KR20100024804A - Animal feed containing powder and extracts of rhus chinensis - Google Patents

Abstract

PURPOSE: Animal feed additives containing powder and extract of Rhus chinensis are provided to improve tastes of meat by reducing fatty acid, and to increase amino acid and unsaturated fatty acid while improving antibacterial effects and immunity promotion effects. CONSTITUTION: A manufacturing method of animal feed additives includes the following steps: extracting Rhus chinensis for 1 hour in 100°C after pulverizing the Rhus chinensis in distilled water for 16 hours in 60°C; concentrating extract with a vacuum condenser; manufacturing the concentrated extract with a spray dryer; extracting the pulverized Rhus chinensis with alcohol for 4 hours in 60°C; and drying and powdering the concentrate. The Rhus chinensis is pulverized to 1.2 mm~1.5mm.

Description

Animal feed containing powder and extracts of Rhus chinensis

The present invention relates to a feed mixture for animals, and relates to an animal feed additive containing a powder and extract of the gall of the strong antibacterial activity.

Meat that humans use for food has generally been treated by feeding antibiotics that show susceptibility to various diseases to treat the animals, or by preparing vaccines to inoculate the animals to obtain immunity to prevent diseases.

Recognizing these problems today, as environmental pollution from livestock wastewater and interest in human health are increasing,

Research on the antimicrobial activity of natural medicinal plants, which are economical and environmentally friendly, is not necessary. The antimicrobial activity tests on the causative agents of bacterial diseases in animals are most efficient. This is the way to reduce the most economic loss for the company.

In addition, the mass production of environmentally friendly meat has emerged as a method of causing resistance to antibiotics and a method for ending antibiotic misuse.

Research on natural medicinal plants is conducted in the laboratory only, and experimental tests on herbal medicines on animal farms have not been carried out yet. There is a big advantage without pollution and toxicity.

On the other hand, the Republic of Korea Patent Publication No. 10-2006-0011239 "Fish mix for fish containing the gallant extract" was extracted from various herbal medicines and carried out the antibacterial activity test by selecting the bacteria of the fish bottle, the extract is fed to cultured fish The antimicrobial test and the evaluation of fish as foods were conducted, but the antimicrobial test on cattle, pigs, chickens and other animals was not implied.

According to the present invention, in the situation of relying on antibiotics for the treatment of major bacterial diseases in animals, antibiotics at the point of increasing consumer's interest in the safety of livestock products, such as residual problems in meat due to misuse and abuse of antibiotics, emergence of resistant bacteria, etc. It is to provide an additive as a substance that can obtain the effect of enhancing the antimicrobial and immunity by adding to the feed of cows, pigs and chickens using natural herbal medicine as a substitute.

Another object of the present invention is to provide a method for preparing an additive,

Another object of the present invention is to prepare a feed additive and auxiliary feed containing the composition material.

The present invention relates to a feed additive for animals, characterized in that it is selected from the herbal medicines that are natural medicinal plants with antimicrobial activity against the causative agent of animal diseases and contained in the feed of the animal.

The present invention provides an animal feed additive comprising 97 to 99.9% of animal feed and a quinine powder or extract in the range of 0.1 to 3%.

The present invention provides from the step of selecting a herbal medicine with high antibacterial activity against the pathogen causing the disease of the animal among the herbal medicines that can be purchased in Korea.

The present invention is to provide a powerful antimicrobial activity and immune system and increase the effect by adding a compound feed to the gall of the natural medicinal plants for disease-causing pathogens in animals,

In addition, by feeding the feed compound containing the gall bladder powder and extract according to the present invention to animals, the rich amino acids and unsaturated fatty acids for meat is increased, crude fat, crude ash, fatty acids are reduced to improve the taste of meat, feed While the demand rate decreases, the weight increases, which contributes greatly to the income increase of livestock farmers.

Furthermore, when feeding the animal feed additives according to the present invention to animals, it is possible to effectively prevent infection of bacterial diseases caused by pathogens without administering antibiotics, and to provide safe and high-quality meat, thereby promoting public health and preference. It has the effect of bringing a change in awareness as a food and greatly contribute to improving meat consumption promotion.

1. Extraction of active ingredients of herbal medicines

1) Selection of herbal medicines

In the present invention, the antimicrobial measurement test for the major diseases of animals was selected by reference to domestic and international literatures, such as 178 natural medicinal plants that can be easily purchased in Korea.

In the present invention, the 178 herbal medicines are natural medicinal plants selected for the test as herbal medicines distributed in Korea are shown in Table 1.

[Table 1. Names of herbal medicines as natural medicinal plants]

Herbal medicine Scholarship Herbal medicine Scholarship Go Terminalia chebula Retzius mint Mentha arvensis l Wolfberry Lycium chinense Macmundong Ophiopogon japonicus Ker-Gaqler Salvia Salvia miltiorrhiza Earl Paeonia japonica Rabokja Raphanus sativus L Whiteness Atractylodes macrocephala A defector Cassia tora Windproof Saposhnikovia divaricata Schisck Gosam Sophora angustifolia Sieb Spoiler Linum usitatissimum L World Cirsii japonici herba Cornus Cornus officinalis Donkey Angelica gigas Seokwicho Pyrrosia lingua (Thunb.) Farwel Hagocho Brunnella vulgaris L Plantain Plantago asiatica Thick Magnolia obovata Thunb White hair root Imperata cylindrica P. Beauviois var Gold Scutellaria baicalensis Ghost Poria cocos Betel nut Areca Catechu L Ginseng Codonopsis pilosula (Fr.) Jimo Anemarrhena asphodeloides Bunge White oyster Fellowship Forsythia suspersa Seokgok Dendrobium moniliforme dermis Citrus Aurantium L Ogapi Acanthopanax sessiliflorus Creation Atractylodes lancea D. C Encyclopedia Ginkgo semen crowd Dryopteris crassirhizoma Nakai Gardenia Gardenia jasminoides Labor Phragmites communis Trinius Circumference Stemona japonica Miquel Gold coins Lonicera japonica Yin Yang Epimedium koreanum Street view Platcodon grandiflorum Thawed blood Kalopanaxii Cortex Kwak Hyang Teucrium veronicoides Arrow Euonymus alatus licorice Glycyrrhiza uralensis Thorn Ogapi Eleuterococcus senticosus Maxim Country Chrysanthemum morifolium Ramart Eoseongcho Houttynia cordat Ephedra Ephedra sinica S Yonggyu Large leaves Isatidis Folium Samneung Scirpus maritimus L

Herbal medicine Scholarship Herbal medicine Scholarship Book Angelica tenuissima Nakai Duckweed Spirodela polyrhiza (L.) schleid Dog Pharbitis Nil Choisy Lump Styphnolobium japonicum Schott Small fennel Foeniculum vulgare Mill Guernsey Rehmannia glutinosa Liboschitz var Yellow wall Phellodendron amurense Golden cherry Rosa laevigate Vigor Ostericum koreanum Gucci Mulberry Cudrania tricuspidata Bureau Fellow ruler Malva verticillata L Pussy willow Salix gracilistyla Miq Cedar Artemisia capillaris Thunb Gaodong Catalpa ovata G. Don Injin Artemisia Messerschmidtiana Besser Dry Rhus verniciflura Stokes Triceps Sophora subprostrata Chun et T Juniper tree Juniperus rigida S. et Z Writing Ixeris dentata Dapsari Kochia scoparia Schrad Four Codonopsis lanceolata An elm bark gallnut Rhus chinensis Cordyceps sinensis cordyceps sinensis Imperial Polygonatum odoratum Druce var Tofu Eucommia ulmoides Oliver motherwort Leonurus sibiricus Rice Cake Mushroom Taxi Alisma plautago Jerk wood Oplopanax elatus Nakai Short axis Polygonum aviculare L A great stroke Arecae pericarpium A dead man Quisqualis indica L Neck skin Paeonia suffruticosa Lake Piper nigrum linne Quince Chaenomeles sinensis mistletoe Viscum album var. coloratum Man Vitex rotundifolia L curcuma Curcuma aromatia Salisbury Ash tree Fraxinus rhynchophylla Hance Goofy magnolia Magnolia kobus DC Bone Crest Drynaria fortunei Neck Akebia quinata Decaisne Rooting Pueraria thunbergiana dandelion Taraxcum platycarpum cinnamon Cinnamomum cassia (Nees) Blume water chestnut Trapa natans L Persimmon leaf Diospyros kaki Wood condition Phellinus linteus

