KR102443495B1 - Novel Strain of Lactobacillus rhamnosus CACC 612, and feed composition using thereof - Google Patents

Abstract

The present invention relates to a novel strain of Lactobacillus rhamnosus CACC 612, and a feed composition for companion animals containing the same as an active component. The Lactobacillus rhamnosus CACC 612 strain and a culture medium thereof of the present invention exhibit antibacterial activity against livestock pathogens, exhibit antibacterial activity against Clostridioides difficile strain, which is a livestock pathogen causing colitis and diarrhea in companion animals, and exhibit excellent effects on enhancing immunity and inhibiting inflammation, thereby being able to be usefully used for antibacterial, immune enhancing, and anti-inflammatory purposes of companion animals.

Description

New strain Lactobacillus rhamnosus CACC 612 and a feed composition for companion animals containing the same as an active ingredient {Novel Strain of Lactobacillus rhamnosus CACC 612, and feed composition using thereof}

The present invention relates to a new strain Lactobacillus rhamnosus CACC 612 and a feed composition for companion animals containing the same as an active ingredient. In Clostridioides difficile strain, it exhibits antibacterial activity, and it relates to a Lactobacillus rhamnosus CACC 612 strain, which has excellent effects in enhancing immunity and inhibiting inflammation, and a feed composition for companion animals using the same.

Currently, the number of single-person and two-person households is rapidly increasing in Korea due to the nuclear family. As a result, the number of people raising companion animals such as dogs and cats to find emotional stability is increasing.

However, since most companion animals in Korea live in houses, they lack exercise and are under a lot of stress, so their digestive and metabolic activities are easily lowered, and their immunity tends to be lowered. Due to this, metabolic diseases or immune-related diseases similar to humans occur, and in the future, it will worsen into more fatal internal and external diseases, resulting in a decrease in the quality of life of companion animals and their guardians.

Currently, interest in animal health and welfare in Korea is increasing, but realistic treatment costs are high and medical expenses are high. the current situation.

On the other hand, living microorganisms, probiotics, promote the health of animals by activating the functions of microorganisms in the intestine (Fuller, 1989). Many researchers have reported that probiotic strains such as Lactobacillus and Bifidobacterium reduce or protect symptoms such as diarrhea (Gilliland, 1990).

These probiotic strains, after being introduced into the intestine, become epithelial cells in the intestine to prevent the colonization of harmful microorganisms and secrete antibacterial substances to suppress the growth of harmful microorganisms and improve diarrhea and constipation, as well as enhance immune activity, anticancer It has effects such as action. Since probiotic strains are introduced into the intestine and secreted through their own metabolic activities, many studies are being conducted using probiotic strains because they provide many benefits to the host. Probiotic strains are normally present in the intestinal tract of most animals, maintain the balance of the intestinal microflora and have a beneficial effect on the health of the host.

Therefore, in order to select a useful intestinal probiotic strain, several important characteristics are identified and selected (Fuller, 1989; Garriga et al., 1998; Martin et al., 2006; Mishra and Prasad, 2005). That is, since the probiotic strain must survive in the digestive tract, it must have acid resistance or bile resistance, and must form colonies in the digestive tract until it reaches the intestine. In addition, it must be safe for food, feed, or clinical use, while at the same time having a beneficial effect on the host. Recently, probiotic strains with many functional properties have been developed, and there are probiotic strains having the function of inhibiting the proliferation of pathogens, BSH activity (Begley et al., 2006; Pereira et al., 2003) or starch hydrolysis. There are probiotic strains having amylolytic activity, Lee et al., 2001; Vizoso Pinto et al., 2006).

However, research on probiotic strains has been mainly focused on improving human health or productivity of livestock, and there are still probiotic strains that can effectively suppress the onset of immune-related diseases in companion animals, especially cats. Development is sluggish.

Accordingly, as a result of intensive research efforts to find a probiotic strain effective for immune-related diseases in cats, the present inventors found that the new strain Lactobacillus rhamnosus CACC 612 isolated from cat feces has excellent acid resistance, bile resistance, and intestinal adhesion. It is suitable for use as a probiotic strain, and at the same time has antibacterial activity against livestock pathogens, and has been confirmed to have an excellent effect on immunity enhancement and inflammation suppression, thereby completing the present invention.

