Food composition, feed composition, and feed additive for improving intestinal health comprising lactobacillus sakei CVL-001 strain as active ingredient, and other uses thereof

Abstract

The present invention relates to: a food composition, a feed composition, and a feed additive, for preventing or ameliorating infectious intestinal diseases or improving intestinal health function, each comprising, as an active ingredient, a Lactobacillus sakei CVL-001 strain isolated from kimchi; and other uses thereof. Specifically, the composition of the present invention exhibits an intestinal infection inhibitory effect in a mouse model in which an infectious intestinal disease was induced by Citrobacter rodentium, which is a mouse-specific intestinal bacterial pathogen, and thus can be used as a composition for preventing or ameliorating infectious intestinal diseases or improving intestinal health, which can be safely used in humans or animals without toxicity and side effects.

Description

Food composition for improving gut health, feed composition, feed additive, and other uses comprising the Lactobacillus sakei CVL-001 strain as an active ingredient

[0001] The present invention was carried out under the support of the Ministry of Science and ICT under project number 2710011602 and sub-project number 2022R1A2C2012287, the research management agency for the above project is the National Research Foundation of Korea, the research project name is "Basic Research Project (Mid-career Researcher Type I)", the research project name is "Study on the disease control mechanism of probiotics through the modulation of node-like receptor signaling", the lead institution is the Industry-Academic Cooperation Foundation of Chonnam National University, and the research period is 2022.03.01 ~ 2027.02.28.

[0002] This patent application claims priority to Korean Patent Application No. 10-2025-0024877 filed with the Korean Intellectual Property Office on February 26, 2025, and the disclosures of said patent application are incorporated herein by reference.

[0003] The present invention relates to a food composition, feed composition, feed additive, and other uses for the prevention or improvement of infectious intestinal diseases or for improving intestinal health function, comprising the Lactobacillus sakei CVL-001 strain as an active ingredient.

[0004] Bacterial infections are diseases caused by pathogenic bacteria that infect various body parts and can lead to serious health problems. As a type of infectious disease, it records high morbidity and mortality rates worldwide. In particular, the emergence of antibiotic-resistant bacteria is increasing the difficulty of treating bacterial infections. In Korea as well, bacterial infections are one of the major causes of both healthcare-associated and community-acquired infections, with a large number of patients diagnosed annually. Specifically, diseases such as pneumonia, urinary tract infections, and tuberculosis are considered major bacterial infections, and their incidence is rising due to the aging population and the increase in chronic diseases. The key characteristics of bacterial infections are that pathogens invade the body and trigger an immune response, and symptoms can progress rapidly. Clinical manifestations vary depending on the site of infection, and in severe cases, it can lead to systemic complications such as sepsis.

[0005] Infectious enteritis is a disease in which pathogenic microorganisms invade the intestinal tract, causing inflammation, diarrhea, and dehydration; it is emerging as a significant public health and veterinary issue in both humans and animals. Infectious enteritis exhibits high morbidity and mortality rates worldwide and is particularly fatal to humans and animals with weakened immune systems. Recently, infectious agents such as pathogenic Escherichia coli and Clostridioides difficile (C. difficile) have been receiving attention in both humans and animals.

[0006] Enteropathogenic E. coli (EPEC) and Enterohemorrhagic E. coli (EHEC) are major pathogens causing infectious intestinal diseases, with cases of infection reported in both humans and animals. EPEC possesses pathological characteristics that involve attaching to intestinal epithelial cells and altering their structure, and is known to be a primary cause of diarrhea in children. Enterohemorrhagic E. coli causes serious complications such as bloody stools and hemolytic uremic syndrome (HUS), and is transmitted through the consumption of contaminated food. Since there are frequent carrier cases not only in humans but also in livestock, particularly ruminants such as cattle, it is a critical target for management in both food and animal hygiene.

[0007] Clostridium difficile (C. difficile) is a major cause of enteritis following antibiotic use and is causing problems not only in humans but also in companion animals and farm animals. Toxin-producing C. difficile causes diseases that can lead to diarrhea, intestinal necrosis, and, in severe cases, death, and treatment is becoming increasingly difficult due to the spread of antibiotic-resistant strains.

[0008] The prevention and management of infectious intestinal diseases are critical tasks in both public health and veterinary medicine, making hygiene management, vaccine development, and monitoring of infection sources essential. However, the emergence of antibiotic-resistant bacteria presents serious challenges to treatment and prevention. As the efficacy of conventional antibiotics declines, treatment is becoming increasingly complex, and there is a growing need for the development of new drugs and alternative treatment strategies to curb the spread of resistant strains. To address these issues, a "One Health" approach encompassing both animal and human health is essential, requiring continuous research and the strengthening of infection control policies.

[0009] Lactic acid bacteria are garnering attention as an alternative solution to the problem of antibiotic resistance in the prevention and treatment of infectious intestinal diseases. By regulating the balance of intestinal microorganisms, lactic acid bacteria inhibit the proliferation of pathogenic microbes and strengthen the immune response, offering a novel approach that does not rely on antibiotics. In particular, given that the spread of antibiotic-resistant bacteria has become a serious issue due to antibiotic overuse, lactic acid bacteria are emerging as an alternative therapy that can be used safely with fewer side effects.

[0010] In actual research, lactic acid bacteria such as Lactobacillus plantarum and Bifidobacterium bifidum play an effective role in the prevention and treatment of infectious intestinal diseases. These lactic acid bacteria are effective in inhibiting the proliferation of resistant bacteria, such as pathogenic E. coli (e.g., EPEC, EHEC) and Clostridioides difficile, protecting the intestinal mucosa, and reducing inflammatory responses. Research results have reported that lactic acid bacteria strengthen intestinal barrier function and prevent pathogens from adhering to the intestines, thereby improving the intestinal environment and limiting the activity of pathogenic microorganisms.

