KR101465568B1 - Feed additive for reducing methane emission from ruminants comprising Sanguisorba offiinalis extract - Google Patents

Abstract

The present invention relates to a process for the preparation of Sanguisorba offiinalis) feed additive composition for cattle containing the extract as an active ingredient, Sanguisorba officinalis (Sanguisorba offiinalis ) as an active ingredient, and a pharmaceutical composition and food for preventing or treating an infectious disease caused by a pathogenic bacterium containing an extract of Sanguisorba offiinalis extract as an active ingredient.

Description

[0001] The present invention relates to a feed additive for reducing ruminants comprising Sanguisorba offiinalis extract,

The present invention relates to a process for the preparation of Sanguisorba offiinalis) feed additive composition for cattle containing the extract as an active ingredient, Sanguisorba officinalis (Sanguisorba offiinalis ) as an active ingredient, and a pharmaceutical composition and food for preventing or treating an infectious disease caused by a pathogenic bacterium containing an extract of Sanguisorba offiinalis extract as an active ingredient.

Antibiotics are all natural antimicrobials, semi-synthetic or synthetic materials, and they are called antibiotics. They are used for the treatment or prevention of diseases caused by microorganisms by oral administration or injection or topically. These antibiotics have been used for over 70 years to treat patients with infectious diseases. Currently, antibiotics are used to treat or prevent diseases of animals as well as human diseases.

On the other hand, antibiotics have been used as growth promoters for promoting the growth of animals in addition to the purpose of treatment or prevention of diseases in animals. The addition of antibiotics to the feed as a growth promoter to the animal promotes animal growth, improves feed utilization, and lowers the prevalence of bacterial infectious diseases in the group of animals raised do. Antibiotics prevent infectious diseases in animals, reduce the prevalence and mortality rate of infectious diseases, and increase the growth rate, thereby bringing about economical effects of lowering prices of meat, eggs and milk, It also has a positive impact on the industry. As a growth promoter, antibiotics stimulate microorganisms beneficial in the gastrointestinal tract of animals and inhibit harmful microorganisms, thereby inhibiting the production of undesirable fermentation products such as lactic acid, volatile fatty acids, ammonia, amines, and methane by intestinal bacteria . In particular, methane is a major cause of global warming, and the amount of methane gas emitted by animals per year is 80 to 115 million tons, accounting for 15 to 20% of global methane emissions. In particular, it has been reported that monenchin, an antibiotic growth promoter in animals, increases fermentation of propionate on rumen to reduce the production of methane gas, a major contributor to global warming.

As such, antibiotics used as feed additives in animals are not only economically beneficial, but can also be a way to prevent global warming from an environmental point of view. However, bacteria will naturally acquire resistance to the antibiotic if they are exposed to small doses of antibiotics for a long period of time that are below the level at which they can completely kill the bacteria. Therefore, there is a concern that the use of antibiotics as a growth promoter, which is added to feed for a long time in animal feeds with small doses, may cause microorganisms resistant to antibiotics to appear. In fact, the use of antibiotics in animals for 30 years has shown resistance to microorganisms And it has been proven by several researchers that this affects humans. For example, in animals fed with antibiotics called avoparcin in Europe, vancomycin-resistant enterococci (VRE-van A) has developed, and this resistant strain remains in the animal after slaughter, These strains were found in meat and meat products. In a study of the European population, it was found that 2 to 17% of people had this resistant strain in the intestinal tract. From this fact, the use of avopacin in animals was the main cause of VRE-van, It was concluded that it spread to humans. Thus, the use of antibiotics as a growth promoter is one of the direct causes of bacterial resistance to antibiotics. Therefore, there is a need for antibiotic substitute for it, and studies have been conducted in order to find a new antimicrobial substance in a plant that has been used for the treatment of diseases in civilian society.

Plants produce secondary metabolites to protect themselves from the environment. These secondary metabolites exhibit a variety of physiological activities and thus can be used as medicines for the prevention or treatment of diseases in humans and animals. The antimicrobial effect against pathogenic microorganisms is also one of the physiological activities that the secondary metabolites of plants can exhibit. There are also reports that plant secondary metabolites alter the fermentation of rumen in rumen to increase the efficiency of feed energy utilization, while reducing methane production. Studies have shown that some plant extracts containing phenolic compounds, essential oils, flavonoids, and sarsaponins, which are representative of the antimicrobial activity of plant-produced secondary metabolites, reduce methane production, Tannin, a secondary metabolite, has also been shown to reduce methane production in rumen.