Accordingly, the main object of the present invention is to provide a Lactobacillus rhamnosus CACC 612 strain, a new probiotic strain having excellent acid resistance, bile resistance, and intestinal adhesion.

Another object of the present invention is to provide a feed composition for companion animals containing the Lactobacillus rhamnosus CACC 612 strain as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a new probiotic strain, Lactobacillus rhamnosus CACC 612 (Accession No.: KACC 92348P) strain.

The term " Lactobacillus rhamnosus CACC 612 (Accession Number: KACC 92348P)" strain of the present invention is a novel Lactobacillus rhamnosus strain newly isolated and identified from the feces of cats by the present inventors, "CACC 612 It can be used interchangeably with "strain" or "strain 612".

The novel strain Lactobacillus rhamnosus CACC 612 (accession number: KACC 92348P) strain of the present invention is characterized in that it has antibacterial activity, immune enhancing activity and anti-inflammatory activity.

The present inventors have investigated and analyzed microorganisms useful for companion animals for many years for the development of probiotics that can be used for companion animals. It was confirmed that it was suitable for use as a probiotic strain due to its excellent properties and intestinal adhesion, and had antibacterial activity against livestock pathogens, and had an excellent effect on immune enhancement and inflammation suppression, and there was no problem with safety during storage.

The novel strain Lactobacillus rhamnosus CACC 612 (Accession No.: KACC 92348P) of the present invention is characterized in that it has the 16s rRNA sequence of SEQ ID NO: 1.

As a result of comparative analysis of the gene sequence with the gene sequences of conventionally known strains, the present inventors confirmed that the strain of the present invention was a new strain belonging to Lactobacillus rhamnosus and named it Lactobacillus rhamnosus CACC 612, National Academy of Agricultural Science It was deposited with the Agricultural Genetic Resources Information Center on June 1, 2021, and was given an accession number KACC 92348P.

According to another aspect of the present invention, the present invention provides a feed composition for companion animals comprising the new strain Lactobacillus rhamnosus CACC 612 (Accession No.: KACC 92348P), or a culture solution thereof as an active ingredient.

The feed composition for companion animals of the present invention is suitable for use as a probiotic strain because of its excellent acid resistance, bile resistance, and intestinal adhesion activity. In a preferred embodiment of the present invention, it was confirmed that the CACC 612 strain of the present invention had very good acid resistance as the survival rate was measured to be 75.9% even under a strongly acidic condition of pH 2.5, and showed a survival rate of 97.9% or more even under a 0.3% bile condition. It was confirmed that it has bile resistance. In addition, it was confirmed that the CACC 612 strain of the present invention exhibited a high intestinal adhesion activity of 78.4% (see FIG. 1).

The feed composition for companion animals of the present invention is characterized in that it exhibits antibacterial activity against livestock pathogens. The CACC 612 strain of the present invention or its culture medium exhibits antibacterial activity by itself, and has antibacterial activity against various livestock pathogens. In one embodiment of the present invention, it was confirmed that the CACC 612 strain culture solution according to the present invention has a growth inhibitory effect against various livestock pathogens. Specifically, the livestock pathogen has antibacterial activity against Salmonella Derby, Salmonella Enteritidis, Salmonella Typhimurium, and Escherichia coli , but is not limited thereto. .

In addition, in one embodiment of the present invention, it was confirmed that the CACC 612 strain culture solution according to the present invention exhibits antibacterial activity against the Clostridioides difficile strain, a livestock pathogen that causes colitis and diarrhea in companion animals. have.

The feed composition for companion animals of the present invention is characterized in that it has storage stability. In a preferred embodiment of the present invention, it was confirmed that there was no significant change in the number of viable cells for a period of 3 months when the CACC 612 strain was stored, thereby confirming that the strain had excellent storage stability (see FIG. 2).

In the feed composition for companion animals of the present invention, the strain, or its culture medium, exhibits anti-inflammatory activity by inhibiting the expression of inflammation-related genes IFN-γ, IL1B, IL-2, IL-4, and TNF-α genes. characterized in that In a preferred embodiment of the present invention, the expression of inflammatory response-related genes (IFN-γ, IL1B, IL2, IL4, TNF-α) induced by poly(I:C) in the group treated with the culture medium of the CACC 612 strain is It was confirmed that there was a significant decrease (see FIGS. 3B and 4 ).

