KR101987321B1 - PET Animal Feed Additive for improving intestinal and brain function - Google Patents

Abstract

The present invention relates to a feed additive for pets, and has an excellent effect of providing a functional feed additive for improving bowel and brain health prepared by combining GABA and essential amino acids in the same ratio (w / w).

Description

PET Animal Feed Additive for improving intestinal and brain function}

The present invention relates to a functional pet food additive for improving bowel and brain health.

Lactic acid bacteria (lactic acid bacteria) is a representative microorganism widely used as a probiotic (probiotic) is a safe microorganism easily found in the gut and fermented foods of humans and animals. Lactobacillus adheres to the intestinal epithelial cells and improves the properties of the intestinal flora, which stabilizes the intestinal flora and reduces the production of decay products by inhibiting the settlement of harmful bacteria and prevents disease, immune activation, anticancer activity, and cholesterol reduction It helps a lot. The lactic acid bacterium has growth inhibition effect on several decaying microorganisms and pathogenic microorganisms because of several metabolic properties. Because lactic acid bacteria are metabolites, they produce organic acid, hydrogen peroxide, reuterin, diacetyl, acetaldehyde and bacteriocin. to be.

Γ-aminobutyric acid, abbreviated as GABA (GABA), is a nonprotein amino acid and is contained in grains such as human brain, vegetables, fruits, rice, and brown rice. GABA is known to act as a neurotransmitter in higher animals, invertebrates and insects, including humans, and to act as a protective mechanism in plants. In addition, as the effects of GABA on stress suppression, concentration enhancement, and blood pressure strengthening are known in the human body, it has been released as various food and animal and animal additives including health functional foods, and its range of application is gradually expanding.

In addition, it is known that there is an effect of suppressing disease occurrence by stress relief, improving productivity by promoting secretion of growth hormone, and improving the quality of livestock products including beef.

Pets live with people and provide psychological stability and intimacy. According to the 2015 statistics, the proportion of households raising pets is 21.8%, which is estimated to be about 4.45 million households. The largest number of pets kept is 19.1% for dogs and 5.2% for cats, and the size of the pet-related market is continuously increasing. Therefore, not only general feed but also healthy functional feed composition market is expected to expand rapidly.

Essential amino acids are taken up by proteins after they are broken down into amino acids in the body. Therefore, the nutritional value of a protein is determined by the type and amount of amino acids contained therein. The amino acids are made from other amino acids in the body of the animal, and those that must be consumed as food without being synthesized in the body.

Even if it is not synthesized in the body or synthesized, the amount is so small that it is insufficient to achieve physiological function and must be supplied from food is called an essential amino acid.

Prior art related to the present invention is known in the Republic of Korea Patent Registration No. 10-1187512, but relates to a feed additive containing gamma aminobutyric acid (GABA) to the L- glutamic acid by enzymatic reaction to grain powder or ocher powder excipient The present invention relates to a method for preparing a feed additive to convert into butyric acid. Korean Patent Laid-Open Publication No. 10-2011-0009638 discloses a feed additive containing lactobacillus lactic acid bacteria complex strain as an active ingredient, and Japanese Patent Laid-Open Publication No. 10-2012-0036787 discloses L-glutamic acid, L-glutamic acid and / or Non-ruminant feed compositions comprising L-tryptophan are known. However, until now, GABA and essential amino acids in biolactic acid bacteria have been formulated in the same ratio (w / w / w) to improve intestinal and brain function.

Accordingly, an object of the present invention is to provide a functional pet food additive for improving the intestinal and brain health and to evaluate it.

The object of the present invention comprises the steps of mixing GABA and essential amino acids in the freeze-dried powder of lactic acid bacteria live bacteria; Preparation of functional animal feed additives for improving intestinal and brain function by adding fructooligosaccharide to increase the recovery rate of Lactic acid bacteria in the mixture obtained in the above step and experiment of animal immunity, stress and cholesterol content using the feed additive as a test material Achieved by evaluating through

It is another object of the present invention to further add a step of blending pH sensitive hydroxy propylmethyl cellulose phthalate 55 (HPMCP 55) in the same ratio (w / w) to enhance the intestinal viability and increase the stability of the probiotics. Provide a method.

The pet food additive of the present invention has an excellent effect on improving the intestinal and brain function health of the animal.

1 is a photographic view of a feed additive powder formulation prepared according to the present invention.
Figure 2 is a photograph of the final product of the feed additive prepared according to the present invention.
Figure 3 is a graph showing innate immune cell activity by administering a feed additive prepared according to the present invention.

The present invention is a feed additive composition for improving the intestinal and brain health of companion animals containing probiotic, GABA, and essential amino acids as an active ingredient in the same ratio (w / w / w) as a feed additive for pets such as dogs and cats. To provide.