Korean Laid-Open Patent Application No. 2012-0057200 discloses feeds for reducing methane production of ruminants and a reducing method, but is different from feed additives for ruminant methane reduction using the oat extract of the present invention.

The antimicrobial activity of Gram-positive bacteria and Gram-negative bacteria was confirmed by the in vitro antibacterial activity test, and in vitro test using bovine gastric juice, Methane production by reducing the production of methane by replacing antibiotics, pharmaceutical compositions for the prevention or treatment of infectious diseases caused by bacteria, and growth promoting the growth of food and rumen and simultaneously reducing methane production in rumen It is an object of the present invention to provide a feed additive composition for livestock which can be used as an accelerator.

In order to solve the above problems, the present invention Sanguisorba officinalis (Sanguisorba offiinalis extract of the present invention as an active ingredient.

The present invention also relates to a process for the preparation, The present invention provides a pharmaceutical composition for preventing or treating an infectious disease caused by a pathogenic bacterium containing an offiinalis extract as an active ingredient.

The present invention also relates to a process for the preparation, The present invention provides a food for preventing or improving an infectious disease caused by a pathogenic bacterium containing an offiinalis extract as an active ingredient.

The present invention also relates to a process for the preparation, offiinalis extract as an active ingredient.

It was confirmed that the oat extract of the present invention has antimicrobial effect against Gram-positive bacteria and Gram-negative bacteria and has an effect of reducing methane production upon fermentation by adding to the gastric juice of rumen. Therefore, in order to promote the growth of livestock and increase the feed efficiency, instead of antibiotics and growth promoters used for livestock, safe growth-promoting agents and antibiotics which do not cause tolerance to bacteria even if they are used for a long period of low- It can also be used as a feed additive to reduce the methane production of livestock. In addition, it can be used as a prophylactic and therapeutic agent against infectious diseases in humans in place of existing antibiotics which cause resistance to bacteria at present.

Fig. 1 shows the change in pH when the oat extract was added to the gastric juice of the rumen depending on the concentration.
Figure 2 compares the dry digestibility of the oat extract after incubation for 24 h with the ruminal juice added at different concentrations.

In order to accomplish the object of the present invention, the present invention provides a feed additive composition for domestic animals containing an extract of Sanguisorba offiinalis as an active ingredient.

In the feed additive composition of the present invention, the oat extract may be an extract obtained by extracting an oat from an oat at room temperature or 50 to 100 ° C using water or an organic solvent such as alcohol, acetone, methanol or a mixture thereof, preferably oat methanol But are not limited thereto.

In the feed additive composition of the present invention, the oat methanol extract may be prepared by mixing 14 to 16 times of methanol with respect to the weight of oat to an oat, extracting the mixture at 45 to 55 ° C for 4 to 6 hours, filtering and concentrating under reduced pressure, Preferably, the mixture of olive oil with 15 times the weight of oleoresin is added to the mixture, and the mixture is extracted at 50 DEG C for 5 hours, filtered and concentrated under reduced pressure.

In the feed additive composition of the present invention, the extract prepared by concentration under reduced pressure may be further subjected to vacuum drying or freeze-drying at a temperature of -50 to -30 ° C and a pressure of 0.1 to 0.5 Torr followed by pulverization, Do not.

In the feed additive composition of the present invention, the feed additive composition is fed to livestock to prevent or ameliorate an infectious disease caused by a pathogenic bacterium, and to reduce methane production. The infectious disease is characterized by abscess, dermatitis, osteoarthritis , Bacteremia, pneumonia, toxic shock syndrome, food poisoning, high fever, sepsis, acute breast cancer, edema, mastitis or intestinal colicosis.

Further, in the feed additive composition of the present invention, the pathogenic bacteria are Bacillus cereus (Bacillus cereus), Clostridium perfringens (Clostridium perfringens , Staphylococcus ( Staphylococcus aureus) aureus , Escherichia coli ), Salmonella entereraitidis ( Salmonella enteritidis , and Campylobacter jejuni . & lt ; / RTI >

In addition, in the feed additive composition for livestock of the present invention, the livestock may be preferably a ruminant, which may be, but is not limited to, cattle, camel, deer or giraffe.

