Phytobacterium plantarum and probiotic composition thereof, and application thereof in relieving dog stress and improving dog enteritis

Abstract

The invention discloses a plant lactobacillus Kimate-F, a plant lactobacillus Kimate-6, a composite probiotic composition composed of the plant lactobacillus Kimate-F and the plant lactobacillus Kimate-6, and an application of the composite probiotic composition in relieving dog transportation stress and improving dog enteritis. The probiotic composition can effectively relieve dog transportation stress and remarkably reduce indexes such as corticosterone and active oxygen. In addition, the probiotic composition can effectively prevent or relieve canine enteritis, promote reproduction of beneficial bacteria in intestinal flora, inhibit proliferation of potential harmful bacteria, repair intestinal mucosal barrier and reduce inflammatory response. The invention has a wide application prospect, is especially suitable for dogs with long-distance transportation, stress susceptibility and poor intestinal function, and can also be used in cooperation with other conventional treatment means.

Description

Technical Field

[0001] The invention belongs to the field of microorganism and animal health care, and particularly relates to Lactobacillus plantarum and a probiotic composition thereof and application thereof in relieving stress and improving enteritis in dogs. Background Art

[0002] Dogs are often exposed to various stress factors during their growth and development and daily feeding, including long-distance transportation, sudden changes in the environment, changes in social relationships, vaccinations, and adjustments in feeding and management methods. These stress factors can cause dramatic fluctuations in the dog's neuro-endocrine system, usually manifested as adverse reactions such as increased cortisol levels, damaged intestinal barriers, and immune imbalance, which in turn lead to clinical symptoms such as decreased appetite and diarrhea, and may even induce or aggravate intestinal inflammation.

[0003] Canine enteritis is a common digestive disease in dogs. Its pathogenesis is relatively complex. It may be caused by infection with pathogenic microorganisms such as bacteria, viruses, and parasites, or by interactions with factors such as stress, dietary factors, and immune system disorders. Enteritis can manifest as diarrhea and loss of appetite in mild cases, or lead to severe dehydration, malnutrition, and even life-threatening conditions in severe cases.

[0004] At present, the main methods for dealing with canine stress and enteritis include drug therapy (such as antibiotics, anti-inflammatory drugs) and symptomatic supportive therapy. However, the use of antibiotics not only has the risk of inducing the emergence of drug-resistant strains, but may also further disrupt the intestinal microecological balance of dogs. Therefore, finding new prevention and control strategies that are safe, effective, and can replace traditional chemical drugs has become an urgent need in the field of animal health.

[0005] Lactobacillus plantarum is a Gram-positive lactic acid bacterium that is widely found in fermented foods and animal intestines and has excellent probiotic properties. A large number of studies have shown that Lactobacillus plantarum can play an important role in maintaining animal intestinal health by regulating the structure of the flora, enhancing intestinal barrier function, regulating immune responses, and alleviating stress responses.

[0006] Dogs often face various stress factors such as long-distance transportation, environmental changes, changes in social relationships, and vaccinations during their growth and development and daily management. These stresses can cause problems such as immune imbalance, damaged intestinal barriers, and oxidative stress in dogs, which in turn lead to symptoms such as loss of appetite and diarrhea, and even develop into enteritis in severe cases. However, due to the complexity of canine intestinal microbiota, the difficulty of strain screening and identification and functional verification, and the complex pathological mechanisms of canine stress and enteritis, it is difficult to obtain Lactobacillus plantarum that can be used to alleviate canine stress and canine enteritis, and there are certain accidental factors. Summary of the invention

[0007] Object of the Invention: The technical problem to be solved by the present invention is to provide two strains of Lactiplantibacillus plantarum, namely Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6.

[0008] Another technical problem to be solved by the present invention is to provide a compound probiotic composition that is safe, effective, can be used to alleviate the adverse effects caused by stress on the canine body, can prevent or improve canine enteritis, and thus avoid or reduce the risk of using antibiotics and other chemical drugs.

[0009] Another technical problem to be solved by the present invention is to provide a culture, lysate, extract, inactivated product or combination thereof containing the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6 or the probiotic composition.

[0010] Use of the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for alleviating the level of oxidative stress in the body.

[0011] Another technical problem to be solved by the present invention is to provide the use of the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for maintaining intestinal homeostasis and the recovery process after stress.

[0012] Another technical problem to be solved by the present invention is to provide the use of the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for preventing, alleviating or treating canine enteritis.