According to the present invention, the mixing ratio of the biolactic acid bacterium, GABA, and essential amino acid is preferably equal to the weight ratio (w / w).

In addition, according to the present invention, any one or more strains selected from among Lactobacillus, Enterococcus, Pediococcus, Bifidobacterium, Leuconostoc and Streptococcus are preferred, but most preferably, the lactic acid bacteria Pediococcus is a lactic acid bacterium.

The daily dosage of the feed additive prepared according to the invention is most preferably 2% by weight of the total feed amount in the composition ratio, below which the effect is significantly lowered, which is undesirable.

In addition, the feed additive prepared according to the present invention may be prepared in any formulation commonly prepared in the art, for example, may be put into a variety of formulations, such as powders, granules or tablets into a variety of formulations, such as glass bottles or plastics 1 to 2. Most preferably, however, purification of the intestinal viability and stability of the lactic acid bacteria is most preferred.

The invention is illustrated in more detail by the following specific examples, experimental examples and formulation examples, but the following examples, experimental examples and formulation examples are intended to illustrate the invention and are not intended to limit the scope of the invention thereto. Does not.

According to the present invention, the powder or granule additive is preferably administered 2% by weight of the daily feed intake. However, in order to increase the intestinal viability or increase the stability of the probiotic, the amount of the probiotic added in the preparation of the powder or granule additive is preferably about 10 times the pH sensitive carrier, that is, the same ratio as the feed additive (w / w). This is the case when the tablet (tablet) by mixing.

Such tablet formulation increases the survival efficiency of lactic acid bacteria in the intestines of pets and increases the storage stability in the tropics. Carriers include HPMCP, HPMCAS, HPMCMC, but most preferably pH sensitive PHMCP.

Hereinafter, the preferable compounding ratio and manufacture example of the feed additive component of this invention are demonstrated.

Example 1 Manufacture of feed additive of the present invention

The most preferred blending ratio of the intestinal and brain health functional feed additive composition of the pet was blended and mixed in the weight unit of Table 1 below. Hereinafter, the experiments of the present invention used a powder formulation and each experiment did not show a significant difference from the over formulation of the efficacy, but the tablet showed a 10 to 20% increase in efficacy than the powder or granules. Silver powder formulation. In addition, the control group 1 without the fructooligosaccharide, the control group 2 without the essential amino acid, and the control group 3 without the GABA were inferior in feed additive suitability, respectively (Table 10).

The feed additive compounding ratio of the present invention ingredient Compounding ratio (%) Mixed Lactobacillus (Probiotics, 10 billion CFU / g) 5-15 Fructo-Oligosaccharides (Prebiotics) 3 ~ 7 GABA (gammaaminobutyric acid) 10-20 Mixed Essential Amino Acids-PET 10-20 Lactic Acid Bacteria Fermented Extract Powder 10-20 Yeast Extract Powder 5-15 Anhydrous Glucose 3 ~ 7 dextrin 10-20 Other Ingredients (Multivitamin, Silicon Dioxide, Staline Magnesium, etc.) Remaining amount Sum 100.0

Experimental Example 1. Immunomarker IgG Analysis in Blood

Animal test by adding 2% by weight of the feed additive of the present invention obtained in Example 1 to the total feed weight of the general feed in order to determine the effect of the feed additive of Example 1 according to the present invention on the immune markers in the blood of the mouse Was performed. In order to measure the immunomarker in blood, blood was collected from the same vein of the same individual before the start of the test and at the end of the test (after 5 weeks), and the serum obtained after centrifugation at 2,000xg for 30 minutes at 4 ° C. Immunomarkers (WBC, RBC, Lymphocyte, IgG, total protein, albumin) were used for analysis.

As a result, as shown in Table 2 below, as a result of IgG measurement in blood of each experimental animal, the amount of IgG in blood was significantly increased in the experimental group compared to the control group.

IgG (mg / mL) Control Experimental group 0 days 331.5 340.3 14 days 320.8 402.5 35 days 342.5 501.1 Difference 11 160.8

Experimental Example 2. Congenital immune cell activity according to the present invention

In order to confirm the effect of enhancing the activity in the innate immune cells according to the present invention, the change of the markers of immune cell activity was investigated. COX-2 and IFNβ are representative immunoproteins that increase when TLR4 is activated by bacteria or viruses in macrophages, which are innate immune cells, and NF-kB and IRF3 are representative transcription factors activated by TLR4.

To this end, mice macrophages were incubated at a concentration of 1 × 10 ⁵ and injected with luciferase reporter plasmids containing COX-2-promoter, IFNβ-promoter, NF-kB binding site and IRF3 binding site, respectively. The feed additive obtained in Example 1 was treated for 8 hours at a concentration of 100 μg / mL and 500 μg / mL. Cell extracts were obtained and measured at the gene level of COX-2 and IFNβ and activation of NF-kB and IRF3, respectively, using a luminescent enzyme gene analysis kit (luciferase reporter assay system TM, Promega Co., Madison, Wis.). .

