KR20110059408A - A process for producing feedstuff additives comprising natural herbs and having a preventing activity of calf's diarrhea - Google Patents

Abstract

PURPOSE: A producing method of a feed additive with the prevention effect for calf diarrhea is provided to improve the disease resistance for various diseases of calf. CONSTITUTION: A producing method of a feed additive with the prevention effect for calf diarrhea comprises the following steps; cutting lonicera japonica, taraxacum platycarpum, geranium nepalense, agastache rugosa, and glycyrrhiza uralensis, and separately boiling with water; mixing all the obtained extracts after diluting with water; crushing atractylodes japonica, citrus unshiu, plantago asiatica, polyporus umbellatus, poria cocos, cinnamomum loureirii, perilla frutescens, angelica dahurica, angelica dahurica, pinellia ternate, platycodon grandiflorus, and alisma plantago into mixed powder; mixing the extract mixture, the mixed powder, and sodium bicarbonate; and adding probiotic including lactobacillus acidophilus, aspergillus oryzae, and saccharomyces cerevisiae into the mixture.

Description

A method for producing a feed additive having a diarrheal disease prevention activity of a calf containing a natural herbal medicine {A PROCESS FOR PRODUCING FEEDSTUFF ADDITIVES COMPRISING NATURAL HERBS AND HAVING A PREVENTING ACTIVITY OF CALF'S DIARRHEA}

The present invention relates to a method for preparing a feed additive having a diarrheal disease prevention activity of a calf containing a natural herbal medicine, and more specifically, gold silver flower (포 花), pogongyeong (현지 公 英), local herb (玄 之 草, 玄 草), Dilute 10 times the hot water extract extracted from five kinds of herbal medicines, Kwakhyang and Licorice, and as another herbal medicine, finely powdered white extract, dermis, and cha-jeon. A mixture of Sichuan, Garyeong, Baekbokryeong, Broiler, Jassop, Purple Paper, Daebokpi, Banja, Gillyeong and Taeksa After mixing well with sodium bicarbonate (sodium bicarbonate, NaHCO ₃ ), it contains Lactobacillus acidophilus , Aspergillus oryzae and Saccharomyces cerivisiae Feed additives for the prevention of calf diarrhea prepared by adding probiotics and its Feed for calves, which comprises a method for producing the feed additive, and to pay it to the calf, to a method for preventing diarrhea in calves.

Calf diarrhea, one of the most important causes of calf mortality, is particularly one of the most economically damaging livestock diseases in livestock farms, and it is known that calf diarrhea is more than 12 to 24 trillion won per year (Bendali). Et al., 1999).

The causes of newborn calf diarrhea are related to a variety of infectious, nutritional, immunological and environmental factors (Kim et al., 1990). Calf diarrhea is a clinical syndrome in which three factors are linked, including pathogenic microorganisms, the breeding environment, and cattle themselves (Acres et al., 1977; Kim et al., 1990). Above all, young calves are known to be highly susceptible to various pathogens (Webster et al., 1985; Lee et al., 1999). Within six months of age, calves are infected with pathogens from a variety of environments and headstocks, resulting in a variety of diseases caused by these pathogens (Martin and Meek, 1986; Kang et al., 1998).

The pathogen-inducing pathogens of calf known to date in Korea include bovine viral diarrhea virus (BVD), bovine rotavirus (BRV) and bovine coronavirus (BCV), and bacterial strains such as E. coli . And Salmonella spp. , Clostridium spp., Campylobacter and others. Parasites also include Eimeria spp., Isospora spp., Cryptosporidium spp. (Britney et al., 1984; Humura et al., 1985; Holland, 1990; Barker et al., 1993).

The rate of detection of rotavirus as the causative agent of diarrhea in newborn calves was 69.2%, which is significantly higher than that of enterotoxic E. coli (14.7%) (Kim et al., 1990). Viral rotavirus at the age of diarrhea occurred at 41.7% before 7 days of age and 48.1% at 8 to 14 days of age, with approximately 90% detected within 2 weeks of age.

The following is a bacterial enterotoxins hemorrhagic E. coli was detected 26.1% to 34.8%, from 8 to 14 days of age prior to 7 days of age, in the report of more than 22 days of age Acres et al. (1977), a 34.8% was detected in E. coli and rotavirus The detection rate was 62% and 10% in diarrhea calves within 5 days (Kim et al., 1990; Acres et al., 1985). There are many ways to treat calves and diarrheal diseases, but currently available drugs on the market have a temporary effect, but if the treatment is stopped, pathogens may be latent and relapse or become chronic. Resistance problems can be serious.