[0013] The last technical problem to be solved by the present invention is to provide the use of the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for preventing, alleviating or treating canine transportation stress.

[0014] Technical Solution: To solve the above technical problems, the present invention provides a strain of Lactiplantibacillus plantarum Kimate-F. The taxonomic name of the Lactiplantibacillus plantarum Kimate-F is Lactiplantibacillus plantarum, and it was deposited at the China General Microbiological Culture Collection Center (CGMCC) on December 25, 2023, with the deposit number: CGMCC No. 29388.

[0015] The present invention also includes a strain of Lactiplantibacillus plantarum Kimate-6, whose taxonomic name is Lactiplantibacillus plantarum, and it was deposited at the China General Microbiological Culture Collection Center (CGMCC) on January 26, 2024, with the deposit number: CGMCC No. 29777.

[0016] The present invention also includes a probiotic composition, which contains the above-mentioned Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6.

[0017] The present invention also includes the application of the above-mentioned Lactiplantibacillus plantarum Kimate-F, Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for alleviating the body's oxidative stress level. Preferably, the application includes increasing the serum SOD level and decreasing the oxidative stress indices ROS, MDA and the stress hormone cortisol level.

[0018] The present invention also includes cultures, lysates, extracts, inactivated products or combinations thereof containing the above-mentioned Lactiplantibacillus plantarum Kimate-F, Lactiplantibacillus plantarum Kimate-6 or the probiotic composition.

[0019] The present invention also includes the application of the above-mentioned Lactiplantibacillus plantarum Kimate-F, Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for maintaining intestinal homeostasis and the recovery process after stress. Preferably, the application regulates the composition of the canine intestinal flora, inhibits the proliferation of potential pathogenic bacteria and promotes the enrichment of beneficial bacteria. Preferably, the application includes promoting the enrichment of beneficial bacterial species such as Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Mogamoclostridium formicinis and Limosilactobacillus reuteri, and inhibiting the genera of bacteria related to inflammation or intestinal disorders such as Sutterella spp., Blautia wexlerae, Collinsella aerofaciens and Phocaeicola plebeius.

[0020] The present invention also includes the application of the above-mentioned Lactiplantibacillus plantarum Kimate-F, Lactiplantibacillus plantarum Kimate-6 or the probiotic composition in the preparation of feeds, drugs, foods or cosmetics for preventing, alleviating or treating canine enteritis.

[0021] Among them, the application includes alleviating weight loss caused by enteritis or increasing the anti-inflammatory level of the body. Preferably, the increase in the anti-inflammatory level of the body specifically includes reducing the LPS level and increasing the IL-10 level.

[0022] The present invention also includes the application of the Lactiplantibacillus plantarum Kimate-F, the Lactiplantibacillus plantarum Kimate-6, or the probiotic composition in the preparation of feeds, drugs, foods, or cosmetics for preventing, alleviating, or treating canine transportation stress.

[0023] Among them, the application specifically includes preventing stress reactions caused during transportation by increasing the antioxidant stress index serum SOD and reducing the oxidative stress indices ROS, MDA, and the stress hormone cortisol level.

[0024] Beneficial effects: Compared with the prior art, the present invention has the following advantages: The Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimete-6 of the present invention and their composite probiotic composition can significantly reduce indicators such as cortisol and reactive oxygen species in dogs caused by stress factors, thereby reducing the behavioral and physiological stress responses of dogs; this composite probiotic composition can also effectively prevent or alleviate canine enteritis, promote the reproduction of beneficial bacteria in the intestinal flora, inhibit the proliferation of potentially harmful bacteria, repair the intestinal mucosal barrier, and reduce the inflammatory response. This composite probiotic composition is suitable for dog populations with different health needs, especially suitable for dogs with long-distance transportation, stress susceptibility, and poor intestinal function, and can also be used in conjunction with other conventional treatment means. The present invention can partially replace or reduce the use of chemical drugs (such as antibiotics), not only ensuring the health and welfare of dogs, but also helping to reduce the risk of drug resistance and environmental pollution problems. Description of the Drawings

[0025] Figure 1 It is a blood routine test result chart of the canine transportation stress experiment of the "combination of Lactiplantibacillus plantarum Kimate-6 and Lactiplantibacillus plantarum Kimate-F from dogs" of the present invention.

[0026] Figure 2 It is a serum test result chart of the canine transportation stress experiment of the "combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention.