[0011] In animals as well, probiotics are being utilized as an important tool in addressing the problem of antibiotic resistance. In companion animals, the use of probiotic supplements to prevent or alleviate C. difficile infections resulted in improved diarrhea symptoms and suppressed the spread of resistant bacteria. In farm animals, cases have been confirmed where probiotics contributed to reducing productivity losses caused by diarrhea by suppressing pathogenic E. coli infections.

[0012] The greatest advantage of probiotic therapy is its ability to reduce antibiotic use and suppress the emergence of antibiotic-resistant bacteria. Since probiotics can be used in combination with conventional antibiotic treatment, they offer the potential to maximize therapeutic effects while mitigating the problem of resistance. Furthermore, probiotics help prevent the recurrence of infectious diseases by improving the balance of the gut microbiome and strengthening the immune system.

[0013] In conclusion, probiotics are regarded as a powerful alternative for addressing the issue of antibiotic resistance in the treatment and prevention of infectious intestinal diseases. It is crucial to optimize the efficacy of probiotic therapy through continuous research and clinical trials, and to expand its applicability in both humans and animals. Strategies utilizing probiotics are expected to present a new paradigm in the management of infectious intestinal diseases.

[0014] Currently, among the research on lactic acid bacteria, no preparation for improving gut health using Lactobacillus sakei CVL-001 isolated from kimchi has been developed yet.

[0015] [Prior Art Literature]

[0016] [Patent Literature]

[0017] (Patent Document 1) KR 10-2441405 B (Publication Date 2022-09-07)

[0018] The inventors completed the present invention by identifying that the Lactobacillus sakei CVL-001 strain exhibits an infection control effect in a mouse infectious intestinal disease model mimicking EPEC (Enteropathogenic E. coli) and EHEC (Enterohemorrhagic E. coli) infections.

[0019] Accordingly, the objective of the present invention is to provide a food composition for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health functions, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0020] Another objective of the present invention is to provide a feed composition for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health functions, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0021] Another objective of the present invention is to provide a feed additive for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health function, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0022] Another objective of the present invention is to provide a use of the Lactobacillus sakei CVL-001 strain for the prevention or improvement of infectious intestinal diseases, or for the improvement of intestinal health functions.

[0023] The present invention relates to a food composition, a feed composition, and a feed additive for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health, comprising a Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient. It has been confirmed that the composition according to the present invention can provide efficacy in preventing or improving infectious intestinal diseases and efficacy in improving intestinal health by demonstrating control of the proliferation of infectious bacteria in an animal model in which bacterial infection is induced in the large intestine.

[0024] The inventors confirmed that the Lactobacillus sakei CVL-001 strain showed excellent effects in reducing the number of Colony Forming Units (CFU) in feces and reducing histopathological inflammatory markers of the large intestine in a mouse model in which bacterial infection was induced in the large intestine.

[0025] The present invention will be described in more detail below.

[0026] One aspect of the present invention is a food composition for preventing or improving infectious intestinal diseases or improving intestinal health, comprising a Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0027] The above food composition may be a food composition for improving intestinal health function that includes the Lactobacillus sakei CVL-001 strain as an active ingredient and exhibits efficacy in reducing pathogens that cause infectious intestinal diseases.

[0028] In the present invention, the strain may include a 16S rRNA base sequence represented by SEQ ID NO. 1.

[0029] In the present invention, the strain may be the one deposited under accession number KCTC 13816BP.

[0030] In the present invention, the strain may be one or more selected from the group consisting of live bacteria, dead cells, lysed products thereof, and extracts thereof.

[0031] In the present invention, the strain may be isolated from kimchi.

[0032] In the present invention, the food composition comprises 5 x 10 strains 4 Up to 5 x 10 14 CFU / ml, 5 x 10 4 Up to 5 x 10 13 CFU / ml, 5 x 10 4 Up to 5 x 10 12 CFU / ml, 5 x 10 4 Up to 5 x 10 11 CFU / ml, 5 x 10 4 Up to 5 x 10 10 CFU / ml, 5 x 10 4 Up to 5 x 10 9 CFU / ml, 5 x 10 5 Up to 5 x 10 14CFU / ml, 5 x 10 5 Up to 5 x 10 13 CFU / ml, 5 x 10 5 Up to 5 x 10 12 CFU / ml, 5 x 10 5 Up to 5 x 10 11 CFU / ml, 5 x 10 5 Up to 5 x 10 10 CFU / ml, 5 x 10 5 Up to 5 x 10 9 CFU / ml, 5 x 10 6 Up to 5 x 10 14 CFU / ml, 5 x 10 6 Up to 5 x 10 13 CFU / ml, 5 x 10 6 Up to 5 x 10 12 CFU / ml, 5 x 10 6 Up to 5 x 10 11 CFU / ml, 5 x 10 6 Up to 5 x 10 10 CFU / ml, 5 x 10 6 Up to 5 x 10 9 CFU / ml, 5 x 10 7 Up to 5 x 10 14 CFU / ml, 5 x 10 7 Up to 5 x 10 13 CFU / ml, 5 x 10 7 Up to 5 x 10 12 CFU / ml, 5 x 10 7 Up to 5 x 10 11 CFU / ml, 5 x 10 7 Up to 5 x 10 10 CFU / ml, 5 x 10 7 Up to 5 x 10 9 CFU / ml, 5 x 10 8 Up to 5 x 10 14 CFU / ml, 5 x 10 8 Up to 5 x 10 13 CFU / ml, 5 x 10 8Up to 5 x 10 12 CFU / ml, 5 x 10 8 Up to 5 x 10 11 CFU / ml, 5 x 10 8 Up to 5 x 10 10 CFU / ml, 5 x 10 8 Up to 5 x 10 9 CFU / ml, 5 x 10 9 Up to 5 x 10 14 CFU / ml, 5 x 10 9 Up to 5 x 10 13 CFU / ml, 5 x 10 9 Up to 5 x 10 12 CFU / ml, 5 x 10 9 Up to 5 x 10 11 CFU / ml, or 5 x 10 9 Up to 5 x 10 10 It can be included at a concentration of CFU / ml, for example, 5 x 10 8 Up to 5 x 10 10 CFU / ml or 5 x 10 9 It may be included at a concentration of CFU / ml, but is not necessarily limited thereto.

