KR100854567B1 - Composition comprising an extract of prunus fructus, schizandrae fructus, coptidis rhizoma showing antibacterial activity - Google Patents

Abstract

An animal feed supplement composition and antibacterial agent having antibacterial activity containing Prunus fruit, Coptis root and Schizandra fruit are provided to reduce toxicity and resistance and to ensure antibacterial effect and immunity boosting effect. An animal feed supplement composition and antibacterial agent contain a mixed extract of Prunus fruit, Coptis root and Schizandra fruit and one or more antioxidants selected from the group consisting of citric acid, fumaric acid, adipic acid, lactic acid, malic acid, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate, polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan and phytic acid. The animal feed supplement composition is fed in an amount of 0.01 to 2 parts by weight, based on 100 parts by weight of feed. The mixed extract of Prunus fruit, Coptis root and Schizandra fruit is obtained by mixing Prunus fruit, Coptis root and Schizandra fruit in a weight ratio of 2 to 5:1 to 3:1 to 2 and extracting in a C1-5 lower alcohol solvent.

Description

COMPOSITION COMPRISING AN EXTRACT OF PRUNUS FRUCTUS, SCHIZANDRAE FRUCTUS, COPTIDIS RHIZOMA SHOWING ANTIBACTERIAL ACTIVITY}

Fig. 1A is a diagram in which normal central vein and its surrounding sinosides are observed in the liver of the mae, nasturtium, and Schizandra administration groups (liver, mae, sulfur, Schizandra administration group, × 100).

Figure 1b is a congestion observed in the same blood vessels as the liver of the control, the diagram showing the necrosis around the blood vessels (liver, control, × 100).

Fig. 2a is a diagram showing normal intralobulr vein, renal corpuscle, convoluted tuble, and the like of the kidneys of the mae, nasturtium, and Schizandra administration groups (height, the falcon, barberry, Schizandra administration group, × 100).

Figure 2b is a severe congestion and necrosis in the kidneys of the control group and the necrosis of the proximal convoluted tuble around the vein is severely observed (height, control, × 100).

Fig. 3a is a diagram showing normal red medulla and white medulla, lymph nodes, and the like as the spleen of the maeru, nasturtium and Schisandra chinensis groups (spleen, hyacinth, biureum, Schisandra chinensis, × 100).

FIG. 3b shows that more red medulla is observed as the spleen of the control group, congestion of the red medulla is observed, more red blood cells are observed, and white medulla and lymph nodes are more clearly seen. (Spleen, control, x100).

FIG. 4A is a diagram showing that no abnormal tissue findings were found in the livers of the non-medicinal herbs, and necrosis findings and congestion findings were not observed around the central vein and portal vein of the liver (control liver, × 100).

Figure 4b is a diagram in which the proximal and tubular distal tubules around vein are normally observed in the kidney of the nonadministrative group of herbs (control kidney, control, × 100).

5A is a diagram in which no abnormal tissue findings were found in the liver of the mae, nasturtium and schizandra administration groups, and necrosis findings and congestion findings were not observed around the central vein and portal vein of the liver (soy, yam, barberry, Schizandra administration group, × 100). ).

5b shows the right side of the kidney, medullary, and Schisandra chinensis treated group as the medullary cone, and the darker portion in the middle is the renal corpuscle. × 100).

The present invention relates to an animal feed additive composition and an antimicrobial agent having an antimicrobial activity, and more particularly, to an animal feed additive composition and an antimicrobial agent containing buckwheat, rhubarb and schisandra chinensis with low toxicity and resistance to living organisms. .