[0027] Figure 3 It is a fecal microbiota analysis result chart of the canine transportation stress experiment of the "combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention.

[0028] Figure 4This is the result diagram of LEfSe analysis of fecal microbiota in the dog transportation stress experiment of the "Combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention.

[0029] Figure 5 This is the result diagram of the body weight change in the dog enteritis experiment of the "Combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention.

[0030] Figure 6 This is the result diagram of the fecal character analysis in the dog enteritis experiment of the "Combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention.

[0031] Figure 7 This is the result diagram of the serum test in the dog enteritis experiment of the "Combination of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 from dogs" of the present invention. Detailed implementation manners

[0032] The present invention will be specifically described below in conjunction with the accompanying drawings and embodiments.

[0033] Example 1 Obtaining, identification and biological characteristic experiments of Lactiplantibacillus plantarum Kimate-F and Kimate-6 1. Strain isolation

[0034] Prepare MRS medium. Preparation method: Pour an appropriate amount of MRS medium (peptone: 10.0 g / L, beef extract: 10.0 g / L, yeast extract: 5.0 g / L, glucose: 20.0 g / L, ammonium citrate: 2.0 g / L, magnesium sulfate: 0.1 g / L, dipotassium hydrogen phosphate: 2.0 g / L, manganese sulfate: 0.05 g / L, sodium acetate: 5.0 g / L, Tween 80: 1.0 g / L, made up to 1 liter with distilled water) into a sterile petri dish, seal and sterilize. Take 0.5 g of fecal sample from a dog (German Shepherd, male, 3 years old, healthy) and add it to a sterile tube. Use 5 mL of sterile 0.9% normal saline to resuspend, mix well with a vortex mixer to form a suspension. After filtration, serially dilute the filtrate with sterile normal saline, dilute to 10 -5 10 -6 10 -7 gradients. Pipette 100 μL of each dilution and inoculate onto MRS medium plates (agarose is added to the MRS medium) by the plate method (coat the diluted sample on the surface of the medium), and incubate in an incubator at 37°C for 24 h. Observe the colonies formed on the petri dish, select individual colonies, and use a sterile pipette in the petri dish and incubate at 37°C for 24 h. Pick a single colony and inoculate it into 10 mL of MRS liquid medium, and incubate statically at 37°C for 20 h to obtain a pure culture solution of a single strain of lactic acid bacteria.

[0035] 2. Molecular biological identification of strains

[0036] The obtained pure culture solution of lactic acid bacteria was streaked on the MRS medium plate with an inoculation needle and cultivated in an incubator at 37 °C for 12 h. A single colony on the plate was picked up with a pipette tip. The purified bacterial single colony was directly used as the DNA template to amplify the 16S rRNA gene of the strain. PCR amplification was carried out using the universal primers for bacterial identification 16S rDNA 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' and 1492R: 5'-TACGGYTACCTTGTTACGACTT-3', which were synthesized by Shanghai Sangon Biotech Co., Ltd. The PCR amplification system was: 25 μL of 2×Taq PCR Master Mix; 19 μL of ddH2O; 2 μL of forward primer 27F (25 μmol / L), 2 μL of reverse primer 1492R (25 μmol / L), and 2 μL of DNA template. The amplification conditions were 95 °C for 5 min; 94 °C for 30 s, 55 °C for 30 s, 72 °C for 2 min, for 30 cycles; 72 °C for 7 min, and the reaction was terminated at 4 °C. The PCR products were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing. After obtaining the 16S rRNA genes of the two strains, sequence alignment was performed with the NCBI database, and it was determined that the two isolated strains were Lactiplantibacillus plantarum. One strain was Lactiplantibacillus plantarum Kimate-F of the present invention, which was deposited in the China General Microbiological Culture Collection Center on December 25, 2023, with the deposit number: CGMCC No. 29388; the deposit address was Beijing, China. The other strain was Lactiplantibacillus plantarum Kimate-6, which was deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms on January 26, 2024, with the deposit number: CGMCC No. 29777, and the deposit address was Beijing, China.

