KR20090054524A - The feed additive containing the mixed extractions of coptidis rhizoma, lonicerae flos and mori folium herbal mixture as an antibiotic and its manufacturing process - Google Patents

Abstract

A feed additive containing Coptidis Rhizoma, Lonicerae Flos and Mori Folium is provided to increase a survival rate and a carcass rate of poultry and to decrease intestinal fat and abdominal fat of the poultry. A feed additive contains 100 parts of Lonicerae Flos extract, 80-120 parts of Mori Folium extract and 1-15 parts of Coptidis Rhizoma extract. The Lonicerae Flos extract, Mori Folium extract and Coptidis Rhizoma extract are extracted by water, C1-C4 low alcohol and their mixture. The mixed solvent contains 60-80wt% of C1-C4 low alcohol and 20-40wt% of water. 0.1-2.0wt% of the feed additive is added on a total weight basis of feed. Coptidis Rhizoma, Lonicerae Flos and Mori Folium are pulverized, added to the mixed solvent and filtered.

Description

The feed additive containing the mixed extractions of Co., Ltd., and antimicrobial extracts, and extracts thereof, and the preparation method of the same, the extract additive containing the mixed extractions of Coptidis Rhizoma, Lonicerae Flos and Mori Folium herbal mixture as an antibiotic and its manufacturing process

The present invention relates to a feed additive having an antibiotic effect including a mixed extract of yellow lotus, gold and silver leaf, and a method for producing the same.

Feed additive antibiotics, which are used for livestock farming productivity through disease prevention, nutrition supplementation, and growth promotion, have been increasing in use and range of use every year. However, many countries, including the EU, are phasing out their use due to side effects of indiscriminate abuse and overuse of antibiotics, the emergence of new strains, the emergence of new pathogens, and the presence of hazardous substances in livestock foods. At the same time, the grounds for adding antibiotics to livestock feeds are threatened by factors such as increased consumer interest in the safety of livestock foods. As a result, the use of synthetic antibiotics is expected to reduce the production of safe livestock products. Therefore, the development of new antibiotic substitutes is required. However, there are currently 300 animal drug raw materials, 80% of which depend on foreign imports. Development is urgent.

In addition, the secondary hazards to the human body derived from the residue of animal medicines in livestock foods, namely, cross-resistant expression with antibiotics used by humans, are emerging as serious social problems. Since 2001, the market size has been shrinking due to the direct handling of the feed industry and the regulation of antibiotics and antimicrobial agents, which are used in vitamins, amino acids, probiotics, enzymes and trace minerals. In addition, due to the government's mandatory designation of KVGMP and the implementation of the Product Responsibility Act, it is urgent to prepare alternative measures to improve productivity and balance the industry. Therefore, in order to prevent various adverse effects that may occur in livestock and human body, it is required to develop a functional feed additive of natural materials as an alternative to using synthetic antibiotics. As a result, it is possible to establish an eco-friendly life industry by supplying a safe market for meat, and to ensure a healthy diet for the people, and to secure international competitiveness as well as the domestic market by improving the quality of domestic livestock products. In addition, the development and commercialization of high value-added functional additive products in the domestic market limited by the contraction of the domestic feed market, the limitation of the domestic market, and the regulation of drug use materials, has stabilized the domestic market and replaced the raw material imports. Can contribute to the development of

Conventionally, probiotics (probiotics) for the purpose of replacing antibiotics have been used, but their efficacy is insignificant. In addition, there is an attempt to amplify the efficacy by using prebiotics together with probiotics, which is a generic name for non-digestible food ingredients that selectively act on the growth and metabolism of enterococci beneficial bacteria and have a beneficial effect on the host. It is still in its early stages of development.

In addition, when looking at the conventional research on the foreign feed additives, the study on the bioactive oils used in the addition of probiotics and aromatherapy has not been applied to the stage of starting the investigation of the ingredients of the bioactive substances for the study of some efficacy or to manufacture the feed additives. It is a reality. In addition, there is a lack of clear mechanism of action of some agonists, insufficient safety test results of the developed materials, and lack of animal test data through field application. Accordingly, the development of antibiotic substitutes for feed additives necessary for the production of safe and clean livestock foods required by consumers in order to reduce the transfer rate of antibiotics for feed additives to livestock foods and to ensure the continuous supply of livestock foods This is urgently needed.