In recent years, as international economic and cultural exchanges have become more active in Korea, the influx of foreign pathogenic bacteria is increasing, and domestic livestock production is expected to accelerate as the import of livestock products increases. Currently, most of the bacteria present in domestic livestock are resistant to existing antibiotics, and the treatment is becoming increasingly difficult. These treatment problems will be seriously transmitted to the human body through livestock products infected with these bacteria. 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 resistant to antibiotics, methicillin-resistant S. aureus ( MRSA), vancomycin-resistant enterococci (VRE), vancomycin-mediated S. aureus . Aureus (vancomycin intermediate S. aureus ( VISA)) and E. coli O157: H7 with at least six antibiotic resistance. In Korea alone, MRSA is a nosocomial infection agent in hospitals, and it is almost 80% in large general 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 needed 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 to effectively treat various bacteria, development of new drugs different from conventional antibiotics (molecular structure, efficacy of drugs, etc.) is urgently required. 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 conditions to be equipped with such a new drug should be a safe drug having 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 will solve this problem. The reason is that the natural product is a new component that is unknown until now, and since each action is a mixture of various single components, the pharmacological action between the complexes may 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 rates of Salmonella and enterococci such as Salmonella and enterococci isolated from cows, pigs and chickens from all over 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 ferricillin in clinical trials in 1941, about 5,500 species have been discovered to date, of which only 100 are fermented and mostly 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 with excellent antibacterial and low toxicity from natural drug sources 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 epithelium and invade host cells and infect almost all mammals, the most common strains of pathogenic bacteria leading cause of food poisoning in people causing zoonotic epidemic Salmonella typhimurium (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 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.

Accordingly, it is an object of the present invention to provide an animal feed additive composition and an antimicrobial agent, including ume, rhubarb and schisandra chinensis, which are less toxic and resistant to living organisms and have antimicrobial activity.

The above object of the present invention,

It is achieved by providing an additive composition for animal feed having an antimicrobial activity, characterized in that it comprises a mixed powder of ume, rhubarb and schizandra or extracts of the maeru, rhubarb and schizandra.

According to a preferred feature of the present invention, the mae, sulfur, and schizandra have a weight ratio of 2 to 5: 1 to 3: 1 to 2.

According to a further preferred feature of the invention, the extract is soluble in lower alcohols of C₁ to C₄.

According to an even more preferable feature of the present invention, the extract is an organic acid such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphate or polyphenol such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymeric phosphate), etc. It further comprises at least one natural antioxidant selected from the group consisting of catechin (catechin), alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, phytic acid.

According to an even more preferable feature of the present invention, 0.01 to 2 parts by weight is added to 100 parts by weight of the whole feed.

According to a still more preferred feature of the present invention, it further comprises any one or more of β-glucan (β-glucan), vitamin E, yeast culture (Yeast culture) and L-carnitin (L-carnitin).

In addition, the above object of the present invention,

It is also achieved by providing an antimicrobial agent comprising 20 to 50 parts by weight of ergots, 10 to 30 parts by weight of rhubarb and 10 to 20 parts by weight of Schizandra chinensis, comprising extracts soluble in water or methanol.

Hereinafter, an additive composition and an antimicrobial agent for animal feed having an antimicrobial activity according to the present invention will be described in detail.

Animal feed additive composition having an antimicrobial activity according to the present invention comprises a mae, rhubarb and Schizandra, may be made by mixing the powder by cutting the mae, rhubarb and Schizandra, may be made of the extract of the have. Here, it is preferable that the said ume, rhubarb, and schizandra have a weight ratio of 2-5: 1-1-3, and it is more preferable to have a weight ratio of 5: 3: 2.

The extract is chopped dried ume, rhubarb and schisandra chinensis to about 1 to 20 times, preferably about 3 to 10 times water, C₁ to C₄ lower alcohol or a mixed solvent thereof, preferably by weight (kg) Preferably, water or methanol, cold, hot water extraction, ultrasonic extraction, refluxing at 20 ° C. to 100 ° C., preferably 50 ° C. to 100 ° C. for about 1 hour to 10 days, preferably about 2 hours to 5 hours. The extract obtained by using a cold extraction method can be obtained by filtration, concentration under reduced pressure or drying.

