for preparing the fermentation supernatant of Pediococcus lactis AUI2505 includes: inoculating Pediococcus lactis AUI2505 into MRS broth medium at an inoculation rate of 1-5% (v / v), culturing at 30-45℃ for 12-36 h for activation, activating for two generations, centrifuging at 5000-7000 r / min and 2-8℃ for 5-20 min, and taking the supernatant as the fermentation supernatant of Pediococcus lactis AUI2505.

[0061] In one embodiment, the method for preparing fermentation precipitate of Pediococcus lactis AUI2505 includes: inoculating Pediococcus lactis AUI2505 into MRS broth medium at an inoculum of 1-5% (v / v), culturing at 30-45℃ for 12-36 h for activation, activating for two generations, centrifuging at 5000-7000 r / min and 2-8℃ for 5-20 min, discarding the supernatant, and taking the precipitate and washing it with physiological saline to obtain the fermentation precipitate of Pediococcus lactis AUI2505.

[0062] A third aspect of the present invention provides a microbial inoculant, comprising the above-mentioned Pediococcus lactis AUI2505 and / or the above-mentioned culture or fermentation product or fermentation supernatant.

[0063] Microbial preparations may include at least one of Pediococcus lactis strain AUI2505, its inactivated form, culture of Pediococcus lactis strain AUI2505, fermentation product, and fermentation supernatant; in addition, they may include physiologically acceptable diluents, fillers, solvents, promoters, carriers, emulsifiers, dispersants, preservatives, thickeners, adjuvants, or any combination thereof.

[0064] Among them, Porphyromonas lactis AUI2505 can exist in the microbial agent as a live bacteria in the form of a bacterial suspension, or as an inactivated bacterial body in the microbial agent in the form of an inactivated bacterial suspension, or in the form of a lyophilized powder prepared from a bacterial suspension or an inactivated bacterial suspension.

[0065] In one embodiment, the method for preparing a bacterial suspension of Pediococcus lactis AUI2505 includes: inoculating Pediococcus lactis AUI2505 into MRS broth medium at an inoculum of 1-5% (v / v), culturing at 30-45℃ for 12-36 h for activation, activating for two generations, centrifuging at 5000-7000 r / min and 2-8℃ for 5-20 min, discarding the supernatant, washing the precipitate with physiological saline and resuspending it to obtain a bacterial suspension of Pediococcus lactis AUI2505.

[0066] In one embodiment, the method for preparing an inactivated bacterial suspension of Pediococcus lactis AUI2505 includes: inoculating Pediococcus lactis AUI2505 into MRS broth medium at an inoculation rate of 1-5% (v / v), culturing at 30-45℃ for 12-36 h for activation, activating for two generations, centrifuging at 5000-7000 r / min and 2-8℃ for 5-20 min, discarding the supernatant, washing the precipitate with physiological saline and resuspending it, and placing the bacterial suspension in a water bath at 60-80℃ for heat inactivation for 30-50 min to obtain an inactivated bacterial suspension of Pediococcus lactis AUI2505.

[0067] The fourth aspect of the present invention provides the application of the above-mentioned *Pediococcus lactis* AUI2505, the above-mentioned culture or fermentation product or fermentation supernatant, and the above-mentioned microbial agent in the preparation of products for weight loss, anti-oxidation, anti-inflammation, hypoglycemia reduction or adjuvant hypoglycemia reduction.

[0068] The *Pediococcus lactis* AUI2505 strain of this invention, along with its culture, fermentation product, fermentation supernatant, and microbial inoculant, holds significant value in the preparation of products for weight loss, anti-oxidation, anti-inflammation, and blood sugar reduction or adjunctive blood sugar reduction. This strain combines multiple health benefits with rapid growth, acid and bile salt tolerance, and high safety, filling a technological gap, reducing industrial production costs, and enabling the development of multifunctional products. Its natural safety and lack of side effects provide novel solutions to metabolic-related health problems, meeting market demands for natural, safe, and multifunctional products, and possessing multiple significances in terms of technology, industry, health, and the market.

[0069] In one specific embodiment, the viable count of *Pediococcus lactis* AUI2505 in the product is 1 × 10⁻⁶. 6 -10 10 CFU / mL or 1×10 6 -10 10 CFU / g has better effects on weight loss, anti-oxidation, anti-inflammation, lowering blood sugar, or assisting in lowering blood sugar. For example, the live bacteria count can be 1×10⁻⁶. 6 CFU / mL, 1×10 7 CFU / mL, 1×10 8 CFU / mL, 1×10 9 CFU / mL, 1×10 10 CFU / mL and any value between any two of the above ranges; viable count can also be 1×10⁻⁶. 6 CFU / g, 1×10 7 CFU / g, 1×10 8 CFU / g, 1×10 9 CFU / g, 1×10 10 CFU / g and any value between any two of the above ranges.