Therefore, the present invention isolates the antimicrobial and anti-inflammatory actives from the herbal medicine used to treat infectious diseases of the human body, and finds whether the herbal extract showing the physiological activity retains the function of prebiotics, the herbal medicine containing antibiotics Extracts are added to feed to livestock to provide safer livestock for humans as well as livestock at a more economical cost.

Accordingly, the present invention has been made to solve the above problems, and is used in the treatment of infectious diseases of humans in traditional oriental medicine efficiently used in the treatment of human infectious diseases, and the clinical safety of the drug that has antimicrobial function from a number of herbs and gold silver coins And the upper leaf was selected and by extracting the antibiotic from the herbal, to prepare a feed additive comprising the extract as an antibiotic to complete the present invention.

Therefore, an object of the present invention is to provide a feed additive having an antibiotic effect, including a mixed extract of rhubarb, sterling and upper leaves.

In addition, an object of the present invention is to provide a feed having an antibiotic effect by containing a feed additive which is an extract extracted from yellow lotus, gold and silver leaf.

Still another object of the present invention is to provide a method for preparing a feed additive comprising the extract by extracting antibiotics from the yellow lotus, sterling and upper leaves.

In order to achieve the above object, the present invention relates to a feed additive having an antibiotic effect, including a mixed extract of sulfur, gold and silver leaf.

When the gold and silver leaf and the extract were mixed with the extract of the yellow lotus and supplied to the livestock such as chicken, the antimicrobial effect was efficient, and the cost of the material can be reduced compared to the use of the yellow lotus alone. Three herbal medicines were selected from gold silver and upper leaf, and a feed additive having an antibiotic effect including a mixed extract of yellow lotus, gold silver and upper leaf was prepared.

The mixed extract is characterized in that the extraction using water and (C1 ~ C4) lower alcohol or a mixed solvent thereof. At this time, the polar organic solvent may be used, such as methanol, ethanol, butanol and acetone, but is not limited thereto.

In the present invention, E. coli ( Esherichia coli ) and Salmonella enteritidis ( Salmonella enteritidis ) were obtained by extracting methanol and hot water extracts from yellow lotus, gold and silver leaf, which have been known to exhibit antimicrobial and anti-inflammatory effects in traditional oriental medicine. ) Was tested for antimicrobial activity. In the above extraction method, when the extract was obtained using the methanol extraction method than the hot water extraction method, the antibacterial effect was superior. Therefore, when obtaining an extract containing antibiotics from the herbal medicine in the present invention, the methanol extraction method will be used.

Therefore, in the present invention, methanol extraction was performed on the selected sulfur yellow gold, silver and silver leaves as described above, and mixed under the following conditions. The herbal extract mixture is 80 to 120 parts by weight of the upper leaf extract, and 1 to 15 parts by weight of rhubarb extract with respect to 100 parts by weight of the sterling extract. Thus, a feed having an antibiotic effect was prepared by containing the feed additive which is an extract extracted from the yellow lotus, the gold and the silver leaf. At this time, the feed is prepared by including a feed additive of 0.1 to 2.0% by weight of the total weight of the feed. When the feed is included when the feed additive is included in less than 0.1% by weight, the antibiotic effect is insignificant, when included in more than 2.0% by weight there is a problem that the flavor of the feed falls.

The rhubarb and gold and silver leaf herbal extract of the present invention can be obtained as follows. Method for producing a feed additive having an antibiotic effect, including a mixed extract of the yellow lotus, gold and silver leaf of the present invention,

1) pulverizing the yellow lotus, gold and silver leaf, respectively to prepare a pulverized product;

2) obtaining each extract by adding an extracting solvent in which 60 to 80% by weight of methanol and 20 to 40% by weight of water are added to each of the milled products; And

3) filtering each of the extracts and concentrating under reduced pressure to obtain each concentrate;

It includes.