The extract, as described above, can be obtained by extracting the mixed ume, rhubarb and schizandra in a weight ratio of 5: 3: 2 by chopping the plum, rhubarb and schizandra, and the extract is an additive for animal feed according to the present invention. It is preferable to contain 0.01 to 99.9% in a composition, and it is more preferable to contain 0.1 to 90%. However, the composition as described above is not necessarily limited thereto. The extracts of the maeol, rhubarb and schizandra may be in the form of 20 to 90% high concentrate, powder or granules.

In addition, the extract may be organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), polyphenols, catechin, alpha- It is preferable to further include at least one natural antioxidant selected from the group consisting of tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, phytic acid and the like.

Animal feed additive composition containing the extract obtained by slicing omega, rhubarb and Schizandra chinensis according to the present invention is an auxiliary ingredient such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal, enzymes, living microbial preparations, etc. Various adjuvants may be added. In addition, the animal feed additive composition containing the extract obtained by slicing the ume, rhubarb and schisandra chinensis according to the present invention is a grain, for example, crushed 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 large number of animal diets, including mammals, poultry and fish, the diet being of commercially important mammals such as pigs, cattle, sheep, goats, experimental rodents (eg rats, mice, hamsters) And gerbilus rats, etc.), fur-bearing animals (such as minks and foxes), and zoo animals (such as monkeys and tailless monkeys), as well as livestock (such as cats and dogs).

In the blending method of the animal feed composition comprising the animal feed additive composition according to the present invention, the animal feed additive composition is incorporated into the animal feed in an amount of about 0.1 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 may be fed as a mesh or formed into a desired discrete shape for further processing and packaging, in which case water is added to the animal feed to prevent separation during storage, followed by conventional pelletization, expansion, Alternatively, it is preferable to go through an extrusion process and excess water may be dried out.

The present invention provides an antimicrobial agent having low toxicity and resistance to antimicrobial activity by cutting the ume, rhubarb and Schizandra chinensis as an active ingredient.

The composition comprising the extract obtained by slicing the ume, rhubarb and schisandra chinensis of the present invention may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. The composition comprising the extract is 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 and 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.

The preferred dosage of the extract or fraction according to the invention depends on the condition and body weight of the animal, the extent of the disease, the form of the drug, the route of administration and the duration, but 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. Animal feed additive composition according to the present invention can be administered to various mammals such as rats, mice, livestock. 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 additive composition for the animal feed or the antimicrobial agent containing the extract obtained by slicing the ume, rhubarb and schisandra chinensis 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.false (50%), goldthread (30%) and the preparation method of Schizandra chinensis (20%)

1-1. Preparation of Plum, Rhubarb and Schizandra Powder

The dried plums, rhubarb and schisandra chinensis purchased at Kyungdong market are dried at room temperature for about 1 week at a cool place, and then the dried sorghum, rhubarb and schisandra chinensis are ground to a particle size of 30 mesh or less using a mill. Powder was prepared in a ratio of 5: 3: 2. The ume, rhubarb and schizandra powder obtained by the above method were used as the main raw material of the present invention.

1-2. Plums, Rhubarb and Schizandra Crude extract  Produce

100% methanol solution corresponding to three times the mass of the mixed powder (300 g) of the dried ume (50%), rhubarb (30%) and schizandra (20%) obtained in Example 1-1 was added thereto at Cooling extraction under reflux for time, filtration and lyophilization yielded 3.2 g of mixed crude extracts of omega, rhubarb and Schizandra chinensis.

1-3. β- Glucan (β- glucan ) Preparation of extract containing

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 dried mushroom powder (300 g) was added at -20 ° C, mixed for 20 minutes, and filtered. The procedure was repeated three times, followed by extracting, adding all the extracted filtrates and adding a small amount of 2M K2HPO4 solution, 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, neutralized the pH to 4.0 by adding 1 L of saturated sodium carbonate solution, 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, injected into Sephadex LH-20 column, and developed with 70% DMFA solution as a developing solvent. After lyophilization, 30 g of β-glucan-containing extract was obtained.