The present invention also relates to a process for the preparation, The present invention provides a pharmaceutical composition for preventing or treating an infectious disease caused by a pathogenic bacterium containing an offiinalis extract as an active ingredient.

The pharmaceutical composition for preventing or treating an infectious disease caused by a pathogenic bacterium of the present invention may comprise the extract in an amount of 0.02 to 80% by weight, preferably 0.02 to 50% by weight, based on the total weight of the pharmaceutical composition.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the preparation of pharmaceutical compositions.

The pharmaceutical dosage forms of the extract of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in suitable aggregates.

The pharmaceutical composition containing the extract according to the present invention can be administered orally in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions And can be used as formulations. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Various compounds or mixtures including silicates, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The preferred dosage of the extract of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the administration route and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The extract of the present invention can be administered to mammals such as rats, mice, livestock, humans and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

The present invention also relates to a process for the preparation, The present invention provides a food for preventing or improving an infectious disease caused by a pathogenic bacterium containing an offiinalis extract as an active ingredient.

The food is not particularly limited as long as it can be ingested to prevent or improve infectious diseases caused by pathogenic bacteria.

When the extract of the present invention is used as a food additive, the extract may be directly added, used in combination with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the extract of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, in the production of food or beverage. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above extract can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the extract of the present invention.

In addition to the above, the extract of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the extract of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also relates to a process for the preparation, offiinalis extract as an active ingredient.

From pathogenic bacteria controlling composition of the present invention, the pathogenic bacteria are Bacillus cereus (Bacillus cereus), Clostridium perfringens (Clostridium perfringens , Staphylococcus ( Staphylococcus aureus) aureus , Escherichia coli ), Salmonella entereraitidis ( Salmonella enteritidis , and Campylobacter jejuni . & lt ; / RTI >

Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Manufacturing example  1: Preparation of oat extract

100 g of shale and finely chopped oat flour was mixed with 15 times the weight of methanol and then extracted at 50 ° C. for 5 hours in a device equipped with a reflux condenser. After filtration using a filter paper (Whatman No 2 filter paper), the filtrate was concentrated under reduced pressure to obtain an extract.

1. Antimicrobial activity measurement of extract of O.

The paper disc method, MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) tests were performed to measure the antibacterial activity of the extract of O. maximus.

Strain used to measure the antimicrobial activity of Sanguisorba officinalis extract of Preparation Example 1, Gram-positive bacteria of Bacillus cereus (Bacillus cereus KCTC1012), Clostridium perfringens (Clostridium perfringens KCTC3269) and Staphylococcus aureus (Staphylococcus aureus KCTC1916) and Gram-negative bacteria is Escherichia coli (E. coli 0157: H7, Veterinary Research and Quarantine Service), Salmonella entera itty display (Salmonella enteritidis ACTC13076) and Campylobacter ( Campylobacter < RTI ID = 0.0 > jejuni subsp. jejuni KCTC5327).

Nutrient agar and nutrient broth were used for Bacillus cereus, Escherichia coli, Staphylococcus aureus and Campylobacter jejuni, and tryptase soy agar and tryptase soy broth for Staphylococcus aureus And reinforced clostridial agar and reinforced clostridial broth were used for Clostridium perfringens.

Each strain was inoculated on each broth medium and cultured overnight at 37 ° C. Then, 1 ml of each broth was cultured in 200 ml of each broth medium at OD ₆₀₀ of 0.4-0.5 And cultured until reaching. The broth having an OD ₆₀₀ of 0.4-0.5 was diluted with each broth medium to a concentration of 1.0 × 10 ⁶ CFU / ml and used as a dilution for the test strain to measure the antimicrobial activity.

2. Paper In the paper disc method  Antimicrobial activity measurement by

The oat extract of Preparation Example 1 was dissolved in methanol so as to have a concentration of 40 mg / ml, and 40 μl of the extract was absorbed into a sterilized paper disk (diameter 8 mm, thickness 1.5 mm, JINSUNG UNITECH) until methanol was completely volatilized . This paper disk was incubated at 37 DEG C for 24 hours on a disk on which each of the above extracts was placed on each agar plate prepared by uniformly spreading the strain diluted with each strain. The diameter (mm) of the inhibition zone around the disc was then measured. Ampicillin and kanamycin were used as controls.

