KR20130099685A - Antimicrobial agent and feed additives comprising the same - Google Patents

Abstract

PURPOSE: An antimicrobial agent and an animal feed additive containing the same are provided to use a medical herb extract. CONSTITUTION: A byproduct of yangkyuksanwha-tang is inoculated with bacillus for fermenting. A hot-water extract of the fermented product is obtained. The hot-water extract, a mixed extract of Caesalpiniae Lignum, Galla Rhois, and Magnoliae Cortex, and starch are mixed and granulated. The hot-water extract, a mixed extract of Caesalpiniae Lignum, Galla Rhois, and Magnoliae Cortex are mixed in a volume ratio of 1:7-10. Extracts of Caesalpiniae Lignum, Galla Rhois, and Magnoliae Cortex are mixed in a volume ratio of 1:1:1. The extract has the antibacterial activity to Staphylococcus genus or Salmonella genus bacteria. [Reference numerals] (AA) Caesalpiniae Lignum 70% ethanol extract; (BB) Galla Rhois 70% ethanol extract; (CC) Magnoliae Cortex 70% ethanol extract

Description

Antimicrobial agent and feed additives comprising the same

The present invention relates to an antimicrobial agent comprising an herbal extract and an animal feed additive comprising the same.

Since the first reported effects of antibiotic addition in poultry feed, many researchers (Boyd et al., 1994; Damron et al., 1991; Jeffries et al., 1997; Krudger. 1983) A study was conducted. Moreover, as the livestock industry grew in the form of group breeding, the rate of disease outbreaks and the spread of infectious diseases in livestock increased, and the intensive use of drugs to prevent and treat diseases increased (Anadon and Martnez-Larranga, 1999). However, the side effects of excessive use of antibiotics are harmful to the human body due to antibiotic resistance, and the economic consciousness of consumers due to economic growth and diversification of food is changed from quantitative to qualitative. The system is changing to an era that demands health functionalities and stability from harmful substances to the human body. Consumption of organic and organic livestock products is on the rise as consumers' awareness of health and health and disease for life and longevity are increasing. The EU, which has proposed regulations for organic food production and labeling, For animal feed production, antibiotics are no longer allowed if 100% organic feed should be used.

The antibiotic regulations of domestic livestock products have increased consumer interest in eco-friendly livestock products with the introduction of the antibiotic-free livestock certification system in April 2007. Consumer awareness of antibiotic resistance found that awareness of antibiotic resistance increased from 28.5% in 2004 to 74.1% in 2008. In order to increase the stability of livestock products and increase the reliability of livestock products, the government has gradually reduced the use of antibiotics for feeds since 2005. Since July 2011, the government has banned the addition of antibiotics to prevent the use of antibiotics. Much research has been done to produce antibiotic livestock products.

In order to produce low-cost, high-quality livestock products, instead of antibiotics, various by-product feed materials are used to develop alternative materials. It is widely used (Sang Ho Kim, 2008). Most of them show the possibility of promoting antibiotic growth as a disease prevention agent, such as promoting growth through health promotion of livestock, regulation of intestinal microflora, activation of digestive function, and anti-pathogenicity.

Chinese herbal medicine is mostly composed of plants, and there are many by-products, so there is a possibility of supplementary feed. The medicinal effects and physiologically active substances in herbal medicines have been used for a long time for the prevention and treatment of various diseases, and there is a lot of interest even now, and some herbal medicines are easily used in life. An example of the use of by-products in farms has been reported to produce and feed feed using gold, a by-product of herbal medicine, to reduce costs and to produce high-quality Korean beef, and to grow mushroom media using herbal by-products from pharmaceutical companies. Compared with the increase in yield by 10% and cost-saving effect such as energy through the simplification of the cultivation process, various experiments using the functionalities of the by-products and various medicinal plants were performed and showed the possibility as an antibiotic replacement material.

One aspect is to provide an antimicrobial agent comprising one or more herbal extracts selected from the group consisting of joiner, gall bladder and groin.

Another aspect is to provide an animal feed additive comprising the antimicrobial agent.

Another aspect is to provide a method of making the feed additive.

One aspect provides an antimicrobial agent comprising one or more herbal extracts selected from the group consisting of joiner, gall bladder and groin.

The antimicrobial agent of the present invention is characterized in that it comprises an extract of one or more herbal medicines selected from the group consisting of joiner, gall bladder and groin as an active ingredient.

Caesalpinia sappan L. is a plant belonging to the legumes (Leguminosae), and the stem and coarse branches of the tree are called joiner. (Kim Chang-min et al., Chinese Medicinal Dictionary, pp. 3130, 1998). The main components of the joiner are hematoxylin and flazioid, a colorless primary pigment brazilin, which is oxidized in air to become brazilein. Brasilin, the main component of joiner, has been reported to have high blood pressure, control of calcium concentration in platelets, and blood glucose lowering action. In addition, sappanchalcone, one of the components of joiner, has been reported to inhibit platelet coagulation.