1-4. Yeast culture ( yeast culture Manufacturing

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 incubation, the resultant was dried in a cool place for one week so that the moisture was 10% or less, and then warm-dried at a low temperature of 40 ° C or lower. The dried yeast was used to prepare an additive composition for animal feed so that the content per kg was 17% or more.

Example  2. Preparation of Animal Feed Composition

In addition to β-glucan-containing extracts and yeasts, the additives for animal feed were prepared by adding E, L-carnitine powder, and oak powder obtained from Example 1 above. The mixed powder of the maeja (50%), rhubarb (30%) and Schisandra chinensis (20%) is 0.1 to 50%, methanol soluble extract of umeru (50%), rhubarb (30%) and schizandra (20%) is 0.1 To 50%, yeast was to be contained 10 to 20% per kg. 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 additive composition of the present invention is illustrated in Table 1.

TABLE 1

Composition example W% Composition example W% Ome (50%), Rhubarb (30%), Schisandra chinensis (20%) Powder 45 Soybean extract (50%), rhubarb (30%), Schisandra chinensis (20%) methanol soluble extract 45 β-glucan 0.5 L-carnitine 0.5 Vitamin E 0.5 Yeast culture 8.5

1-1. Disclosure strain  And culture with culture medium

Strains used to measure antimicrobial activity are isolated outdoor strains from humans or animals and ATCC standard strains (see Table 2). Strains were inoculated with Tryptic Soy broth® (TSB; Difco, USA) and incubated for 24 hours in a 37 ° C thermostat. It was.

Table 2 List of Bacterial Strains

Table 1. List of the bacterial strains used in this study

Strain describe origin JOL375 Vibrio Cholera 01 (V. Cholerae) Person JOL376 Vibrio Cholera 139 (V. Cholerae) JOL377 Shigella dysenteriae Person JOL378 Shigella flexeneri Person JOL379 E.Coli O157 ATCC 43890 MRSA104 MRSA 104 Person JOL284 E. Coli O157  animal

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

[Table 3] Salmonella List

Strain describe origin JOL380 Salmonella typhi S. Typhi Person JOL381 Salmonella paratyphi Person JOL386 Salmonella Enteritidis pig JOL387 Salmonella typhimurium pig JOL388 Salmonella typhimurium pig JOL389 Salmonella typhimurium pig JOL394 Salmonella gallinarium Chicken (splenic) JOL395 Salmonella gallinarium Chicken liver JOL398 Salmonella typhimurium pig JOL399 Salmonella typhimurium pig JOL400 Salmonella typhimurium pig JOL401 Salmonella typhimurium chicken JOL402 Salmonella typhimurium chicken JOL403 Salmonella typhimurium chicken JOL404 Salmonella typhimurium chicken JOL405 Salmonella Enteritidis chicken JOL407 Salmonella Enteritidis small JOL408 Salmonella typhimurium small JOL409 Salmonella Dublin small JOL410 Salmonella Derby small JOL411 Salmonella Cholerasis ATCC 7001 pig JOL419 Salmonella gallinarium chicken JOL420 Salmonella gallinarium chicken JOL421 Salmonella gallinarium chicken JOL422 Salmonella gallinarium chicken JOL423 Salmonella gallinarium ATCC 9184 chicken

1-2. Scanning disk method ( 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 sensitivity of bacteria, and the sensitivity to each antibiotic is sensitive (S), intermeditely sensitive (I), and resistant (resistant, Divide by R)

1-2-1. Hawk (50%), goldthread (30%) and Schisandra chinensis (20%) Extracts

After treatment with 0.5 mg / ml concentrations of ume (50%), rhubarb (30%) and schizandra (20%), the antimicrobial activity was investigated by disk diffusion method. %) Showed susceptibility to all of the following bacteria (see Table 4).