Herbal medicine Scholarship Herbal medicine Scholarship elecampane Inula Helenium L Osuyu Evodia rutaecarpa Hooker fil blank Angelica dahurica Bentham et Hooker Journeyman Lily Lilium speciosum Thunb Dew Ringworm Dictamnus dsycarpus Turez Paper Polygala tenuifolia L Whiteness Ampelopsis japonica Makino Turmeric Curcuma aromatia Salisbury Rock brush Orostachys japonicus A. Berger Yanggang Alpinia officinarum Hance Non-repair Lespedeza cuneata G. Don Rule Coix lachryma-jobi Bokbunja Rubus coreanus Root skin Ejection Curcuma zedoaaria Journey Room Ligustrum japonicum Thunb Bangui Sinomenium acutum Indong Lonicera japonica Thunb Snake Strawberry Duchesnea chrysantha (Zoll. Et Morr.) Miq cranesbill Geranium nepalense subsp. thunbergii Baekduong Pulsatilla chinensis Alder Alnus japonica Steudel White flower Althaea rosea Cam Fingering Coriolus versicolor Quel Wealthy Aconitum carmichaeli Debx Bed Adenophora triphylla White porcelain Thuja orientalis fist Taxus cuspidata sign Amomum xanthioides Wallich War Anthriscs aemula Schisch Shiho Bupleurum falcaum L Silk tree Albizzia julibrissin Durazz Sounder Rumex crispus Oil Sanguisorba officinalis pine needles Pinus densiflora Low root skin Ailanthus altissima Swingle Hawthorn fruit Crataegus pinnatijida Jogu-dong Situation Mushroom Phellinus linteus Birch Trees Betula platyphylla var Sancho Zanthaxylum schinifolium Three hundred seconds Saururus chinensis Baill Leaflets Perilla frutescens Haegeumsa Temple Lygodium japonicum Thistle Cirsium japonicum Bamboo leaf Sasamorpha purpurascen Gentian Gentiana scabr Engraver Gleditschia horrida Makino

Herbal medicine Scholarship Herbal medicine Scholarship Bark tree Pagoji Psoralea corylifolia L Grafting Sambucus latipinna Nakai Huang Typha latifolia Lenne Jeoldae Echinops setifer Iljin Glyph root Rosa rugosa Natural flower Trichosantes Kirilowii Maxim Brother-in-law Schizonepeta tenuifolia Brig Changza Trigonella foenum-graecum L figwort Scrophularia buergeriana Miq Range Cichorium intybus L Ho Jang Geun Reynoutria japonica Houtt Astronomical building Asparagus lucidus Lindley juniper Sabina chinensis Antoine Cheonma Gastrodia elata enjoyment Lycoppus Lucidus Turcz Tosa Cuscuta chinensis Nasturtium Eclipta Prostrata L Talc Local grass Gokseongcho March health Self one-way Yellow root Excess Astragalus Astragalus membranaceus (AM)

2) Treatment of herbal medicines

The experimental material of the herbal medicine as a natural medicinal plant was washed three times or more with clean water, and then used as a material after hot air drying or natural drying.

3) Hot water extraction method and concentration

The present invention to measure the antimicrobial activity of the 178 herbal medicines,

After grinding the medicinal herbs, 50g of crushed medicinal herbs and 500ml of distilled water were added, followed by extraction at 60 ° C. for 16 hours, followed by extraction at 100 ° C. for 30 minutes.

The extracted extract was first filtered using Advantec No. 1 filter paper, centrifuged at 5,000 rpm using a centrifuge (Hanil Supra22K ^TM ) for 30 minutes, and the precipitated supernatant was filtered using an Advantec 1.00 μm Membrane filter, followed by a vacuum concentrator. (EYELA SB1000 ^™ ) was concentrated under reduced pressure 10-fold in an 80 ° C. hot water bath.

4) Ethanol Extraction Method and Concentration

The present invention to measure the antimicrobial activity of the 178 herbal medicines,

After pulverizing the herbal medicine, 50g of crushed herbal medicine and 500ml of 70% ethyl alcohol were added thereto, and extracted at 60 ° C. for 4 hours.

The extracted extract was first filtered using Advantec No. 1 filter paper, centrifuged at 5,000 rpm using a centrifuge (Hanil Supra22K ^TM ) for 30 minutes, and the precipitated supernatant was filtered using an Advantec 1.00 μm Membrane filter, followed by a vacuum concentrator. (EYELA SB1000 ^™ ) was concentrated under reduced pressure 10 times in a 60 ° C hot water bath.

2. In herbal medicine  Antimicrobial Activity Measurement

1) Pathogen selection for antimicrobial activity test

The main object of the present invention was to test the antimicrobial activity of the herbal components in the bacterial disease causing diseases in animals, the antimicrobial measurement experiments were carried out targeting the pathogens of diseases causing economic losses to livestock farms, The strain used is as follows.

 Bovine bacterial disease

Escherichia coli (calf)

Escherichia coli (k99)

  Salmonella typhimurum

  -Mannheimia haemolytica

  Pastuellar multocida (A)

 B) Swine bacterial disease

Escherichia coli (k88ab)

  Escherichia coli (k88ac)

  Escherichia coli (987P)

  Escherichia coli (F41)

  Escherichia coli

  Salmonella (outdoor segregation)

  Actinobacillus pleuropneumoniae (S2)

  Actinobacillus pleuropneumoniae (S5)

  -Streptococcus suis

  Staphylococcus hyicus

  Hemophilus parasuis

  Hemophilus parasuis

  -Mycoplasma hyopneumoniae

 Chicken bacterial disease

  Salmonella pullorum

  Salmonella gallinarium

  Salmonella enteritidis

  Escherichia coli

  Staphylococcus aureus

  -Mycoplasma gallisepticum

2) Paper disk used for antimicrobial activity test

The 10-fold concentrated solution extracted with hot water and ethanol, respectively, was used to measure the antimicrobial activity against the herbal medicines using a paper disk.

The solution was aspirated by 20 μl of the herbal extracts concentrated on paper discs (ADVANTEC AD49005040 ^™ , 6 mm), dried sufficiently in a 50 ° C. dryer, and then stored in a -40 ° C. freezer, and used for antimicrobial activity experiments.

3. Antimicrobial test for bovine disease

The antimicrobial activity of the 178 herbal medicines were used five strains of bovine pathogens presented above.

In order to conduct antimicrobial activity tests on herbal medicines,

The strains stored at -80 ℃ were incubated for 24 to 48 hours in Agar plate medium, and colonies were selected and inoculated into Brain Heart Infusion medium, which is a culture medium, and cultured at 37 ℃ for 18 to 24 hours. The OD value was measured at a wavelength of 410 nm by using a spectrophotometer (Beckman DU650 ^TM ), and the bacterial growth medium was adjusted to an OD value of 1.2 to 1.3.

Spread 100 μl of bacteria solution on a plate medium to which 15 ml of Mueller Hinton Agar medium was added to a Petri dish (87 × 15㎜), and place each herbal paper disk on it and incubate in a 37 ℃ incubator for 24 hours. The diameters of the clear zones (zones in which the growth of bacteria was suppressed and formed transparently) were measured.

As a result of measuring the antibacterial activity of each herbal medicine by the above method, the diameter of the clear zone was weaker than that of ethanol extraction in hot water extraction, and the test results were not listed. The test results of the herbal medicines showing the antibacterial activity in ethanol extraction are shown in Table 2. .

[Table 2. Antibacterial Activity Test Results for Bovine Disease]

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm Esherichia coli (calf) Local grass, Cornus, Seokwicho, Eoseongcho, Gucci mulberry tree, Journeyman, Journey room, Jiyu (8 kinds) Fivefold (23㎜) Esherichia coli ( k99 ) Let's go, wolfberry, Gucci mulberry, lily, journey room (5 kinds) Cornus, Journeyman Fivefold (22㎜)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm Salmonella typhimurum Wolfberry, Journeyman's Rice Cake, Oyster Mushroom, Bark Peel, Hawthorn Fruit, Sewage Milk, Journey Room, Root Flower, Root (10 kinds) Fivefold (23㎜) Mannheimia haemolytica Revitalization, short axis, lump, tree, white paper, inner bird (6 species) Cornus, red ginseng, wild boil, hard pepper, bone print, dried lacquer, large pill, baekryeom, lobules, sperm, pagoji, licorice, Gaza (13 kinds) 5 times (20㎜) Pastuellar multocida (A) Gaza, Macmundong, Health, Sohyangyang, Daegye, Baekmogeun, Machi Prefecture, Dongguk, Jimo, Sagunja, Ungeun, Baekeungeun, Gilgyeong, Motherwort, Windproof, Baeksunpi, Gardenia, Man Hyungja, Non-Suri, Yonggyu, Samneung, Thistle, Strong Bow, Pagoji , Imperial family, jukdae, right, changja, taksa, grafting alley, cheonhwa, white paper, juniper, yanggang, earthenware, dabsari, jeonho, rice cake mushroom, cloud, chigeori, astronomical dong, indong, cedar, signature, original paper, Exudation, phoenix, squirrel, turmeric, silk tree, stalk, low root skin, hyeonsam, bark tree (54 species) Absolute, Excess, Grain, Beetroot, Labokja, Excess, Betel nut, Dermis, Spectator, Gold Silver, Wedge, Gardenia, Euphorbia, Sassam, Mistletoe, Squash, Decaxed, Turmeric, Bonebone, Cordyceps sinensis, Tofu, Neck tree, Daebokpil, Gobon, Ash Tree, Rock Brush, Bangui, Rich, Baekduong, White Flower, Siho, Baekjain, Pine Needle Hawthorn, Situation Mushroom, Persimmon Leaf, Cultivation, Small Leaf, Yongdamcho, Jojeongsil, Hyungae, Yew, Jogu-dong, Bamboo Leaf, Triticale, Hannyeoncho , Licorice, Talc (48 types) Wolfberry, red pepper, licorice, gamguk, ephedra, baekchul, yin and yang guk, horseshoe, bitter wheel, brown root, jinjin, gucci mulberry, dry lacquer, barrel, elm bark, bark skin, wood condition, lily, baekrye, bokbunja, genus, alder , Birch, Ho Jang Geun (24 species) Sweet Ginseng (30㎜), Golden, White Peony, Yellow Wall (30㎜), Barrel (37㎜), Root Bark, Dried Grass (7 kinds)