Flaw is a rhododendron belonging to the sumac ( Rhus javanica L. ) Aphis chinensis J. Bell on the leaves of the insects caused by the stag beetle, distributed throughout the country. In the form of gall, the outer surface is grayish brown with light hairs, 3 ~ 7cm long, 2 ~ 5cm wide and 2mm thick, and hard and easily broken. The genus is usually empty or gray-white dead worms and secretions remain, and there is a disgusting smell. The collected dwarf is dried, boiled or steamed in the sun. Its main ingredient is 50 ~ 70% of pyrogalloltannin, and it includes malated assets and gallic acid. The drug is used for astringents, branches, hemostatic diarrhea, sputum, diabetes, bleeding, anemia.

Hug ( Magnolia officinalis Rehd. et Wils.) is a perennial herb belonging to the Magnolia family, which has been dried and used as a health agent for a long time in Korea, Japan, and China. Hupak has been used for food in Korea and Japan, and it is known to have the effects of keynote moisture and anti-pacific peace. In oriental medicine, it is used for the treatment of directional stomach and stomach pain, abdominal bloating, acute enteritis, seawater, etc., as a gastric digestive tract, lower extremity, and underground sea expectorant, Wiryngtang, overhanggigi, Soseunggitang, Parkhahubaktang, Pyeongwisan, Mazainhwan, etc. Used as a formulation component of Known components of the hummus are β-eudesmol, β-eudesmol, α-βpinene, magnool and honokiol. honokiol, alkaloid, magnocurarine, magnoflorine, magnoflorine, michelarbine, anonaine, liriodenine, tannin, etc. Among them, magnolol and honokiol are known to have a safe and excellent antibiotic effect.

In the present invention, the extract of the joiner, gall bladder or hoobac is a variety of extraction solvents, for example, water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), the lower alcohol and water A mixed solvent, acetone, ethyl acetate, chloroform or 1,3-butylene glycol can be obtained as an extraction solvent. On the other hand, it will be apparent to those skilled in the art that the extract of the present invention can be obtained in addition to the above-described extraction solvents, extracts having substantially the same effect using other extraction solvents. Preferably, the extract is an extract obtained using water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, more preferably an extract obtained using water, methanol or ethanol, and most preferably using ethanol Obtained extract.

In addition, the extract of the present invention includes not only the extract obtained by the above-mentioned extraction solvent, but also an extract obtained through a conventional purification process. For example, through various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). The obtained fractions may also be included in the extract of the present invention.

According to one embodiment, the extract is Staphylococcus Genus or Salmonella May exhibit antimicrobial activity against the bacteria of the genus. The bacterium, for example, causes disease in broilers Staphylococcus epidermidis , Salmonella enteritidis or Staphylococcus It can be aureus . On the other hand, the extract may exhibit an antimicrobial effect on each of the bacteria at a concentration of 25 to 100 mg / ml.

Another aspect provides an animal feed additive comprising an antimicrobial agent comprising one or more herbal extracts selected from the group consisting of joiner, gall bladder and groin.

According to one embodiment, the animal may be poultry, for example, most preferably a chicken.

According to one embodiment, the feed additive may further comprise a hydrothermal extract of by-products of the yangyangtang.

Yuyang Yutang is composed of 8: 8: 8: 4: 4: 4: 4: 4% by weight As a medicinal herb, it is widely used as a prescription for fish blood in Chinese medicine. The feed additive may include an extract extracted by hot water (by distilled water of 70 to 90 ℃) of the by-product of the yangyangtang. By-product of the two-sided oxidation bath means the remaining portion after removing all the liquid after decoupling the two-sided oxidation bath.

Yanggyuktangtang of the present invention is characterized in that it comprises an extract of herbal medicine consisting of Yeongyo, Indongung, Saenghwanghwang, Gardenia, Peppermint, mold opening, windproof, gypsum, and hair.

Forsythiae Fructus is a fruit extract of the forsythia, asteraceae, mainly treating Nag power, carbuncle, swelling, permanent, cracking, loneliness, lanceolate and aching pain, detoxification of urinary tract and pain, decongestion It is used for anti-inflammatory, and in modern medicine, it is used to inhibit c-AMP phosphodiesterase action, antibacterial action against Staphylococcus aureus. Lignans, Pinoresinol, Pinoresinol Glucoside Matairesinoside, Matairesinol, Philyrin, Arctigenin ), Actinin, Caffeoyl glycoside, Forthiaside, Susensoside, Acteoside, Viridissimaside, Acetyloleanolic acid acid), Beturin, Viridissimic acid, Orleanolic acid, Betulinic acid, Rutin, Ursolic acid, and the like.

Indong lanterns ( Lonicera japonica Thunberg ) is a vine plant of the dicotyledonous locust tree, which is a vine plant. In the oriental medicine, the leaves and stems of the inflorescences are seen, and the flowers are white and yellow flowers at the same time. Its main ingredients include leucine, flavonoids, tannins, alkaloids, luteurins, inositol, saponins, roniserine, and luteolin, which are known to act on menopause, parenteral and heart, and have antipyretic and detoxifying effects. have. In addition, it is known that it is applied to fungi, immune revive, anti-inflammatory, analgesic, diuretic, anticancer, and antiviral. It is used as a medicine for antipyretics and healing.