[0070] Furthermore, the product also includes a carrier and / or physiologically acceptable excipients; the carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes; the excipients include at least one of fillers, flavoring agents, diluents, wetting agents, binders, disintegrants, lubricants, color and flavor modifiers, solvents, solubilizers, cosolvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

[0071] In one specific implementation, the product includes at least one of food, functional food, health food, and medicine.

[0072] Among them, food can include at least one of snacks, complementary foods, food additives, dietary supplements, and nutritional fortifiers.

[0073] Functional foods may include at least one of the following: foods for special dietary purposes, foods for special medical purposes, infant formula, children's formula, adolescent formula, adult formula, functional powders, functional granules, functional capsules, functional beverages, and sports nutrition foods.

[0074] Health foods may include at least one of the following: those for weight loss, anti-oxidation, and blood sugar reduction.

[0075] The medication may include at least one of the following: medication for the prevention or treatment of hyperglycemia, diabetes, obesity, or diseases caused by overweight.

[0076] The technical solution of this application will be further explained below with reference to specific embodiments. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise specified, all reagents used are commercially available or obtained through public channels.

[0077] Example 1: Sample Preparation

[0078] (1) Pediococcus lactis AUI2505:

[0079] This embodiment provides an isolated Pediococcus acidilactici AUI2505, with the following preservation information: Preservation number: GDMCC NO.67956; Classification name: Pediococcus acidilactici; Preservation date: March 16, 2026; Preservation institution: Guangdong Provincial Center for Microbial Culture Collection; Address of the preservation institution: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou.

[0080] (2) Fermentation supernatant of Pediococcus lactis AUI2505:

[0081] This embodiment provides a method for preparing the fermentation supernatant of *Pediococcus lactis* AUI2505, comprising: inoculating *Pediococcus lactis* AUI2505, which is frozen at -80℃ using the glycerol preservation method, into MRS broth medium at an inoculation rate of 2% (v / v), culturing at 37℃ for 24 h, activating for two generations, centrifuging at 6000 r / min and 4℃ for 10 min, and filtering the supernatant through a 0.22 mm aqueous microfiltration membrane to obtain the fermentation supernatant of *Pediococcus lactis* AUI2505.

[0082] (3) Bacterial suspension of Pediococcus lactis AUI2505:

[0083] This embodiment provides a method for preparing the bacterial suspension of *Pediococcus lactis* AUI2505, comprising: inoculating *Pediococcus lactis* AUI2505, which is cryopreserved at -80℃ using the glycerol preservation method, into MRS broth medium at an inoculation rate of 2% (v / v), culturing at 37℃ for 24 h, activating for two generations, centrifuging at 6000 r / min and 4℃ for 10 min, discarding the supernatant, washing the precipitate three times with 0.85% physiological saline, resuspending it, and then adjusting the OD with 0.85% physiological saline. 600nm The concentration was 0.5, and the density of the bacterial suspension was 1×10⁻⁶. 8 A bacterial suspension of *Pediococcus lactis* AUI2505 was obtained by dispersing cfu / mL.

[0084] (4) Inactivated bacterial suspension of Pediococcus lactis AUI2505:

[0085] This embodiment provides a method for preparing an inactivated bacterial suspension of *Pediococcus lactis* AUI2505, comprising: inoculating *Pediococcus lactis* AUI2505, which has been frozen at -80℃ using the glycerol preservation method, into MRS broth medium at an inoculation rate of 2% (v / v), culturing at 37℃ for 24 h, activating for two generations, centrifuging at 6000 r / min and 4℃ for 10 min, discarding the supernatant, washing the precipitate three times with 0.85% physiological saline, resuspending it, and then adjusting the OD with 0.85% physiological saline. 600nm The concentration was 0.5, and the density of the bacterial suspension was 1×10⁻⁶. 8 The concentration of cfu / mL was increased, and the bacterial suspension was placed in a 65°C water bath for 40 min to inactivate it, thus obtaining an inactivated bacterial suspension of Pediococcus lactis AUI2505.

[0086] (5) Fermentation supernatant, bacterial suspension and inactivated bacterial suspension of LGG probiotics:

[0087] This embodiment provides a method for preparing fermentation supernatant, bacterial suspension and inactivated bacterial suspension of LGG probiotic (Lactobacillus rhamnosus GG strain), which can be referred to in Example 1 (2)-(4).

[0088] (6) A bacterial suspension of Staphylococcus aureus ATCC 25923:

[0089] This embodiment provides a method for preparing a bacterial suspension of Staphylococcus aureus ATCC 25923, which can be referred to in Example 1 (3), except that the MRS broth medium is replaced with LB liquid medium.