[0037] 3. Acid tolerance detection of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 strains

[0038] Probiotic acid tolerance experiment: Pipette the activated Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 (1×10 7CFU) bacterial solution, each with 10 mL, centrifuged and discarded the supernatant to obtain bacterial precipitate. Add 10 mL of sterile PBS (sodium chloride (NaCl): 8.0 g, sodium dihydrogen phosphate (NaH2PO4·H2O): 0.144 g, sodium dihydrogen phosphate (Na2HPO4·2H2O): 1.44 g, potassium chloride (KCl): 0.2 g, add distilled water to 1 liter, adjust the pH to 7.4) to each portion, resuspend and mix, and wash away the residual culture medium. Then centrifuge and discard the supernatant to obtain bacterial precipitate. Add 10 mL of sterile PBS with a pH value of 2 to three of the bacterial precipitates, and add 10 mL of untreated sterile PBS to the other three portions. The six bacterial solutions were incubated in an incubator at 37°C. After 3 hours, two bacterial solutions were taken out and centrifuged, and the supernatant was discarded. Each was resuspended with 10 mL of sterile PBS to wash away the residual treatment solution. The two tubes of bacterial solution were diluted and applied, and the dilution gradient was 10 6 , 10 7 , 10 8 After incubation at 37°C for 12 hours, the colonies were counted and the bacterial concentration of the culture was calculated. The survival rate of bacteria at pH 2 was calculated using the following formula: survival rate of bacteria at pH 2 = (pH = 2 concentration of the culture after PBS treatment / concentration of the culture after ordinary PBS treatment)%. Bifidobacterium BB-12, a commonly used probiotic strain for dogs, was selected as a reference strain and the acid resistance test was performed using the same method.

[0039] Through the acid resistance test, the acid resistance test results are shown in Table 1. The survival rate of Lactobacillus plantarum Kimate-F and Lactobacillus plantarum Kimate-6 exposed to a pH 2 environment for 3 hours is still higher than 25%, which is significantly higher than the reference strain Bifidobacterium BB-12 (product sold by Difule Biotechnology Co., Ltd.: Xiaoan Jiabao BB-12), and has strong acid resistance.

[0040] Table 1 Acid resistance test results of Lactobacillus plantarum Kimate-F and Lactobacillus plantarum Kimate-6

[0041]

[0042] 4. Bile salt tolerance test of Lactobacillus plantarum Kimate-F and Lactobacillus plantarum Kimate-6 strains

[0043] The test steps are basically the same as those of the probiotic acid resistance test, except that the pH=2 PBS needs to be replaced with 0.3% swine bile salt concentration PBS. The formula for calculating the bacterial survival rate under 0.3% swine bile salt saline treatment is: bacterial survival rate under 0.3% bile salt conditions = (bacterial solution concentration after 0.3% swine bile salt PBS treatment / bacterial solution concentration after ordinary PBS treatment)%.

[0044] Through the bile salt tolerance test, the results of the bile salt tolerance test are shown in Table 2. The survival rate of Lactiplantibacillus plantarum Kimate-6 was still higher than 45% after 3 hours of exposure to a high concentration of bile salt environment, which was similar to the survival rate of the reference strain Bifidobacterium BB-12, with no significant difference, indicating its strong bile salt tolerance performance.

[0045] Table 2 Results of bile salt tolerance test of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6

[0046]

[0047] Example 2 Study on the combined use of Lactiplantibacillus Kimate-F and Kimate-6 to relieve stress in dogs

[0048] Fourteen beagle dogs of 18 months old with similar body weights were randomly divided into 2 groups, namely the probiotic intervention group and the control group, with 7 dogs in each group. Both groups were fed the same commercial dog food (Myfoodie, barf 3.0), 60 g / dog / day. Diet and water were not restricted during the experiment. The probiotic intervention group was additionally fed 2×10 9 CFU of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 daily. After 4 weeks, transportation stress was carried out. All dogs were placed in a closed truck and transported at 60 km / h for 3 hours. After the transportation ended, 3 mL of blood was immediately collected with a vacuum blood collection tube. After the collected blood was left standing at room temperature for 15 min, it was centrifuged at 4000 rpm for 10 min to collect the serum for detecting stress and inflammation indexes. At the same time, fresh feces of dogs (within 15 minutes after defecation) were collected, aliquoted and stored at -80 °C for microbiota detection and analysis.