In the feed composition for companion animals of the present invention, the strain or its culture medium activates immune cells and increases the number of eosinophils within a normal range to have immune enhancing activity. In a preferred embodiment of the present invention, the culture medium of the CACC 612 strain significantly activates Fcwf-4 cells, a cat immune cell line, and increases the ratio of eosinophils in leukocytes (WBC) within a normal range, thereby enhancing immunity , and through these results, it was confirmed that the CACC 612 strain of the present invention had the effect of increasing the immune function by increasing the leukocytes that play a major role in the cellular immune response in the normal range (FIG. 3a). and Figure 5).

The feed composition of the present invention may mean any natural or artificial diet, meal, snack, or ingredient of the meal for companion animals to eat, ingest, and digest or suitable for, and is known in the art. It can be manufactured in various forms.

The type of the feed is not particularly limited, and a feed commonly used in the art may be further used, and in this case, the feed composition of the present invention serves as a feed additive. Non-limiting examples of the feed that can be added include plant feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, gourds or grain by-products; and animal feeds such as proteins, inorganic materials, oils and fats, minerals, oils and fats, single cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

As another example, the feed composition may be appropriately used depending on the type of companion animal to be administered, age, and other types of feed components.

The feed composition of the present invention may further contain substances exhibiting various effects such as nutrient supplementation and weight loss prevention, enhancement of digestibility of fiber in feed, improvement of oil quality, prevention of reproductive disorders and improvement of fertility rate, prevention of high temperature stress in summer. For example, mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, trace minerals such as zinc, copper, cobalt, selenium, kerotene, vitamin E, vitamins A, D, E, nicotinic acid , vitamins such as vitamin B complex, protective amino acids such as methionine and lyic acid, protective fatty acids such as fatty acid calcium salts, live bacteria such as yeast cultures and mold fermented products, yeast agents, etc. may be further included.

In another aspect, the present invention provides a method for enhancing immunity and inhibiting inflammation of a companion animal, comprising administering the feed composition to the companion animal.

The administration means introducing the composition of the present invention to the companion animal by any suitable method, and the composition administration route of the present invention may be administered through various routes, either oral or parenteral, as long as it can reach the target tissue. The present inventors do not show acute toxicity even when the feed composition of the present invention is orally administered.

The features and advantages of the present invention are summarized as follows:

(1) The present invention provides a new strain Lactobacillus rhamnosus CACC 612 derived from cat feces and a feed composition for companion animals containing the same as an active ingredient.

(2) The Lactobacillus rhamnosus CACC 612 strain of the present invention has excellent acid resistance, bile resistance and intestinal adhesion, so it is suitable for use as a probiotic strain, and exhibits excellent antibacterial activity against livestock pathogens, and is used to enhance immunity and suppress inflammation. It has an excellent effect.

1 is a graph showing the measurement of acid resistance, bile resistance and intestinal adhesion of the Lactobacillus rhamnosus CACC 612 strain of the present invention.
2 is a graph showing the change in the number of live cells during long-term storage of the Lactobacillus rhamnosus CACC 612 strain of the present invention.
3 is a graph showing the effect of measuring the immune cell activity of the Lactobacillus rhamnosus CACC 612 strain of the present invention [FIG. 3a: (-) poly(I:C), FIG. 3b: (+) poly(I:C) ].
4 is a graph showing changes in gene expression of inflammatory response-related genes (IFN-γ, IL1B, IL2, IL4, TNF-α) according to the treatment of the culture supernatant of the Lactobacillus rhamnosus CACC 612 strain of the present invention.
5 is a diagram summarizing the blood test results of cats according to the treatment of the culture supernatant of the Lactobacillus rhamnosus CACC 612 strain of the present invention.
Figure 6 is a schematic diagram showing a schematic diagram of the phylogenetic tree of the Lactobacillus rhamnosus CACC 612 strain of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention by these examples.

Example 1. Verification of acid resistance, bile resistance and intestinal adhesion activity of microorganisms derived from cat feces

In order to select a candidate group of probiotics having antibacterial and immune-enhancing activity against livestock pathogens, the probiotic properties of microorganisms collected under anaerobic conditions from cat fecal samples were verified.