[0090] Example 2: Strain Identification

[0091] (1) Morphological identification:

[0092] A suspension of *Pediococcus lactis* AUI2505 was inoculated into sterile MRS broth at a 2% (v / v) inoculation rate and incubated at 37°C for 24 h. The culture was then serially diluted with sterile phosphate-buffered saline (PBS) (pH=7.0), and each diluted solution was plated onto sterile MRS solid medium and incubated at 37°C for 48 h. Colony morphology was observed. (See attached image for details.) Figure 1 Select a small number of bacterial strains, stain them according to the instructions in the Gram staining kit, and then observe the individual morphology of the strains under an oil immersion microscope. See the attached image for details. Figure 2 .

[0093] The experimental results showed that after 48 hours of incubation, the colonies were white, round, raised, smooth, moist, with neat edges, and a soft texture that was easily picked up. Under a microscope, the strain stained purple with Gram stain, indicating it was a Gram-positive bacterium. The strains were spherical, arranged in pairs or clusters, and did not form chains. These results are consistent with the morphological characteristics of *Pediococcus lactis*, and no other contaminating bacteria were found throughout the field of view, making them suitable for subsequent experimental studies.

[0094] (2) Identification of 16S rDNA gene:

[0095] The 16S rDNA gene identification was conducted by Beijing Aoke Dingsheng Biotechnology Co., Ltd. The sequencing results of Pyotrophic Lactococcus AUI2505 were compared with the NCBI gene database of the National Center for Biotechnology Information in the United States to obtain the identification results of the species relationship of the strain, as detailed in Table 1.

[0096] Table 1

[0097]

[0098] The experimental results showed that the homology between Pediococcus AUI2505 and Pediococcus Acidilactici DSM 20284 (the type strain of Pediococcus Acidilactici DSM 20284) was as high as 99.38%, indicating that Pediococcus AUI2505 belongs to Pediococcus Acidilactici.

[0099] (3) Growth rate measurement:

[0100] A suspension of *Pediococcus lactis* AUI2505 was inoculated into sterile MRS broth at a 2% (v / v) inoculation rate and incubated at 37°C. During the incubation period, OD values ​​were measured every 1 hour using a microplate reader. 600nm The growth rate of the strain was measured in triplicate, and the OD values ​​obtained were used. 600nm The values ​​were used to plot the growth curve, and the results are visible. Figure 3 .

[0101] Experimental results showed that, with the increase of time, *Pediococcus lactis* AUI2505 did not have a significant lag period and had a short incubation period, indicating that *Pediococcus lactis* AUI2505 has rapid metabolic activity and strong growth ability in MRS broth medium, and can quickly enter the fermentation state. The metabolites of *Pediococcus lactis* AUI2505 accumulated in large quantities in the fermentation system.

[0102] (4) Study on carbohydrate metabolism characteristics:

[0103] Microorganisms vary in their carbohydrate metabolism capabilities and require different nutrients. Therefore, a carbohydrate identification strip (API 50 CH) was used to study carbohydrate metabolism characteristics. This strip contains 49 carbon sources that can be detected. The determination of carbohydrate metabolism was then made in conjunction with factors such as price and edibility. The specific steps are as follows:

[0104] A suspension of *Pediococcus lactis* AUI2505 was diluted and inoculated into sterile MRS solid medium. The culture was anaerobic at 37°C for 24 h. The bacterial strain was collected from the MRS solid medium using cotton swabs and a bacterial suspension was prepared. This suspension was then added to the wells of an API 50 CH test strip using a sterile pipette. The strips were incubated for 24 h and 48 h before being read a second time. (See details...) Figure 4 And Table 2.

[0105] Table 2

[0106]

[0107] Experimental results show that the optimal carbon source for *Pediococcus lactis* AUI2505 can be L-arabinose, D-ribose, D-galactose, D-glucose, D-maltose, D-lactose, D-melibiose, D-sucrose, or D-raffinose.