[0049] After exposure to transportation stress, compared with the control group, significant changes occurred in multiple inflammation and stress-related indexes in the probiotic intervention group ( Figure 1 and Figure 2 ). As Figure 1 shown, after transportation stress, the total number of white blood cells (WBC) in the control group increased significantly, while the WBC level in the probiotic intervention group was significantly lower (P<0.05) ( Figure 1 A). In addition, probiotic intervention significantly increased the number of lymphocytes (P<0.05) ( Figure 1 B), suggesting that probiotics may relieve the systemic inflammatory response caused by transportation through immunomodulation. Figure 2 shows the changes in stress hormones and oxidative stress-related indexes. The serum cortisol level in the probiotic intervention group was significantly lower than that in the control group (P<0.05) ( Figure 2(A), indicating its role in alleviating transportation stress response. Meanwhile, the content of reactive oxygen species (ROS) in the probiotic group decreased significantly (P<0.001). Figure 2 (B), indicating its strong antioxidant potential. The activity of superoxide dismutase (SOD), an antioxidant enzyme, increased significantly in the probiotic group (P<0.05). Figure 2 (C), while the content of malondialdehyde (MDA), a lipid peroxidation product, decreased significantly (P<0.05). Figure 2 (D), further verifying the potential protective effect of probiotics in alleviating oxidative damage induced by transportation stress.

[0050] Analysis of fecal microbiota found that there were significant differences in β-diversity between the intestinal microbiota of the two groups of dogs (P = 0.043), and the samples in the probiotic intervention group were significantly deviated from the control group in terms of spatial distribution. Figure 3 (A), suggesting that the combined intervention of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 can significantly regulate the composition and structure of the intestinal flora. Further shown by the box plot of Bray-Curtis distance, the differences among the intestinal flora in the probiotic group were significantly higher than those in the control group (P<0.05). Figure 3 (B), indicating that probiotic intervention increased the community diversity and dynamic changes of the intestinal flora. The results of LEfSe analysis further revealed the differences in characteristic bacteria between the two groups. Figure 4 ( ). Compared with the control group, the probiotic intervention group was significantly enriched in beneficial bacterial species such as Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Mogamoclostridium formicinis, and Limosilactobacillus reuteri, showing strong probiotic colonization ability; while the control group was enriched in genera related to inflammation or intestinal disorders such as Sutterella spp., Blautia wexlerae, Collinsella aerofaciens, and Phocaeicola plebeius.

[0051] The above results indicate that the combined intervention of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 can effectively relieve the immune disorder and oxidative stress response caused by transportation stress in dogs. At the same time, it can effectively regulate the composition of the intestinal flora of dogs, inhibit the proliferation of potential pathogenic bacteria, and promote the enrichment of beneficial bacteria, thus possibly contributing to maintaining intestinal homeostasis and the recovery process after stress.

[0052] Study on the combined use of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 to alleviate canine enteritis in Example 3

[0053] After the stress test, the probiotic intervention group continued to be fed 2×10 9 CFU of Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 daily, and the control group was not given probiotics. The experiment lasted for 14 days. From the 7th day, 5% concentration of DSS was added to the drinking water of all dogs to induce enteritis, and the body weight was recorded daily from the 7th day until the end of the experiment. Blood samples were collected on the 14th day, and serum was separated to detect inflammatory indicators (LPS, IL-10), and the body weight changes, diarrhea degree and serum inflammation levels of the two groups were compared to evaluate the alleviating effect of probiotics on DSS-induced canine enteritis.

[0054] To evaluate the alleviating effect of the combined intervention of Lactiplantibacillus plantarum Kimate-6 and Lactiplantibacillus plantarum Kimate-F on DSS-induced canine enteritis, evaluations were carried out from aspects such as body weight changes, fecal traits, and inflammatory factors. As Figure 5 shown, DSS treatment significantly induced a continuous decrease in the body weight of dogs. Since the 11th day, the decrease in body weight in the control group was significantly greater than that in the probiotic group, and the difference was statistically significant (P<0.05). By the 14th day, the body weight loss in the probiotic group was controlled at about 12%, while that in the control group had exceeded 16% (P<0.01). The results showed that probiotic intervention could effectively alleviate the body weight loss caused by DSS. Fecal trait analysis further verified the alleviating effect of probiotics. As Figure 6 shown in A, the feces of dogs in the DSS group were thin and accompanied by obvious bleeding, while the feces in the probiotic group were better formed and the bleeding phenomenon was significantly reduced. The fecal score showed that the probiotic group was significantly lower than the control group (P<0.05). At the same time, the results of the occult blood test also showed that the fecal occult blood score in the probiotic group was significantly reduced (P<0.05)( Figure 6 B), suggesting its certain hemostatic and protective effect. In addition, the results of serum inflammatory factor detection showed that DSS induction significantly increased the lipopolysaccharide (LPS) level, while LPS in the probiotic intervention group decreased significantly (P<0.001), indicating that it could alleviate the endotoxin transfer caused by intestinal barrier damage( Figure 7 A). At the same time, the anti-inflammatory factor interleukin-10 (IL-10) increased significantly in the probiotic group (P<0.01)( Figure 7 B), suggesting that Lactiplantibacillus plantarum Kimate-6 and Lactiplantibacillus plantarum Kimate-F have the potential to activate the anti-inflammatory pathway and restore intestinal immune balance.