1-1. acid and bile resistance

For the study of acid resistance and bile resistance characteristics of strains isolated from cat feces, the isolated strains were subjected to anaerobic conditions (90% N ₂ , 5% CO ₂ , 5% H ₂ ) using an anaerobic chamber (Coy Laboratories, Grass Lake, MI). ) in MRS broth (BD Difco™, USA) was used in the experiment by culturing at 37° C. for 24 hours.

Acid resistance is inoculated into MRS medium adjusted to pH 2.5 using 1N HCl, and bile resistance is 0.3%, 1% (w/v) Oxgall (BD Difco™, USA) is added to MRS medium and cultured for 2 hours. , the number of viable cells after 0 and 2 hours was measured and the survival rate (%) is shown in [Table 1] below.

[Table 1]

As a result of the experiment, it was confirmed that the CACC 612 strain from the isolated candidate group had very good acid resistance as the survival rate was measured to be 75.9% even under the strongly acidic condition of pH 2.5, and it was confirmed that it had a survival rate of 97% or more even in the bile condition, showing very good bile resistance. could confirm that

1-2. intestinal adhesion activity

The intestinal epithelial HT-29 cell line is DMEM (Gibdo-BRL, USA) supplemented with 10% fecal calf serum (Gibdo-BRL, USA) and 1% penicillin/streptomycin (10,000U/ml, Gibdo-BRL, USA). The culture medium was used at 37° C., 5% CO ₂ conditions. The intestinal adhesion activity test was also performed under anaerobic conditions (90% N ₂ , 5% CO ₂ , 5% H ₂ ).

Candidate strains of 1.0×10 ⁸ CFU/mL or more were inoculated into a 24-well plate in which a HT-29 cell monolayer was formed, and after 2 hours of incubation, HT-29 cells and the strain were used using 0.1% triton X 100. After removing the , the number of viable cells was measured. For comparative experiments, Lactobacillus rhamnosus GG strain, which is known to have excellent intestinal adhesion activity, was used as a control strain, and the experimental results are summarized in [Table 2] below.

[Table 2]

As a result of the experiment, the intestinal adhesion capacity of the CACC 612 strain among the isolated candidates was measured to be 78.40%, and the intestinal adhesion activity of the CACC 612 strain was Lactobacillus rhamnosus GG strain, which is known to have excellent intestinal adhesion among conventional probiotic strains. It was also confirmed that it was even better compared with .

Example 2. Antimicrobial activity against livestock pathogens

To verify the antimicrobial activity against livestock pathogens, the antimicrobial activity was verified by the disk diffusion method.

As pathogens, 4 strains of Salmonella Derby NCCP 12238, Salmonella Enteritidis NCCP 14546, Salmonella Typhimurium NCCP 10328, and Escherichia coli KCTC 2617 were purchased from the National Hospital Resources Bank and the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center and used. The four pathogens were cultured at 37° C. for 24 hours using LB (BD Difco™, USA) medium.

After inoculating the strains isolated from cat feces in MRS broth, incubate at 37°C for 24 hours, and inoculate 80 μl of the culture supernatant sterilized with a 0.22㎛ pore size membrane filter on an 8mm paper disc, and measure the diameter of the clear zone to measure the antibacterial activity. evaluated.

In addition, Clostridioides difficile JCM 1296 ^T strain, a pathogenic strain causing colitis and diarrhea in companion animals, was used by culturing at 37° C. for 48 hours using EG medium (Becton, Dickinson and Company). 80 μl of the culture supernatant of the strains isolated from cat feces was inoculated on an 8 mm paper disc, and the diameter of the clear zone for Clostridioides difficile JCM 1296 ^T strain was measured to evaluate the antibacterial activity, and are summarized in [Table 3] below.

[Table 3]

As a result of the experiment, among the isolated candidates, the CACC 612 strain formed a clear zone of 9-10 mm against the livestock pathogens Salmonella Derby NCCP 12238, Salmonella Typhimurium NCCP 10328 and Clostridioides difficile JCM 1296 ^T strain, Salmonella Enteritidis NCCP 14546 and Escherichia coli KCTC It was confirmed to have very good antibacterial activity by forming a clear zone of 11-12mm for the 2617 strain.