[0108] Example 3: Safety evaluation of the strain

[0109] (1) Simulated gastrointestinal fluid tolerance test:

[0110] The environment of the human gastrointestinal tract can disrupt the integrity of bacterial cells, effectively preventing bacteria from entering the body and maintaining their activity. The pH in the stomach is generally between 2.5 and 3.5, while food remains in the stomach for 2-4 hours. The intestines are another barrier for probiotics to enter the human body, containing bile salts and trypsin at a concentration of 0.15-0.30% (m / v), with a stable pH of around 8.0; meanwhile, food typically transits through the intestines for 1-4 hours. Therefore, for probiotics to exert their beneficial functions in the human body, they must be tolerant to the above conditions and maintain good activity in the human gastrointestinal tract. To investigate the tolerance of *Pediococcus lactis* AUI2505 in simulated gastric or intestinal fluids, the following experiment was conducted:

[0111] Pepsin was dissolved in PBS to a final concentration of 0.3% (m / v), and the pH was adjusted to 3.0 with 1 mol / L hydrochloric acid (HCl) solution. The solution was then filtered through a 0.22 μm aqueous microfiltration membrane to obtain sterile simulated gastric fluid. 1.0 mL of a suspension of *Pediococcus lactis* AUI2505 or a suspension of *LGG* probiotics was inoculated into 9 mL of simulated gastric fluid. After mixing, the mixture was incubated at 37°C for 3 h. The viable cell counts at 0 h and 3 h were recorded using the dilution plating method, as detailed in Table 3.

[0112] Trypsin and bovine bile salts were dissolved in PBS to final concentrations of 0.1% (m / v) and 0.3% (m / v), respectively. The pH was adjusted to 8.0 with 1 mol / L sodium hydroxide (NaOH), and then filtered through a 0.22 μm aqueous microfiltration membrane to obtain sterile simulated intestinal fluid. 1.0 mL of a suspension of *Pediococcus lactis* AUI2505 or a suspension of *LGG* probiotics was inoculated into 9 mL of simulated intestinal fluid, mixed thoroughly, and incubated at 37°C for 3 h. The viable cell counts at 0 h and 3 h were recorded using the dilution plating method, as detailed in Table 4.

[0113] In Tables 3 and 4, the survival rate is calculated using the following formula: N1 / N0 × 100%, where N1 is the number of viable bacteria after 3 hours and N0 is the number of viable bacteria after 0 hours; different letters in the same column represent significant differences (p < 0.05).

[0114] Table 3

[0115]

[0116] Table 4

[0117]

[0118] The experimental results showed that the survival rate of *Pediococcus lactis* AUI2505 in simulated gastrointestinal fluid was significantly higher than that of strain LGG (p<0.05), demonstrating good acid and bile salt tolerance.

[0119] (2) Drug sensitivity test:

[0120] Antibiotic resistance is a crucial safety indicator for probiotics, as the transfer of resistance genes to pathogenic bacteria can threaten the host's safety. Therefore, to investigate the drug susceptibility of *Pediococcus lactis* AUI2505, the following experiment was conducted:

[0121] Staphylococcus aureus ATCC 25923 was used as the quality control strain, and LGG probiotics were used as the control strain. 100 μL of Pediococcus lactis AUI2505 bacterial suspension was evenly spread on MRS solid medium. Antimicrobial susceptibility test strips were placed on the surface of the MRS solid medium, with 3 strips evenly spaced on each petri dish. After incubation at 37°C for 48 h, the diameter of the inhibition zone was measured and recorded. The drug susceptibility was determined according to the instructions of the antimicrobial susceptibility test strips (see Table 5). The specific results are shown in Table 6.

[0122] Table 5

[0123]

[0124] Table 6

[0125]

[0126] Experimental results showed that *Pediococcus lactis* AUI2505 was sensitive to most antibiotics and had a high safety profile. Furthermore, its resistance to some clinically important antibiotics allows *Pediococcus lactis* AUI2505 to be used as an effective strain in combination with antibiotics to treat bacterial infections.

[0127] (3) Hemolytic test:

[0128] Hemolysis is an important indicator for assessing the safety of bacterial strains. Pathogenic bacteria produce hemolysin during their growth, which causes red blood cell rupture and resulting in hemolysis. Therefore, to investigate the hemolytic activity of *Pediococcus lactis* AUI2505, the following experiment was conducted:

[0129] Staphylococcus aureus ATCC 25923 was used as the quality control strain, and LGG probiotics were used as the control strain. A suspension of Pediococcus lactis AUI2505 was streaked onto Columbia blood agar plates and incubated upside down at 37°C for 48 hours. Color changes around individual colonies were then observed. A greenish-yellow ring around a single colony indicated α-hemolysis, a clear ring indicated β-hemolysis, and no color change indicated γ-hemolysis (i.e., no hemolysis). Specific results can be found in [link to results]. Figure 5 In the results, A represents the quality control bacteria result, B represents the result of Pseudococcus lactis AUI2505, and C represents the result of LGG probiotics.

[0130] Experimental results showed that the hemolysis result of Pyorrhococcus lactis AUI2505 was γ-hemolysis (i.e., non-hemolysis).