[0033] When the content of the strain in the food composition according to the present invention is within the above content range, it exhibits excellent effects of preventing and improving infectious intestinal diseases and improving intestinal health, and when it falls outside the above content range, these effects may decrease.

[0034] In the present invention, the daily dosage of the strain is 5 x 10 4 Up to 5 x 10 14 CFU, 5 x 10 4 Up to 5 x 10 13 CFU, 5 x 10 4 Up to 5 x 10 12 CFU, 5 x 10 4 Up to 5 x 10 11 CFU, 5 x 10 4 Up to 5 x 1010 CFU, 5 x 10 4 Up to 5 x 10 9 CFU, 5 x 10 5 Up to 5 x 10 14 CFU, 5 x 10 5 Up to 5 x 10 13 CFU, 5 x 10 5 Up to 5 x 10 12 CFU, 5 x 10 5 Up to 5 x 10 11 CFU, 5 x 10 5 Up to 5 x 10 10 CFU, 5 x 10 5 Up to 5 x 10 9 CFU, 5 x 10 6 Up to 5 x 10 14 CFU, 5 x 10 6 Up to 5 x 10 13 CFU, 5 x 10 6 Up to 5 x 10 12 CFU, 5 x 10 6 Up to 5 x 10 11 CFU, 5 x 10 6 Up to 5 x 10 10 CFU, 5 x 10 6 Up to 5 x 10 9 CFU, 5 x 10 7 Up to 5 x 10 14 CFU, 5 x 10 7 Up to 5 x 10 13 CFU, 5 x 10 7 Up to 5 x 10 12 CFU, 5 x 10 7 Up to 5 x 10 11 CFU, 5 x 10 7 Up to 5 x 10 10 CFU, 5 x 10 7 Up to 5 x 10 9 CFU, 5 x 10 8 Up to 5 x 10 14 CFU, 5 x 10 8Up to 5 x 10 13 CFU, 5 x 10 8 Up to 5 x 10 12 CFU, 5 x 10 8 Up to 5 x 10 11 CFU, 5 x 10 8 Up to 5 x 10 10 CFU, 5 x 10 8 Up to 5 x 10 9 CFU, 5 x 10 9 Up to 5 x 10 14 CFU, 5 x 10 9 Up to 5 x 10 13 CFU, 5 x 10 9 Up to 5 x 10 12 CFU, 5 x 10 9 Up to 5 x 10 11 CFU, or 5 x 10 9 Up to 5 x 10 10 CFU, for example, 5 x 10 9 It may be CFU, but is not necessarily limited to this.

[0035] When the daily dosage of the strain in the food composition according to the present invention is within the above range, it exhibits excellent effects of preventing and improving infectious intestinal diseases and improving intestinal health, and when the content falls outside the above range, these effects may decrease.

[0036] In the present invention, the food composition comprises one or more intestinal pathogenic microorganisms selected from the group consisting of Citrobacter rodentium, Enteropathogenic E. coli (EPEC), Enterohemorrhagic E. coli (EHEC), Clostridioides difficile, Salmonella species (Salmonellaspp.), Campylobacter species (Campylobacterspp.), Yersinia enterocolitica, Shigella species (Shigellaspp.), Brucella species (Brucellaspp.), Listeria monocytogenes, Mycobacterium avium subspecies paratuberculosis (MAP), and Clostridium perfringens The number may be reduced, but is not necessarily limited thereto, and the above pathogenic microorganisms may include, without limitation, various bacteria that cause disease in animals, including humans.

[0037] The food composition according to the present invention can achieve the effects of preventing and treating infectious intestinal diseases and improving intestinal health by controlling, reducing, or inhibiting pathogenic microorganisms in the intestines.

[0038] In the present invention, the food composition can increase the expression of one or more genes selected from the group consisting of Reg3β (regenerating islet-derived 3β), IL-17A (Interleukin-17A), and IL-22 (Interleukin-22).

[0039] In the present invention, the food composition can increase neutrophils in the lamina propria of the colon mucosa.

[0040] In the present invention, the food composition can increase the length of the colon or decrease the length of the intestinal crypt.

[0041] In one embodiment of the present invention, the strain may be pre-cultured for 10 to 24 hours, 10 to 22 hours, 10 to 20 hours, 10 to 18 hours, 12 to 24 hours, 12 to 22 hours, 12 to 20 hours, 12 to 18 hours, 14 to 24 hours, 14 to 22 hours, 14 to 20 hours, 14 to 18 hours, 16 to 24 hours, 16 to 22 hours, or 16 to 20 hours, for example, 16 to 18 hours, but is not necessarily limited thereto.

[0042] In one embodiment of the present invention, the strain may be pre-cultured at 25°C to 35°C, 25°C to 34°C, 25°C to 32°C, 25°C to 30°C, 27°C to 35°C, 27°C to 34°C, 27°C to 32°C, 27°C to 30°C, 29°C to 35°C, 29°C to 34°C, 29°C to 32°C, or 29°C to 30°C, for example, 30°C, but is not necessarily limited thereto.

[0043] In one embodiment of the present invention, the strain may be a pre-cultured strain diluted to 2 to 50 times, 2 to 40 times, 2 to 30 times, 2 to 20 times, 2 to 10 times, 4 to 50 times, 4 to 40 times, 4 to 30 times, 4 to 20 times, 4 to 10 times, 6 to 50 times, 6 to 40 times, 6 to 30 times, 6 to 20 times, 6 to 10 times, 8 to 50 times, 8 to 40 times, 8 to 30 times, 8 to 20 times, or 8 to 10 times, for example, 10 times, and then cultured.

