KR100777833B1 - composition comprising an extract of Schizandra chinensis showing antibacterial activity - Google Patents

Abstract

The present invention relates to a composition for animal feed comprising Schizandra chinensis extract and feed additives, and more particularly, to an antimicrobial agent, animal feed composition or animal feed additive comprising a Schizandra chinensis extract, a polar solvent and a non-polar solvent soluble extract. will be. The composition of the present invention may have an antimicrobial effect and an immune enhancing effect, thereby contributing to increasing farm household income.

Schisandra chinensis, animal feed composition, animal feed additives, antibacterial agents, antibacterial, boost immunity.

Description

Composition comprising an extract of Schizandra chinensis showing antibacterial activity

1A is a diagram showing ischemic necrosis of the intestinal wall and atrophy of the intestinal wall as a control of intestinal tissue, and FIG. 1B is a diagram showing that there is no significant difference from normal tissue in the group of Schisandra chinensis extract treated in the intestinal tissue.

Figure 2a is a control group of the spleen tissue showing severe congestion and bleeding accompanied by some necrosis, Figure 2b is a diagram showing the congestion is weak in the group of Schisandra chinensis extract treatment,

Figure 3a is a control of the liver tissue showing a severe congestion in the central vein or portal vein, Figure 3b is a group showing the relatively weak congestion in the group treated with Schisandra chinensis extract in the liver tissue,

4A is a diagram showing congestion, ischemic necrosis, and the like as a control group of kidney tissue, and FIG. 4B is a diagram showing weak tissue findings such as congestion and ischemic necrosis in a group treated with Schisandra chinensis extract.

In order to cope with the 15% market share of the drug market and the 1.5 trillion won antibiotic market, it is urgent to develop new antibacterial drugs. In addition, in Korea, economic and cultural international exchanges are active, foreign pathogenic bacteria are more likely to be introduced, and livestock imports are increasing. Currently, most of the bacteria present in domestic livestock are resistant to conventional antibiotics, and the treatment is becoming increasingly difficult. Due to these treatment problems, the transmission of these bacteria through livestock products will be severe. According to recent reports, more than 50% of human infectious diseases appear to be common infectious diseases, so safe livestock supply is a prerequisite for public health. Antibiotic resistance of bacteria is expressed in almost all drugs currently in use, rather than in some localized antibiotics, and bacterial physiological and genetic characterization is expected to have the ability to resist any drug. Among the various bacteria that are antibiotic resistant, methicillin-resistant S. aureus ( MRSA), vancomycin-resistant enterococci (VRE), vancomycin-mediated S. Aureus (vancomycin intermediate S. aureus; VISA) and E. coli O157: H7 with at least six antibiotic resistances. In Korea alone, MRSA is a nosocomial infection agent in hospitals, and it is almost 80% in large hospitals. VRE and VISA have also been rampant since the 1990s and have spread throughout the country. Escherichia coli O157: H7 is also resistant to multiple antibiotics due to multiple agent resistance and there is no complete therapeutic antibiotic yet. E. coli has already been a frequent social problem in the United States, Europe, and nearby Japan, and has occurred sporadically in humans and livestock in Korea. Since the import of cattle has been liberalized since 2001, it is clear that E. coli is widespread in Korea when livestock products or live cattle are imported from the E. coli Sangje region. These resistant bacteria do not have special antibiotic selection system in Korea, and the factor of rapid increase in resistance is much larger than in other countries. Increasing antibiotic resistance is a vicious cycle of increasing use of new expensive antibiotics, increasing treatment duration, and mortality, which in turn increases economic and social problems. Therefore, in order to solve this problem, it is urgently necessary to understand the mechanism of resistance that is occurring and to develop a new antibiotic having a structure that does not cause resistance.