[0131] Example 4: Evaluation of the strain's auto-aggregation ability and surface hydrophobicity

[0132] The autoaggregation ability of bacterial strains refers to the phenomenon where bacteria multiply rapidly and massively during culture, causing them to spontaneously aggregate into clusters. The surface hydrophobicity of bacterial strains refers to the unstable state of the strain in polar water, leading to a series of redistribution and arrangement changes in the bacterial cells. The autoaggregation ability and surface hydrophobicity of bacterial strains are related to various adhesion phenomena, and the ability to adhere to the intestinal epithelial mucus layer is one of the main criteria for selecting probiotics: the autoaggregation ability of probiotics can increase their survival rate and residence time in the gastrointestinal tract, thereby exerting a positive health effect; the surface hydrophobicity of probiotics is often determined by assessing their affinity for hydrocarbon solvents. Generally, the higher the adhesion of probiotics to hydrocarbons, the higher the hydrophobicity and the stronger the adhesion ability. The hydrophobicity of bacterial strains can be divided into three categories: strongly hydrophobic (>50%), moderately hydrophobic (20%-50%), and hydrophilic (<20%). The strength of hydrophobicity mainly depends on the expression of polysaccharides and proteins on the cell surface.

[0133] (1) Evaluation of automatic aggregation capability:

[0134] LGG probiotics were used as the control strain. The OD 600 nm of the *Pediococcus lactis* AUI2505 bacterial suspension was adjusted to 1.00. 3 mL of the suspension was vortexed for 10 s and incubated at 37℃ for 0 h and 2 h. The absorbance of the upper layer at 600 nm was measured, as detailed in Table 6. The formula for calculating the autoaggregation ability (%) is as follows: (A0-A1) / A0×100%, where A0 is the absorbance measured after 0 h of incubation and A1 is the absorbance measured after 2 h of incubation. Different letters in the same column represent significant differences (p<0.05).

[0135] (2) Evaluation of surface hydrophobicity:

[0136] LGG probiotics were used as the control strain. The OD 600 nm of the *Pediococcus lactis* AUI2505 bacterial suspension was adjusted to 1.00. 3 mL of the suspension was mixed with 1 mL of xylene, vortexed for 2 min, and then allowed to stand at room temperature for 15 min. Layering occurred, and the aqueous phase was carefully sampled. The OD value of the aqueous phase was measured at 600 nm. Details are shown in Table 7. The surface hydrophobicity (%) was calculated using the following formula: (B0-B1) / B0×100%, where B0 is the initial absorbance value (i.e., OD). 600nm =1.00), B1 is the absorbance value of the aqueous phase; different letters in the same column represent significant differences (p<0.05).

[0137] Table 7

[0138]

[0139] Experimental results showed that the autoaggregation ability and surface hydrophobicity of Pediococcus lactis AUI2505 were significantly higher than those of LGG probiotics, demonstrating good autoaggregation ability and surface hydrophobicity, indicating that Pediococcus lactis AUI2505 may have good adhesion ability to the intestinal epithelial mucus layer.

[0140] Example 5: Evaluation of the antioxidant properties of the strain

[0141] (1) Ability to remove DPPH

[0142] 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a synthetically produced stable free radical. Its contained electronic domains give it a deep purple color, and it has a maximum absorption peak at 517 nm. Antioxidants have the ability to capture and neutralize free radicals and can react with DPPH, leading to a decrease in the absorbance of DPPH-containing solutions. Therefore, to investigate the ability of *Pediococcus lactis* AUI2505 to scavenge DPPH, the following experiment was conducted:

[0143] LGG probiotics were used as the control strain. 4 mg of DPPH was accurately weighed, dissolved in anhydrous ethanol, and diluted to a final volume of 50 mL in a volumetric flask to obtain a 0.2 mmol / L DPPH anhydrous ethanol solution. This solution was stored at 0-4℃ in the dark and used immediately after preparation. 2 mL of the DPPH anhydrous ethanol solution was mixed with 2 mL of fermentation supernatant, bacterial suspension, or inactivated bacterial suspension of *Pediococcus lactis* AUI2505. 2 mL of distilled water was then mixed with 2 mL of the DPPH anhydrous ethanol solution. The mixture was reacted in the dark for 30 min, and the absorbance was measured at 517 nm. Three parallel experiments were conducted. Details are shown in Table 8. The formula for calculating the DPPH free radical scavenging rate (%) is as follows:

[0144]

[0145] Cr represents the absorbance value of the mixture of DPPH anhydrous ethanol solution and fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; Cs represents the absorbance value of the mixture of anhydrous ethanol solution and fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; Ct represents the absorbance value of the mixture of distilled water and DPPH anhydrous ethanol solution; different letters in the same column indicate significant differences (p < 0.05).