As a result, as shown in Figure 3, the feed additive of the present invention increases the expression of COX-2, IFNβ and NF-kB, IRF3 activation in a concentration-dependent manner at 100μg / mL and 500 μg / mL, respectively, These results confirm that the feed additive of the present invention increases the activation of innate immune cells by increasing the activity of transcription factors related to innate immunity and increasing the expression of immune proteins.

Experimental Example 3. Immune Enhancing Effects of Animal Models Inducing E. Coli Infection

To determine the immune enhancing effect of the feed additive of the present invention obtained in Example 1, E. coli was administered intraperitoneally in mice to confirm the protective effect against pathogenic bacteria through the prevention of death by E. coli bacterial infection.

To this end, E. coli was treated with 30 6-week-old male ICR mice. In the test, five mice were used in each group, as an untreated control group, E. coli inoculation positive control group, and three experimental groups inoculated with E. coli after feeding additives. Feed additives were tested by adding 2% by weight of the total weight of the feed to the feed. E. coli was inoculated by intraperitoneal administration of 0.5 mL of the culture medium. The mortality was recorded for 4 days after inoculation, and the pathogenic bacterial defense rate of the feed additives was examined.

Experimental results, as shown in Table 3 below, the animals of the E. coli inoculation positive control group died at 10 days after inoculation with E. coli, and showed a mortality rate of 100%. At the end of the final test, there were two surviving mice at the end of the first and third days, and as a result, 30% survival rate was confirmed in the protective effect against Escherichia coli inoculation.

E.coli
Compund
No. of death Surcical rate (%) D1 D2 D3 D4 Negative Control x x 0 0 0 0 100 Positive control group o x 10 - - - 0 Experimental group o The present invention
Feed additives 5 One One 0 30

Experimental Example 4. Contrast Shift Experiment

Contrast movement experiments are based on the premise that the mouse innately dislikes bright light and spontaneously seeks in a new environment. In other words, mice with stress anxiety will have fewer movements from the darkroom to the darkroom without spontaneous exploration. The effect of stress inhibition can be confirmed by measuring the number of times the mice move from the dark room to the clear room.

Male ICR mice were used for this purpose in the experiment. It was bred in an acrylic cage (45 × 60 × 25 cm), kept a constant temperature of 20-24 ° C. and was used for an experiment after one week of adaptation. The experimental animals were divided into a negative control group fed only feed, a test group fed by adding 2% by weight of the feed additive of the present invention to the general feed, and a positive control group administered diazepam.

As a result, as shown in Table 4, it was confirmed that the number of shifts from the dark room to the bright room increased in the experimental group fed the feed additive of the present invention as compared to the negative control group.

Laboratory animal population Number of moves from darkroom to bright room Negative Control 5 5.0 ± 4.38 Positive control group 5 12.6 ± 4.89 Experimental group 5 11.5 ± 4.81

Experimental Example 5. Hormonal concentration changes

Experimental animals and experimental groups were treated in the same manner as in Experiment 4. The animals were freely fed for 4 weeks. After 4 weeks, each mouse was induced with an electric foot shock by applying 0.6 mA per second at 5 second intervals for 30 minutes. Blood- and brain tissues (cerebral cortex, hippocampus and striated body) were extracted by autopsy of stress-induced mice, and dopamine, corticosterone and noradrenaline secreted from the extracted blood and brain tissues were enzyme-linked immunosorbents. analysis).

As a result, as shown in Table 5, it was confirmed that the amount of dopamine, noradrenaline and corticosterone was decreased in the experimental group fed 2 wt% of the feed additive of the present invention compared to the general feed as compared to the negative control group.

Dopamine Concentration (%) Noradrenaline
density(%) Corticosterone Concentration (%) Negative Control 100 100 100 Positive control group 43.2 37.5 16.1 Experimental group 62.5 60.0 33.2

Experimental Example 5. Blood cholesterol

Spontaneous hypertension rats (SHR) and rats of normal blood pressure (WKY) were used as control animals to confirm the effect on blood cholesterol according to the feed additive of the present invention of Example 1. The experiment was carried out with the negative control group fed the normal feed to the WKY rats, the experimental group 1 fed the normal feed and the present feed additive to the WKY rats, the positive control group fed the normal feed to the SHR rats, and the normal feed and the present invention to the SHR rats. Feed additives were divided into experimental group 2, which was fed to general feed for 4 weeks, and then blood pressure was measured. The feed additive of the present invention was fed by adding 2% by weight relative to the total weight of the general feed.