   In order to cope with the rapidly increasing bacterial drug resistance and effectively treat various bacteria, new drugs different from the existing antibiotics (molecular structure, efficacy of drugs, etc.) are urgently needed. In particular, due to the emergence of super bacteria that are resistant to antibiotics, the development of new antibiotics is urgently required. In addition, the condition of the new drug should be a safe drug with no side effects on human and animal cells as well as bactericidal effect. No matter how good the antibiotic is, resistance develops over time, and natural medicines are sure to solve this problem. The reason for this is that natural products are new components that have not been known so far, and since each action is a mixture of various single components, it is convinced that the pharmacological action between the complexes will reduce side effects such as resistance.

   Many antibiotics currently used for livestock are at severe levels of resistance and some are close to 100%. In livestock, antibiotics are mainly used as feed additives or for cleaning purposes regularly between 7 and 10 days for broilers. According to the report, the antibiotic resistance rate of food poisoning bacteria such as Salmonella and enterococci isolated from cattle, pigs, and chickens from all slaughterhouses in the country showed that the resistance of E. coli from chicken feces was more than 57% to quinolone antibiotics. As such, currently used for livestock raising, most antibiotics are resistant to bacteria, so it is urgent to develop new antibiotics that can be effectively used for livestock raising.

   Recently, the research trend of medicines are getting serious side effects, toxicity and resistance, so the research on the development of medicines from natural products is actively being carried out worldwide. On the other hand, in the East, including Korea, it has a background that has promoted the steady development of traditional medicine, which is one of natural medicines, through long periods of time. Since the use of penicillin in clinical trials in 1941, about 5,500 species have been discovered so far, of which only 100 are fermented and produced, most of which are chemically synthesized. In order to be effectively used for the treatment of diseases, the therapeutic dose should not be toxic to the host and the antibacterial effect to the infectious agent should be very large, but it has poisoning or unusual side effects. to be. Therefore, research and development of new antibiotics that are less toxic and effective against resistant bacteria are needed, and studies to reduce the appearance and toxicity of resistant bacteria by methods such as co-administration are required. It is possible to develop new drugs by searching for new antibiotics that are excellent in antimicrobial activity and less toxic from natural drug resources and new antibiotics that are effective against resistant bacteria.

The most common incidence of bacterial diseases in Korea is Salmonellosis, which is especially mediated in food for humans. More than 2,300 serotypes have been reported, and their symptoms are similar. Salmonella (Salmonella) can cause a variety of clinical symptoms and there is a variety of toxins such as endotoxins, cell toxin, enterotoxin cells exist. In particular, the pathogen can be transmitted through the intestinal epithelial cells invade host cells, and almost being infected by all mammals typically a bacterium of the most common microorganisms of the pathogens causing food poisoning in people causing zoonotic infectious diseases flock Salmonella typhimurium Stadium (Salmonella typhimurium) . It is spread to humans mainly through contaminated meat and usually causes symptoms such as acute enteritis, high fever and sepsis.

   Antibiotics have been continuously used everywhere for the control of bacteria, and antibiotic-resistant bacteria have recently emerged due to the misuse and abuse of such antibiotics and antimicrobial substances, which has become a big problem in society. As awareness of these problems spreads, societal atmospheres that prevent the use of antibiotics or antimicrobial agents are being created. In addition, research on substances that can replace antibiotics has been actively conducted, and plants and probiotics (which have been used as folk remedies for a long time) ( Probiotics have been actively used to obtain antimicrobial and immune enhancing effects.

An object of the present invention is to provide a composition for animal feed, animal feed additives or antimicrobial agents containing Schizandra chinensis extract of Schizandra chinensis with low toxicity and antimicrobial activity as an active ingredient.

In order to achieve the above object, the present invention provides a composition or animal feed additives for animal feed comprising Schizandra chinensis extract having an effective antimicrobial activity and low toxicity and resistance to a living body.

The extract of Schizandra chinensis may further include additives well known in the art, for example β-glucan, vitamin E, yeast culture or L-carnitin. .

The extract comprises an extract soluble in water, a mixed solvent selected from lower alcohols having C ₁ to C ₄ carbon atoms or a mixed solvent thereof, preferably methanol.

The animal may be a mammal such as a pig, a cow or a goat; Fish such as carp and goldfish; And poultry such as pheasants, chickens, ducks, turkeys and the like.