[0146] Table 8

[0147]

[0148] The experimental results showed that the DPPH scavenging rates of fermentation supernatant, bacterial suspension and inactivated bacterial suspension of Pediococcus lactis AUI2505 were significantly higher than those of LGG probiotics (p<0.05), indicating that Pediococcus lactis AUI2505 has strong in vitro antioxidant capacity.

[0149] (2) Ability to scavenge hydroxyl radicals:

[0150] The ability of a bacterial strain to scavenge hydroxyl radicals is an important indicator of its antioxidant activity. Hydroxyl radicals are a type of free radical; while free radicals at normal concentrations have physiological functions, excessive amounts can lead to chronic oxidative damage. Excessive free radicals can damage nucleic acids, lipids, proteins, polyunsaturated fatty acids, and carbohydrates, ultimately causing cell dysfunction and even death. Furthermore, hydroxyl radicals can induce lipid peroxidation, exacerbating oxidative stress. Therefore, to investigate the ability of *Pediococcus lactis* AUI2505 to scavenge hydroxyl radicals, the following experiment was conducted:

[0151] LGG probiotics were used as the control strain. 40 μL of 2.5 mmol / L o-phenanthroline was added to 0.02 mol / L PBS (pH 7.4), 40 μL of fermentation supernatant, bacterial suspension, or inactivated bacterial suspension of *Pediococcus lactis* AUI2505. After sufficient reaction, 40 μL of 2.5 mmol / L ferrous sulfate was added and mixed thoroughly. Then, 40 μL of 20 mmol / L hydrogen peroxide was added. The mixture was incubated at 37℃ for 1.5 h, and the absorbance (Ds) was measured at 536 nm. Three parallel experiments were set up, as detailed in Table 9. The formula for calculating the hydroxyl radical scavenging rate (%) is as follows:

[0152]

[0153] In the formula, Db is prepared in the same way as Ds, except that distilled water is used instead of hydrogen peroxide, and distilled water is used instead of the fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; D0 is prepared in the same way as Ds, except that distilled water is used instead of the fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; different letters in the same column represent significant differences (p < 0.05).

[0154] Table 9

[0155]

[0156] The experimental results showed that the hydroxyl radical scavenging capacity of the fermentation supernatant, bacterial suspension and inactivated bacterial suspension of Pediococcus lactis AUI2505 was better than that of LGG probiotics (P<0.05), indicating that Pediococcus lactis AUI2505 has good in vitro antioxidant capacity.

[0157] Example 6: Evaluation of the inhibition rate of the strain against α-glucosidase

[0158] α-Glucosidase inhibition rate refers to the degree to which the sample inhibits the catalytic activity of α-glucosidase, expressed as a percentage (%). α-Glucosidase is responsible for hydrolyzing intestinal carbohydrates (such as starch and sucrose) into glucose. Therefore, inhibiting the activity of this enzyme can delay glucose absorption and reduce postprandial blood glucose peaks, making it a key in vitro indicator for screening probiotics with hypoglycemic or adjunctive hypoglycemic effects. To investigate the α-glucosidase inhibition rate of *Pediococcus lactis* AUI2505, the following experiment was conducted:

[0159] Acarbose (0.001 M) was used as the drug control group, and LGG probiotics were used as the strain control. A 96-well microtiter plate was prepared, and 25 μL of 2.5 mmol / L nitrophenyl-β-D-glucopyranoside (PNPG) solution and 25 μL of fermentation supernatant, bacterial suspension, or inactivated bacterial suspension of *Pediococcus lactis* AUI2505 were added to the reaction wells and incubated at 37°C for 10 min. Then, 50 μL of 0.2 U / μL α-glucosidase solution was added, and the reaction was continued at 37°C for 15 min. Finally, 100 μL of 0.2 mol / L sodium carbonate (Na2CO3) solution was quickly added to terminate the reaction. The absorbance (E) was measured at 405 nm using a microplate reader. Three parallel experiments were set up, as detailed in Table 10. The formula for calculating the α-glucosidase inhibition rate (%) is as follows:

[0160]

[0161] In the formula, F can be prepared in the same way as E, except that PBS (pH 6.8) is used instead of α-glucosidase solution; G can be prepared in the same way as E, except that PBS (pH 6.8) is used instead of fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; H can be prepared in the same way as E, except that PBS (pH 6.8) is used instead of α-glucosidase solution, and PBS (pH 6.8) is used instead of fermentation supernatant, bacterial suspension or inactivated bacterial suspension of Pediococcus lactis AUI2505; different letters in the same column represent significant differences (p < 0.05).