4. Antimicrobial Activity Test on Swine Disease

The antimicrobial activity of the 178 herbal medicines was carried out with seven strains of pathogens of swine diseases (types).

In order to conduct antimicrobial activity tests on herbal medicines,

Actinobacillus pleuropneumoniae , Hemophilus parasuis The experiment was carried out by incubating the strain stored at -80 ° C in β-NAD-added Agar plate for 24 to 48 hours, and selecting Colony. 5 µg of β-NAD (Nicotinamideade-ninedinucleotide) was added to the Brain Heart Infusion medium. / Ml was added and cultured, and the bacterial growth confirmation of the culture medium was measured using a spectrophotometer at the wavelength of 410nm, and the bacterial growth culture medium was adjusted to an OD value of 1.2 to 1.3.

In addition, the strain was incubated in the Agar plate medium for 24 to 48 hours in the strain stored in -80 ℃ inoculated into the colony Brain Heart Infusion medium and cultured the strain 18 to 24 hours at 37 ℃, culture medium To check the growth of bacteria, the OD value was measured at 410 nm using a spectrophotometer.

Spread 100 μl of bacteria solution on a plate medium to which 15 ml of Mueller Hinton Agar medium was added to a Petri dish (87 × 15㎜), and place each herbal paper disk on it and incubate in a 37 ℃ incubator for 24 hours. The diameters of the clear zones (zones in which the growth of bacteria was suppressed and formed transparently) were measured.

Also, For Actinobacillus and Hemophilus strains, add 100 µl of bacterial solution to a plate medium in which 5 µg / ml β-NAD was added to a Brain Heart Infusion medium and 15 ml of Petri dish (87 × 15 mm). The paper disk was placed and incubated in a 37 ° C. incubator for 24 hours, and then the diameters of the clear zones formed around the disks were inhibited from growth of bacteria.

As a result of measuring the antibacterial activity of each herbal medicine by the above method, the diameter of the clear zone was weaker than that of ethanol extraction in hot water extraction, and the test results were not listed. The test results of the herbal medicines showing the antibacterial activity in ethanol extraction are shown in Table 3.

[Table 3. Antimicrobial Activity Test Results for Swine Disease]

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Esherichia coli ( k88ab ) Let's go, wolfberry, cornus, lily, wild root, gucci mulberry, bark skin, tree, hawthorn fruit Journeyman, Jiyu (2 types) Fivefold (22㎜) Esherichia coli ( k88ac ) Wolfberry, Sweet Ginseng, Golden, Fellowship, Cornus, Dried, Shelled, Lily, Sewage (9 types) Let's go, wild boil, Gucci mulberry tree, large pill, leaflet, hawthorn fruit, journeyman, fat milk (8 kinds) 5 times (20㎜) Esherichia coli (987P) Wolfberry, Golden, Jimo, Fellowship, Creation, Labor, Gilkyung, Mcmun-dong, Cornus, Jiyu, Journeyman, Journey Room (12 types) Let's go, health, spectators, gold and silver coins, Kwakhyang, ephedra (7 kinds) Fivefold (22㎜) Esherichia coli (F41) Goji, golden, yeonkyo, cornus, hawthorn, wild boil, turmeric, gucci mulberry, bark skin, mokyang, cedar mugwort (11 kinds) Let's go, Journeyman, Journey Room, Jiyu (4 types) Fivefold (22㎜) Esherichia coli ( Pathogenic isolates ) Let's go, wolfberry, gojicho, fat milk, ephedra, earl pear, local herb, Mokyang bark bark, journeyman, journey room (11 kinds) Yonkyo, Dansam (2 types) Cornus 5 times (30㎜) Salmonella (Outdoor segregation) Yeongyo, Count, Gucci Mulberry, Journeyman, Journey Room (5 types) Gaza, Hagocho, Golden, Seokcho-cho, Ephedra, Local vinegar (6 kinds) Cornus  Fivefold (32㎜)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Actinobacillus pleuropneumoniae ( S2 ) Gugija, red ginseng, spectators, labokja, large blue leaf, white gulchae, yonggyu, peppermint, ogapi, yinyanggok, turmeric, samreung, haedongpi, goofy, cinnamon, mistletoe, bone print, gucci mulberry, dabsari seeds, elm bark, tofu, Wood barrel, large pencil, dandelion, cedar, lily, ringworm, rock brush, baekryeom, yew, non-recovery, bonsai, chrysanthemum, ridiculous, hawthorn, journey room, Jogu-dong, bamboo leaf, Banggi, nasturtium, triticale, seokwicho, plantain, Yellow wall, evergreen mugwort, situation mushroom, taxan (47 types) Nameless, Thick, Golden, Betel nut, Licorice, Let's go, Ephedra, Ginseng, Peanuts, wild boiled, persimmon, Injin, Bark skin, Dried fruit, Persimmon leaf, Snakeberry, Ash, Bokbunja, Pine needles, Rich, Osuyu, Journeyman, Lichen grass , Leaflets, alder trees, birch trees, Ho keun root (28 species) Sweet Ginseng, Hagocho, Jimo, Earl, Local Herb, Dried, Yugeunpi, Jiyum, Dried Root (9 types) Cornus 5 times (28㎜)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Actinobacillus pleuropneumoniae ( S5 ) Health, roots, large blue leaves, excess, cinnamon, grafting alley, fingering, wedge, dermis, baekrye, gardenia, samreung, motherwort, mistletoe, bone print, dapsali, rice cake mushrooms, sapphire, dandelion, wood situation, lily, Hyunsam, baekseonpi, bud, stalk, baekduong, baekhwa, sancho, phoenix, genus, Jeonho, thistle, dew, eonji, jeongsil, Jogu-dong, low root baekpi, jaepi, jeokdae, cheonhwabun, changja, chichi, earthenware (43 types) Rabokja, Gosam, Creation, Spectators, Root, Geumhwahwa, Licorice, Gamgukcho, Seokwicho, Yanggang, Bokryeong, Baekmogeun, Mansam, Indong, Baekgulchae, Ogapi, Yinyanggak, Yew, Ulgeum, Haedongpi, Gaogapi, Eoseongcho, Yonggyu, Kangbyeol , Yellow wall, moth, texas, rosacea, monarch, persimmon leaf, tofu, gucci mulberry tree, bark of bark, tree trunk, baekryeok, rock brush, non-repair, talan, banggi, bokbunja, sine, pine needles, hawthorn, sorghum, lump, bamboo leaf, Silk tree, juniper tree, nasturtium, Yanggang (50 kinds) Let's go, wolfberry, defector, gourd, betel nut, fellowship, ephedra, peppermint, earl, local herb, plantain, dongjakja, injin, wild boiled, shredded, lake, dried, bark skin, ash, moth, snakeberry, rich man , Leaflet, Climber, Cranberry, Alder, Birch, Fatty Milk, Corvus root, Hungae, Hojang-geun, Dried, Three hundred seconds Sweet Ginseng, Golden, Jimmy, Cornus, Peanuts, Rhizome. Gochocho (7 kinds) Fivefold (37㎜)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Streptococcus suis Wolfberry, Angelica, Gokjeongcho, Hubak, Rabokjak, Creation, Betel nut, Gamkook, Peppermint, Macmun-dong, Baekchul, Local Cho, Plantain, Mansam, Baekmo-geun, Yonggyu, Baekgulchae, Injin, Haedongpi, Samsam, Gyogapi, Donggyuja, Ganghuang, Wormwood mugwort, root, moth, cinnamon, old mulberry, pussy willow, chrysanthemum, elm bark, Cordyceps sinensis, manza, cedar, ash, dandelion, tree, white pine, bud, non-repair, signature, white flower, sancho, situation mushroom, Osuyu, Wonji, Low root skin, Bamboo leaf, Jelgukdae, Astronomical dong, Indong, Tosa, Hyungae, Juniper (54 species) Nameless, health, red ginseng, golden, gold and silver, gwakyang, earl, ephedra, ephemera, spectators, seokwicho, kyobo, ogapi, yellow root, yinyanggwa, samreung, gobon, small hoeyang, vigor, deflation, mistletoe, bone chain, dry, Dapsari, Big Bok, Mokyang, Wood, Situation, White Paper, Lily, Baiyun, Rock Brush, Bokbunja, Bangui, Snakeberry, Pine needles, White porcelain, Sea Lake, Lobster, Thistle, Journey, Yanggang, Yu Geunpi, Sosae, Journey Room, Stall , Alder tree, jeonho, triticale, japonica, Jogu-dong, birch, branch oil, cheonhwabun, takran, pagoji, perilla root, Hyunsam, hojang-geun (60 species) Sweet Ginseng, Red Pepper, Licorice, Cornus, Ridge Root, Bark Skin, Dry, Sounder (8 kinds) Let's go, large blue leaves (2 types) Mallow (30㎜) Yellow Wall (2 types) Staphylococcus hyicus Health, Rabokjak, Hakbak, Creation, Spectators, Kwakhyang, Peppermint, Local Herbs, Baekgulchae, Root, Ogapi, Bangui, Lobber, Snakeberry, Stall, Gentian, Phu Quoc, Journey Room, Dried roots (19 species) Gaza, red ginseng, red pepper, golden, betel nut, yeonkyo, count, peony, windproof, cornus, dried lacquer, bark, dry, cedar, bokbunja, algae grass, fat milk, pagoji, hojanggeun, yinyangkok (19 species) ZIMO, licorice, large blue leaf, wild rump, white porcelain (5 types) Salvia, Yellow Wall (2 types) Fivefold (37㎜)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Hemophilus parasuis ( Outdoor separator ) Gokjeongcho, Gosam, Daegye, Excess, Nogeun, Gwakhyang, Baekchul Macmun-dong, Baekmogeun, Giogapi, Samreung, Gobon, Sohoehyang, Motherwort, Mistletoe, Turmeric, Taxa, Goofy, Cinnamon, Guernsey, Geumjaja, Dapsari, Dungchung, Mokwa, Gulf Brother, woody situation, bamboo leaf, magnolia, lily, bud, white pine, sea lake, pine needles, situation mushroom, sesame oil, raw paper, turmeric, trekking room, dongdong, Yanggang, seaweed, yeast, Jeonho, Jogu-dong, sculptor, graft alley, jeokdae , Astronomical dong, Pohang, Licorice, Hyunsam, Humpback, Juniper (53 types) Clan, Labokja, Yellow Root, White Root, Hocho, Brown Root, Duckweed, Gochu, Gucci Mulberry, Juniper Tree, Cordyceps Sinensis, Rice Cake Mushroom, Large Bok Chok, Keguk, Rock Brush, Non-Suri, White Flower, Pleurotus , 300 seconds, Cheonma, earthenware, gardenia, yonggyu, donggyuja, samsam. Cheonhwa (29 types) Wolfberry, Health, Angelica, Dermis, Creation, Spectator, Large Blue Leaf, Ogapi, Yin-Yang, Eoseongcho, Root, Evergreen, Bone, Bellied Bark, Ash, Dandelion, Bangui, Baekduong, Wealthy, Sign, Hawthorn, Leaflet , Changzai, Chichi, Taeglan, Pagoji, Gypsum roots. Fat milk (28 kinds) Thick, betel nut, gold and silver, licorice, ephedra, peppermint, stone, pruning, dried, cedar, white, bokbunja, white porcelain, fingering (14 kinds) Injin, moth, persimmon leaf, birch, Haedongpi, Byeonja, Earl, Salvia, Golden, Jimmo, Gumi, Plantain, Seokwicho, Korean Dokdogi, Hagocho, Dried, Falcon, White Paper, Snakeberry, Journey, Egret, Alder Root-back skin, Hojang-geun, Hannyeoncho, Bark skin, Local herb, Cornus, White oyster, Hedgehog, Thistle, Yellow wall (50㎜), Gaza (53㎜), Gall (54㎜) (34 species)