TABLE 4 Bacterial Growth Inhibitory Activities of Extracts of Ome (50%), Sulfur (30%), and Schisandra chinensis (20%)

 Botanical designation Diameter of clean area (mm) JOL 375 JOL 376 JOL 377 JOL 378 JOL 379 MRSA 104 JOL 284 Omae, Rhubarb, Schizandra Extract 27 25 15 17 20 12 10

1-2-2. Hawk on salmonella (50%), goldthread (30%), Antibacterial activity test of Schisandra chinensis (20%) extract

Antimicrobial Activity of Salmonella Extracts Treated with 0.5mg / ml Concentrations of Herb Extracts It showed high antimicrobial activity in omega (see Table 5).

[Table 5] Salmonella growth inhibitory activity of medicinal plant extracts

Sample Diameter of clean area (mm) Ome (50%), Rhubarb (30%) and Schisandra (20%) JOL380 13 JOL381 13 JOL386 11 JOL387 12 JOL388 12 JOL389 10 JOL407 8 JOL408 11 JOL409 7 JOL410 10 JOL411 12 JOL419 9 JOL420 10 JOL421 12 JOL422 10

1-3. Minimum inhibitory concentration ( MIC )

1-3-1. Experiment process

Muller-Hinton agar medium was also used to measure the minimum inhibitory concentration of the herbal medicine showed antibacterial activity. 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, followed by visual observation to determine the concentration at which the population was less than 100 as the minimum inhibitory concentration.

1-3-2. Experiment result

The minimum inhibitory concentration (MIC) of Salmonella strains was tested against 100% methanol extracts of three Chinese herbs, antibacterial (50%), rhubarb (30%) and Schisandra chinensis (20%). It was shown that antimicrobial activity ranged from 0.49 mg / ml up to 7.8 mg / ml in the OH (50%), rhubarb (30%) and Schisandra chinensis (20%) groups. (See Table 6).

Table 6 Minimum Inhibitory Concentrations of Salmonella Strains of Ome (50%), Huang Yan (30%), and Schisandra chinensis (20%)

Jol NO. species  (mg / ml) Jol NO. species  (mg / ml) Standard strain Salmonella.spp 3.9 403 S.Typhimurium 1.9 380 S.Typhi 1.9 404 S.Typhimurium 1.9 381 S.Paratyphi 0.49 405 S.Enteritidis 1.9 386 S.Enteritidis 3.9 407 S.Enteritidis 1.9 387 S.Typhimurium 1.9 408 S.Typhimurium 3.9 388 S.Typhimurium 1.9 409 S.Dublin 1.9 389 S.Typhimurium 1.9 410 S.Derby 0.49 394 S.Gallinarum 1.9 411 S.Cholerasuis 7.8 395 S.Gallinarum 3.9 419 S.Gallinarum 1.9 398 S.Typhimurium 1.9 420 S.Gallinarum 7.8 399 S.Typhimurium 1.9 421 S.Gallinarum 1.9 400 S.Typhimurium 7.8 422 S.Gallinarum 1.9 401 S.Typhimurium 1.9 423 S.Gallinarum 1.9 402 S.Typhimurium 1.9

1-4. Chicken Inoculation Experiment

1-4-1. Laboratory animals

In this experiment, chickens for three chickens of 7-10 days of age were used. There was a three-day adaptation period prior to use in the experiment. Experimental animals were divided into three groups of 30 animals in each group.

Test group-Provided with 2% of omega (50%), rhubarb (30%) and omija (20%) to feed

Group 2 (positive control)-Provide tetracycline as feed in 0.05% recommended amount

Negative control-feed only

Salmonella was tested before microbiological testing It was confirmed that it is not infected with spp . Sufficient amount of feed and drinking water were supplied throughout the duration of the experiment. Also, to reduce re-infection in the kennel, the daily feed bucket, bucket and bottom were changed.

1-4-2. Chicken inoculation experiment method

Mouse inoculation experiments included tetracycline susceptible strain S. Gallinarum (JOL 420) was inoculated in Tryptic Soy broth (TSB; Difco, USA) and cultured for 20 hours at 37 ℃ shaking incubator and centrifuged at 2,500rpm for 30 minutes to recover the cells. The cells were washed once with 1X phosphate buffered saline (PBS), resuspended in 20% sucrose in 1X PBS and inoculated orally using zondae at 1x106cfu / 2ml.