Hereinafter, a method of obtaining the Schizandra extract is described in detail.

Schizandra chinensis extract of the present invention, the root or ground portion (leaf, stem) of dried Schizandra chinensis, about 1 to 20 times of the weight (kg), preferably about 3 to 10 times methanol, 20 ℃ to An extract obtained by using a cold, hot water extraction, ultrasonic extraction, reflux cooling extraction method for about 1 hour to 10 days, preferably about 2 hours to 5 hours at 100 ° C, preferably 50 ° C to 100 ° C extraction temperature Filtration, concentration under reduced pressure, or drying may yield the Schizandra chinensis extract of the present invention.

The present invention provides a composition for animal feed having an antimicrobial activity using the Schizandra chinensis extract obtained by the above production method as an active ingredient.

The composition of the present invention preferably contains 0.01 to 99.9% of Schizandra chinensis extract, more preferably 0.1 to 90%. However, the composition as described above is not necessarily limited thereto.

The Schizandra chinensis extract may be in the form of 20 to 90% high concentrate, powder or granules.

The extract of Schizandra chinensis extract includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polyphosphate), polyphenols, catechin, and alpha-tocopherol. It may further include any one or more of natural antioxidants such as rosemary extract, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, phytic acid.

Animal feed composition containing Schizandra chinensis extract of the present invention can be added as a supplementary components, such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal, enzymes, live microbial preparations and the like.

Animal feed compositions containing Schisandra chinensis extract of the present invention also include grains such as milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed, such as rape, soybeans and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugars and dairy products, for example, a dry ingredient consisting of various powdered milk and whey powder, and a drying additive, all mixed together with a liquid component, and a component which becomes a liquid after heating, that is, a lipid, for example, an animal liquefied by heating. In addition to the main components such as fats and vegetable fats and the like can be used with substances such as nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents and the like.

The invention is applicable to a number of animal diets, including mammals, poultry and fish. More specifically, the diet is commercially important mammals such as pigs, cows, sheep, goats, laboratory rodents (eg rats, mice, hamsters and gerbils), furry-owned animals (eg mink and foxes). Etc.), and zoo animals (eg monkeys and tailless monkeys), as well as livestock (eg cats and dogs, etc.).

In the method of blending the animal feed composition comprising the composition according to the present invention, the composition of the present invention is incorporated into an animal feed in an amount of about 1 g to 100 g per kg of feed on a dry weight basis.

In addition, the feed mixture is thoroughly mixed and then a viscous granulated or granular material is obtained depending on the degree of grinding of the components. It can be fed as a mesh or formed into the desired discrete shape for further processing and packaging, in which case water is added to the animal feed and then conventional pelletization to prevent separation during storage. , Expansion, or extrusion is preferred, and excess water may be removed to dryness.

The present invention provides an antimicrobial agent having a low toxicity and resistance to antimicrobial activity using Schizandra chinensis extract.

The composition comprising the Schizandra chinensis extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Compositions comprising extracts according to the invention are formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents which may be used in combination with the extract, and which may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts or fractions of the present invention vary depending on the condition and weight of the animal, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be administered to mammals such as mice, mice, livestock, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections.

In the above, the composition for animal feed, animal feed additive or antimicrobial agent containing Schisandra chinensis extract according to the present invention has been described. Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Examples.

Example 1. Preparation of Schizandra chinensis Extract

1-1. Preparation of Schizandra Powder

Schisandra chinensis powder purchased from Gyeongdong market was dried in a cool place at room temperature for about 1 week, and then dried schisandra chinensis powder was pulverized to a particle size of 30 mesh or less using a mill. Schisandra chinensis powder obtained by the above method was used as the main raw material of the present invention.

1-2. Preparation of Schizandra chinensis Extract

Add 100% methanol solution corresponding to three times the mass of dried Schizandra chinensis powder (300g) obtained in Example 1-1, reflux cooling extraction at 100 ℃ for 24 hours, and 3.2g of Schizandra chinensis through filtration and freeze-drying Crude extract was obtained.