Name Clear zone Less than 10mm 11-15 mm 16-20 mm 21-25mm 26mm or more Hemophilus parasuis (Standard stock) Taxi, keg, monarch, turmeric, white paper, large wreck, ash tree, bud, pine needles, sancho, vinegar, gentian, dew, grass, turmeric, twig, silk tree, sculptor, cheonmundong, huang, hyunsam, earthenware, balm, Toe Chung, three hundred seconds, Eoseongcho, Man-shaped, lily, ringworm, non-repair, chidori (31 species) Red ginseng, Kwakhyang, Kangbyeong, Hoe, Mistletoe, Juniper, Daffodil, White Flower, Sea Lake, Cactus, Situation Mushroom, Milk Thistle, Yanggang, Honeysuckle, Journey Room, Yew, Bamboo Leaf, Low Root Skin, Jeoldae, Cheonwha, Taklan, Hyeonggae, Juniper, 300 sec (24 species) Large blue leaves, ginseng, gardenia, gyogapi, baekgulchae, ogapi, yonggyu, persimmon leaf, cinnamon, chrysanthemum, duckweed, gucci mulberry, pussy willow, Gaodong, dried, Bangui, dandelion, woody situation, Baekduong, white porcelain, squirrel. Thick, Skewer, Root, Sine, Journeyman, Jogu-dong, Changiza, Hanryoncho (29 types) Angelica, betel nut, licorice, plantain, yin and yang guk, dog, jinjin, wild boar, bitter gourd, golden gingko, gaodong, arbor bark, tree and tree, cedar, rich, hawthorn, birch, hojanggeun (18 kinds) Let's go, Dansam, Labojak, Hagocho, Golden, Jimo, Seokgok, Jiyu, Gold and Silver Coins, Gums, Peppermint, Local Herbs, Seokwicho, Yellow Wall, Celadon, Bone, Black Bark, Dried, Baekbaek, Rock Brush, Bokbunja, Snakeberry, Small leaf, hyssop, sesame oil, root skin, dysentery grass, perilla root, gall (46㎜) (29 species)

5. Antibacterial test for chicken disease

The antimicrobial activity of the 178 natural herbal medicines was carried out with six strains of the pathogen of chicken disease.

In order to conduct antimicrobial activity tests on herbal medicines,

The strains used in the experiment were cultured at -80 ℃ for 24 to 48 hours in Agar plate medium, Colony was selected and inoculated in Brain Heart Infusion medium, which is a bacterial culture medium, strains were cultured for 24 to 48 hours at 37 ℃, The bacterial growth confirmation of the culture medium was measured by using a spectrophotometer at an OD wavelength of 410 nm, and the bacterial growth culture medium was adjusted to an OD value of 1.2 to 1.3.

Spread 100 μl of bacteria solution on a plate medium to which 15 ml of Mueller Hinton Agar medium was added to a Petri dish (87 × 15㎜), and place each herbal paper disk on it and incubate in a 37 ℃ incubator for 24 hours. The diameters of the clear zones (zones in which the growth of bacteria was suppressed and formed transparently) were measured.

As a result of measuring the antibacterial activity of each herbal medicine by the above method, the diameter of the clear zone was weaker than that of ethanol extraction in hot water extraction, and the test results were not listed. The test results of the herbal medicines showing the antibacterial activity in ethanol extraction are shown in Table 4.