1-4-3. Chicken Survival and In feces  Effect measurement by detection of Salmonella

S to chicken. After administering Gallinarum, Salmonella spp . The number of was investigated. Survival was a daily survey for 5 days after the administration of medicines, the bacteria are excreted in feces spp Salmonella bacteria through the changes of the previous vaccination. 1, 3, and 5 days after the inoculation of the bacterium, it was subdivided into 1x10-3 and 1x10-5 using 1X PBS. The diluted samples were inoculated into Salmonella & Shigella (Difco, Sparks, USA) agar at 100 μl and incubated in 37 ° C incubator for 18-20 hours.

Salmonella detected in feces after fungal administration The number of colonies and the survival rate of chickens were investigated. Table 3 shows the survival rate of the chickens. There were no significant differences in the number of fecal collapses in each group. However, in survival, 63% (19) out of 30 survived in the mae (50%), rye (30%) and schizandra (20%) feeding groups until the 5th day after the fungal administration, while 43% (13) in the tetracycline group. ) Survived and only 20% (6) survived in the control group. Also, until the end of the experiment, the chicks (50%), rhubarb (30%), and Schisandra chinensis (20%) feeding chicks were very active and healthy, while group 2 and especially group 3 were very depressed. Had symptoms. Thus, ume (50%), rhubarb (30%) and schisandra (20%) seem to have an excellent effect on improving the survival of chicks.

Table 7 Survival rate of chicken after infection of S. Gallinarum

Group Number of dead chicken after S. Gallinarum infection 0day 1day 2day 3day 4day 5day Total death Ome (50%), Rhubarb (30%) and Schisandra (20%) 3 5 3 11 Tetracycline 4 6 7 17 Control 3 5 8 8 24

1-4-4. Pathological investigation

The liver, spleen, small intestine, and kidney tissue of the dead chicken is cut off after the chicken dies and placed in 10% NBF for 4 days at 4 ° C. Observe pathological changes after H & E staining.

1-4-5. Pathological findings

1) soy sauce

Liver of normal group (50%), rhubarb (30%) and Schisandra chinensis (20%) was normal. Normal central vein and its surrounding sinosides are observed (1a). In the liver of the control group, congestion is observed in the same blood vessel, and necrosis (1b) is around the blood vessel.

2) height

Kidney (50%), rhubarb (30%) and Schisandra chinensis (20%) showed normal height (2a) and normal intralobulr vein, renal corpuscle, and convoluted tuble. However, the renal corpuscle and convoluted tuble were not easily observed because the kidneys of the control group had congestion and necrosis. In particular, necrosis of proximal convoluted tubles around vein was severely observed (2b).

3) spleen

The group treated with ume (50%), rhubarb (30%) and Schisandra chinensis (20%) showed normal findings, and normal red and white medulla and lymph nodes were observed (3a). In the control group, more red medulla was observed, indicating congestion of red medulla (3b). In addition, more erythrocytes were observed in the control group, and white medulla and lymph nodes were more clearly seen, indicating that the inflammatory response due to inoculation was strong.

1-5. Chicken Specification

1-5-1. Laboratory animals

 (1) Broiler Specification Test

Test 1 was carried out for 5 weeks using a broiler 1-day-old Cove male (average 44g). Oysters, rhubarb and schisandra chinensis were added to broiler feed at levels of 0, 0.25, 0.5, 1, 2%, and a total of 320 waters, 4 repetitions per treatment and 16 per repetition, were disclosed. The nutrient content in the feed was divided into 2 weeks before and 3 weeks after breeding, and CP 22, 20% and ME were 3,100 kcal / kg, respectively. Other nutrient contents were based on NRC specification standard (1994).

(2) Spectrometer Specification Test

In Test 2, a four-week feeding test was conducted using a laying hen highline brown system (86 weeks old). Four treatments were added to the laying hens at levels of 0, 0.05, 0.1 and 0.2%, and 4 repetitions per treatment and 18 per repetition were published.