Rehmannia glutinosa Libschitz var . purpurea Makino) uses fresh roots of perennial herbaceous plants belonging to Hyunsam family, which is called Jihwang. Fresh root roots are either raw or sulfur, dried or dried, dried or dried, and then steamed and dried in the sun 9 times. Bamboo leaf is good for diabetes, and it stops bleeding, nosebleeds and uterine bleeding, and it is good for menstrual irregularity and constipation. As for the components, beta cystosterol (β-sistosterol), alkaloids and fatty acids were identified. Pharmacologically lowering blood sugar, acting on the circulatory system, has a strong heart and diuretic effect, liver function protection, antibacterial action.

Gardenia ( Gardeniae Fructus ) is the fruit of gardenia or other related plants belonging to the family Maddera, and has been used as a food coloring agent since ancient times. In one shot, anti-inflammatory, diuretic, hemostatic. It is used for antipyretic, soothing, antibacterial, bile secretion, hepatitis, jaundice, hemostasis, reducing serum triglycerides, phospholipids, lipid peroxides, blood sugar, free fatty acids, suppress stomach movements, reduce tension, and It is reported that it suppresses an increase in the amount of bilirubin and has an effect of preventing atherosclerosis.

Peppermint ( Mentha arvensis var . piperascens Makinv.) has a peculiar smell and it is spicy and cold if you put it in your mouth and give it a cool feeling. Efficacy is used for symptoms of fever, headache and sweating caused by colds of the eyes, eye congestion, sore throat, early measles itching, liver disease, abdominal pain, diarrhea and vomiting. Pharmacological actions include fever, anti-inflammatory, dry stomach, bile secretion and gastrointestinal smooth muscle suppression, increased mucus secretion in the respiratory tract, central nervous excitability, and uterine contractions.

Hyungae ( Schizonepeta) tenuifolia var . japonica KITAGAWA) uses sun-dried grasses of annual herb belonging to Lamiaceae. The oil contains 1.8% of essential oils, and d-menthone, dl-menthone and a small amount of d-limonene are included in the essential oil. It is used for sweating, antipyretic, cramping, analgesic, cough, fever, cold, headache, limb stiffness, stroke, postpartum dizziness, lymphadenitis, sore throat, tonsillitis, hemostasis, anemia, gallstones, rubella, margins.

Windstorm ( Ledebouriella seseloides ) is a dicotyledonous plant and is a perennial herbaceous plant that grows in dry sandy grasses. It is native to northeastern China, Huabei and Inner Mongolia, and is grown as a medicinal crop. In oriental medicine, two-year-old roots are used as medicine for colds, headaches, sweating, and expectoration.

Anemarrhena asphodeloides ) is a plant belonging to the family Liliumaceae. It is native to China and is a perennial plant grown in the central part of Korea. Usually, it has been used as a medicine by drying the rhizome. It is known to have anti-inflammatory, antipyretic, branch, diuretic and low back pain relief and soothing effects. In addition to 6% asphonin as an active ingredient of the hair, steroids such as salsasaponin (Sarsasaponin) and markogenin (2-hydroxy sarsasapogenin) have been reported to contain sapongenin, flavonoids and tannins. .

According to one embodiment, the feed additive may include a mixture of the mixture of the joiner, gall bladder and hulky extract and hot water extract of the by-product of the yangyangtang in a volume ratio of 7: 1 to 10: 1. Most preferably, the mixed solution of the joiner, gall bladder and the pear extract and the hydrothermal extract of the by-product of the oxidized tang can be mixed at 9: 1. In addition, according to one embodiment, the mixed solution may be a mixture of joiner, gall bladder and hulky extract in a volume ratio of 1: 1: 1.

According to one embodiment, the feed additive may be granular. The feed additive may include corn starch to produce granular feed additives.

Another aspect includes the step of inoculating Bacillus bacteria in the by-product of the yangyangtang by fermentation; Obtaining a hydrothermal extract of the fermentation result; And it provides a feed additive manufacturing method comprising the step of mixing and granulating the mixed solution and starch of the hot water extract, joiner, gall bladder and thick gourd extract.

The method of preparing the feed additive may first include inoculating Bacillus bacteria in by-products of the yangyangtang by fermentation.

According to one embodiment, the Bacillus bacteria Bacillus It may be licheniformis , rice bran may be added in a proportion of about 1% by weight to the by-product of the yangyangtang to create fermentation conditions. The Bacillus bacteria is preferably inoculated by 1% by weight of the by-pass oxidized water by-product. Fermentation may be performed at room temperature (25-30 ° C.) for about 4-7 days in a closed state.

Thereafter, the method may include obtaining a hydrothermal extract of the fermentation result.

The hot water extract may be obtained by adding distilled water of 3 to 7 times, most preferably about 5 times to the fermentation result of the fermentation is completed in the fermentation step, by heating for 20 to 40 minutes. According to one embodiment, the obtaining of the hydrothermal extract may further comprise the step of removing the precipitate and Bacillus cells through the filtration, after the heating.

Finally, the method may comprise the step of mixing and granulating the mixed solution and starch of the hydrothermal extract, joiner, gall bladder and hulky extract.

According to one embodiment, the hot water extracts of the mixed solution of the joiner, gall bladder and hulky extract and by-product of the yangyangtang may be mixed in a volume ratio of 7: 1 to 10: 1, most preferably at 9: 1 It may have been. In addition, according to one embodiment, the mixed solution may be a mixture of joiner, gall bladder and hulky extract in a volume ratio of 1: 1: 1. Feed additives may be granulated by mixing starch (for example, corn starch) in a mixture of the hydrothermal extract, joiner, gall bladder, and pear extract, and starch is a mixture of the mixture of the hydrothermal extract, joiner, quince and pear extract. Pear to 10 times, most preferably 9 times, may be added. After the granulation step, the feed additive may be obtained through a drying step.