[Table 4. Antimicrobial Activity Test Results for Chicken Disease]

Name Clear zone Less than 10mm 11-15 mm 16-19 mm 20 mm or more Salmonella pullorum Tonsil, neck, neck, mountain root, bark skin, rock brush, snakeberry, traveler, journey room, fingering, grass, Jiyu, Ho Jang Geun (13 kinds) Cornus, rich man (2 kinds) 5 times (20㎜) Salmonella gallinarium Wolfberry, Missed, Golden, Fellowship, Small leaf, Betel nut, Local herb, Seokwicho, Yew, Mansam, Persimmon Leaf, Mokhyang, Arbor bark, Osuyu, Journeyman, Journey room, Jiyu (17 kinds) Cornus, triceps, tofu, neck skin (4 types) 5 times (30㎜) Salmonella enteritidis Wolfberry, Peel, Traveller, Journey, Persimmon Leaf, Alder (6 types) Earl of Peony, Cornus, Wealthy (3 kinds) 5 times (25㎜) Esherichia coli Cornus, Journeyman, Mokyang, Licorice (4 types) 5 times (20㎜) Staphylococcus aureus Thick, ephedra, rosacea, betel nut, gwakyang, yellow wall, tofu, mistletoe, cedar, cordyceps, twigs, larvae, mantis, reeds, ash, dandelion, cinnamon, woody condition, huang, white flower, dry White porcelain, inner ear, journey room, pine needles, Jogu-dong, autograph, taklan, Hyunsam (29 species) Sweet Ginseng, Red Ginseng, Golden, Fellowship, Turmeric, Persimmon Leaf, Count, Local Herbs, Ogapi, Eumyanggok, Sandugung, Bone Crush, Gucci Mulberry, Daffari Seeds, Bark Peel, Rock Brush, Trees, Tree Bark, Lily, Non-Suri, Bokbunja, Banggi, Snakeberry , Squirrel, hedgehog, hawthorn, lobule, sesame oil, itinerary, turmeric, yanggang, baekryeom, yugeunpi, algae grass, yew, birch, fat milk, pagoji, perilla root (39 species) Daecheongyeop, Seokwicho, Sohwa, Dried (4 types) Licorice, Cornus, Ginseng (23㎜), Alder, Ho Jang Geun (5 kinds)

6. Mycoplasma Antimicrobial activity test

Mycoplasma stored at -80 ℃ The growth of hyopneumoniae bacteria was incubated for more than 7 days at 37 ° C by adding reagents and serum necessary for the growth of bacteria to Hank's BSS medium, and the growth of bacteria was measured at 410 nm using a Spectrophotometer (Beckman DU650 ^TM ). The bacterial growth culture was made by dispensing the bacterial solution with an OD value of 1.35 in a test tube, adding 0.1% of the extracted herbal medicines, and incubating at 37 ° C for 7 days. The bacterial growth was indirectly compared with the control. The figures are shown in the table.

Mycoplasma stored at -80 ℃ gallisepticum growth of bacteria by the addition of 1% β-NAD, 1% L-cystein, 20% D-glucose, Penicillin G (200,000IU), Horse serum in basal medium were cultured Myco over 2 days at 37 ℃, the culture medium To check the growth of bacteria, measure the OD value at the wavelength of 410 nm using a spectrophotometer. Dispense the bacteria growth culture solution with an OD value of 1.35 in a test tube, add 0.1% of the extracted herbal medicine, and incubate at 37 ° C for 2 days, and then grow the bacteria. Indirectly compared with the control, the OD value measured at the wavelength of 410nm using the spectrophotometer is shown in the table.

The OD value is high when the growth of bacteria is good at 410 nm using the spectrophotometer, and the OD value is low when the growth of bacteria is suppressed.

As a result of measuring the antimicrobial activity of 178 herbal medicines by the above method, in the case of gall bladder, the fungus cultured with extracts inhibited the growth of bacteria and killed the germs. .

[Table 5. Antimicrobial Activity Test Results for Mycoplasma]

Medicine Mycoplasma gallisepticum Mycoplasma hyopneumoniae Fungal growth O.D value Fungal growth O.D value Go - 1.3401 + 1.3302 Salvia - 1.3420 + 1.3317 A defector - 1.3453 + 1.3336 Gold - 1.3427 + 1.3299 Fellowship - 1.3428 + 1.3311 licorice ++ 1.3087 ++ 1.3212 Local grass + 1.3310 + 1.3295 Seokwicho + 1.3334 + 1.3301 Gardenia ++ 1.3068 - 1.3472 March + 1.3303 - 1.3469 Eoseongcho + 1.3347 - 1.3397 Dog - 1.3471 ++ 1.3181 gallnut +++ 1.2853 +++ 1.2974 Paper ++ 1.3120 + 1.3289

* +,-Indirectly confirmed the growth of bacteria in the culture medium,

  Same as control:-, intermediate growth inhibition: ++, full growth inhibition: +++.

The inventors of the present invention showed the antimicrobial activity of the gall bladder extract in various experimental agents of animal diseases than other herbal medicines,

In addition, it was confirmed that the herbal medicines such as yellow wall, red ginseng, Gaza, golden, licorice, cornus, hachigo, jimo, bokbunja also showed excellent antimicrobial activity in the experiments on the causative agent of animal diseases.

In addition, the present inventors concluded that it is most preferable to use a gall bladder extract as a natural antimicrobial agent against animal pathogens, and based on this, gall bladder extract is used as a natural herbal medicine that can effectively prevent and treat infection of bacterial diseases in animals. It was added to animal feed and tested.

7. Experimental materials

Experimental materials of the powder and extract of the gall of the five used in this experiment

In the case of powder, the herbal medicines were washed three times or more with clean water, and then completely dried by using a hot air dryer, and then the particle size was ground using a grinder as follows.

Bovine feed additives were ground to a powder size of 1.2 mm to 1.5 mm, and then used as pellets for testing.

Pig feed additives were used in the test by grinding the powder particle size of the gall bladder into the size of 1.5-2.0 mm for fresh pigs, 2.0-2.5 mm for lactating pigs, 3.0-3.5 mm for growing pigs, mammal pigs, and 3.5 mm for candidate pigs.

Broiler feed additives were used in the test by grinding the powder particle size of the gall bladder into the size of 2.0 mm for the first season (for first feeding), 3.6 mm for the first time, and 4.5 mm for the latter.

Feed additives for laying hens are ground to a size of 2.0mm for herbivorous lettuce (for first feeding), 2.8mm for chicks, 3.0mm for chicks, 3.2mm for chicks, 3.5mm for before and after laying hens and 3.7mm for broiler breeders. Used.

In the case of the extract, extract the active ingredient by extracting at 60 ℃ for 4 hours using 70% ethanol as an ethanol extraction method, and then concentrated under reduced pressure 10 times using a vacuum concentrator in a 60 ℃ hot water bath, and then lyophilized or sprayed Powder dried using a spray dryer was used for the experiment (extracted as a extract for convenience).

8. Efficacy test on feed additive

Digestive diseases and respiratory diseases, which are caused by viral and bacterial diseases in animals, are a highly contagious disease. Some of the pathogens, including diarrhea and chronic respiratory disease, have been tested.

In order to select a substance that can prevent and treat these bacterial diseases, the antimicrobial activity of 178 natural medicinal plants, herbal medicines, was tested.

As a result of the experiment, it was confirmed that the natural medicinal plant is a gallant with strong antimicrobial activity, and whether it has antimicrobial action, immunity enhancement and increase effect on the animal and its function as a feed additive is effective. Animal experiments were conducted.

end. small

1) Antimicrobial Effect on the Minimum Inhibitory Concentration of Feed Additives

Investigation of the minimum inhibitory concetration (MIC) of bovine pathogenic bacteria showed that E. coli , Salmonella , and M. haemolytica had antimicrobial effects at concentrations of 0.5% or higher and 1% or higher for P. multocida (A) . Respectively.

Test strain MIC (%) 2.0 1.0 0.5 0.25 0.10 E. coli - ^* - - + + S. typhimurum - - - + + P. multocida - - + + + M. haemolytica - - - + +

*-: do not form colony, +: form colony

2) Effect of Feed Additives on Intestinal Escherichia Coli

The feed additives were mixed homogenously (powder and extract) 1% each in the feed, and then fed to the calf for 1 month, and the separation rate of E. coli and Salmonella from the feces was investigated. 22.8% and Salmonella spp. Ranged from 23.7% to 24.4%, and the control group had 35.8% E. coli isolates and Salmonella spp. 33.8%.

division Powder payroll Extract salary Control Test two numbers 127 118 145 Two isolates Escherichia coli 29 26 52 Salmonella 31 28 49 Fungal separation rate (%) Escherichia coli 22.8 22.0 35.8 Salmonella 24.4 23.7 33.8

I. pig

1) Antimicrobial Effect on the Minimum Inhibitory Concentration of Feed Additives

Investigation of the minimum inhibitory concetration (MIC) of porcine pathogenic bacteria revealed that Salmonella was found to be at least 0.25% for A. pleuropneumoniae , H. parasuis and S. hyicus , and at 0.5% or higher for E. coli and S. suis . The antimicrobial effect was shown at a concentration of more than 1%.

Test strain MIC (%) 2.0 1.0 0.5 0.25 0.10 E. coli - ^* - - + + P. multocida - - - + + Salmonella - - + + + A. pleuropneumoniae - - - - + H. parasuis - - - - + S. hyicus - - - - + S. suis - - - + +

*-: do not form colony, +: form colony

2) Fecal separation rate of E. coli administered feed additives.