1-5-2 Survey Items

1) Productivity of broilers and laying hens

The body weight, feed intake and feed demand of broilers were measured by the first and last stages. Feed demand was calculated by dividing feed intake by weight gain. Laying eggs and egg weight of the laying hens were examined daily, and feed intake was examined at the end of the test. The spawning rate was expressed as the number of eggs divided by the number of spawning and breeding (Henday's spawning rate), and the average egg weight was calculated excluding eggs and malformed eggs. The daily egg production was calculated by multiplying the average egg yield and the average egg weight per day, and the feed requirement was calculated by dividing the average feed intake by the daily egg production.

2) the appearance of blood

Serum total protein, albumin, total cholesterol, triglyceride, HDL-cholesterol and the like were measured by ADVIA 1650 (JEOL, JAPAN).

3) Eggshell Strength, Eggshell Thickness, Haugh Unit, Egg Yellow Color

Eggshell strength, eggshell thickness, arc unit, and egg yolk were examined at 8-week intervals and measured at 30 replicates per treatment. The eggshell strength was measured using an eggshell strength meter (FHK, Japan), and the eggshell thickness was measured using a micrometer (FHK, Japan) after removing the inner eggshell membrane. Hounit and egg yellow were measured using an egg quality tester (QCM +, TSS, England).

1-5-3 Specification Experiment Result

end. When broiler broilers were fed omega, rhubarb, and schizandrae extracts by level, there was no significant difference in weight gain between 1-2, 3-5, and 1-5 weeks. Increased more than the control. In the case of feed intake, there was no difference in 2, 5 and 5 weeks of omega, rhubarb, and Schisandra chinensis extracts. Decreased. Feed demand rate was significantly improved in the whole group of o medium, yellow lotus and Schisandra chinensis supplemented group compared to the control group at 3-5 weeks and whole test period (1-5 weeks).

I. The broiler broiler fed mae, rhubarb and schisandra chinensis extracts according to their levels, and the results showed that the total protein, albumin and glucose levels in blood were not different from those of the control. However, cholesterol and triglyceride concentrations were significantly decreased in the 1% and 2% diets of Omae, Rhubarb and Schisandra chinensis extracts compared to the control.

All. As a result of comparing the productivity and egg quality by feeding the ume, rhubarb and Schisandra chinensis extracts in the laying hens, the addition of the buckwheat, rhododendron and Schisandra chinensis extract tended to improve compared to the control, but it was not significant.

Table 1. Effect of Herbal Supplementation on Broiler Productivity (Test 1)

Treatment Zone (%) Weight gain (g) Feed Intake (g) Feed rate 1-2 weeks 3-5 weeks all 1-2 weeks 3-5 weeks 1-5 weeks 1-2 weeks 3-5 weeks 1-5 weeks 0 384 1,294 1,679 495 2,362 2,857a 1.287 1.826a 1.702a 0.25 390 1,353 1,743 500 2,375 2,874a 1.285 1.755b 1.649b 0.50 391 1,291 1,682 494 2,283 2,777ab 1.266 1.767b 1.651b 1.00 382 1,277 1,659 501 2,229 2,731ab 1.315 1.747b 1.647b 2.00 376 1,244 1,619 492 2,171 2,663b 1.310 1.746b 1.645b SEM 3.47 13.56 14.62 4.02 25.91 25.52 0.01 0.01 0.01

Table 2 Effect of Herbal Supplementation on Blood Characteristics of Broilers (Test 1)

Treatment Zone (%) albumin protein Glucose cholesterol Triglyceride 0 1.11 2.77 200.1 171.3ab 20.3a 0.25 1.18 2.90 217.9 182.0a 20.3a 0.50 1.18 2.84 219.7 182.9a 20.9a 1.00 1.13 2.81 211.9 153.3c 15.3b 2.00 1.10 2.65 212.3 159.6bc 15.5b SEM 0.01 0.03 3.33 2.67 0.73