3. Of the extract of the oat extract MIC  And MBC  Measure

The oiula extract of Preparation Example 1 was dissolved in methanol and ethanol to a concentration of 40 mg / ml, and used as a test solution for measuring antimicrobial activity by MIC. The above oat extract was placed in a well of a 96 well plate to a final concentration of 0.125, 0.25, 0.5, 1, 2, 4, 6, 8, 16 mg / ml and methanol was used as a solvent until it was completely volatilized. After the solvent was completely volatilized, 20 μl of water, 100 μl of the prepared diluted strains and 80 μl of 0.025 w / v% TTC (2,3,5-triphenyltetrazolium chloride) were added to adjust the total volume to 200 μl per well . After incubation at 37 ° C for 20 hours, MIC was defined as the minimum concentration at which the growth of bacteria was inhibited and no red color was observed. At the same time, 100 μl of each of the wells exhibiting a MIC or higher concentration was added to the corresponding agar medium used for culturing each strain. When cultured at 37 ° C. for 24 hours, the minimum concentration at which colony growth did not occur was expressed as MBC Respectively. In order to investigate the effect of solvent methanol on the growth of bacteria, 20 ㎕ of methanol was added in place of oat extract and tested by the same method to confirm that methanol did not inhibit the growth of bacteria.

4. Of the extract of oat extract Within the rumen  Fermentation property

To investigate the effect of oat extract on the fermentation status in rumen, in vitro test was performed using gastric juice in rumen of bovine. Specifically, the oat extract prepared in Preparation Example 1 was used, and 0.2 g of substrate (TDN 70%, CP 9.6%) was added to a serum bottle and the extract was applied at a final concentration of 3.125, 6.25, 12.5, 25 and 50 mg, respectively, and dried until the methanol completely volatilized (Table 1). Twenty milliliters of rumen juice diluted with McDougall buffer mixture (pH 6.8) was added to a serum bottle containing substrate and extracts and incubated at 37 ° C. PH, dry digestibility, total gas production, methane production, volatility Fatty acid production, and ammonia production of the rumen were investigated. The ANOVA test was performed using a SAS statistical analysis program (Release 7.2, SAS Institute). The results were significant at P <0.05.

Concentration of oat extract in serum bottle (mg / L) Control group Oisal extract treated group T1 T2 T3 T4 T5 0 3.125 6.25 12.5 25 50

Example  1: Paper In the paper disc method  Antimicrobial activity of extracts from O.

As shown in Table 2, the oat extract showed 12 to 14 mm inhibition of all the strains used in the test, which is similar to the inhibitory effect of 10 μg of kanamycin, which is currently used as a broad-spectrum antibiotic, It was confirmed that it has antibacterial effect.

Antimicrobial activity of oat extract by Agar disc diffusion method Strain  Retaining ring size (mm) extract Ampicillin (10 ug) Kanamycin (10 ug) Bacillus cereus 13 - 13 Clostridium perfringens 13 28 11 Staphylococcus aureus 12 23 11 E. coli  0157: H7 12 17 14 Salmonella enteritidis 14 18 13 Campylobacter jejuni subsp. jejuni 12 21 10

Example  2: Extract of oat extract MIC  And MBC

The Minimum Inhibitory Concentration (MIC) of oat extracts ranged from 0.125 to 0.5 mg / ml and the Minimum Bactericidal Concentration (MBC) ranged from 0.5 to 16 mg / ml. The amount of ampicillin used was similar to or even lower than that of the purified antibiotic diluted 1/1000. Especially when the active ingredient is purified by measuring the vitality of crude extract before purification, antibiotic resistance can be doubled.

MIC and MBC of oat extract Strain Eucalyptus extract (mg / ml) Ampicillin ([mu] g / ml) MIC MBC MIC MBC Bacillus cereus 0.125 4 4 1.6 Clostridium perfringens 0.5 8 One 8 Staphylococcus aureus 0.5 4 0.5 8 E. coli 0157: H7 0.25 16 0.5 0.5 Salmonella enteritidis 0.25 16 0.25 0.25 Campylobacter jejuni subsp. jejuni 0.125 0.5 0.5 6.4

Example  3: pH  And dry digestibility change

As shown in FIG. 1, pH was decreased with increasing incubation time in both the control group to which no oat extract was added and the oat extract-treated group, but there was no significant difference between the control and treatment groups.