Hereinafter, the present invention will be described in detail.

At this time, if there is no other definition in the technical terms and scientific terms used, it has a meaning commonly understood by those of ordinary skill in the art.

In addition, repeated description of the same technical configuration and operation as in the prior art will be omitted.

The present invention produces a pulverized product by grinding the yellow lotus, gold and silver leaf and the upper leaf, the dry weight is not limited. Water and (C1 to C4) lower alcohols or a mixed solvent thereof were added to each of the pulverized powders, and the extracts were run at 50 to 500 rpm for 18 to 24 hours using a large-capacity extractor and filtered 2 to 10 times with gauze and filter paper. This is concentrated under reduced pressure using a vacuum concentrator. The mixed solvent is a mixture of 60 to 80% by weight of (C1 to C4) lower alcohol and 20 to 40% by weight of water. At this time, if the lower alcohol (C1 ~ C4) is less than 60% by weight, the amount of extract to extract is not enough, if it is more than 80% by weight it takes a long time to remove the (C1 ~ C4) lower alcohol from the extract There is this.

Chromatography is used for the separation of herbal extracts, which is mainly used for silica gel column chromatography and reverse phase chromatography using equilibrium according to the polarity of fixed and mobile phases.In addition, gel filtration chromatography, ion exchange chromatography, and preparative silica gel thin layer are used. Chromatography and the like can be used, but are not limited to these.

In the present invention, when the final extract extracted as described above is added to the feed, the upper leaf extract is 80 to 120 parts by weight, the rhubarb extract is mixed with 1 to 15 parts by weight with respect to the final extract of the sterling silver extract is used. The concentrate includes dissolving again in 0.5 to 2 L of an extraction solvent mixed with 60 to 80% by weight of methanol and 20 to 40% by weight of water. Finally, the extract is prepared in powder form by evaporating methanol, and the feed additive, which is the final extract prepared by the manufacturing method as described above, may be mixed with a feed for livestock to prepare a feed including a herbal extract. The feed additive may be provided in an oral dosage form including powder, granule, capsule, suspension, emulsion, syrup or aerosol form.

As described above, the present invention provides a feed additive having an antibiotic effect and a method for preparing the same, including a mixed extract of rhubarb, sterling silver and upper leaf, and the continuous administration of the herbal extract containing antibiotics causes the onset and death of diarrhea. Salmonella is controlled to increase the survival rate of poultry and increase the carcass rate by reducing the ratio of visceral and abdominal fat in broilers, raising egg production rate by 5-6% in laying hens and unsaturated fatty acids in fatty acid composition It raises the effect of lowering the content of cholesterol.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the present invention is not limited to the following examples, and it will be apparent to those skilled in the art that various changes or modifications can be made within the spirit and scope of the present invention.

Preparation Example 1 Methanol Extract of the Target Herbal Medicine

Methanol extracts of these were prepared for sulfur, gold and silver leaf. Methanol extract was extracted by using an extractor (COBIOTECH, internal capacity 500L, maximum extraction capacity 325L) for 24 hours at room temperature by adding 2L of 75% methanol solution to each 1kg dry weight of the herbal medicine used in the experiment. Filtration was performed five times, and the resultant was concentrated under reduced pressure using a vacuum concentrator and redissolved in 10 mL of 75% methanol solution.

[Comparative Example 1] Preparation of hydrothermal extract of the target herbal medicine

A hydrothermal extract was prepared for the herbal medicine of Preparation Example 1. The hot water extract was extracted with hot water extracted twice with 100 mL distilled water at 100 ° C. for 3 hours, and then concentrated under reduced pressure using a rotary vacuum evaporator. Redissolved in distilled water.