Table 3 Effects of Herbal Medicines on Productivity of Laying Hens (Test 2)

Treatment Zone (%) Spawn rate Lanzhou Scattering Feed intake Feed rate (feed / egg) (%, Henday) (g) (g / Wed / 1 day) (g / 1 day) 0 82.85 68.6 56.8 124.0 2.181 0.05 83.43 69.1 57.6 123.4 2.142 0.10 83.58 69.4 58.0 125.0 2.156 0.20 84.85 68.8 58.4 128.5 2.203 SEM 0.40 0.29 0.39 0.81 0.02

Table 4 Effect of Herbal Supplementation on Egg Quality of Laying Hens (Test 2)

Treatment Zone (%) Eggshell strength Eggshell thickness Egg white Ho Unit Egg yolk (kg / cm2) (Μm) (mm) 0 4.89 332 7.15 81.4 7.4 0.05 5.09 349 7.19 81.4 7.5 0.10 4.99 335 7.15 81.5 7.9 0.20 4.75 343 7.18 81.2 7.6 SEM 0.10 2.85 0.12 0.78 0.07

1-5-4. Pathological investigation

The liver and kidney tissue of the dead chicken is cut off after the chicken dies and placed in 10% NBF for 1 day at 4 ° C. Observe pathological changes after H & E staining.

1-5-5. Pathological findings

1) soy sauce

The livers of the o medium (50%), rhubarb (30%), and Omija (20%) and non-administered groups were all normal. Necrosis and congestion were not observed around normal central vein and portal vein (4ab).

2) height

The livers of the o medium (50%), rhubarb (30%), and Omija (20%) and non-administered groups were all normal. Proximal and distal tubules around the vein of the kidney were normally observed (5ab). The right side of 5b is normal as a medullary cone. The darker part in the middle is the renal corpuscle, which is also normal. Therefore, the drug appears to be a safe composition that does not show toxicity in body organs. No abnormal tissue findings were found in both the drug and non-administrative groups.

Feed composition example 1. (See Table 1)

Crude (50%), rye (30%) and Schisandra chinensis (20%) crude extracts of Example 1-2. 100 mg

Omega (50%), rhubarb (30%) and Omija (20%) powders of Example 1-1 ... 100 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 Example 1-2 (50%), rhubarb (30%) and Schisandra chinensis (20%). 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. Pilled  Produce

Omega (50%), rhubarb (30%) and Omija (20%) powders of Example 1-2.

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 Tablets

Omega (50%), rhubarb (30%) and Omija (20%) powders of Example 1-2.

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 Capsules

Crude extract of Example 1-2 (50%), rhubarb (30%) and Schisandra chinensis (20%). 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 animal feed additive composition comprising the ume, rhubarb and schisandra chinensis powder or extract according to the present invention is less toxic and resistant and has antimicrobial activity, thus contributing to the increase of farm income due to the antimicrobial or immune enhancing effect. Can be.

Claims (7)

delete delete delete In the animal feed composition having an antimicrobial activity, characterized in that the mixed extract of maeja, rhubarb and Schisandra chinensis as an active ingredient, The mae, halyeon and Schisandra chinensis has a weight ratio of 2 to 5: 1 to 3: 1, 2, The extract is a solvent of a lower alcohol of C₁ to C₄, Citric acid, fumaric acid, adipic acid, lactic acid, malic acid, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate, polyphenols, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, Licorice extract, chitosan, phytic acid has an antimicrobial activity animal feed additive composition further comprises one or more natural antioxidants selected from the group consisting of. A method of using the animal feed additive composition according to claim 4, wherein the composition is added in an amount of 0.01 to 2 parts by weight based on 100 parts by weight of the whole feed. The method of claim 4, wherein Animal feed additive composition having an antimicrobial activity, characterized in that it further comprises any one or more of β-glucan (β-glucan), vitamin E, yeast culture (yeast culture) and L-carnitin (L-carnitin). delete

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