Also, there was no significant difference in the digestibility of dry matter between the control (C) and the treated group after 24 hours of culture (FIG. 2). From the above results, it was confirmed that the addition of oat extract to rumen juice did not affect pH and dry digestibility in rumen.

Example  4: Depending on incubation time Total gas  Changes in production and methane production

After 24 hr incubation, total gas production was measured. As the concentration of added oat extract increased, the amount of generated gas tended to decrease. In addition, the amount of methane was analyzed and it was confirmed that methane production was reduced in all treated groups compared with the control group. In particular, when 50 mg / L of oat extract was added for 12 hours, the methane production was reduced by 47% compared to the control (Table 4).

Comparison of total gas production and methane production after 12 hours incubation Control T1 T2 T3 T4 T5 Gas (ml) 27.2 26.2 24.4 23.6 21.6 18.6 Gas (mM) 1.07 ^a 1.03 ^ab 0.96 ^bc 0.93 ^c 0.85 ^d 0.73 ^a Gas (mM / g DM incubated) 5.35 ^a 5.15 ^ab 4.8 ^bc 4.64 ^c 4.25 ^d 3.66 ^a Gas (mM / gDM digested) 10.92 ^a 10.43 ^ab 10.05 ^b 9.49 ^b 8.28 ^c 8.83 ^c Fermentation efficiency (mg DM digested / ml gas) 3.6 ^d 3.77 ^d 3.91 ^cd 4.15 ^bc 4.75 ^a 4.45 ^ab CH4 (mM) 0.04 ^a 0.03 ^b 0.03 ^bc 0.03 ^bc 0.02 ^cd 0.02 ^d CH4 (mM / gDM incubated) 0.21 ^a 0.16 ^b 0.14 ^bc 0.13 ^bc 0.11 ^cd 0.08 ^d CH4 (mM / g DM digested) 0.43 ^a 0.32 ^b 0.29 ^bc 0.28 ^bc 0.22 ^cd 0.19 ^d CH4 (mmol / mol gas) 0.040 ^a 0.030 ^b 0.029 ^b 0.029 ^b 0.026 ^bc 0.021 ^c CH4 decrease% - 24% 28% 28% 35% 47%

Comparison of total gas production and methane production after 24 hours incubation Control T1 T2 T3 T4 T5 Gas (ml) 37.0 35.4 34.4 34.6 32.6 31.8 Gas (mM) 1.46 ^a 1.39 ^ab 1.35 ^abc 1.36 ^abc 1.28 ^bc 1.25 ^c Gas (mM / g DM incubated) 7.28 ^a 6.96 ^ab 6.77 ^abc 6.8 ^abc 6.41 ^bc 6.26 ^c Gas (mM / gDM digested) 24.49 ^a 24.29 ^a 23.10 ^ab 22.27 ^bc 20.97 ^cd 19.71 ^d Fermentation efficiency (mg DM digested / ml gas) 3.8 ^d 4.03 ^cd 4.11 ^cd 4.01 ^bc 4.26 ^ab 4.3 ^a CH4 (mM) 0.07 ^a 0.07 ^ab 0.06 ^b 0.06 ^ac 0.05 ^cd 0.05 ^c CH4 (mM / gDM incubated) 0.36 ^a 0.33 ^ab 0.31 ^b 0.3 ^bc 0.27 ^cd 0.26 ^d CH4 (mM / g DM digested) 1.21 ^a 1.15 ^ab 1.06 ^bc 0.98 ^cd 0.89 ^da 0.82 ^a CH4 (mmol / mol gas) 0.049 ^a 0.047 ^a 0.046 ^bc 0.044 ^bcd 0.043 ^cd 0.041 ^d CH4 decrease% - 4% 8% 11% 14% 16%

From the above results, it was confirmed that the extract of Oi extract had an effect of inhibiting methane production in the rumen.

Example  5: Comparison of Amount of Volatile Fatty Acid and Ammonia

Analysis of volatile fatty acids produced during the culture by gas chromatography showed no significant difference in the amount of volatile fatty acid production between the control group and the treatment group as shown in Tables 6 and 7. The amount of ammonia nitrogen source was increased in all treatment groups to which the oat extract was added for 12 hours compared with the control group to which no oat extract was added. Especially, when the oat extract was added at 25 mg / L or more, the amount of ammonia nitrogen Respectively. However, when cultured for 24 hours, the amount of ammonia nitrogen source decreased in the treated group compared to the control group. Therefore, it is considered that the extract of oat extract affects the degradation of the protein contained in the matrix and the protein synthesis of the microorganism.