[0044] In one embodiment of the present invention, the strain may be cultured for 1 to 12 hours, 1 to 10 hours, 1 to 8 hours, 1 to 6 hours, 2 to 12 hours, 2 to 10 hours, 2 to 8 hours, 2 to 6 hours, 4 to 12 hours, 4 to 10 hours, or 4 to 8 hours, for example, 4 to 6 hours, but is not necessarily limited thereto.

[0045] In one embodiment of the present invention, the strain may be cultured at 25°C to 35°C, 25°C to 34°C, 25°C to 32°C, 25°C to 30°C, 27°C to 35°C, 27°C to 34°C, 27°C to 32°C, 27°C to 30°C, 29°C to 35°C, 29°C to 34°C, 29°C to 32°C, or 29°C to 30°C, for example, 30°C, but is not necessarily limited thereto.

[0046] In one embodiment of the present invention, the strain may have an absorbance (OD) at a wavelength of 600 nm of 0.1 to 1.2, 0.1 to 1, 0.1 to 0.8, 0.1 to 0.6, 0.2 to 1.2, 0.2 to 1, 0.2 to 0.8, 0.2 to 0.6, 0.4 to 1.2, 0.4 to 1, 0.4 to 0.8, or 0.4 to 0.6, for example, 0.6, but is not necessarily limited thereto.

[0047] In the present invention, the term "included as an active ingredient" means including a sufficient amount of the Lactobacillus sakei CVL-001 strain in the composition to achieve the desired effect. At this time, the "desired effect" in the present invention may be the improvement of intestinal health functions, such as the prevention or improvement of infectious intestinal diseases, and the sufficient amount to achieve such desired effect may be appropriately adjusted by the target of administration, the form of administration, the route of administration, etc.

[0048] In the present invention, the strain may be a live cell, a dead cell, or a mixture thereof. The dead cell may be obtained by killing the strain with heat or radiation, for example, by killing it by heat treatment, but is not limited thereto. The strain killed by heat treatment may contain cytoplasm, a cell wall, antimicrobial active substances such as bacteriocin, polysaccharides, organic acids, etc.

[0049] In the present invention, the dead cell may be a strain obtained by heat-treating the strain, a freeze-dried product thereof, or a diluted product thereof.

[0050] In the present invention, the food composition may include a culture solution of the Lactobacillus sakei CVL-001 strain. The culture solution may be a culture supernatant obtained after culturing the strain and removing the strain, a concentrate thereof, a fraction thereof, a filtered liquid thereof, a frozen product thereof, or a freeze-dried product thereof, and may be, for example, a culture supernatant, but is not necessarily limited thereto.

[0051] In the present invention, the term "object" refers to an animal. The animal may be, but is not limited to, a human, a non-human primate, a mouse, a rat, a hamster, a chicken, a duck, a goose, a turkey, a camel, a pig, a cow, a horse, a goat, a sheep, a dog, a cat, or a rabbit. The object may be an animal including a human, or an animal excluding a human.

[0052] The above food composition may be a health functional food composition or a general food composition.

[0053] When the food composition of the present invention is used as a food additive, the food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods.

[0054] The food composition according to the present invention may be various foods, beverages, food additives, etc.

[0055] There are no specific restrictions on the types of the above-mentioned foods. Examples of foods to which the above-mentioned substance may be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all foods in the conventional sense.

[0056] The above-mentioned beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the above-mentioned natural carbohydrates may be appropriately determined by the choice of a person skilled in the art.

[0057] In addition to the above, the food composition of the present invention may contain various nutritional agents, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, the food composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit beverages, and vegetable beverages. These ingredients may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.

[0058] The above food composition may be selected from the group consisting of capsules, tablets, powders, granules, liquids, pills, syrups, and bars.

[0059] The content of the Lactobacillus sakei CVL-001 strain, its dead cells, or its culture solution as an active ingredient contained in the food composition according to the present invention is not appropriately limited depending on the form of the food, the desired use, etc., and can be added in an amount of, for example, 0.01 to 15 weight% of the total weight of the food, and the health beverage composition can be added in a ratio of 0.02 to 10 g, for example, 0.3 to 1 g, based on 100 mL.

[0060] The health beverage composition according to the present invention contains, in the indicated proportions, the Lactobacillus sakei CVL-001 strain, its dead cells, or its culture medium as essential components, and there are no particular limitations on the liquid components, and it may contain various flavoring agents or natural carbohydrates as additional components, as in conventional beverages. Examples of the natural carbohydrates mentioned above include monosaccharides, such as glucose and fructose; disaccharides, such as maltose and sucrose; polysaccharides, such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (taumatin, stevia extract, such as rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used. The proportion of natural carbohydrates is generally about 1 to 20 g, for example, about 5 to 12 g per 100 mL of the composition of the present invention.

[0061] The food composition according to the present invention may contain various nutritional agents, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the compositions of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination. Although the proportion of these additives is not particularly important, it is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

[0062] Another aspect of the present invention is a feed composition for preventing or improving infectious intestinal diseases or improving intestinal health, comprising a Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0063] The feed composition of the present invention may be for use as feed for companion animals and livestock, and may be for use as feed applicable to various animals.

[0064] The above pets may be dogs, cats, rabbits, hamsters, birds, horses, turtles, guinea pigs, goldfish, guppies, carp, ferrets, lizards, parrots, iguanas, frogs, spiders, chameleons, miniature pigs, or chinchillas, but are not necessarily limited thereto, and any pet with a bowel may be included without restriction.

[0065] The above livestock may be cattle, horses, donkeys, pigs, sheep, goats, chickens, ducks, geese, turkeys, llamas, alpacas, deer, quails, or camels, but are not necessarily limited thereto, and may be included without restriction as long as they have intestines.

[0066] The above animals may be non-human primates, monkeys, gorillas, mice, rats, elephants, tigers, dolphins, penguins, koalas, kangaroos, lions, hippos, giraffes, wolves, bears, pandas, crocodiles, snakes, turtles, raccoons, or cranes, but are not necessarily limited thereto, and may be included without restriction as long as they have intestines.