1-3. Preparation of β-glucan-containing extract

Sichuan mushrooms were collected and washed with water, and then powdered to a size of 30 mesh using a grinder. 70% methanol solution corresponding to three times the mass of dried situation mushroom powder (300 g) was added at -20 ° C, mixed for 20 minutes, and filtered. The procedure was repeated three times, and then extracted. The combined filtrates were combined, and a small amount of 2M K ₂ HPO ₄ solution was added thereto, followed by centrifugation at 3000 rpm for 20 minutes. The remaining part after centrifugation was washed with methanol, dissolved in 1 L of 1.2N hydrochloric acid solution, 1 L saturated sodium carbonate solution was added to neutralize the pH to 4.0, and dialyzed in dialysis solution for 36 hours. The same amount of saturated solution of caffeine was added to the dialysate and centrifuged again at 3000 rpm for 20 minutes. The remaining portion was dissolved in 1% hydrochloric acid-dimethylformamide solution and injected into Sephadex LH-20 column to develop 70% DMFA solution as a developing solvent. After lyophilization, 30 g of β-glucan-containing extract was obtained.

1-4. Preparation of yeast culture

10 g of BactoYeast extract was dissolved in 1000 ml of distilled water, followed by autoclave. 20 g of agar (Agar) was added to the autoclaved extract to dissolve, and then cultured in a YPD incubator at 37 ° C. After the incubation period was dried in a cool place for one week so that the moisture is less than 10%, and then warm air dried at a low temperature of less than 40 ℃ again. The dried yeast was used to prepare a composition for animal feed so that the content per kilogram of 17% or more.

Example 2. Preparation of Animal Feed Composition

In addition to the β-glucan-containing extract and yeast in the Schizandra chinensis powder obtained from Examples 1-1 to 1-4 and Schizandra chinensis extract, vitamin E and L-carnitine were added to prepare a composition for animal feed. Schisandra chinensis powder was 5 to 15%, Schisandra chinensis soluble extract 50 to 80%, yeast 10 to 20% per kg contained. Small amounts of L-carnitine, β-glucan and vitamin E were added to prepare the animal feed composition of the present invention. The ratio of the animal feed composition of the present invention is illustrated in Table 1.

Composition example W% Composition example W% Schizandra Powder 11.5 Vitamin E 0.5 Schisandra chinensis soluble extract 70 L-carnitine 0.5 β-glucan 0.5 Yeast culture 17

Experimental Example 1. Antimicrobial Test

1-1. Disclosure strains and medium and strain culture

Strains used for the determination of antimicrobial activity were isolated from humans or animals and ATCC standard strains (see Table 2). The strains were inoculated with Tryptic Soy broth ^ⓡ (TSB; Difco, USA) and incubated for 24 hours in a 37 ° C thermostat. For the visual observation, Muller-Hinton (Difco, USA) agar medium was used. It was.

List of Bacteria Strains Strain describe origin JOL 375 Vibrio cholera (Vibrio cholera) O1 Person JOL 376 Vibrio cholera (Vibrio cholera) O139 ATCC 51394 JOL 377 Shigella dysenteriae Person JOL 378 Shigella flexeneri Person JOL 379 Escherichia coli O157: H7 ATCC 43890 JOL 380 Salmonella typhi Person JOL 381 Salmonella typhimurium parameters (Salmonella paratyphi) A Person MRSA 104 Methicillin resistance Staphylococcus Aureus Person MRSA 6 Staphylococcus aureus Animals (chicks) ATCC Salmonella gallinarium ATCC JOL 284 Escherichia coli O157 animal

Salmonella used to measure antibacterial activity are natural strains and ATCC strains isolated from humans or animals (see Table 3). The strains were inoculated in Tryptic Soy broth ^ⓡ (TSB; Difco, USA) and incubated for 24 hours in a 37 ° C thermostat and used for the experiment. Muller-Hinton agar medium was used for visual observation.