Example 3. Storage stability

The CACC 612 strain was cultured anaerobically at 37° C. for 48 hours using MRS broth, centrifuged at 13,000 rpm for 5 minutes, and only the cells were collected.

The collected cells were freeze-dried by adding lactic acid bacteria probiotic powder freeze-drying protectant (5% fructo-oligosaccharide, 10% skim milk, 15% trehalose, 0.5% glycerin, 1% NaCl). After the freeze-dried cells were pulverized and powdered, 0.2 g (5.3 x 10 ⁹ CFU/g) per bag was packaged in small packages, and stored at 5° C. for 3 months. Formulation stability was confirmed by measuring the number of viable cells during storage.

As a result of the experiment, the CACC 612 strain was observed to have no change in the number of viable cells during the storage period, as the initial number of viable cells of 5.3×10 ⁹ CFU (one package, 0.2g small package) was maintained at 5×10 ⁹ CFU or more even after 3 months of storage, It was confirmed that the strain had very good storage stability (see FIG. 2).

Example 4. Increased immune cell activity and inhibition of inflammatory response

4-1. activation of immune cells

In order to evaluate the effect of the CACC 612 strain on the activity of immune cells, Fcwf-4 cells, a cat macrophage cell line, were cultured in a cell culture medium (10% FBS, 1% anitibiotics in DMEM) at 37° C. and 5% CO ₂ conditions. did.

The CACC 612 strain was cultured anaerobically at 37° C. for 24 hours using MRS broth, and after centrifugation at 13,000 rpm for 5 minutes, only the supernatant was collected.

The culture supernatant of the CACC 612 strain was added to the Fcwf-4 cells in culture at a level of 10% of the cell culture volume, and after culturing for 24 hours at 37° C. and 5% CO ₂ conditions, Fcwf using WST-1 assay. The immune cell activity of -4 cells was evaluated in comparison with the negative control group, which was a condition in which the culture medium was not added.

Referring to the experimental results [Fig. 3a], it was confirmed that the culture supernatant of the CACC 612 strain significantly activated Fcwf-4 cells, a cat macrophage cell line.

On the other hand, after inducing an inflammatory response by treating the Fcwf-4 cells in culture with poly(I:C), an immune-inducing substance, for 24 hours, the culture supernatant of the CACC 612 strain was added at a level of 10% of the cell culture solution volume. . After culturing for 24 hours at 37 ° C., 5% CO ₂ condition, the Fcwf-4 cell inflammation inhibitory activity was evaluated by using WST-1 assay compared to the negative control, which is a condition in which the culture medium is not added.

Referring to the experimental results [Fig. 3b], it was confirmed that the culture supernatant of the CACC 612 strain partially restored the activity of the cat macrophage line decreased by the inflammatory response (P < 0.05).

4-2. Inflammatory response-related gene expression analysis

Additionally, by extracting RNA from Fcwf-4 cells, gene expression patterns related to the inflammatory response were confirmed.

In the experiment, after inducing an inflammatory response by treating the Fcwf-4 cells in culture with poly(I:C), an immune-inducing substance, for 24 hours, the culture supernatant of the CACC 612 strain was added at a level of 10% of the cell culture medium volume. . After culturing for 24 hours at 37° C., 5% CO ₂ conditions, RNA of Fcwf-4 cells was extracted and the inflammatory response-related gene expression pattern was analyzed. Gene expression analysis was normalized to the GAPDH gene, which is a housekeeping gene, and the primers in Table 4 below were used.

[Table 4]

Referring to the experimental results [Fig. 4], the culture supernatant of the CACC 612 strain is poly(I:C)-induced inflammatory response-related genes (IFN-γ, IL1B, IL-2, IL-4, TNF-α) was observed to significantly suppress their gene expression (P < 0.05).

Through the above results, it was confirmed that the CACC 612 strain or its culture medium could be usefully utilized for enhancing immune function or anti-inflammatory purposes by activating immune cells and suppressing the expression of inflammation-related genes.

Example 5. Clinical trial on immune enhancing activity

A clinical test was conducted on the immune-enhancing effect of the CACC 612 strain whose probiotic activity was verified through Examples 1 and 2 on 5 cats under the consent of the guardian.