Example 1 Antibacterial Activity of Herbal Extracts

In order to measure the antimicrobial activity of the methanol extract and hot water extract extracted from the yellow lotus, gold silver and upper leaf in Preparation Example 1 and Comparative Example 1, Escherichia coli and Salmonella enteritidis stored in the laboratory were used. The antibacterial activity was measured, and the results are shown in Table 1 below. Antimicrobial activity was determined by paper disc diffusion. E. coli and Salmonella entertains on a plate of LB solid medium (10 g / L sodium chloride, 5 g / L yeast extract, 10 g / L tryptone, 7.5 g / L agarose, 121 ° C., sterilized for 15 minutes) to perform paper disk diffusion. After inoculating the bacterial growth medium of the DIS (brain-myocardial leaching medium, Brain Heart Infusion Medium), respectively, the paper disk was placed on the medium with sterile tweezers. In order to absorb the herbal extract in the paper disk was loaded 50μL and incubated in a 37 ℃ incubator for 24 to 48 hours. As a result, where the antimicrobial activity is indicated, an inhibitory zone is generated, and the presence and effect of the antimicrobial activity are determined according to the size of the inhibitory ring.

As shown in Table 1, as a result of searching for herbal extracts having antimicrobial activity against Escherichia coli and Salmonella enteritidis, it was confirmed that methanol extract exhibited higher antimicrobial activity than hydrothermal extract. It showed strong antimicrobial activity against two kinds of bacteria.

Table 1. Result of Antibacterial Activity of Herbal Extracts

 -: Crush, +: inhibition size 1-3mm, ++: inhibition size 4-5mm

Preparation Example 2 Preparation of Herbal Extracts of Selected Herbal Medicines

Herbal medicines of high antibacterial activity, sulfur and gold and silver leaf were selected, and in order to prepare the antimicrobial extract, each 100 kg of dry weight of each herbal was pulverized and 100 mesh size particles were selected as a standard sieve. 200L of 75% methanol solution was added to the particles, and the mixture was extracted at 125 rpm for 18 hours using a large-capacity extractor at room temperature, and then filtered 3 to 5 times with gauze and filter paper. This was concentrated under reduced pressure using a vacuum condenser, and then dissolved in 1 L of 75% methanol solution. The methanol was completely volatilized to obtain a final extract in powder form.

Preparation Example 3 Preparation of Feed Containing Herbal Extracts

Poultry feed additive according to the present invention was prepared in the composition shown in Table 2. The ratio of each extract prepared by the preparation method of Preparation Example 2 was used in the following experimental example by combining the ratio of gold silver: upper leaf: yellow lotus = 48.5: 48.5: 3.0. Feed preparations containing herbal extracts were prepared by mixing with 50 kg of rice bran.

Table 2. Feed Ingredients Containing Poultry Feed Additives

[ Experimental Example  1] Effects of Herbal Extracts on Broilers

In this experimental example, the control broiler (0-21 days old) and late (22-35 days old) feed containing 0% of the herbal extract prepared by the present invention in the test feed was used as a control, and the control and dynamic energy- Test feed was prepared using T1 as the feed containing the herbal extract 0.3% and T2 as the feed containing the herbal extract 1.0% to be the same protein, and the components are shown in Tables 3 and 4 below. The feed was formulated to be 3100 kcal / kg metabolic energy and 21.0% crude protein, and the late feed was formulated to be 3100 kcal / kg metabolic energy and 19.0% crude protein.

Table 3. Preparation of Test Feeds for Broiler Electric Feeds (0 to 21 Days of Age)

¹ control: 0% herbal extract

      T1: 0.3% herbal extract

      T2: 1.0% herbal extract

Content per ² feed / kg: vitamin A, 5,500 IU; Vitamin D ₃ , 1,100 IU; Vitamin E, 11 IU; 0.0066 mg of vitamin B ₁₂ ; Riboflavin, 4.4 mg; Niacin, 44 mg; Pantothenic acid, 11 mg (calcium-pantonate, 11.96 mg); Choline, 190.96 mg (choline cholide 220 mg); Menadione, 1.1 mg (menadione sodium bisulfate complex, 3.33 mg); Folic acid, 0.55 mg; Pyridoxine, 2.2 mg (pyridoxine hydrocollide, 2.67 mg); Biotin, 0.11 mg; Thiamine, 2.2 mg (thiamin mononitrate, 2.40 mg); Ethoxyquine, 125 mg.