[0162] Table 10

[0163]

[0164] The experimental results showed that the fermentation supernatant and inactivated bacterial suspension of *Pediococcus lactis* AUI2505 had higher inhibition rates against α-glucosidase than LGG probiotics, and the fermentation supernatant also showed higher inhibition rates than the hypoglycemic drug acarbose. The bacterial suspension of *Pediococcus lactis* AUI2505 had a slightly lower inhibition rate against α-glucosidase than LGG probiotics and acarbose. These results indicate that *Pediococcus lactis* AUI2505 has the ability to inhibit α-glucosidase, and its fermentation supernatant has a better inhibitory effect.

[0165] Example 7: Evaluation of the hypoglycemic effect of the strain on a zebrafish hyperglycemia model

[0166] (1) Construction of a zebrafish hyperglycemia model:

[0167] Eight tanks of sexually mature wild-type zebrafish were naturally paired for breeding. The resulting eggs were disinfected, and 300 mL of embryo culture medium was added to the fry hatching tank, with the medium changed every 24 hours. Well-developed and highly active fry, 5 days post-fertilization (5 dpf), were randomly selected and placed in newly opened 6-well plates, with 30 fry per well. They were divided into a blank group, a model group, a positive control group, and an AUI2505 group, with the time, date, and group clearly marked. The experimental groupings and intervention methods are shown in Table 11. The intervention period was 5 days, and each group was fed algae feed normally at 9:00 AM daily.

[0168] Table 11

[0169]

[0170] Zebrafish with a growth factor of 10 dpf were treated with the drug for 4 h from 10:30 to 14:30. After the intervention, blood glucose was measured. The glucose concentration in the tissue fluid of the zebrafish in the model group was significantly higher than that in the blank group (P < 0.001), which indicates that the zebrafish hyperglycemia model was successfully constructed.

[0171] (2) Mortality rate:

[0172] During the zebrafish model construction, the number of dead fish was counted every day at 9:00 AM. After the count, the dead fish were removed. The mortality rate statistics of zebrafish juveniles can be seen in the graph. Figure 6 .

[0173] The experimental results showed that the mortality rate of zebrafish juveniles in all groups was normal, indicating that Pietrococcus lactis AUI2505 has good biosafety and will not cause significant damage to zebrafish.

[0174] (3) Weight:

[0175] The successfully constructed zebrafish model was washed three times with distilled water, and then three times with PBS. After washing, the PBS in the centrifuge tubes was thoroughly removed using 200 μL and 10 μL pipettes, and then the water was thoroughly absorbed using filter paper strips. The centrifuge tubes were weighed on a 0.01% balance, and the weight of the zebrafish fry was obtained by subtracting the weight of the empty centrifuge tube from the measured value. The weight statistics of the zebrafish fry can be seen in the graph. Figure 7 .

[0176] The experimental results showed that the weight of the blank group was significantly lower than that of the model group (p<0.001), and the weight of the positive control group and the AUI2505 group was significantly lower than that of the model group (p<0.05). This indicates that Pyorrhizococcus lactis AUI2505 can produce a good weight loss effect by lowering blood sugar or assisting in lowering blood sugar, and the effect is similar to that of the positive control metformin.

[0177] (4) Blood glucose concentration:

[0178] Add 20 μL of PBS to centrifuge tubes containing weighed zebrafish larvae, add one magnetic bead to each tube, and grind thoroughly at 50 Hz for 300 s. Centrifuge at 12000 rpm for 3 min. The supernatant is the tissue fluid of the zebrafish larvae. Immediately pipette 2 µL of the tissue fluid onto a blood glucose test strip to obtain the blood glucose concentration of the zebrafish larvae. The blood glucose concentration statistics of the zebrafish larvae can be seen in the graph. Figure 8 .

[0179] The experimental results showed that the blood glucose concentration in the blank group was 0.91 mmol / L, the blood glucose concentration in the model group was 2.63 mmol / L, the blood glucose concentration in the positive control group was 1.25 mmol / L, and the blood glucose concentration in the AUI2505 group was 1.22 mmol / L. Compared with the blank group, the blood glucose concentration in the model group increased significantly (P < 0.001), indicating that the zebrafish model was successfully established. Compared with the model group, the blood glucose concentration in the positive control group decreased significantly (P < 0.001), with a hypoglycemic efficacy of 52.47%, indicating that metformin has a significant hypoglycemic or adjunctive hypoglycemic effect on hyperglycemic zebrafish juveniles under the experimental concentration conditions. Compared with the model group, the blood glucose concentration in the AUI2505 group decreased significantly (P < 0.001), with a hypoglycemic efficacy of 53.61%, indicating that under the experimental concentration conditions, the bacterial suspension of *Pediococcus lactis* AUI2505 has a significant hypoglycemic or adjunctive hypoglycemic effect on hyperglycemic zebrafish juveniles, and its effect is similar to that of the positive control drug metformin.