List of Salmonella Strain describe Origin Standard strain Salmonella gallinarium ATCC JOL 380 Salmonella typhi Person JOL 381 Salmonella paratyphi A Person JOL 386 Salmonella enteritidis chicken JOL 387 Salmonella typhimurium (Salmonella typhimurium) pig JOL 388 Salmonella typhimurium (Salmonella typhimurium) pig JOL 389 Salmonella typhimurium (Salmonella typhimurium) chicken JOL 407 Salmonella enteritidis Sal-36 Quarantine JOL 408 Salmonella typhimurium Sal-13 Quarantine JOL 409 Salmonella dublin Sal-37 Quarantine JOL 410 Salmonella derby Sal-9 Quarantine JOL 411 Salmonella cholerae-suis ATCC 7001 JOL 419 Salmonella gallinarium chicken JOL 420 Salmonella gallinarium chicken JOL 421 Salmonella gallinarium chicken JOL 422 Salmonella gallinarium chicken

1-2. Bauer-Kirby test

Inoculating bacterial culture on Muller-Hinton agar medium and injecting a drug extracted from natural substances onto the disc creates an antibiotic gradient in the medium. At this time, the inhibitory ring around the disc shows antibacterial activity. The inhibitory ring is sized according to the solubility of antibiotics and the degree of sensitivity of bacteria, and the sensitivity of each antibiotic is sensitive (S), intermeditely sensitive (I), and resistant (R, R) depending on the size of the inhibitory ring. Divide by)

1-2-1. Antimicrobial Activity of Schizandra chinensis Extract

After the Schizandra chinensis extract of Example 1-2 was treated at a concentration of 0.5 mg / ml, the antimicrobial activity was examined by the disk diffusion method and showed susceptibility to all bacteria (see Table 4).

Bacteria Growth Inhibitory Activity of Schizandra chinensis Extract Botanical name Diameter of clean area (mm) JOL375 JOL376 JOL377 JOL378 JOL379 MRSA104 MRSA6 ATCC JOL284 Schizandra chinesis 28 24 16 24 17 18 16 18 16

1-2-2. Antimicrobial Activity of Schizandra chinensis Extract against Salmonella

   After treating the Schisandra chinensis extract of Example 1-2 to Salmonella at a concentration of 0.5 mg / ml, the antimicrobial activity of the disc diffusion method showed high antibacterial activity (see Table 5).

 Salmonella Growth Inhibitory Activity of Schizandra chinensis Extract Sample Diameter of clean area (mm) Schizandra chinensis Standard strain 11 JOL380 19 JOL381 17 JOL386 14 JOL387 19 JOL388 12 JOL389 12 JOL407 14 JOL408 13 JOL409 16 JOL410 14 JOL411 23 JOL419 23 JOL420 25 JOL421 21 JOL422 20

1-3. Minimum inhibitiry concentration (MIC)

1-3-1. Experiment process

Muller-Hinton agar medium was also used to measure the minimum inhibitory concentration of Schizandra chinensis extract. The bacteria enriched in TSB were re-sterilized using the same medium, and the absorbance at 590 nm was measured to calculate CFU (dog / ml), and then diluted to 1 × 105 / ml using 0.85% physiological saline.

   The measured concentrate is dissolved in Muller-Hinton broth at 55-60 ° C, in the same amount as the diluted bacteria and incubated in a 37 ° C thermostat for 24 hours. The cultured bacteria are diluted 100-fold using sterile distilled water. 100 μl dilutions were spread on Muller-Hinton agar medium and incubated for 18 hours at 37 ° C. incubator.

1-3-2. Experiment result

As a result of testing the minimum inhibitory concentration (MIC) of Salmonella strains against the 100% methanol extract of Schisandra chinensis, the antimicrobial activity showed antimicrobial activity in the low concentration range. (See Table 6).