[0067] The feed composition of the present invention can prevent or improve the intestinal health of companion animals and livestock by reducing pathogens that cause infectious intestinal diseases.

[0068] The feed composition of the present invention may further include functional substances.

[0069] The above functional substances may use one or more selected from DHA (Docosahexaenoic acid), EPA (Eicosapentaenoic acid), aloe, squalene, corn silk, Garcinia cambogia, hibiscus, bamboo leaves, chlorella, spirulina, chives, green tea, Solomon's seal, persimmon leaves, mulberry leaves, red sesame, brown rice, barley, rooibos, Pu-erh, and crystallizers, but are not limited thereto.

[0070] The feed composition of the present invention may further include components added to conventional feed compositions. Examples of components added to such feed compositions may include cereal powder, meat powder, and legumes.

[0071] In addition to the cereal powder, meat powder, and legumes added to the feed composition mentioned above, the feed composition of the present invention may include one or more selected from nutrients and minerals to increase the nutritional value of the feed, and may include one or more selected from antifungal agents, antioxidants, anticoagulants, emulsifiers, and binders to prevent deterioration of feed quality.

[0072] The components of the feed composition mentioned above may be appropriately selected and implemented by those skilled in the art, and since they differ somewhat from the essential components of the present invention, detailed information regarding this invention will be omitted below.

[0073]

[0074] In the present invention, the strain may include a 16S rRNA base sequence represented by SEQ ID NO. 1.

[0075] In the present invention, the strain may be the one deposited under accession number KCTC 13816BP.

[0076] In the present invention, the strain may be one or more selected from the group consisting of live bacteria, dead cells, lysed products thereof, and extracts thereof.

[0077] In the present invention, the feed composition comprises 5 x 10 strains 4 Up to 5 x 10 14 It may be included at a concentration of CFU / ml, but is not necessarily limited thereto.

[0078] In the present invention, the feed composition comprises one or more intestinal pathogenic microorganisms selected from the group consisting of Citrobacter rodentium, Enteropathogenic E. coli (EPEC), Enterohemorrhagic E. coli (EHEC), Clostridioides difficile, Salmonella species (Salmonellaspp.), Campylobacter species (Campylobacterspp.), Yersinia enterocolitica, Shigella species (Shigellaspp.), Brucella species (Brucellaspp.), Listeria monocytogenes, Mycobacterium avium subspecies paratuberculosis (MAP), and Clostridium perfringens The number may be reduced, but is not necessarily limited thereto, and the above pathogenic microorganisms may include, without limitation, various bacteria that cause disease in animals, including humans.

[0079] In the present invention, the feed composition can increase the expression of one or more genes selected from the group consisting of Reg3β, IL-17A, and IL-22.

[0080] In the present invention, the feed composition can increase neutrophils in the lamina propria of the colon mucosa.

[0081] In the present invention, the feed composition can increase the length of the colon or decrease the length of the intestinal crypt.

[0082] In the present invention, the feed composition may be administered orally, but is not necessarily limited thereto.

[0083] A feed composition for the prevention or improvement of infectious intestinal diseases or for improving intestinal health, comprising the Lactobacillus sakei CVL-001 strain of the present invention as an active ingredient, may be manufactured by additionally including additives that are conventionally added during feed production. For example, it may additionally include various nutrients such as vitamins, amino acids, and minerals, antioxidants, antibiotics, antimicrobial agents, and other additives, and its form may include powder, granules, pellets, or suspension.

[0084] The feed composition of the present invention may be fed to animals alone or mixed with other feeds. Other feeds mixed with the feed composition of the present invention include, but are not limited to, powdered feed, solid feed, moist pellet feed, dry pellet feed, EP (Extruder Pellet) feed, raw feed, etc.

[0085] Methods for adding the feed composition of the present invention to other feeds include mechanical mixing, adsorption, and adsorption, but are not limited thereto.

[0086] In the present invention, the term "feed" refers to any natural or artificial prescribed food, single meal, etc., or the components of said single meal, for animals to eat, consume, and digest, or suitable for such purposes. The feed composition containing the Lactobacillus sakei CVL-001 strain of the present invention may be a concentrated feed, roughage, and / or special feed.

[0087] The above concentrated feed includes seed fruits including grains such as wheat, oats, and corn; bran including rice bran, wheat bran, and barley bran as by-products obtained by refining grains; oilseed meal as by-products obtained by extracting oil from soybeans, rapeseed, sesame, flaxseed, and coconut; residues such as residual starch, which is the main component of starch residue remaining after removing starch from sweet potatoes, potatoes, etc.; animal feeds such as fish meal, fish residue, fish soluble which is a concentrated fresh liquid obtained from fish, meat meal, blood meal, feather meal, skim milk powder, cheese from milk, and dried whey which is the residue when producing casein from skim milk; yeast, Chlorella, and seaweed.

[0088] The above forage includes fresh forage such as wild grass, pasture grass, and green cuts; root vegetables such as feed turnips, feed beets, and a type of turnip called root vegetable; silage, which is a stored feed produced by filling a silo with fresh grass, green cut crops, and grains and fermenting them with lactic acid; hay made by cutting and drying wild grass and pasture grass; straw of crops; and leaves of legumes.

[0089] The above special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivatives such as diuretic isobutane, and feed additives, which are substances added in small amounts to compound feed to supplement components that are prone to being lacking when only natural feed ingredients are mixed, or to increase the shelf life of the feed.

[0090] The feed composition of the present invention can be appropriately used depending on the type of animal to be administered, age, and other types of feed ingredients.