³ content per feed / kg; Manganese, 120; Zinc, 100; Iron, 60; Copper, 10; Iodine, 0.46; Calcium, min: 150, max: 180.

Table 4. Preparation of Test Feed for Late Broiler Feed (22-35 Days Old)

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

Content per ² feed / kg: vitamin A, 5,500 IU; Vitamin D ₃ , 1,100 IU; Vitamin E, 11 IU; 0.0066 mg of vitamin B ₁₂ ; Riboflavin, 4.4 mg; Niacin, 44 mg; Pantothenic acid, 11 mg (calcium-pantonate, 11.96 mg); Choline, 190.96 mg (choline cholide 220 mg); Menadione, 1.1 mg (menadione sodium bisulfate complex, 3.33 mg); Folic acid, 0.55 mg; Pyridoxine, 2.2 mg (pyridoxine hydrocollide, 2.67 mg); Biotin, 0.11 mg; Thiamine, 2.2 mg (thiamin mononitrate, 2.40 mg); Ethoxyquine, 125 mg.

³ content per feed / kg; Manganese, 120; Zinc, 100; Iron, 60; Copper, 10; Iodine, 0.46; Calcium, min: 150, max: 180.

The broilers used in the experiments were randomly placed with 480 freshly hatched broilers (Cobb 500) in 40 pens in 12 pens (3.0m × 1.0m). The litter used sawdust and lit for 24 hours. Feed was fed from 0 to 21 days of age, and late feed was added between 22 and 35 days of age. Feed and water were freely available. Body weight was measured once every 8 days, 22 days, 36 days. Feed intake was measured for electric and late feed intake respectively. Feed demand was calculated by dividing total feed intake by total weight. The effect of adding the herbal extract to the feed is shown in Table 5 below. The herbal extracts were added 0.3% and 1.0% in the early and late broiler feeds, but no significant differences were found in the growth rate, feed intake and feed efficiency compared to the control. However, in the experimental group to which the herbal extract was added, the survival rate was increased by 1 to 3% compared to the control group.

Table 5. Effect of Medicinal Herb Extracts on Fodder Growth

¹ control: 0% herbal extract

      T1: 0.3% herbal extract

      T2: 1.0% herbal extract

² means ± SD.

When the herbal extracts were added to the feed, the effects on the ratio and carcass ratios of the viscera and abdominal fat, which are not planted, are shown in Table 6 below. In the visceral ratio, the addition of herbal extracts was lower than the control, and the abdominal fat ratio was significantly lower than the control, and the carcass ratio was higher than the control. When the herbal extract used in this study was added to the feed and added to the broiler, the ratio of visceral and abdominal fat, which is non-planting site, was reduced, which resulted in an increase in carcass rate. It is believed that it can help to improve the conductor quality.

Table 6. Effects of Herbal Extracts on Broiler, Abdominal Fat and Carcass Rates in Broilers

¹ control: 0% herbal extract

      T1: 0.3% herbal extract

      T2: 1.0% herbal extract

^ab Means with different superscripts within the same column are significantly different (P <0.05).

Experimental Example 2 Effects of Herbal Extracts on Laying Hens

In this Experimental Example, the control diets were made from chicken broiler (0 to 21 days old) and late (22 to 35 days old) feed containing 0% herbal extract prepared by the present invention as a control, and control and dynamic energy (TMEn 2,780 kcal). / kg)-the same protein (17.0%) to the feed containing 0.3% herbal extract to T1, the feed containing 1.0% herbal extract to T2 to prepare a test feed, the ingredients are shown in Table 7 Indicated.

Table 7. Preparation of Test Feed Added with Herbal Extracts in Laying Hens

¹ control: 0% herbal extract

      T1: 0.3% herbal extract

      T2: 1.0% herbal extract

² feed / kg content: copper, 10 mg; Iron, 80 mg; Manganese, 80 mg; Zinc, 80 mg; Iodine, 0.9 mg; Selenium, 0.2 mg; Cobalt, 0.5 mg.