In order to investigate the effect of feed additives on the suppression of intestinal colonization of Escherichia coli by inoculation, 1.0 ml of 8 × 10 ⁸ CFU / mL was inoculated and feces were collected one day before the inoculation and 5 weeks after the inoculation to investigate the isolation rate of E. coli. As a result, in the feces before inoculation, it was 2.3 × 10 ^{4 to} 2.5 × 10 ⁴ MPN / g, but in the test group fed the feed additive, it was 4.2 × 10 ^{8 to} 4.6 × 10 ⁸ MPN / g after one day of inoculation, but it decreased significantly after one week. After 5 weeks, it was 2.0 × 10 ^{2 to} 2.3 × 10 ² MPN / g.

division Test Methods 1 day before inoculation 1 day after inoculation 1 week after vaccination 2 weeks after vaccination 3 weeks after vaccination 4 weeks after vaccination 5 weeks after vaccination Powdered Salary 2.3 × 10 ⁴ 4.6 × 10 ⁸ 1.5 × 10 ⁴ 4.3 × 10 ³ 2.5 × 10 ³ 2.5 × 10 ² 2.3 × 10 ² Extract salary 2.5 × 10 ⁴ 4.2 × 10 ⁸ 1.9 × 10 ³ 3.9 × 10 ³ 2.3 × 10 ³ 2.2 × 10 ² 2.0 × 10 ² Control 2.5 × 10 ⁴ 1.5 × 10 ⁶ - - - - -

* MPN: Change of feces in piglet infected with E. coli

3) Effect of Feed Additives on Intestinal Escherichia Coli

In order to investigate the effect of suppressing intestinal colonization after feeding the feed additive, 1% of five ginseng (powder and extract) were homogeneously mixed in the feed, and then fed for 6 months. In the test group, the number of Escherichia coli continuously decreased compared to the control group.

division Test Methods 1 month after salary 2 months after salary 3 months after salary 4 months after salary 5 months after salary 6 months after salary Powdered Salary 1.8 × 10 ⁸ 4.1 × 10 ⁴ 2.1 × 10 ⁴ 8.5 × 10 ³ 4.7 × 10 ³ 4.1 × 10 ³ Extract salary 1.9 × 10 ⁸ 3.6 × 10 ⁴ 1.9 × 10 ⁴ 5.9 × 10 ³ 4.3 × 10 ³ 3.9 × 10 ³ Control 1.9 × 10 ⁸ 4.3 × 10 ⁶ 2.6 × 10 ³ 9.8 × 10 ⁴ 8.7 × 10 ⁶ 7.2 × 10 ⁶

* MPN: Change of small intestine in piglet

4) Effect of feed additives on fecal coliform excretion

In order to investigate the effects of feed additives on the excretion of E. coli in feces, homogenously mix 5% (powder and extract) 1% in the feed, feed for 6 months, and collect feces to investigate the separation rate of E. coli. In the test group, the relative excretion rate of E. coli excreted in the feces was higher than the control group.

division Test Methods 1 month after salary 2 months after salary 3 months after salary 4 months after salary 5 months after salary 6 months after salary Powdered Salary 3.5 × 10 ⁸ 5.3 × 10 ⁴ 2.1 × 10 ⁴ 3.0 × 10 ³ 6.8 × 10 ³ 6.5 × 10 ³ Extract salary 3.2 × 10 ⁸ 5.1 × 10 ⁴ 1.9 × 10 ⁴ 2.6 × 10 ³ 6.3 × 10 ³ 6.1 × 10 ³ Control 2.6 × 10 ⁷ 6.7 × 10 ⁶ 9.4 × 10 ⁵ 9.3 × 10 ⁵ 3.7 × 10 ⁶ 5.7 × 10 ⁵

* MPN: Change feces in piglet

All. chicken

The chickens used in the experiment were tested in a sterile experimental animal breeding farm using two-week-old chickens hatched with SPF eggs and one-week-old broilers raised in farms.

1) Antimicrobial Effect on the Minimum Inhibitory Concentration of Feed Additives

Results of testing the Minimum inhibitory concetration (MIC) of the pathogenic bacteria in the chicken E. coli, at least 0.25% concentration, S. enteritidis, S. aureus is more than 0.5% in concentration, S. pullorum, S. gallinarium is At concentrations of more than 1% each showed an antimicrobial effect.

Test strain MIC (%) 2.0 1.0 0.5 0.25 0.10 E. coli - ^* - - - + Sal . pullorum - - + + + Sal . gallinarium - - + + + Sal . enteritidis - - - + + Staphylococcus aureus - - - + +

*-: do not form colony, +: form colony

2) Salmonella Stagnation inhibitory effect on enteritidis

① Diarrhea prevention and weight gain

Salmonella enteritidis in order to examine the diarrhea and weight change for Salmonella infected flocks in Salmonella in 10 ⁷ CFU / ml Two-week-old chicks were administered orally twice a day at 0.2 ml each for 2 weeks of age. Diarrhea in the test and control groups was examined after feeding 1% of the 5% (powder and extract) into the feed. Three of the animals and 8 of 10 animals in the control group had diarrhea, but after 14 days, they did not show any symptoms.

In the test group, 430.20 ± 6.90g was increased to 904.34 ± 13.55g on the 14th day, and the increase rate was increased compared to the control group.

division Onset of diarrhea Weight gain (g) 1 day 7 days 14 days 1 day 7 days 14 days Test group 0/10 3/10 0/10 430.20 ± 6.90 825.54 ± 25.11 904.34 ± 13.55 Control 0/10 8/10 0/10 381.81 ± 14.20 753.20 ± 22.30 798.50 ± 58.75

* Infected / Tested

② Salmonella in the contents of the cecum Distribution of bacteria

Salmonella from Caecum Contents of Infected Family As a result of isolates, the test group was reduced to log 5.87 ± 0.24 CFU / g on day 1 and log 5.65 ± 0.62 CFU / g on day 7, but not separated on day 14.

In the control group, it increased to log 5.91 ± 0.36 CFU / g at 1 day of inoculation and log 7.15 ± 0.61 CFU / g at 7 days, but decreased at 14 days.

division Inoculation day 1 day 7 days 14 days Test group 5.87 ± 0.24 (5/5) ^* 5.65 ± 0.62 (5/5) 0 (0/5) Control 5.91 ± 0.36 (5/5) 7.15 ± 0.61 (5/5) 3.25 ± 1.22 (5/5)

* Infected / Tested

③ Salmonella from organs infected Separation frequency

The bacteria were separated from the nasal cavity, liver and caecum on the 7th and 14th day of inoculation, and 2 out of 5 mice and 3 out of the spleen were separated on the 7th day of the bacterial infection. ,

In the control group, all 5 animals were separated from the liver and spleen at 7 days of bacterial infection, and after 14 days, they were separated from the 2 mice.

division Test system Infectious organs spleen liver Caecum Test group 7 days 3/5 ^* 2/5 5/5 14 days 0/5 0/5 0/5 Control 7 days 5/5 5/5 5/5 14 days 2/5 2/5 5/5

* Infected / Tested

9. Physicochemical after Feed Additives Constellation test

Feeding gall (powder and extract) as feed additives to the animals, weight gain, lethality, and physicochemical tests (Cholesterol, moisture, crude protein, fatty acids, ash, feed requirement, weight, fatty acids, amino acids), etc. were performed. The results obtained were analyzed by variance analysis and Duncan's multiple test method using SAS / PC (SAS, 1988).

end. small

1) Effects on protein, fat, batch feed requirement and weight

In order to investigate the effect of feed additives on crude fat, crude ash, and feed demand in beef, the test group prepared five gall (powder and extract) into pellets, mixing 1% homogeneously into the feed, and then receiving the salary for six months. After that, the effects of slaughtered cows on crude fat, crude meal and feed demand were examined.The group fed the gallant (powder and extract) decreased crude fat (17.2 ± 0.57 → 14.2 ± 0.97: 15.3 ±). 0.75), decreased Cholesterol (48.25 ± 0.24 → 43.54 ± 0.32: 45.43 ± 0.43), and decreased feed demand (9.79 → 8.75: 8.94), respectively.

division Fat (100 g) Cholesterol (mg / fatg) protein (%) Feed requirement Daily weight increase (Kg / day) Powdered Salary 14.2 ± 0.97 43.54 ± 0.32 10.75 ± 0.13 8.75 0.650 Extract salary 15.3 ± 0.75 45.43 ± 0.43 9.69 ± 0.15 8.94 0.652 Control 17.2 ± 0.57 48.25 ± 0.24 7.72 ± 0.71 9.79 0.598

2) Evaluation of safety on calves

In order to evaluate the safety of calf in feed additives, blood was collected from the calf and centrifuged at 3,000 rpm for 20 minutes at 4 ° C. Serum was analyzed by biochemical analyzer (RV-X7, Techmmicon, USA). Glucose, Bilirubin, LDH and others were within the normal range.