[0091] The feed composition of the present invention may additionally include substances exhibiting various effects, such as supplementing nutrients and preventing weight loss, enhancing the digestibility and utilization of fiber in the feed, improving milk quality, preventing reproductive disorders and improving conception rates, and preventing high-temperature stress during the summer. For example, it may additionally include mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamin preparations such as carotene, vitamin E, vitamins A, D, E, nicotinic acid, and vitamin B complex; protected amino acid preparations such as methionine and lysic acid; protected fatty acid preparations such as calcium salts of fatty acids; live bacteria such as yeast cultures and mold fermentation products; yeast preparations, etc.

[0092] Another aspect of the present invention is a feed additive for preventing or improving infectious intestinal diseases or improving intestinal health, comprising a Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

[0093] In the present invention, the strain may include a 16S rRNA base sequence represented by SEQ ID NO. 1.

[0094] In the present invention, the strain may be the one deposited under accession number KCTC 13816BP.

[0095] In the present invention, the strain may be one or more selected from the group consisting of live bacteria, dead cells, lysed products thereof, and extracts thereof.

[0096] In the present invention, the feed additive comprises 5 x 10 strains 4 Up to 5 x 10 14 It may be included at a concentration of CFU / ml, but is not necessarily limited thereto.

[0097] In the present invention, the feed additive comprises one or more intestinal pathogenic microorganisms selected from the group consisting of Citrobacter rodentium, Enteropathogenic E. coli (EPEC), Enterohemorrhagic E. coli (EHEC), Clostridioides difficile, Salmonella species (Salmonella spp.), Campylobacter species (Campylobacter spp.), Yersinia enterocolitica, Shigella species (Shigella spp.), Brucella species (Brucella spp.), Listeria monocytogenes, Mycobacterium avium subspecies paratuberculosis (MAP), and Clostridium perfringens The number may be reduced, but is not necessarily limited thereto, and the above pathogenic microorganisms may include, without limitation, various bacteria that cause disease in animals, including humans.

[0098] The feed additive according to the present invention can achieve the effects of preventing and treating infectious intestinal diseases and improving intestinal health by controlling, reducing, or inhibiting pathogenic microorganisms in the intestines.

[0099] In the present invention, the feed additive can increase the expression of one or more genes selected from the group consisting of Reg3β, IL-17A, and IL-22.

[0100] In the present invention, the feed additive can increase neutrophils in the lamina propria of the colon's intestinal mucosa.

[0101] In the present invention, the feed additive can increase the length of the colon or decrease the length of the intestinal crypt.

[0102] In the present invention, the term "feed additive" refers to a substance added to feed to improve animal productivity, promote and / or maintain animal health, or increase body weight.

[0103] The content of the above feed additive included in the feed is not particularly limited thereto, but may be 0.01 to 20.0% (w / w), and preferably 0.05 to 10.0% (w / w).

[0104] The feed additive of the present invention may further include excipients. The excipients refer to components added to provide a suitable hardness or shape to the active ingredient, or to provide a certain volume and weight in cases where the amount of the main ingredient is small, for the purpose of making it easy to handle. The excipients may include those commonly used in the manufacture of feed additives, but are not limited thereto, and may include one or more of wheat bran, corn powder, cereal starch, silica powder, and diatomaceous earth, and may include excipients recognized in the Standards and Specifications for Feed, etc. (Ministry of Agriculture, Food and Rural Affairs Notification No. 2014-106).

[0105] The feed additive of the present invention may be manufactured in a conventional formulation in the art and may be implemented as a powder or in the form of pellets having a predetermined shape, but is not limited thereto. The feed additive may be powdered to enhance absorption in the body or manufactured as a fat-coated formulation to enhance activity in the small intestine.

[0106] The above excipient may be included in an amount of 50 to 99 parts by weight per 100 parts by weight of the total feed additive.

[0107] Another aspect of the present invention is a pharmaceutical composition for the prevention or treatment of infectious intestinal diseases comprising the Lactobacillus sakei CVL-001 strain as an active ingredient.

[0108] Another aspect of the present invention is a method for preventing, improving, or treating infectious intestinal disease, comprising the step of administering a composition containing the Lactobacillus sakei CVL-001 strain to an animal.

[0109] The above administration means introducing the composition of the present invention to an animal by any appropriate method, and the administration route of the composition of the present invention may be administered through various routes, such as oral or parenteral, as long as it can reach the target tissue.

[0110] Another aspect of the present invention is the use of the Lactobacillus sakei CVL-001 strain for the prevention, improvement, or treatment of infectious intestinal diseases, or for the improvement of intestinal health function.

[0111] The present invention relates to a food composition, feed composition, feed additive, and other uses for the prevention or improvement of infectious intestinal disease or for improving intestinal health function, comprising a strain of Lactobacillus sakei CVL-001 isolated from kimchi as an active ingredient. Since the composition of the present invention exhibits an infection-inhibiting effect and an effect of reducing the pathogen causing infectious intestinal disease in a mouse model in which infectious intestinal disease is induced using Citrobacter rodentium, an intestinal infection bacterium native to mice, it can provide an effect for the prevention or improvement of infectious intestinal disease and an effect for improving intestinal health.

[0112] Figure 1 is a figure showing the schedule for an animal experiment on infectious intestinal disease using the Lactobacillus sakei CVL-001 strain according to one embodiment of the present invention.

[0113] FIGS. 2a to 2d are the results of an animal experiment on the inhibition of infectious intestinal disease by the Lactobacillus sakei CVL-001 strain according to one embodiment of the present invention.

[0114] Figure 2a shows the result of quantifying the change in length due to an inflammatory response by measuring the length of the colon according to one embodiment of the present invention.

[0115] FIG. 2b shows the results of quantitative analysis of the genetic expression of the antibiotic Reg3β produced in the colon and the colony forming units (CFU) of Citrobacter rodentium detected in feces according to one embodiment of the present invention.

[0116] FIG. 2c is a graph showing an H&E (Hematoxylin and Eosin) stained photograph of colon tissue and a histopathological evaluation based thereon according to one embodiment of the present invention.