Content per ³ feed / kg: vitamin A, 12,000 IU; Vitamin D3, 3,000 IU; Tocopherol 15 mg; Vitamin K3, 2 mg; Thiamine, 2.0 mg; Riboflavin, 6.0 mg; Pyridoxine, 2 mg; Vitamin B12, 0.03 mg; Folic acid, 1.0 mg; Biotin, 0.15 mg; Niacin, 45 mg; D-calcium pantotinate, 15 mg; Antioxidant, 0.5 mg.

⁴ Grains dried with corn distiller.

⁵ Calculated value.

The clock used in the test was a 108-week-old broiler egg of Roman Brown, which was bred for six weeks at Chungnam National University. The experimental design was completely randomly placed in 3 treatments, 6 repetitions, 6 numbers per repetition, and managed by lighting for 17 hours per day. The feed intake was measured by subtracting the remaining amount from the total amount in 7-day increments, and the egg laying rate was measured daily by Hen-day egg laying rate by dividing the number of eggs collected at 1 pm daily by the number of breeding eggs. . The results on the egg productivity of laying hens by the addition of herbal extracts are shown in Table 8. As a result, in laying hens with added herbal extracts, the egg production rate increased by 5 to 6%, and egg weight and total egg weight increased slightly. However, there was a significant difference in feed intake, egg production and feed efficiency according to herbal treatment. Was not found.

Table 8. Laying hens by adding herbal extracts ^One Of productivity

^One Targeting Roman Brown Laying Hens

² control: 0% herbal extract

      T1: 0.3% herbal extract

      T2: 1.0% herbal extract

Weekly spawn rate of ³ hens

⁴ Feed Intake / Egg Weight

⁵ means ± SD.

Fatty acid and cholesterol analysis was performed according to the method of Folch et al. (1957) and fat was extracted by gas chromatography (Gas Chromatography). The effect of the herbal extracts on the fatty acid composition and cholesterol content of egg yolk is shown in Table 9 below. Linolenic acid (18: 2), an essential fatty acid among unsaturated fatty acids, increased 1.05 ~ 1.6% in the herbal extract addition group, and the content of linolenic acid (18: 3) also showed a high increase in the herbal extract additive family. The herbal extract added group showed a result of higher unsaturated fatty acid content of about 1.5-2% of the fatty acids of egg yolk compared to the control group. In addition, the cholesterol content of yolk extract added was lowered around 1.5% compared to the control, which was found in palmitic acid (16: 0), which is known to increase blood cholesterol concentration in saturated fatty acids. It appears to be due to a significant increase in the content of linolenic acid (18: 3), which is known to possess a function of lowering blood cholesterol levels with a decrease in content.

Table 9. Effect of Herbal Extracts on Fatty Acid Composition and Cholesterol Content in Egg Yolk

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

^* Saturated fatty acids

^** Unsaturated Fatty Acid

Experimental Example 3 Effect of Herbal Extracts on Intestinal Microflora

In this experimental example, we tried to verify the effects of mixing the herbal extracts on the intestinal microflora of broilers and laying hens. The broilers and laying hens used in this experiment were selected from three-day-old 36-day-old broiler (Cobb 500) and Roman-Brown-based 34-week-old hens. . This was washed several times with physiological saline and diluted step by step, followed by SS medium (Salmonella Shigella agar, peptone 10.0 g / L, lactose 10.0 g) for the measurement of Salmonella sp., Escherichia coli and total bacteria and anaerobic bacteria. / L, ox bile 8.5g / L, sodium citrate 10.0g / L, sodium thiosulfate 8.5g / L, iron (III) chloride citrate 1.0g / L, brilliant green 0.0003g / L, neutral red 0.025g / L, loot 12.0 g / L), MacConkey agar, casein peptone 17.0 g / L, meat peptone 3.0 g / L, sodium chloride 5.0 g / L, lactose 10.0 g / L, bile salt mixture 1.5 g / L, Neutral Red 0.03g / L, Crystal Violet 0.001g / L, Locust 13.5g / L, 4-methylumbelliferyl-bD-glucuronide (0.1g / L), Nutrient agar, broth extract 3.0g peptone 5.0g, pH 6.8) and bacteria growth medium (Brain Heart Infusion medium agar (Difco, product number: 4203290) 26g, distilled water 500ml) using Analyzed.