As a result, as shown in Table 6, the WKY rat had no effect, but it was confirmed that the blood pressure was significantly reduced in the hypertensive rat SHR rat.

Blood pressure (mmHg) WKY (Normal Feed) 128.38 WKY (Normal Feed + Invention Feed Additive) 125.19 SHR (General Feed) 202.67 SHR (General Feed + Invention Feed Additive) 159.36

Preparation Example 1. Container products

The container product was formulated as shown in Table 6, and then manufactured in powder or granule form and packaged in a container as shown in the photograph to produce a product.

Raw material name Input amount (g) Mixed Lactobacillus (Probiotics, 100 billion CFU / g) 3.00 Fructooligosaccharide or galactooligosaccharide (prebiotics) 1.50 GABA (gammaaminobutyric acid) 4.50 Mixed Essential Amino Acids-PET 4.50 Lactic Acid Bacteria Fermented Extract Powder 4.50 Yeast Extract Powder 3.00 Anhydrous Glucose 1.50 dextrin 3.00 Other Ingredients (Multivitamin, Silicon Dioxide, Staline Magnesium, etc.) Remaining amount Sum 30

Preparation Example 2. Stick cloth products

The necessary feed additives per day were formulated in powder or granule form as shown in Table 8 below, and then packaged in a small packaged stick cloth product to provide portability and convenience.

Raw material name Input amount (g) Mixed Lactobacillus (Probiotics, 100 billion CFU / g) 0.2 Fructooligosaccharide or galactooligosaccharide (prebiotics) 0.1 GABA (gammaaminobutyric acid) 0.3 Mixed Essential Amino Acids-PET 0.3 Lactic Acid Bacteria Fermented Extract Powder 0.3 Yeast Extract Powder 0.2 Anhydrous Glucose 0.1 dextrin 0.3 Other Ingredients (Multivitamin, Silicon Dioxide, Staline Magnesium, etc.) Remaining amount Sum 2

Preparation Example 3 Tablet Product

The necessary feed additives per day were formulated as a tablet product by mixing as shown in Table 9, and then packaged in a container as shown in FIG.

Raw material name Input amount (g) Mixed Lactobacillus (Probiotics, 100 billion CFU / g) 3.00 Fructooligosaccharide or galactooligosaccharide (prebiotics) 1.50 GABA (gammaaminobutyric acid) 4.50 Mixed Essential Amino Acids-PET 4.50 Lactic Acid Bacteria Fermented Extract Powder 4.50 Yeast Extract Powder 3.00 Anhydrous Glucose 1.50 dextrin 3.00 Other Ingredients (Multivitamin, Silicon Dioxide, Staline Magnesium, etc.) Remaining amount Sum 30 pH sensitive hydroxypropylmethyl cellulose phthalate 55 (HPMCP 55) 30 sum 60

Comparative Example 1. Comparison of Effects of the Feed Additives of the Present Invention

On the other hand, the feed additive (control group 1) prepared by the method of the present invention prepared by the method of Example 1 and the prebiotic (control group 1), the feed additive (control group 2) and GABA prepared except the essential amino acid In order to perform a comparative experiment of the prepared feed additives (control group 3) except for the comparison was carried out by adding 2% by weight to the total weight in the general feed in the same manner as in Experimental Examples 1 to 5.

Experimental results, as can be seen in Table 10 below, the feed additive of the present invention (experimental group) was confirmed that the superior intestinal and brain function improvement effect than the control 1 to 3.

IgG
(35 days) Immune enhancing effect
(% Survival) stress Hormone Concentration (%) Blood
cholesterol
(SHR rat) Dopamine Nord
Adrenaline Corti
Costeron Experimental group 501.1 30 11.54.81 100 100 100 159.36 Control group 1 421.6 10 9.84.25 135.2 137.1 138.6 179.57 Control 2 452.5 20 10.84.11 116.5 119.2 116.8 165.35 Control group 3 426.2 10 7.73.87 134.5 135.5 135.7 178.92

As described above, the feed additive of the present invention, in which the biolactic acid bacterium, GABA, and essential amino acids are added at a predetermined ratio according to Example 1, has an excellent effect on improving intestinal and brain health, and thus, it may be a very useful invention in the pet health functional feed industry. .

Claims (6)

delete delete delete Mixing the lactic acid bacteria freeze-dried powder and GABA and L-glutamic acid in the same ratio of 1: 1: 1 (w / w / w); To the mixture obtained in the above step, the addition of the flactooligosaccharide and pH sensitive hydroxypropylmethyl cellulose phthalate 55 (HPMCP 55) is characterized in that the pet food additive composition for pets.
Pet food for which the feed additive composition of claim 4 is added at least 2% by weight.
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