According to the antimicrobial agent and the feed additive including the same according to one embodiment, it is possible to secure a safe food by using a natural antimicrobial agent, there is an effect to improve the quality of broilers.

1 is a Staphylococcus of the antimicrobial agent according to one embodiment Photograph showing antimicrobial activity against aureus KCTC 3881.
Figure 2 Salmonella of the antimicrobial agent according to one embodiment Enteritidis KCTC 2054 showed antimicrobial activity.
3 is Staphylococcus of the antimicrobial agent according to one embodiment epidermidis Pictures show antimicrobial activity against KCTC 1917.

Hereinafter, one or more embodiments will be described in more detail by way of examples. However, these embodiments are intended to illustrate one or more embodiments, and the scope of the present invention is not limited to these embodiments.

Example 1 Preparation of Joiner Extract

The seedlings were prepared by purchasing raw materials in a dry state on the market and preparing them into powder using a grinder, and then extracting refluxed at 80 ° C for 3 hours by adding 250g of 70% ethanol to 50g of the seedling powder. Then, the extraction solution was collected by filtration using filter paper, and 250 g of 70% ethanol was further added to the residue, followed by heat extraction, followed by filtration three times. In addition, all the extracts were combined using a 1 μm pore size paper filter to remove polymer precipitates and cells, and the solution was concentrated under reduced pressure at 47 ° C using a distillation apparatus equipped with a cooling capacitor. Joiner 70% ethanol extract was obtained.

Example 2: Preparation of Mellitus Extract

Gallja was prepared by purchasing the raw material in the dry market commercially prepared to make a powder using a grinder, and extracted with reflux at 80 ° C for 3 hours by adding 250g of 70% ethanol to 50g gallola powder. Then, the extraction solution was collected by filtration using filter paper, and 250 g of 70% ethanol was further added to the residue, followed by heat extraction, followed by filtration three times. In addition, all the extracts were combined using a 1 μm pore size paper filter to remove polymer precipitates and cells, and the solution was concentrated under reduced pressure at 47 ° C using a distillation apparatus equipped with a cooling capacitor. A gall bladder 70% ethanol extract was obtained.

Example 3: Method of Making Thick Extract

Thick gourd was prepared by purchasing a raw material in a commercially available dry state to make a powder using a grinder, and then extracted by refluxing at 80 ° C for 3 hours by adding 250 g of 70% ethanol to 50 g of the gourd powder. Then, the extraction solution was collected by filtration using filter paper, and 250 g of 70% ethanol was further added to the residue, followed by heat extraction, followed by filtration three times. In addition, all the extracts were combined using a 1 μm pore size paper filter to remove polymer precipitates and cells, and the solution was concentrated under reduced pressure at 47 ° C using a distillation apparatus equipped with a cooling capacitor. Thick 70% ethanol extract was obtained.

Example  4: method of preparing feed additives containing herbal by-products

Chinese herbal by-products were obtained from yeonkyo, Indongung, Saenghwanghwang, Gardenia, Mint, mold opening, windbreak, gypsum and hair from the formulation as follows. The blend was used at a ratio of 8: 8: 8: 4: 4: 4: 4: 4: 4% by weight, and the mixture was sterilized at 121 ° C. for 20 minutes, and then 30 L of A sterile container and a plastic pack were prepared, containing 20 kg of the mixture in the plastic pack. At this time, in order to make the fermentation conditions, rice bran was put together at a ratio of 1% by weight of the mixture. Hot water at 80 ° C. was poured into the plastic pack until the mixture was submerged and then inoculated with Bacillus licheniformis KCCM10885P by 1% by weight of the total weight. After sealing the plastic pack, it was fermented at room temperature (25 to 30 ℃) for 5 days.

Thereafter, the fermented product was collected and 5 times the weight of distilled water was added, followed by heat extraction at 121 ° C. for 20 minutes, and the extraction solution was collected by filtration using filter paper, and then the filtrate was 1 μm pore size. Fermented medicinal herb by-product extract was obtained by removing the polymer precipitate and the microbial cells using a paper filter. Then, joiner, gall bladder, and grommets in 70% ethanol at a ratio of 1: 1: 1, and the ethanol mixture and the fermented medicinal herb by-product extract are mixed at a ratio of 9: 1, where 9 times the weight of 100% Corn starch was mixed and granulated and then dried for 3 days to obtain granules.

Test Example  1: Disk diffusion method for measuring antimicrobial activity paper disk diffusion assay )

Strains used in the present invention cause disease in broilers Staphylococcus epidermidis KCTC 1917, Salmonella enteritidis KCTC 2054, Staphylococcus A total of three microorganisms of aureus KCTC 3881 were selected, and the strains were distributed from the Korea Institute of Bioscience and Biotechnology. Staphylococcus epidermidis KCTC 1917, Salmonella enteritidis KCTC 2054, Staphylococcus Culture medium of three aureus KCTC 3881 microorganisms was used Nutrient medium (Difco), respectively, and activated at 2 days before the experiment while stored at 4 ° C. The bacteria were inoculated in the culture medium and incubated for 1 to 2 days in a 37 ° C incubator.