Addition of herbal extracts was shown to significantly reduce the number of Salmonella genus, E. coli and total bacteria in broilers and laying hens as shown in Table 10 below. This proved that the herbal extract used in this experiment possessed an antagonistic activity against various types of pathogenic bacteria in the process of passing through the intestinal tract, while reducing the total number of microorganisms.

Table 10. Effects of herbal extracts on intestinal microflora of broilers and laying hens

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

Experimental Example 4 Effects of Herbal Extracts on the Excretion of Gas

In this experimental example, ammonia and hydrogen sulfide gas were measured in the broilers collected in the middle (3 weeks) and late (5 weeks) of broilers in order to investigate the effects of the addition of herbal extracts on the generation of noxious gases generated from feces. Hazardous gas emissions were measured in the middle and late broiler seasons respectively. To measure the ammonia and hydrogen sulfide gas emissions, 100 g of each excreted within the same time was weighed in a sealed container and left for 24 hours at room temperature. After that, it was measured using Gastec (GV-100S, Japan).

The results measured as described above are shown in Table 11 below. In general, the amount of ammonia gas produced was significantly lower than that of hydrogen sulfide, and the amount of harmful gas was decreased in the herbal extract added group as the broiler was added more lately. As shown in Table 11, the decrease in the intestinal microbial population was associated with the propagation of ammonia-producing bacteria that secrete urea by the addition of herbal extracts, thereby reducing the amount of harmful gases.

Table 11. Effect of Herbal Extracts on Harmful Gas Emissions from Broiler Manure

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

^ab Means with different superscripts within the same column are significantly different (P <0.05) ..

Experimental Example 5 Inhibitory Effect of Salmonella Enteritidis on Intestinal Settlement by Addition of Herbal Extract

1) Influence of Diarrhea with Herbal Extracts and Effect of Salmonella Enteritidis in Caecum

In this experiment, we investigated the distribution and the incidence of diarrhea in the organs after artificially infecting Salmonella enteritidis strains to investigate whether Salmonella infection can be suppressed in broilers when mixed with herbal extracts. We reviewed the effects of intestinal settlement. Two-week-old broiler chickens (Cobb 500) were used, and the experimental design consisted of 3 treatments, 3 repetitions, 6 randomly placed per repetition, and the feeds were 0.3% and 1 herbal extracts from commercial broilers. Each was added in percentage and control was used only commercial broiler electric feed. For artificial infection of Salmonella bacteria, Salmonella enteritidis ATCC 13076 (1 ㅧ 107 cfu / ml) in liquid culture in Tryptic soy broth (Product No. BD-9061-73) once daily In total, three oral administrations were performed. The incidence of diarrhea was examined at intervals of one week for two weeks after inoculation, and the distribution of Salmonella enteritidis bacteria in the cecum was measured simultaneously. The serum antibody was separated from the serum at 1 week intervals for 2 weeks after Salmonella enteritidis inoculation, and then subjected to the Microplate Agglutination Test (MAT) using an antigen prepared using Salmonella enteritidis ATCC 13076 as a standard strain. It was measured by.

As a result of the above experiment, the results of diarrhea incidence caused by artificial infection of Salmonella enteritidis are shown in Table 12 below. The control group showed diarrhea symptoms in 5 waters (2 deaths) in 6 waters at 7 days of infection and the 2 extracts (T1) or 3 waters (T2, 1 water deaths) in 6 waters at 7 days of infection. Only 1 out of 6 cases showed diarrhea.

Table 12. Effect of Herbal Extracts on the Development of Diarrhea in Broiler Chickens

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

^a Tested with 6 broilers.