[0180] (5) Biochemical indicators:

[0181] The levels of insulin and glucagon in the tissue fluid of the zebrafish juveniles were detected using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. For details, please refer to [link to kit]. Figure 9 and Figure 10 .

[0182] The experimental results showed that the insulin and glucagon levels in the model group were significantly different from those in the blank group (P<0.01); compared with the model group, the insulin levels in the positive control group and the AUI2505 group were significantly decreased (P<0.001), indicating that the AUI2505 group has a certain improving effect on diabetes-related biochemical indicators and can alleviate the metabolic disorder in hyperglycemic zebrafish.

[0183] (6) Inflammatory cytokines:

[0184] The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the tissue fluid of the above-mentioned zebrafish juveniles were detected using a commercially available ELISA kit (relative expression levels divided by the internal reference actin). For details, please refer to [link to kit]. Figure 11 and Figure 12 .

[0185] The experimental results showed that the levels of IL-6 and TNF-α in the model group were significantly different from those in the blank group (P < 0.001); compared with the model group, the levels of IL-6 and TNF-α in the zebrafish AUI2505 group were significantly different (P < 0.01), indicating that the AUI2505 group has a certain effect on alleviating inflammation in hyperglycemic zebrafish.

[0186] (7) OGTT & AUC glucose :

[0187] An oral glucose tolerance test (OGTT) was conducted on zebrafish with a blood glucose level of 10 dpf at 10:30. Blood glucose levels were measured at 0, 1, 2, 3, and 4 h. Blood glucose change curves were plotted, and the area under the blood glucose concentration-time curve (AUC) was calculated to explore the feasibility and stability of the blood glucose model. (See details...) Figure 13 and Figure 14 .

[0188] Zebrafish with a blood glucose level of 10 dpf had been immersed in culture water containing 3% (m / v) glucose for 17 hours before 10:30. Blood glucose monitoring was started at 10:30 for a total duration of 4 hours, with monitoring every 1 hour. Figure 13 The results showed that the blood glucose levels of zebrafish juveniles in all groups initially increased and then decreased. Figure 14The results showed that the AUC value of the AUI2505 group was significantly lower than that of the model group (P < 0.001), specifically 9.31, similar to the effect of the positive drug control. Based on the OGTT test results and the AUC value, it can be concluded that strain AUI2505 can significantly alleviate the glucose load capacity of hyperglycemic zebrafish and improve their symptoms.

[0189] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A type of *Pediococcus lactis* AUI2505, characterized in that, The *Pediococcus acidilactici* strain AUI2505 was deposited on March 16, 2026, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCC NO.67956 and classification name Pediococcus acidilactici.

2. A culture, fermentation product, or fermentation supernatant of Pediococcus lactis AUI2505 as described in claim 1.

3. A microbial inoculant, characterized in that, Includes the *Pediococcus lactis* AUI2505 as described in claim 1 and / or the culture, fermentation product, or fermentation supernatant as described in claim 2.

4. The use of the *Pediococcus lactis* AUI2505 of claim 1, the culture or fermentation product or fermentation supernatant of claim 2, and the microbial agent of claim 3 in the preparation of products that help with weight loss, anti-oxidation, anti-inflammation, blood sugar reduction, or blood sugar reduction.

5. The application according to claim 4, characterized in that, In the product, the viable count of the *Pediococcus lactis* AUI2505 is 1 × 10⁻⁶. 6 -10 10 CFU / mL or 1×10 6 -10 10 CFU / g.

6. The application according to claim 4 or 5, characterized in that, The product also includes a carrier and / or physiologically acceptable excipients; The carrier includes at least one of microcapsules, microspheres, nanoparticles, and liposomes; The excipients include at least one of the following: fillers, flavoring agents, diluents, wetting agents, binders, disintegrants, lubricants, color, flavor and aroma modifiers, solvents, solubilizers, co-solvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local analgesics, pH adjusters, and isotonic or isotropic modifiers.

7. The application according to claim 4 or 5, characterized in that, The products include at least one of the following: food, functional food, health food, and medicine.

8. The application according to claim 7, characterized in that, The functional foods include at least one of the following: special dietary foods, special medical purpose foods, infant formula milk powder, children's formula milk powder, adolescent formula milk powder, adult formula milk powder, functional powders, functional granules, functional capsules, functional beverages, and sports nutrition foods.

9. The application according to claim 7, characterized in that, The health food products include at least one of the following: those that help with weight loss, anti-oxidation, anti-inflammation, and blood sugar reduction.

10. The application according to claim 7, characterized in that, The drug includes at least one of the following: drugs for the prevention or treatment of hyperglycemia, drugs for the prevention or treatment of diabetes, drugs for the prevention or treatment of obesity, and drugs for the prevention or treatment of diseases caused by overweight.

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