[0055] In summary, the combined use of Lactiplantibacillus plantarum Kimate-6 and Lactiplantibacillus plantarum Kimate-F can significantly alleviate the DSS-induced colitis phenotype in dogs, improve body weight, relieve diarrhea, reduce bleeding, and play a protective role by inhibiting pro-inflammatory factors and enhancing anti-inflammatory factors.

Claims

1. A Lactiplantibacillus plantarum Kimate-F, characterized in that, The taxonomic name of the Lactiplantibacillus plantarum Kimate-F is Lactiplantibacillus plantarum, and it was deposited in the China General Microbiological Culture Collection Center (CGMCC) on December 25, 2023, with the deposit number: CGMCC No. 29388.

2. A Lactiplantibacillus plantarum Kimate-6, characterized in that, The taxonomic name of the Lactiplantibacillus plantarum Kimate-6 is Lactiplantibacillus plantarum, and it was deposited in the China General Microbiological Culture Collection Center (CGMCC) on January 26, 2024, with the deposit number: CGMCC No. 29777.

3. A probiotic composition, characterized in that, The composition comprises the Lactiplantibacillus plantarum Kimate-F and Lactiplantibacillus plantarum Kimate-6 as claimed in claims 1 and 2.

4. Use of the Lactiplantibacillus plantarum Kimate-F according to claim 1, the Lactiplantibacillus plantarum Kimate-6 according to claim 2, or the probiotic composition according to claim 3 in the preparation of feed, medicine, food or cosmetics for alleviating the body's oxidative stress level. Preferably, the use includes increasing the serum SOD level and reducing the oxidative stress indices ROS, MDA and the stress hormone cortisol level.

5. A culture, lysate, extract, inactivated product or combination thereof containing the Lactiplantibacillus plantarum Kimate-F according to claim 1, the Lactiplantibacillus plantarum Kimate-6 according to claim 2, or the probiotic composition according to claim 3.

6. Use of the Lactiplantibacillus plantarum Kimate-F according to claim 1, the Lactiplantibacillus plantarum Kimate-6 according to claim 2, or the probiotic composition according to claim 3 in the preparation of feed, medicine, food or cosmetics for maintaining intestinal homeostasis and the recovery process after stress. Preferably, the use regulates the composition of the canine intestinal flora, inhibits the proliferation of potential pathogenic bacteria, and promotes the enrichment of beneficial bacteria. Preferably, the use includes promoting the enrichment of beneficial bacterial species such as Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Mogamoclostridium formicinis and Limosilactobacillus reuteri, and inhibiting the genera of bacteria related to inflammation or intestinal disorders such as Sutterella spp., Blautia wexlerae, Collinsella aerofaciens and Phocaeicola plebeius.

7. Use of the Lactiplantibacillus plantarum Kimate-F according to claim 1, the Lactiplantibacillus plantarum Kimate-6 according to claim 2, or the probiotic composition according to claim 3 in the preparation of feed, medicine, food or cosmetics for preventing, alleviating or treating canine enteritis.

8. The use according to claim 7, characterized in that, The application includes alleviating weight loss caused by enteritis or increasing the anti-inflammatory level of the body. Preferably, the increase in the anti-inflammatory level of the body specifically includes reducing the LPS level and increasing the IL-10 level.

9. Use of Lactiplantibacillus plantarum Kimate-F according to claim 1, Lactiplantibacillus plantarum Kimate-6 according to claim 2, or the probiotic composition according to claim 3 in the preparation of feed, medicine, food or cosmetics for preventing, alleviating or treating transportation stress in dogs.

10. The use according to claim 9, characterized in that The application specifically includes preventing stress reactions caused during transportation by increasing the antioxidant stress index serum SOD and reducing the oxidative stress indexes ROS, MDA and the stress hormone cortisol level.

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