division Test group Control Remarks (normal range) Powder payroll Extract salary SGOT 32.7 (I.U / l) 31.8 (I.U / l) 22.9 (I.U / l) 8.5-93.0 (I.U / l) SGPT 25.1 (I.U / l) 28.5 (I.U / l) 22.8 (I.U / l) 20.0-76.8 (I.U / l) Glucose 82.4 (I.U / l) 79.3 (I.U / l) 78.4 (I.U / l) 45-130 (I.U / l) Bilirubin 0.20 (I.U / l) 0.21 I.U / l) 0.1 (I.U / l) 0.1-0.7 (I.U / l) LDH 16.5 (I.U / l) 15.8 (I.U / l) 17.3 (I.U / l) 8.0-302 (I.U / l) CPK 75.4 (I.U / l) 74.9 (I.U / l) 76.1 (I.U / l) 66.0-120 (I.U / l) Alkaline phosphate 153.5 (I.U / l) 155.1 (I.U / l) 157.3 (I.U / l) 94.0-170 (I.U / l) Amylase 129 (I.U / l) 128 (I.U / l) 119.6 (I.U / l) 126-250 (I.U / l)

I. pig

Select healthy feed pigs at 4 weeks of age, select commercially available feed, homogenously mix 5 g powder and extract with 1% of feed, feed for 4 months, and then perform physicochemical test using general ingredient analysis method. Was carried out.

1) Effects on protein, fat, batch feed requirement and weight

In order to investigate the effects of feed additives on crude fat, crude ash and feed demand of pork, the test group mixed 1% gall (powder and extract) into the feed homogeneously and then added crude fat, Investigation of the effect on crude ash and feed demand showed that the group fed the five embryos (powder and extract) decreased crude fat (6.20 ± 0.62 → 3.71 ± 0.42: 3.74 ± 0.38) and decreased Cholesterol (60.20 ± 0.32 → 55.43 ± 0.51: 55.43 ± 0.57) and feed demand (2.85 → 2.73: 2.75), respectively.

division Fat (%) Cholesterol (mg / fatg) protein (%) View% (%) Feed requirement weight Powder payroll 3.71 ± 0.42 55.43 ± 0.51 20.85 ± 0.13 3.85 ± 1.10 2.73 108.15 Extract salary 3.74 ± 0.38 55.43 ± 0.57 20.89 ± 0.15 3.88 ± 1.16 2.75 108.20 Control 6.20 ± 0.62 60.20 ± 0.32 20.72 ± 0.71 3.90 ± 1.08 2.85 106.60

2) Effect on Fatty Acids

To investigate the effects of feed additives on the fatty acids in pigs, the five gall (powder and extract) were mixed homogeneously in 1% feed, and the effects on the fatty acids in pork after 6 months of feeding were examined. Palmitic acid and Stearic acid decreased in the fed group compared with the control group, and unsaturated fatty acids were increased in the test group fed the five embryos compared to the control group by the increase of Palmitoleic acid, Oleic acid, Linoleic acid, Dihomo-γ-linolenic acid and Arachidonic acid. An increase and a decrease in saturated fatty acids were shown, respectively.

Fatty acids Test group Control Powder payroll Extract salary Myristic acid 13.80 ± 0.01 13.90 ± 0.02 14.0 ± 0.01 Palmitic acid 20.80 ± 0.05 21.02 ± 0.06 26.00 ± 0.60 Palmitoleic acid 4.20 ± 0.05 4.25 ± 0.05 3.25 ± 0.14 Stearic acid 11.00 ± 0.15 11.09 ± 0.07 15.30 ± 0.10 Oleic acid 45.20 ± 0.65 45.29 ± 0.66 42.22 ± 0.01 Linoleic acid 17.50 ± 0.07 17.60 ± 0.07 15.70 ± 0.13 Dihomo-γ-linolenic acid 1.65 ± 0.01 1.68 ± 0.01 0.82 ± 0.02 Arachidonic acid 3.20 ± 0.04 3.19 ± 0.03 1.45 ± 0.03 MUFA ^* 49.50 ± 0.75 49.58 ± 070 45.50 ± 0.15 PUFA ^* 21.80 ± 0.12 22.50 ± 0.10 18.04 ± 0.20

* MUFA: Monounsaturation fatty acid

* PUFA: Polyunsaturation fatty acid

3) Effect on Amino Acid Composition

In order to investigate the effect of feed additives on the amino acid composition of pork, the mixture of five gall (powder and extract) was homogeneously mixed in 1% of the feed, and then the effect on the amino acid composition of pork was six months after feeding. Asparatic acid, Glutamate, Histidine, Proline, valine, Lysine, Isoleucine and Leucine were increased in the group fed the extract compared to the control group.

Amino acids Test group Control Powder payroll Extract salary Asp 1005.2 ± 18.15 1030.2 ± 18.20 805.5 ± 26.05 Ser 680.5 ± 19.20 750.5 ± 15.10 685.5 ± 15.20 Glu 1750.2 ± 38.20 1870.2 ± 32.20 1628.1 ± 85.53 Gly 895.2 ± 50.52 ^* 980.40 ± 12.20 ^* 334.5 ± 13.20 His 294.3 ± 10.54 ^* 350.1 ± 20.54 ^* 133.5 ± 045 Thr 430.5 ± 18.20 450.1 ± 13.12 306.8 ± 5.46 Arg 710.3 ± 45.10 576.5 ± 38.20 755.2 ± 40.20 Ala 480.5 ± 10.16 ^* 489.6 ± 14.12 ^* 458.2 ± 32.80 Pro 832.1 ± 52.12 ^** 851.2 ± 59.20 ^** 730.1 ± 32.95 Tyr 234.2 ± 13.31 232.5 ± 23.10 304.2 ± 5.60 Val 825.2 ± 35.38 ^** 875.4 ± 23.20 ^** 741.1 ± 11.45 ^** Met 526.6 ± 12.35 ^* 542.4 ± 26.02 ^* 531.3 ± 3.28 ^* Lys 923.3 ± 23.42 ^** 910.8 ± 10.12 ^** 824.4 ± 56.23 Isoleu 723.3 ± 13.47 ^* 729.2 ± 11.20 ^* 471.7 ± 18.94 ^* Leu 1252.3 ± 22.41 ^* 1175.5 ± 12.81 ^* 1013.2 ± 3.86 ^* Phe 411.2 ± 33.81 418.1 ± 20.76 544.9 ± 10.24

* P <0.05, ** P <0.01

4) Evaluation of safety on pigs

In order to perform a safety evaluation of the feed additives in pigs, blood was collected from the external vein of pigs and centrifuged at 3,000 rpm for 20 minutes at 4 ° C., and serum was analyzed by a biochemical analyzer (RV-X7, Techmmicon, USA). As a result, SGOT, SGPT, Glucose, Bilirubin and LDH were in the normal range.

division Test group Control Powder payroll Extract salary SGOT 34-56 33-54 31-60 SGPT 10.0-41.0 10.0-41.0 10.0-44.0 Glucose 66-116 65-110 62-125 Bilirubin 0.01-0.14 0.01-0.12 0.00-0.58 LDH 37-86 36-87 32-98 CPK 85-148 86-146 83-160 Alkaline phosphate 43-92 45-90 35-105 Amylase 78-125 78-120 65-130

All. chicken

Seven-day-old broilers were mixed homogeneously to obtain 1% of each of the gall bladder powder and extract in the commercial feed, and then fed until just before shipment and subjected to physicochemical test.

1) Effect of Chicken Meat on Crude Fat, Crude Meal and Whole Moisture

The effects of feed additives on crude fat, crude ash, and starch in broiler broilers were investigated. Decrease (0.92 ± 0.00 → 0.90 ± 0.00: 0.89 ± 0.00) and increase in total moisture (74.61 ± 1.01 → 76.23 ± 0.13: 78.14 ± 0.16).

division Crude fat View minutes Starch Powder payroll 10.11 ± 0.48 0.90 ± 0.00 76.23 ± 0.13 Extract salary 12.22 ± 0.438 0.89 ± 0.00 78.14 ± 0.16 Control 9.61 ± 0.62 0.92 ± 0.00 74.61 ± 1.01

2) Effect of Chicken Meat on Fatty Acids

The effects of feed additives on fatty acids in broilers were investigated. acid: 34.52 ± 1.00 → 35.41 ± 0.45: 37.13 ± 0.73, Linoleic acid: 17.18 ± 0.12 → 18.32 ± 0.17: 18.15 ± 0.21, Linolenic acid: 0.88 ± 0.02 → 0.92 ± 0.04: 0.96 ± 0.06).