[0117] FIG. 2d is a graph showing the result of quantifying the length of the crypt within the colon tissue according to one embodiment of the present invention.

[0118] Figure 3 is a figure showing the schedule for animal experiments to investigate the core mechanism of inhibition of infectious intestinal diseases by Lactobacillus sakei CVL-001 according to one embodiment of the present invention.

[0119] FIGS. 4a and 4b are the results of animal experiments to investigate the core mechanism of inhibition of infectious intestinal disease by Lactobacillus sakei CVL-001 according to one embodiment of the present invention.

[0120] Figure 4a shows the results of quantitatively analyzing the ratio of neutrophils in the colon through flow cytometry according to one embodiment of the present invention.

[0121] FIG. 4b presents the results of quantitatively evaluating changes in gene markers IL-17A and IL-22 expressed in the colon using Real-Time Polymerase Chain Reaction according to one embodiment of the present invention.

[0122] The present invention will be described in more detail below through examples. These examples are intended solely to explain the invention more specifically, and it will be obvious to those skilled in the art that the scope of the invention is not limited by these examples according to the gist of the invention.

[0123] Throughout this specification, "%" used to indicate the concentration of a particular substance is (weight / weight)% for solid / solid, (weight / volume)% for solid / liquid, and (volume / volume)% for liquid / liquid, unless otherwise noted.

[0124]

[0125] Example 1: Culture of Lactobacillus sakei CVL-001

[0126] Lactobacillus sakei CVL-001, isolated from kimchi and deposited under accession number KCTC13816BP, was used as the lactic acid bacteria. The lactic acid bacteria stock was plated onto MRS agar, a selective lactic acid bacteria medium, and cultured in a 30°C incubator for 18 hours. A single colony was extracted, released into MRS broth, a liquid medium, and pre-cultured for 16 to 18 hours at 30°C and 150 rpm. To obtain the bacteria in optimal condition, the pre-cultured bacteria were diluted 10-fold in fresh MRS broth and main cultured for 4 to 6 hours at 30°C and 150 rpm. Subsequently, the sample was harvested when the optical density (OD) value was 0.6 at an absorbance of 600 nm, which is the point of highest bacterial activity. Under these conditions, the Lactobacillus sakei CVL-001 strain was 8.9 x 10 8 It is present in CFU / ml. Live lactic acid bacteria were washed with PBS by centrifugation (3,000 rpm, 15 min) and then administered using PBS as a solvent according to the administration concentration. The 16S rRNA coding sequence (total 1,439 bp) of the above Lactobacillus sakei CVL-001 strain is shown in Table 1 below.

[0127]

[0128]

[0129] Example 2: Effect of using Lactobacillus sakei CVL-001 on controlling infectious intestinal disease

[0130] 2-1. Schedule for Mouse Animal Experiments on Infectious Intestinal Disease Using Citrobacter rodentium

[0131] As can be seen in Fig. 1, 1x10 of Citrobacter rodentium (C. rodentium), an endemic mouse intestinal pathogen, were in 8-week-old female C57BL / 6 mice 9 Infection was induced by oral administration of CFU / 200 μL. Lactobacillus sakei CVL-001 1x10 was administered daily for a total of 17 days, starting 7 days prior to the date of infection and ending 10 days after infection. 9 CFU / 200 μL was administered orally. On the 10th day of infection, euthanasia was performed followed by autopsy. Citrobacter rodencium and PBS administration groups were used as controls.

[0132] 2-2. Evaluation of Disease Suppression in Infectious Intestinal Disease Mouse Animal Experiments Using Citrobacter rodentium

[0133] Prior to necropsy, mouse feces were plated on MacConkey selective medium to quantitatively measure the number of Colony Forming Units (CFU) of Citrobacter rodentium. Subsequently, the mice were euthanized, and necropsy was performed. The colon was excised to measure changes in colon length due to inflammation. The excised colon was subjected to histological analysis using hematoxylin and eosin (H&E) staining to evaluate inflammatory findings, such as changes in crypt length, a decrease in goblet cell count, and inflammatory cell infiltration.

[0134] As can be seen in Figure 2a, the colon length was significantly reduced in the PBS-administered group, which was infected with Citrobacter rodencium and administered PBS, compared to the control group, which was not infected with Citrobacter rodencium, whereas the colon length was significantly increased in the Lactobacillus sakei CVL-001-administered group compared to the PBS-administered group.

[0135] In addition, as can be seen in the left graph of Figure 2b, it was observed that the gene expression of Reg3β, an antimicrobial peptide (AMP) that helps eliminate infectious bacteria in the intestinal environment, was significantly increased.

[0136] As a result, as can be seen in the right graph of Figure 2b, the number of Colony Forming Units (CFU) of Citrobacter rodensium in feces was significantly reduced.

[0137] As can be seen in Figures 2c and 2d, the results of histopathological analysis evaluated by microscopy after H&E staining showed that the length of the intestinal stratum, which had increased due to Citrobacter rodensium infection, was significantly inhibited (Figure 2d), and the inflammatory response caused by the infection was significantly reduced, including inhibition of goblet cell loss and inhibition of inflammatory cell infiltration (Figure 2c).

[0138] 2-3. Experiment Schedule for Analyzing the Mechanism of Infection Inhibition by Lactobacillus sakei CVL-001 in a Mouse Model of Infectious Intestinal Disease Using Citrobacter rodentium

[0139] As can be seen in Fig. 3, Citrobacter rodentium, a bacterium endemic to the mouse gut, 1x10 9 Infection was induced by oral administration of CFU / 200 μL. Lactobacillus sakei CVL-001 1x10⁻¹⁰ was administered daily for a total of 12 days, starting 7 days prior to the date of infection and ending 5 days after infection. 9 CFU / 200 μL was administered orally. On the 5th day of infection, euthanasia was performed followed by a necropsy.