Single colonies of each strain were taken and incubated in Nutrient Broth (Difco) for 1 to 2 days, and then the bacterial suspension was diluted to a constant concentration and uniformly plated onto an agar plate using a sterile swab. After drying the plate, a paper disk (Φ 8 mm, Adventec. USA) was placed on the surface of the plate inoculated with the strain, and the extracts obtained in Examples 1, 2 and 3 were diluted by concentration in DMSO (dimethyl sulfoxide, sigma). 50 μl / disk was absorbed into the paper disk. After incubating the plate treated with the sample for 1-2 days, the antimicrobial activity was measured by the size of the growth zone around the disc.

Experimental sample Treatment concentration Restraining ring (mm) S. epidermidis S. enteritidis S. aureus control DMSO - - - Example 1 25mg / ml 20mm 11 mm 18mm 50mg / ml 22mm 11 mm 20mm 100 mg / ml 25 mm 12mm 21mm Example 2 25mg / ml 12mm 12mm 13mm 50mg / ml 14mm 15mm 16mm 100 mg / ml 16mm 16mm 20mm Example 3 25mg / ml 16mm 10mm 15mm 50mg / ml 17mm 10.5mm 16mm 100 mg / ml 18mm 12mm 17mm

As shown in Table 1 and Figures 1 to 3, the extracts obtained in Examples 1, 2 and 3 were all grown with respect to three species S. epidermidis , S. enteritidis and S. aureus at each concentration of 25 to 100 mg / ml. Inhibitory ring was measured and it was confirmed that the double joiner extract showed the best antibacterial effect. Therefore, it is confirmed that the joiner, gall bladder, and buckwheat extract of the present invention contain an antimicrobial effect ingredient that inhibits the growth of S. epidermidis , S. enteritidis and S. aureus microorganisms , which are disease-causing bacteria in broilers.

Test Example  2: Confirmation of broiler productivity according to feed additives

In Experiments 2 to 5, animal experiments were conducted to determine the effects of feed additives using by-products of herbal medicines on broiler feeds to replace the antibiotics on broiler productivity, blood composition, carcass quality, and immune system.

The test revealed 300 broilers (Ross species), and the control group (C), antibiotics (antibiotics, Tylosin phosphate 0.05% = + C), 0.5% feed additive treatment (T1), 1% feed additive treatment (T2) , 5% of 2% feed additive treatment (T3). The test was carried out for 35 days for each of 35 days of 5 treatments and 3 repetitions were randomly placed in a kennel (1m horizontal, 1.5m vertical).

The effect of the feed additives prepared in Example 4 on the weight gain, feed intake and feed demand in broilers is shown in Table 2 below.

The gains over the 0-3 weeks were + C 1012.64g, T2 996.25g, T3 991.73g, T1 967.55g and C 942.52g. During the period, weight gain of + C was significantly higher than that of C, but there was no statistically significant difference from the treatment. Feed intake of T3 was 1284.66g, + C 1275.44g, T1 1265.69g, T2 1261.43g, C 1216.87, and T3 was the lowest, but C was not significant. Feed efficiency was T1 1.37, T3 1.35, C 1.35, T2 1.32, + C 1.32. The feed efficiency was the lowest in + C and T2, and the highest in T1, but not significant.

The weight gain was the highest with C 758.26g and low with T2 676.72g and there was a significant difference, but the intake was highest with T3 1543.74g and low with C 1477.26g, but there was no significant difference. Feed efficiency was as low as C 2.02g and as high as T3 2.22, but no significant difference. On the other hand, there was no significant difference in weight gain, intake and feed efficiency during the whole period.

Initial weight
(g / bird) Final weight
(g / bird) Weight gain
(g / bird) Feed intake
(g / bird) feed / gain
(g / g) Grower (0-3rd week)  C 42.17 ± 0.31 _a 942.52 ± 42.22 _b 900.35 ± 42.47 _b 1216.87 ± 69.07 _a 1.35 ± 0.02 _ab + C 42.33 ± 1.61 _a 1012.64 ± 37.16 _a 970.30 ± 38.14 _a 1275.44 ± 28.00 _a 1.32 ± 0.03 _b T1 42.92 ± 1.28 _a 967.55 ± 14.55 _ab 924.64 ± 15.70 _ab 1265.69 ± 16.81 _a 1.37 ± 0.01 _a T2 43.08 ± 0.14 _a 996.25 ± 23.96 _ab 953.17 ± 23.91 _ab 1270.77 ± 32.75 _a 1.32 ± 0.00 _ab T3 41.58 ± 0.52 _a 991.73 ± 24.79 _ab 950.14 ± 24.40 _ab 1284.66 ± 14.59 _a 1.35 ± 0.02 _ab Grower (4 ~ 5th week) C 942.52 ± 42.22 _b 1695.45 ± 78.44 _a 754.26 ± 36.28 _a 1477.26 ± 62.48 _a 1.99 ± 0.05 _c + C 1012.64 ± 37.16 _a 1665.54 ± 37.15 _a 732.90 ± 1.16 _b 1522.72 ± 42.21 _a 2.33 ± 0.08 _a T1 967.55 ± 14.55 _ab 1717.20 ± 39.32 _a 722.28 ± 25.95 _a 1508.65 ± 25.54 _a 2.01 ± 0.04 _bc T2 996.25 ± 23.96 _ab 1644.14 ± 93.41 _a 676.72 ± 69.46 _b 1488.57 ± 2.44 _a 2.32 ± 0.32 _a T3 991.73 ± 24.79 _ab 1687.06 ± 27.77 _a 695.33 ± 25.09 _ab 1543.74 ± 33.94 _a 2.24 ± 0.07 _ab Grower (0 ~ 5th week) C 42.17 ± 0.31 _a 1695.45 ± 78.44 _a 1653.28 ± 78.69 _a 2717.73 ± 105.52 _b 1.64 ± 0.01 _a + C 42.33 ± 1.61 _a 1665.54 ± 37.15 _a 1623.21 ± 39.15 _a 2798.16 ± 57.46 _ab 1.72 ± 0.03 _a T1 42.92 ± 1.28 _a 1717.20 ± 39.32 _a 1674.29 ± 40.23 _a 2774.35 ± 16.46 _ab 1.66 ± 0.02 _a T2 43.08 ± 0.14 _a 1644.14 ± 93.41 _a 1601.06 ± 93.36 _a 2759.33 ± 4.04 _ab 1.72 ± 0.10 _a T3 41.58 ± 0.52 _a 1687.06 ± 27.77 _a 1645.48 ± 27.27 _a 2842.76 ± 48.39 _a 1.73 ± 0.01 _a