In addition, the results of the population of Salmonella enteritidis present in the cecum of each test zone is shown in Table 13 below. In the first week of inoculation, a slight decrease in the number of bacteria was observed in the control group compared to the first day of the inoculation, but there was a significant decrease in the number of herbal extracts. Salmonella enteritidis in the cecum increased more than 89% in the cecum extract 1%, T1 in the cecum extract 0.3% added Salmonella enteritidis significantly more than 77%.

Table 13. Salmonella Enteritidis Population in Broiler Caecum

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

^a mean ± SD 3 to 6 measures tested on broilers.

2) Changes in Broiler Organ Tissue according to Salmonella Enteritidis Addition of Herbal Extracts

In this experimental example, the broiler was slaughtered two weeks after the inoculation in order to observe changes in broiler organ tissues due to artificial infection of Salmonella enteritidis, and the cecum was extracted. L, sodium monophosphate 3.2g / L, sodium chloride 90g / L, pH 7.2) several times and observed with a scanning electron microscope (SEM). Observation results of broiler organs were shown in FIG. 1. In the control group, the inflammatory response was observed on the intestinal mucosal epithelium, whereas the intestinal epithelial tissue was relatively healthy in the T1 supplemented with 0.3% herbal extract and T2 supplemented with 1% herbal extract.

3) Analysis of serum antibody titer according to the addition of herbal extracts at the time of inoculation of Salmonella enteritidis

In the present experimental example, blood was collected from the heart immediately after the cervical dislocation method at 1, 1, and 2 weeks after artificial infection of Salmonella enteritidis, blood was coagulated at room temperature, and centrifuged. Serum antibody was stored in the cryogenic refrigerator at 80 ℃ was used for analysis. The results are shown in Table 14 below. In this experimental condition, the antibody titer in serum was lower than the control group.

Table 14. Salmonella Entertainers  Broiler in artificial infection Serum Antibody Analysis

¹ control: 0% herbal extract

       T1: 0.3% herbal extract

       T2: 1.0% herbal extract

1 is a result of the change in broiler organ tissues caused by artificial infection of Salmonella enteritidis

Figure 1a is a scanning electron microscopic observation of the control,

Figure 1b is a scanning electron microscopic observation of the herbal extract 0.3% added T1,

Figure 1c is a scanning electron microscopic observation of the 1% herbal extract T1.

Claims (9)

Feed additive having an antibiotic effect, including a mixed extract of rhubarb, gold and silver leaf. The method of claim 1, The mixed extract is a feed additive, characterized in that the extraction using water and (C1 ~ C4) lower alcohol or a mixed solvent thereof. The method of claim 2, The mixed solvent is a feed additive, characterized in that the extraction using the extraction solvent mixed 60 to 80% by weight (C1 ~ C4) lower alcohol and 20 to 40% by weight of water. The method of claim 1, The mixed extract is a feed additive mixed with 80 to 120 parts by weight of upper leaf extract and 1 to 15 parts by weight of rhubarb extract with respect to 100 parts by weight of gold silver extract. Feed having an antibiotic effect by containing a feed additive which is an extract extracted from the yellow lotus, gold and silver leaf according to any one of claims 1 to 4. The method of claim 5, Feed comprising a feed additive of 0.1 to 2.0% by weight of the total weight of the feed. 1) pulverizing the yellow lotus, gold and silver leaf, respectively to prepare a pulverized product; 2) obtaining each extract by adding an extracting solvent in which 60 to 80% by weight of methanol and 20 to 40% by weight of water are added to each of the milled products; And 3) filtering each of the extracts and concentrating under reduced pressure to obtain each concentrate; Feed additive manufacturing method comprising a. The method of claim 7, wherein Feed each of the concentrates obtained in step 3) is dissolved again in the extraction solvent mixed with 60 to 80% by weight of methanol and 20 to 40% by weight of water, respectively, further comprising the step of obtaining each final extract Additive manufacturing method. The method of claim 7, wherein The final extract is a feed additive manufacturing method, characterized in that the mixture of 80 to 120 parts by weight of the upper leaf extract and 1 to 15 parts by weight of rhubarb extract with respect to 100 parts by weight of sterling extract.

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