[0140] 2-4. Analysis of the mechanism of infection inhibition by Lactobacillus sakei CVL-001 in a mouse model of infectious intestinal disease using Citrobacter rodentium

[0141] After autopsy, the colon was excised, and immune cells were isolated from the lamina propria of the intestinal mucosa. Immune cell isolation from the lamina propria was achieved through mechanical fragmentation and enzymatic digestion, and the isolated cells were used as a lamina propria immune cell population for subsequent analysis. Subsequently, flow cytometry was performed to quantitatively measure the proportion of neutrophils within the lamina propria immune cell population.

[0142] In addition, Real-Time Polymerase Chain Reaction (Real-Time PCR) was performed on excised colon to quantitatively analyze changes in the expression levels of specific genes associated with infection control. To this end, RNA was extracted and reverse transcribed into cDNA to evaluate gene expression levels, and changes in expression levels were quantified using the ΔCt method. This analysis focused on identifying the regulatory mechanisms of key genes related to the inflammatory response induced by Citrobacter rodentium infection.

[0143] As can be seen in Figure 4a, flow cytometry analysis confirmed that the proportion of neutrophils, which play an important role in phagocytizing and eliminating intestinal pathogens within the colonic lamina propria, was statistically significantly increased in the Lactobacillus sakei CVL-001 administration group compared to the PBS administration group.

[0144] Additionally, as can be seen in the left graph of Figure 4b, Real-Time Polymerase Chain Reaction (Real-Time PCR) analysis showed that the expression level of IL-17A (Interleukin-17A), a major cytokine that induces neutrophil recruitment, was significantly increased in the Lactobacillus sakei CVL-001 administration group compared to the PBS administration group.

[0145] In addition, as can be seen in the graph on the right in Figure 4b, the expression level of the cytokine IL-22 (Interleukin-22), which plays a role in inducing the secretion of antimicrobial peptides (AMPs) that protect the intestinal epithelium and promote the elimination of infectious bacteria in intestinal bacterial infections, also showed a significant increase in the Lactobacillus sakei CVL-001 administration group compared to the PBS administration group. These results suggest that Lactobacillus sakei CVL-001 contributes to the elimination of infectious bacteria through the activation of the intestinal immune system and can play an important role, particularly in improving intestinal health.

[0146]

[0147] Sintering

[0148] From the above results, the inventors were able to confirm that when Lactobacillus sakei CVL-001 was orally administered to a mouse animal model of infectious intestinal disease using Citrobacter rodensium, a native mouse intestinal pathogen, the length of the colon, which had been reduced by Citrobacter rodensium infection, was significantly increased, the gene expression of the antimicrobial peptide Reg3β was significantly increased, the number of Citrobacter rodensium in the feces was significantly reduced, and histopathological analysis results showed that the length of the intestinal fibrosis, which had been increased by Citrobacter rodensium infection, was significantly inhibited, and the inflammatory response caused by infection was significantly reduced, including the inhibition of goblet cell loss and the inhibition of inflammatory cell infiltration.

[0149] Furthermore, the inventors confirmed that when the Lactobacillus sakei CVL-001 strain was orally administered to a mouse animal model of infectious intestinal disease caused by Citrobacter rodensium, the proportion of neutrophils in the lamina propria of the colon increased and the cytokines IL-17A and IL-22 were significantly increased, thereby confirming that the Lactobacillus sakei CVL-001 strain contributes to the elimination of infectious bacteria through a mechanism that activates the intestinal immune system.

[0150] Accordingly, the inventors have confirmed that the Lactobacillus sakei CVL-001 strain can be usefully used not only for the prevention or improvement of infectious intestinal diseases but also for the improvement of intestinal health functions.

[0151]

[0152] [Consignment Number]

[0153] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resources Center (KCTC)

[0154] Trustee Number: KCTC13816BP

[0155] Date of entrustment: 20190219

[0156]

Claims

1. A food composition for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

2. A food composition according to claim 1, wherein the strain comprises a 16S rRNA base sequence represented by SEQ ID NO.

1.

3. A food composition according to claim 1, wherein the strain is the one deposited under accession number KCTC 13816BP.

4. A food composition according to claim 1, wherein the strain is one or more selected from the group consisting of live bacteria, dead cells, crushed products thereof, and extracts thereof.

5. In claim 1, the food composition comprises 5 x 10 strains 4 Up to 5 x 10 14 A food composition containing at a concentration of CFU / ml.

6. In claim 1, the food composition comprises one or more intestinal pathogens selected from the group consisting of Citrobacter rodentium, Enteropathogenic E. coli (EPEC), Enterohemorrhagic E. coli (EHEC), Clostridioides difficile, Salmonella species (Salmonella spp.), Campylobacter species (Campylobacter spp.), Yersinia enterocolitica, Shigella species (Shigella spp.), Brucella species (Brucella spp.), Listeria monocytogenes, Mycobacterium avium subspecies paratuberculosis (MAP), and Clostridium perfringens A food composition that reduces the number of pathogenic microorganisms.

7. The food composition of claim 1, wherein the food composition increases the expression of one or more genes selected from the group consisting of Reg3β, IL-17A, and IL-22.

8. The food composition of claim 1, wherein the food composition increases neutrophils in the lamina propria of the colon.

9. The food composition of claim 1, wherein the food composition increases the length of the colon or decreases the length of the intestinal crypt.

10. A feed composition for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

11. A feed composition according to claim 10, wherein the strain comprises a 16S rRNA base sequence represented by SEQ ID NO.

1.

12. A feed composition according to claim 10, wherein the strain is the one deposited under accession number KCTC 13816BP.

13. A feed additive for the prevention or improvement of infectious intestinal diseases or for the improvement of intestinal health, comprising the Lactobacillus sakei CVL-001 strain or a culture solution thereof as an active ingredient.

14. A feed additive according to claim 13, wherein the strain comprises a 16S rRNA base sequence represented by SEQ ID NO.

1.

15. In paragraph 13, the above strain is a feed additive deposited under accession number KCTC 13816BP.

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