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

C = control, + C = antibiotic 0.05%, T1 = 0.5%, T2 = 1.0%, T3 = 2.0%

Test Example  3: Blood characteristics of broilers according to feed additives

The results of blood appearance are shown in Tables 3 and 4 below. Plasma analysis showed no significant differences between treatments in T. Protein, Albumin, BUN, AST (sGOT), ALT (sGPT), T. Cholesterol and Triglyceride, but not in HDL-C and LDL-C. appear. In HDL-C, T1 was the highest, C was the lowest, and there was a significant difference (p <0.05), and there was no significant difference in the other treatment groups. In the case of LDL-C, C was the highest and there was a significant difference (p <0.05) from other treatment groups.

As a result of serum analysis, white blood cells (WBC), which are known to increase during initial inflammation, and eosinophils (EO), which increase during parasitic infections or immune hypersensitivity reactions, and basophils (BA), which show similar reactions, are more active than control. Although there was a tendency to be high, there was no significant difference, especially in the case of monocytic monocytes (MO) which increased during purulent disease or tissue necrosis, antibiotics were significantly higher (P <0.05) than the other groups. Was lower than the control and there was a significant (P <0.05) difference. Routine CBC-Related Levels Leukocytes (WBC), Red Blood Cells (RBC), Hemoglobin (Hb), Erythrocyte Volume (HCT or PCV) and Average Erythrocyte Volume (MCV), Average Erythropoietin (MCH) and Average Erythrocyte Pigment Concentration (MCHC) There were significant differences in the addition intervals in some of the HCTs, with the highest T1 and the lowest T2 for MCH and MCHC, showing a significant difference (P <0.05). However, RBC and erythrocyte volume (HCT or PCV) were high in antibiotics and low in addition, but not significantly different. The concentration of immune cells increased as the level of feed additives containing herbal by-products increased. T3 showed similar effects as antibiotics, suggesting the possibility of antibiotic replacement.

C + C T1 T2 T3 T.Protein
(g / dL) 4.17 ± 0.06 3.83 ± 0.12 4.37 ± 0.32 4.33 ± 0.06 4.73 ± 0.21 Albumin
(g / dL) 2.2 ± 0.1 2.03 ± 0.06 2.57 ± 0.21 2.27 ± 0.06 2.47 ± 0.06 BUN
(mg / dL) 3.2 ± 0.3 3.13 ± 0.38 2.9 ± 0.2 2.83 ± 0.06 3.43 ± 0.06 AST (sGOT)
(IU / L) 261.67 ± 10.79 337.33 ± 35.12 284 ± 11.36 297.33 ± 26.86 280.33 ± 6.66 ALT (sGPT)
(IU / L) 3.67 ± 0.58 4 ± 1 3.67 ± 0.58 4.33 ± 0.58 3.67 ± 0.58 T.Cholesterol
(mg / dL) 231 ± 8.54 224.67 ± 27.59 215.67 ± 2.52 192.33 ± 24.99 205 ± 20.66 Triglyceride
(mg / dL) 91.33 ± 26.5 73 ± 18.73 61.33 ± 3.79 54.33 ± 8.39 64.33 ± 16.77 HDL-C
(mg / dL) 117.33 ± 7.64 _b 106.33 ± 6.66 _c 142.33 ± 1.53 _a 126 ± 6.08 _b 126.33 ± 5.51 _b LDL-C
(mg / dL) 127.67 ± 1.53 _a 107.33 ± 7.64 _b 86.33 ± 2.89 _c 82 ± 8.66 _c 111.67 ± 6.81 _b Glucose
(mg / dL) 290.33 ± 4.04 284.67 ± 10.69 301.67 ± 11.5 280.67 ± 4.04 283.33 ± 2.31