Comparison of volatile fatty acid and ammonia production after 12 hours incubation control T1 T2 T3 T4 T5 Total VFA, mM 49.232 55.278 50.027 55.008 46.566 48.683 Acetate 30.242 33.496 30.499 33.228 28.769 29.956 Propionate 14.203 16.311 14.565 16.456 13.401 14.050 Isobutyrate 0.439 ^ab 0.512 ^a 0.457 ^ab 0.470 ^ab 0.386 ^b 0.393 ^b Butyrate 3.113 3.573 3.262 3.567 2.941 3.164 Isovalerate 0.684 ^ab 0.788 ^a 0.703 ^ab 0.735 ^ab 0.591 ^b 0.616 ^ab Valerate 0.551 0.598 0.542 0.553 0.478 0.502 A / P 2.129 2.054 2.094 2.019 2.147 2.132 NH3-N (mg / 100 ml) 0.0637 ^c 0.0892 ^bc 0.0566 ^c 0.1488 ^a 0.1554 ^a 0.1350 ^ab

Comparison of volatile fatty acid and ammonia production after 24 hour incubation control T1 T2 T3 T4 T5 Total VFA, mM 76.773 73.849 70.719 81.428 70.463 73.502 Acetate 45.807 43.528 41.931 48.183 41.345 42.852 Propionate 23.390 23.101 21.852 25.338 22.017 23.225 Isobutyrate 0.708 0.697 0.657 0.750 0.657 0.674 Butyrate 4.960 4.774 4.607 5.236 4.689 4.924 Isovalerate 1.095 1.057 1.007 1.124 1.014 1.048 Valerate 0.811 0.690 0.666 0.797 0.741 0.780 A / P 1.958 ^a 1.884 ^ab 1.919 ^ab 1.902 ^ab 1.878 ^ab 1.845 ^b NH3-N (mg / 100 ml) 0.4126 0.3093 0.3329 0.3000 0.2954 0.2869

In conclusion, the oat extract of the present invention has antimicrobial effects against Gram-negative bacteria and Gram-positive bacteria, and proves that it has an effect of reducing methane production in rumen.

Thus, it has been confirmed that the extract of the present invention has antibacterial effect and can be used as an environmentally friendly feed additive for reducing the amount of methane production, which is the main cause of global warming in animals. Also, it is confirmed that the extract of Gram- Which is useful for the treatment and prevention of &lt; RTI ID = 0.0 &gt;

Formulation example  1: Preparation of injection

The following ingredients were mixed, prepared by a conventional method, filled in ampoules of 2 ml capacity and sterilized to prepare an injection.

The oat extract of Preparation Example 1 100 mg pH adjusting agent Suitable amount Dissolution aid Suitable amount

Formulation example  2: Preparation of tablets

The following ingredients were mixed and tableted according to the conventional preparation method of tablets.

The oat extract of Preparation Example 1 50 mg Lactose  125 mg Microcrystalline cellulose 75 mg Magnesium stearate Suitable amount

Formulation example  3: Preparation of capsules

The following ingredients were mixed and filled into capsules according to a conventional preparation method of capsules to prepare capsules.

The oat extract of Preparation Example 1 100 mg Lactose 125 mg Starch 125 mg Talc Suitable amount Magnesium stearate Suitable amount

Formulation example  4: Preparation of feed additive

The following ingredients were mixed and prepared according to the conventional method for preparing feed additives.

The oat extract of Preparation Example 1 100 g Excipient Suitable amount

Formulation example  5: Preparation of feed containing oat extract

The following ingredients were mixed and prepared according to the conventional method for preparing feed.

The oat extract of Preparation Example 1 50 g Mushroom badge 200 g Wheat flour 30 g Beet pulp 50 g Rice anchoring 220 g Corn flakes 200 g Battery soybean 40 g Starch powder 100 g Concealed 200 g Corn cob 180 g Beji 400 g Ryegrass 323 g Geolite 14 g tapioca 40 g

Claims (1)

A mixture of methanol of 14-16 times the weight of the oat flour was mixed with the oat flour, and the resulting mixture was extracted at 45 to 55 ° C for 4 to 6 hours, filtered and concentrated under reduced pressure to obtain a methanol extract of Sanguisorba officinalis , Feed additive composition for reducing production.

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