Salmonella Inhibitory Concentration of Schizandra chinensis Extract Inhibitory concentration (㎍ / μl) Schizandra chinensis Standard strain - JOL380 500 JOL381 15.6 JOL387 - JOL388 31.3 JOL389 - JOL408 125 JOL409 - JOL410 125 JOL411 62.5 JOL419 31.3 JOL420 31.3 JOL421 31.3 JOL422 31.3

1-4. Mouse Inoculation Experiment

1-4-1. Laboratory animals

 Seventeen-week-old male ICR mice were used in this experiment, and microscopic examinations were performed by visual observation after confirmation to confirm that they were non-infected with Salmonella before being used in the experiment. Five negative control groups and two 6 drug administration groups were randomly selected and bred in cages, and experiments were performed after three days of adaptation. During the acclimation period, distilled water containing 5 g / ℓ of streptomycin was used as a negative water, and feed and litter were changed daily.

1-4-2. Mouse inoculation experiment method

Mice inoculated experiments, the bacteria were recovered by centrifugal separation 30 minutes at 2,500rpm after 24 hours incubation at 37 ℃ vibration incubator by inoculating Salmonella typhimurium (Salmonella typhimurium) in TSB containing streptomycin (Sigma, USA). The cells were washed once with phosphate buffered saline (PBS), resuspended in 20% sucrose in 1X PBS, and adjusted to 1X 10 4 cfu / ml, and 0.5 ml of the bacterial solution was orally inoculated using a zondae.

   The experiment was divided into three groups. The negative control group was inoculated with 20% sucrose in 1X PBS, not the test strain, and provided only distilled water containing 5 g / L of streptomycin during the experiment. The drug administration group was inoculated with the strain strain and provided with a negative dilution of Schisandra chinensis extract in distilled water containing 5g / L of streptomycin. Herbal medicine extract Schisandra chinensis was methanol extract at a concentration of 5 mg / ml. During the experiment, cages and litter were bred daily.

1-4-3. Therapeutic Effect of Schizandra chinensis Extract in Mice

Feces were collected from each group of mice to analyze the pattern of Salmonella excreted in the feces. Four times of each strain, once before inoculation, 1 day, 3 days, and 6 days after inoculation, fecal dilutions were made in each group of mice with 0.85% physiological saline and 100 μl in SS ^ⓡ (Difco, USA) agar medium. The diluted solution is plated and incubated for 24 hours in a 37 ° C thermostat to measure the number of Salmonella.

1-4-4. Effect of Salmonella on the Survival and Feces of Mice

After inoculating Salmonella typhimurium, the amount of Salmonella was measured by mouse feces. Survival rate of the extract-treated mice was much higher than that of the control group, and the number of bacteria showed a significant decrease (see Table 7).

 group Fecal Experiment Day 0 days 1 day 3 days 4 days Control group-1 0 0 2.7 X 10 ⁴ Death Control-2 0 3.0 X 10 ⁵ 2.0X10 ⁵ Death Control-3 0 5.0 X10 ⁸ Death Control-4 0 2.0X10 ³ 9.0X10 ⁵ Death Control-5 0 6.0X10 ⁷ Death Schisandra-1 0 3.0 X 10 ² 3.0 X 10 ³ 0 Schisandra-2 0 2.5 X 10 ² 3.0 X 10 ² 2.0X10 ² Schisandra-3 0 0 4.7 X 10 ³ Death Schisandra-4 0 0 0 0 Schisandra-5 0 0 30 4.4 X 10 ²

Experimental Example 2. Investigation of the safety of the drug

2-1. Establishment of experimental animal model and investigation of effect safety

To investigate the efficacy and safety of the drug in vivo, the bactericidal and antimicrobial mechanisms for infection of various bacteria (E. coli, Salmonella spp, Staphylococcus aureus ; MRSA, etc.) were investigated in mice (SPF ICR, female, 5-6 weeks) or pigs were set as experimental animals. After infection with bacteria, Schisandra chinensis extract was administered, and clinical symptoms and histological examinations were performed to investigate pathogenic changes, residual bacteria, and mortality (survival rate). Survival rate is determined by oral administration of 50 or more (up to 40 times) lethal bacteria to 8 or more animals per group, and left for 24 hours, followed by oral or intraperitoneal injection of Schisandra chinensis for 20 days. .