In the experiment, the CACC 612 strain was formulated in powder form as in Example 3, and 0.2 g (5 x 10 ⁹ CFU/g or more) was fed once a day for 45 days, and then blood was collected before and after feeding the microorganism and tested for blood. was carried out.

Referring to [Fig. 5], which summarizes the blood test results, pH, K ⁺ (mmol/L), BE-ecf (mmol/L), BE-b, SBC, HCO ₃ (mmol/L), Hemoglobin [Hb ](g/dL), Hematocrit[Hct](%), MCV(fL), MCH(pg), MCHC(g/dL), MPV(fL), WBC-Eos(%) are all normal. was able to confirm that it was in the range.

In addition, when comparing before and after the clinical trial, it was confirmed that all index values increased significantly. In particular, it was confirmed that the ratio of eosinophils among leukocytes (WBC) significantly increased within the normal range, and hemoglobin It was confirmed that the levels of (Hemoglobin, Hb) and hematocrit (Hct) were also increased (P < 0.05).

Through the above results, it was confirmed that the CACC 612 strain was effective in strengthening the immunity of healthy cats by increasing the leukocytes that play a major role in the cellular immune response in the normal range.

Example 6. Identification of strains

16S rRNA gene sequencing analysis was performed to identify the CACC 612 strain whose probiotic activity was verified in Examples 1 to 5 above.

27F and 1492R primers were used as primers, and after amplifying the 16S rRNA gene using a PCR device, a nucleotide sequence of about 1,400 bp or more was secured with inter-primer 785F and 907R primers.

[Table 5]

The obtained nucleotide sequence was analyzed by 16S rRNA through the 16S-based ID of Ezbiocloud (https://www.ezbiocloud.net).

As a result of the analysis, the CACC 612 strain was identified as a novel strain showing 99.40% homology with the Lactobacillus rhamnosus JCM 1136 ^T (accession number, BALT01000058) strain. The phylogenetic tree is shown in [Fig. 6] based on the neighbor joining method using the Mega 7 program.

The strain of the present invention having excellent probiotic activity identified as described above was named Lactobacillus rhamnosus CACC 612, and on June 01, 2021, it was deposited with the Agricultural Genetic Resource Information Center of the National Academy of Agricultural Sciences as an accession number (KACC 92348P).

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Agricultural Biotechnology Research Institute KACC92348 20210601

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Claims (9)

It is characterized in that it exhibits antibacterial activity, immunostimulating activity and anti-inflammatory activity,
It is characterized in that it exhibits antibacterial activity against the Clostridioides difficile strain, a livestock pathogen that causes colitis and diarrhea in companion animals,
Characterized in that it has immune enhancing activity by activating immune cells and increasing the number of eosinophils,
New strain Lactobacillus rhamnosus CACC 612 (Accession No.: KACC 92348P). delete According to claim 1,
The strain is a new strain Lactobacillus rhamnosus CACC 612 (accession number: KACC 92348P), characterized in that it has the 16s rRNA sequence of SEQ ID NO: 1. It is characterized in that it exhibits antibacterial activity, immunostimulating activity and anti-inflammatory activity,
It is characterized in that it exhibits antibacterial activity against the Clostridioides difficile strain, a livestock pathogen that causes colitis and diarrhea in companion animals,
Characterized in that it has immune enhancing activity by activating immune cells and increasing the number of eosinophils,
A feed composition for companion animals comprising the new strain Lactobacillus rhamnosus CACC 612 (accession number: KACC 92348P), or a culture solution thereof as an active ingredient. 5. The method of claim 4,
The strain, or its culture medium, is a feed composition for companion animals, characterized in that it has acid resistance, bile resistance, and intestinal adhesion activity. 5. The method of claim 4,
The strain, or its culture medium, is livestock pathogens Salmonella Derby ( Salmonella Derby), Salmonella Enteritidis ( Salmonella Enteritidis), Salmonella typhimurium ( Salmonella Typhimurium), and Escherichia coli characterized in that it exhibits antibacterial activity against A feed composition for companion animals. delete 5. The method of claim 4,
The strain, or its culture medium, is an inflammation-related gene IFN-γ, IL1B, IL-2, IL-4, and a companion animal feed composition, characterized in that by suppressing the expression of TNF-α genes to exhibit anti-inflammatory activity. delete

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