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

C = control, + C = antibiotic 0.05%, T1 = 0.5%, T2 = 1.0%, T3 = 2.0%

C + C T1 T2 T3 Leukocyters WBC (× 10 ³ cells / ul) 1.3 ± 0.37 1.48 ± 0.5 1.44 ± 0.5 1.09 ± 0.48 1.21 ± 0.27 Mono (× 10 ³ cells / ul) 134.38 ± 7.56 _b 157.62 ± 8.91 _a 125.64 ± 4.17 _bc 109.84 ± 14.42 _c 128.5 ± 13.19 _bc Base (× 10 ³ cells / ul) 0.08 0.02 0.09 ± 0.02 0.12 ± 0.04 0.09 ± 0.02 0.15 ± 0.09 Erythrocytes RBC (× 10 ³ cells / ul) 3.34 ± 0.13 3.30 ± 0.35 3.16 ± 0.11 3.26 ± 0.25 3.33 ± 0.19 HGB (g / dl) 12.7 ± 0.26 _b 12.3 ± 1.3 _b 12.93 ± 0.84 _b 10.7 ± 2.23 _b 16.53 ± 1.03 _a HCT (%) 33.63 ± 1.78 _b 36.1 ± 2.39 _ab 37.4 ± 0.3 _a 33.07 ± 2.27 _b 38 ± 1.75 _a MCV (fl) 111.03 ± 3.82 _b 111.13 ± 4.72 _b 116.43 ± 2.20 112.43 ± 1.17 118.03 ± 1.33 _a MCH (pg) 42.03 ± 2.96 _a 40.3 ± 3.3 _ab 40.83 ± 1.91 _ab 32 ± 0.44 _c 35.33 ± 5.60 _bc MCHC (g / dl) 37.83 ± 1.93 _a 33.47 ± 3.25 _ab 35.8 ± 2.52 _a 28.8 ± 0.75 _b 39.67 ± 5.65 _a

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

C = control, + C = antibiotic 0.05%, T1 = 0.5%, T2 = 1.0%, T3 = 2.0%

Test Example  4: Measurement of carcass quality in broiler according to feed additives

The carcass quality results of poultry are shown in Table 5. There was a significant difference in carcass quality, which was significant in water retention and protein content.

In the case of protein, the control group was 23.27 ± 0.25, and in the addition group, T1 24.12 ± 0.10%, T2 24.26 ± 0.30%, and T3 24.24 ± 0.30%, respectively. Moisture and fat shear force were lower in the addition group than the control group, but there was no significant difference.

C + C T1 T2 T3  heating loss (%) 15.16 ± 1.26 14.6 ± 2.2 15.59 ± 1.57 15.04 ± 0.89 14.27 ± 1.56 WHC ² (%) 59.12 ± 1.31 _b 60.55 ± 1.40 59.94 ± 0.97 60.84 ± 1.61 _a 59.27 ± 0.69 Moisture (%) 74.93 ± 0.23 _ab 75.15 ± 0.16 74.31 ± 0.41 74.64 ± 0.29 73.94 ± 1.01 _c Crude fat (%) 0.36 ± 0.22 0.30 ± 0.14 0.23 ± 0.18 _a 0.21 ± 0.10 _a 0.50 ± 0.29 _a Crude protein (%) 23.27 ± 0.25 _b 23.43 ± 0.14 _b 24.12 ± 0.10 24.26 ± 0.30 24.24 ± 0.30 Crude ash (%) 1.00 ± 0.06 1.01 ± 0.08 1.00 ± 0.1 0.97 + - 0.12 1.03 ± 0.08 shearing stress (/0.5 inch) 2.10 ± 0.37 _a 1.82 ± 0.43 _ab 1.58 ± 0.26 _b 1.66 ± 0.34 _ab 1.84 ± 0.34 _ab CIE L 58.25 ± 2.20 _a 53.6 ± 1.68 55.82 ± 1.13 _b 53.81 ± 0.61 _bc 53.64 ± 2.27 _c a 1.25 ± 0.31 _c 1.76 ± 0.25 _bc 2.05 ± 0.35 _abc 2.46 ± 0.76 _ab 2.77 ± 1.21 _a b 4.97 ± 1.06 _a 2.76 ± 1.56 _b 5.59 ± 0.59 _a 3.78 ± 0.76 _b 3.47 ± 0.64 _b

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

C = control, + C = antibiotic 0.05%, T1 = 0.5%, T2 = 1.0%, T3 = 2.0%

Test Example  5: Determination of broiler immunity according to feed additives

Immediately after the end of the specification for each of Weeks 4 and 6, 5 mL of blood from the heart of broilers was collected using EDTA-treated vacutainer and vacutainer needle & holder, and then leukocytes and erythrocytes were analyzed by a blood analyzer within 24 hours. After centrifugation with g for 20 min, plasma was separated and stored frozen (-50 ° C.) until IgG, IgA, IgM analysis. Serum IgG, IgA, IgM concentration was measured by absorbance 450 nm in ELISA reader according to the single immuno-diffusion test (RID test) developed by Mancini (1965). IgG, IgA, and IgM standard reference values were determined using Chicken IgG, IgA, and IgM (Koma Biotech. Co. Ltd .: ELISA chicken IgG, IgA, and IgM core kit) 1,000, 200, 100, 50, 25, 12.5, 6.25, 3.12 Each dilution was measured at ng / mL and the value was calculated using each rare equation for each analysis. Serum IgG, IgA, IgM analysis results at 4 and 6 weeks are shown in Tables 6 and 7, respectively.