2-2. Pathological investigation method

   The liver, spleen, small intestine and kidney tissues of the deceased mice are cut off after the mouse dies and placed in 10% NBF for 4 days at 4 ° C. Observe pathological changes after H & E staining.

2-3. Execution Result

2-3-1. Intestine

    The control group had severe atrophy of the barrier with ischemic necrosis of the barrier (FIG. 1A). However, in the Schisandra chinensis extract treatment group, there was no obvious tissue finding and there was no significant difference from normal tissue (FIG. 1B).

2-3-2. spleen

   In the control group, congestion and hemorrhage was severe on the red pulp side, and necrosis was observed in some cases (FIG. 2A). (FIG. 2B).

2-3-3. liver

   In the control group, except for the second control group II-2, the central vein or the portal vein was severely congested (FIG. 3a), but the congestion was relatively weak in the Schizandra extract treatment group (FIG. 3b).

2-3-4. kidney

   The control group (FIG. 4A) had severe congestion of blood vessels compared to the Schizandrae extract treatment group (FIG. 4B). As a result, it was recognized that the Schisandra chinensis extract treatment group was weak in histological findings such as congestion and ischemic necrosis compared to the control group.

Feed composition example 1. (See Table 1)

Crude extract of Schisandra chinensis of Example 1-2 ............ 100 mg

Schizandra Powder of Example 1-1 ................................. 16 mg

Β-glucan-containing extract of Examples 1-3 ... 0.7 mg

Yeast culture of Examples 1-4 ... 24 mg

Vitamin E ........................................ 0.7 mg

L-Carnitine ......................................... 0.7 mg

Formulation Example 1 Preparation of Injection

Crude Extract of Schizandra chinensis Bacillus of Example 1-2 ..... 100 mg

Sodium metabisulfite .................. 3.0 mg

Methylparaben ...................................... 0.8 mg

Propylparaben ......................................... 0.1 mg

Sterile Distilled Water for Injection ...

The above ingredients are mixed and prepared in a conventional manner to have a final volume of 2 ml, and then filled into 2 ml ampoules and sterilized to prepare an injection.

Formulation Example 2 Preparation of Pills

Schizandra chinensis extract of Example 1-2 ... 120 mg

Corn starch ... 100 mg

Sterilized Distilled Water ...............

The above components are mixed and refilled in a 0.3 cm diameter by a conventional pill manufacturing method to prepare pills.

Formulation Example 3 Preparation of Tablet

Schizandra chinensis extract of Example 1-2 ... 200 mg

Lactose ... 100 mg

Starch ..................... 100 mg

Magnesium Stearate ...............

A tablet is prepared by mixing and tableting the above components according to a conventional tablet manufacturing method.

Formulation Example 4 Preparation of Capsule

Crude Extract of Schizandra chinensis Bacillus of Example 1-2 ..... 100 mg

Lactose ... .50 mg

Starch ..................... .50 mg

Talc .................. ..2 mg

Magnesium stearate .....................................

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

As described above, the Schizandra chinensis extract of the present invention is less toxic and resistant and has antimicrobial activity, thereby contributing to the increase of farm income due to the antimicrobial or immune enhancing effect.

Claims (5)

delete delete delete delete Vibrio cholerae ATCC, Salmonella typhimurium (Vibrio cholera) 01, Vibrio cholera (Vibrio cholera) 0139 ATCC 51394, Shigella di center riae (Shigella dysenteriae), Shigella flat rekse Nourishing (Shigella flexeneri), Salmonella Galina Solarium (Salmonella gallinarium) ( Salmonella typhi), Salmonella typhimurium parameters (Salmonella paratyphi) A, salmonella enterprise disk utility (Salmonella enteritidis), Salmonella Enterprise disk utility to buy (Salmonella enteritidis Sal) -36, Salmonella dublin two years old (Salmonella dublin Sal) -37, Salmonella live deolbi ( Salmonella derby Sal) -9, Salmonella cholerae -suis Antimicrobial agent using methanol extract of Schizandra chinensis , which has antimicrobial activity against ATCC 7001 or Salmonella gallinarium .

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