Fatty acids Test group Control Powder pay group Extract pay group Myristic acid 0.75 ± 0.03 0.75 ± 0.06 0.76 ± 0.04 Palmitic acid 24.48 ± 0.86 24.75 ± 068 24.12 ± 0.71 Palmitoleic acid 6.40 ± 0.06 6.39 ± 0.03 6.31 ± 0.13 Stearic acid 9.54 ± 0.12 9.44 ± 0.43 11.41 ± 0.71 Oleic acid 35.41 ± 0.45 37.13 ± 0.73 34.52 ± 1.00 Linoleic acid 18.32 ± 0.17 18.15 ± 0.21 17.18 ± 0.12 Arachidonic acid 0.92 ± 0.04 0.96 ± 0.06 0.88 ± 0.02 SFA ^* 34.52 ± 0.83 34.65 ± 0.89 37.15 ± 0.78 UFA ^* 63.72 ± 076 65.37 ± 0.82 62.82 ± 077 MUFA ^* 36.53 ± 0.86 39.68 ± 0.57 34.52 ± 0.38 PUFA ^* 24.32 ± 0.83 24.06 ± 033 24.02 ± 0.23

* SFA: Saturation fatty acid

* UFA: Unsaturation fatty acid

* MUFA: Monounsaturation fatty acid

* PUFA: Polyunsaturation fatty acid

3) Effect of Chicken Meat on Amino Acid Composition

The effects of feed additives on broiler amino acid composition of broilers were increased in Methionine, Asparatic acid, Threonine, Leucine, Phenylalanine, Histidine, Methionine, and Valine in the five-fold powder and extract supplemented groups.

Amino acids Test group Control Powder pay group Extract pay group Val 1053.2 ± 48.15 ^** 1064.3 ± 56.20 ^** 942.5 ± 13.45 ^** Met 480.2 ± 19.52 ^* 486.5 ± 25.24 ^* 437.9 ± 4.58 ^* Lys 1750.2 ± 56.20 1766.6 ± 56.29 1728.0 ± 65.78 Isoleu 1023.2 ± 52.27 ^* 980.40 ± 12.20 ^* 902.5 ± 21.47 ^* Leu 1600.2 ± 35.54 1668.8 ± 53.80 ^** 1430.3 ± 4.00 ^** Phe 810.2 ± 34.15 ^* 815.1 ± 43.12 ^* 730.0 ± 20.49 ^* Asp 1847.3 ± 64.10 1876.5 ± 58.17 ^* 1755.8 ± 76.05 ^* Ser 770.8 ± 22.36 780.6 ± 32.12 795.7 ± 32.23 Glu 2632.1 ± 142.1 2611.2 ± 121.22 2980.0 ± 110.07 Gly 933.6 ± 53.94 942.2 ± 38.98 854.6 ± 83.12 His 672.2 ± 36.13 ^* 680.1 ± 33.50 ^* 574.8 ± 26.90 ^* Thr 825.2 ± 35.38 ^* 868.4 ± 23.20 ^* 798.1 ± 11.45 ^* Arg 1338.6 ± 50.35 1342.4 ± 46.02 1356.3 ± 85.13 Ala 1472.6 ± 53.42 ^* 1490.8 ± 71.20 ^* 612.4 ± 46.23 ^* Pro 781.3 ± 63.47 ^** 790.2 ± 57.20 ^** 603.7 ± 46.94 ^** Tyr 67.3 ± 72.41 701.5 ± 47.81 630.2 ± 9.36

* P <0.05, ** P <0.01

4) Effect on broiler color of broilers

In order to investigate the effect of feed additives on the broiler color of broiler chickens, the effects of nutrients were mixed homogeneously to 1% in the feed, and the effects of broiler color on broiler color were examined at 30 days of feeding. Showed a decrease.

division Color Lightness Redness Yellowness Powder payroll 49.14 ± 1.63 3.25 ± 1.52 9.75 ± 0.79 Extract salary 48.96 ± 1.52 3.28 ± 1.49 9.69 ± 0.81 Control 50.62 ± 2.41 2.76 ± 1.41 9.86 ± 2.72

5) Influence on broiler specifications

Investigation of the effects of feed additives on the broiler specifications showed that the average weight gain (1.68kg → 1.74kg), growth rate (91% → 95%), and mortality were lower in the five-german powder and extract-feeding groups than the control group. 9% → 5%), and the decrease in feed demand (1.85% → 1.74%).

division Test group Control Results Etc. Average weight 1.74 kg 1.68 kg + 0.06kg Growth rate 95% 91% + 4% + 4% Fassa Rate 5% 9% -4% -4% Feed rate 1.74% 1.85% -0.11%

6) Impact on safety

In order to evaluate the effects on the safety of broilers for feed additives, the test group and control group fed with 5% of the 5% fed to the feed were subjected to safety evaluation on the 15th and 30th days. .

division Test Items ALP (I.U / L) ALT (I.U / L) AST (I.U / L) LDH (I.U / L) Salary 15th day Test group 36.42 ± 3.01 27.22 ± 2.16 86.62 ± 1.45 91.34 ± 3.74 Control 37.30 ± 2.21 22.43 ± 2.32 88.72 ± 4.31 91.32 ± 3.22 30 days of salary Test group 36.26 ± 3.32 23.72 ± 1.79 81.71 ± 4.11 89.75 ± 4.54 Control 37.11 ± 2.88 26.12 ± 3.14 74.03 ± 1.44 90.54 ± 2.14

7) Effect of protein expression in broiler body

In order to investigate the effects of feed additives on broiler protein in the broiler, homogenous 5% homogenous extract was mixed with feed to the feed, blood was collected at 30 days of feeding, serum was separated and two-dimensional electrophoresis (SDS + Isoelectric focusing) As a result, three spots were found to be thicker in the group fed the five-fold extract than in the control group, and the results were in the position similar to the separation by two-dimensional electrophoresis of other herbal preparations.

 spot1: pI4.13 ± 0.1, molecular weight 25.13 ± 4.47kD,

 spot2: pi 4.36 ± 0.09, molecular weight 24.99 ± 4.36kD,

 spot3: pI4.64 ± 0.09, molecular weight 24.89 ± 4.32) (Fig. 3-1, 3-2).

      3 <-------- PI ------------> 10

The test group was more heavily expressed than the control group.

Claims (8)

In the hot water extraction method, Grinding gallza, putting ingredients in distilled water, extracting at 60 ° C for 16 hours and then extracting at 100 ° C for 1 hour to collect the active ingredients and using a vacuum concentrator to concentrate under reduced pressure in a hot water bath at 80 ° C. Animal feed additives containing powders and extracts. The method of claim 1, Animals containing gallant powder and extracts, characterized in that the concentrated solution concentrated under reduced pressure by hot water extraction and dried into a powder using a freeze-drying method and a spray dryer using a spray method in the form of a disk and a nozzle. Feed additives. In the ethanol extraction method, Grind the gallant and add the ingredients to 70% ethyl alcohol to extract the active ingredient at 60 ° C for 4 hours to collect the active ingredients and to concentrate the powder and extract of the gallant by vacuum concentrating in a 60 ℃ hot water bath. Animal feed additives containing. The method of claim 3, wherein The extracted extract is concentrated under reduced pressure to freeze-dried the concentrate, and dried using a spray method in the form of a disk and a nozzle using a spray dryer to prepare a powder, and to prepare the concentrate as a liquid formulation. Animal feed additives containing the powder and extract of the gall bladder. In the cattle feed additives and auxiliary feed manufacturing method, The size of the gall bladder powder is 1.2 mm to 1.5 mm, and the powder and extract of the gall bladder is characterized by a method of granulating the additive prepared from the pellet into 0.1% to 3% of the total weight of the feed and granulated with the gall bladder powder particles. Animal feed additives containing. In the pig feed additives and auxiliary feed manufacturing method, The size of the gallnut powder is 1.5 ~ 2.0mm for gutting pigs, 2.0 ~ 2.5mm for lactating pigs, 3.0 ~ 3.5mm for growing pigs and mammals, and 3.5mm for candidate pigs. Animal feed additives containing the powder and extract of the gall bladder characterized in that the manufacturing method of the size of the powder particles. In the feed additive and poultry manufacturing method for poultry, The size of the gall nut powder is 2.0mm for herbivorous weight (for first feeding), 3.6mm for electricity, 4.5mm for the latter, and additives for laying hens are 2.0mm for herbaceous weight (for first feeding), 2.8mm for chicks and 3.0mm for middle chicks. Grow-up powder, characterized in that the manufacturing method of grinding the chicks 3.2mm, chicks before and after laying hens, 3.5mm, broiler breeders to 3.7mm size to the size of 0.1% to 3% of the blast powder particles relative to the total weight of the feed And animal feed additives containing the extract. Animal feed additives containing the powder and extract of the gall bladder, characterized in that using the gall bladder powder and extract to prepare a feed additive and supplementary feed for the purpose of preventing or treating animal diseases.