Item Treatments (mg / mL) C + C T1 T2 T3 IgG 7.65 ± 0.51 _b 7.85 ± 0.48 _b 7.72 ± 0.40 _b 8.84 ± 0.05 _a 8.69 ± 0.16 _a IgA 1.67 ± 0.18 _v 2.33 ± 0.17 _b 2.94 ± 0.08 _a 2.30 ± 0.22 _b 2.24 ± 0.02 _b IgM 0.96 ± 0.12 _a 0.90 ± 0.02 _ab 0.98 ± 0.04 _a 1.03 ± 0.21 _a 0.72 ± 0.09 _b

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

As a result of week 4 blood analysis, the concentrations of IgG, IgA, and IgM were increased in addition groups T1, T2, and T3, respectively, compared to the control group. In other words, it was confirmed that the immunity obtained through the feed additive is higher than the antibiotic addition group.

Item Treatments (mg / mL) C + C T1 T2 T3 IgG 7.94 ± 0.21 _c 8.28 ± 0.07 _bc 8.78 ± 0.14 _a 8.86 ± 0.28 _a 8.40 ± 0.20 _b IgA 1.42 ± 0.14 _b 2.16 ± 0.3 _a 2.05 ± 0.19 _a 2.10 ± 0.16 _a 2.38 ± 0.15 _a IgM 0.44 ± 0.01 _b 0.58 ± 0.08 _a 0.51 ± 0.08 _ab 0.63 ± 0.04 _a 0.57 ± 0.10 _ab

^{a ~ c} mean with different superscripts differ significantly (p <0.5).

As a result of blood analysis at week 6, except for T3, the concentration of IgG was higher than that of week 4, and especially T1 and T2 were significantly higher than antibiotic treatment. Week 6 IgA tended to increase with higher treatment concentration, but did not show significant difference from antibiotic treatment group. At week 6, IgM showed no significant difference between T1, T2, and T3 in the antibiotic treatment group.

In order to determine whether the feed additive including the herbal by-product prepared in Example 4 as an additive to broiler feed in Test Examples 1 to 5 can replace antibiotics, the effect on productivity, blood composition, and carcass quality. An experiment was conducted to confirm. The experimental group had five treatment groups: control, antibiotics (antibiotics, Tylosin phosphate 0.05%), feed additive 0.5% treatment (T1), 1.0% treatment (T2), 2.0% treatment (T3) containing the herbal by-products. In the test, 300 broilers (Ross species) were disclosed, and 5 treatments, 3 repetitions, and 20 animals per repetition were randomly arranged in a kennel (1 m in length and 1.5 m in length), respectively, for 35 days. Weight gain and feed efficiency were higher in the control group than in the control, but there was no significance. Among the plasma components, HDL-C showed a 10 ~ 20% higher level in the treated group than the control group, and LDH-C showed a significant difference (P <0.05) with 10-30% less than the control group. HCT had the highest T1 and MCH and MCHC had the lowest T2, showing a significant difference (P <0.05). In T1, carcass quality was significantly different (P <0.05) compared to control and antibiotic treatment groups. In the case of T3, the immune material showed a similar or higher level than antibiotics.

As the concentration of the feed additive containing the herbal by-products was higher than that of the antibiotic, it was confirmed that the feed additive containing the herbal by-products of the present invention has the possibility of antibiotic replacement.

Claims (13)

Antimicrobial agent comprising one or more herbal extracts selected from the group consisting of joiner, gall bladder and groin. The antimicrobial agent of claim 1 wherein the extract is an ethanol extract. The method of claim 1, wherein the extract is Staphylococcus Genus or Salmonella Antibacterial agent that exhibits antimicrobial activity against bacteria of the genus. Animal feed additive comprising the antimicrobial agent of any one of claims 1 to 3. The feed additive of claim 4 wherein the animal is poultry. The feed additive of claim 4, wherein the feed additive further comprises a hydrothermal extract of a by-product of the yangyangtang. The feed additive according to claim 4, wherein the feed additive comprises a mixture of a mixture of joiner, gall bladder and pear extract and hot water extract of the by-product of the yangyangtang in a volume ratio of 7: 1 to 10: 1. The feed additive of claim 7, wherein the mixed solution is a mixture of joiner, gall bladder, and pear extract in a volume ratio of 1: 1: 1. The feed additive of claim 4, wherein the feed additive is granular. Fermenting by inoculating Bacillus bacteria to by-products of the yangyangtang;
Obtaining a hydrothermal extract of the fermentation result; And
Feed additive manufacturing method comprising the step of mixing and granulating the mixed solution and starch of the hot water extracts, joiner, gall bladder and thick gourd extract. The method of claim 10, wherein the Bacillus bacteria Bacillus how is licheniformis . The method according to claim 10, wherein the mixture of the joiner, gall bladder, and pear extract and hot water extract are mixed in a volume ratio of 7: 1 to 10: 1. The method of claim 10, wherein the mixed solution is a mixture of the joiner, gall bladder and bakbak extract in a volume ratio of 1: 1: 1.

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