Herbal herbal medicines have been used to treat livestock diseases by farmers' long-term livestock experiences. The use of herbal medicines has particular strengths in the preventive aspects of preventing livestock diseases. First of all, natural herbal medicines used in herbal medicine are characterized by low toxicity or non-toxicity, do not remain in the body of livestock, and do not cause drug resistance.

Recently, calf diarrheal diseases caused by pathogens such as Salmonella are increasing due to the enlargement of cattle and cow breeding, and the use of drugs is also increasing. The use of chemical drugs has a temporary effect on infectious diseases. However, discontinuation of the medications presents a problem of re-infection, and the increase in the appearance of resistant bacteria following long-term use, as well as the retention of drugs, impedes the safe production of livestock products.

Recently, consumers are demanding the production of livestock products that are safe for the human body, so livestock producers will have to reduce the use of antibiotics (drugs) they have used so far and be prepared to ban drug use in the near future.

The present invention develops a feed additive that combines the prevention and treatment of calf diarrhea using natural herbal medicines, and feeds the feed containing the same to the calf, thereby reducing farming difficulties and calf mortality caused by calf diarrhea and safe livestock products. By increasing the consumption of livestock products through production, the government tried to contribute to raising incomes of farmers and improving public health.

An object of the present invention for solving the above problems is to prevent diarrhea of the calf to reduce diarrhea, as well as to improve the anti-competence against various diseases to increase the productivity of livestock farms, natural herbal medicines having a diarrheal disease prevention activity It is to provide a method for producing a feed additive comprising a feed additive prepared by this method.

Another object of the present invention is to provide a feed comprising a feed additive having a diarrhea prevention effect of the calf containing the natural herbal medicine.

In addition, another object of the present invention is not only to prevent the diarrhea of the calf containing the natural herbal medicines, but also to feed the calf with a feed additive having a gastrointestinal reinforcement and efficacy as a digestive agent to healthy without infection of the disease It is to provide a way to raise calves that can grow and improve their growth rate.

Hereinafter, the present invention will be described in detail.

The present invention

1) finely cut gold silver, pogongyoung, local vinegar, kwakhyang and licorice, and boiled by adding water in a weight ratio of 1: 10 to evaporate water to obtain respective hot water extracts;

2) diluting each of the hot water extracts of step 1) with 10 times of water and then mixing them in the same volume ratio;

3) Grind the white powder, dermis, chajeon, seonbi, baekbokyeong, broiler, pomegranate, white paper, daebokpi, halves, gilkyung, and tack with fine powder of 0.3 mm or less and take each fine powder in the same weight ratio and mix powder Obtaining step;

4) the hot water extract mixture of step 2): the mixed powder of step 3): sodium bicarbonate prepared in a weight ratio of 1: 2: 0.2 by first adding the mixed mixture of the hot water extract solution and step 3) to the automatic mixer After stirring, mixing sodium bicarbonate there; And

5) The probiotic consisting of Lactobacillus acidophilus , Aspergillus oryzae and Saccharomyces cerevisiae was added to the mixture obtained in step 4). It is a method of preparing a feed additive having a diarrheal disease prevention activity of a calf containing a natural herbal medicine, characterized in that the step of mixing in a weight ratio.

Hereinafter, the present invention will be described in more detail.

In preparing a feed additive having a diarrheal disease preventive activity of a calf according to the present invention, a mixture of hot water extract of Geum, Hwagong, Pogongyoung, Haechocho, Kwakhyang and Licorice Is the first herbal ingredient.

In order to prepare the first herbal ingredient described above, the five kinds of herbal medicines are cut into appropriate sizes, preferably 3 to 5 cm in size. Fine herbal medicines evaporate moisture by heating with the addition of individual herbs and water. Add water to the shredded medicinal herbs in a weight ratio of 1:10 and heat it to a high temperature. When the amount of water remaining is about 20%, stop the heating and filter out three layers of gauze to obtain the final hot water extract. The main components are gold and silver, Lonicerin, pogongyoung, taraxasterol, local herbaceous ellagitannin, methyl vicvicol (esdragol) and liquorice glycyrrhizin.

Each hot water extract obtained by the above method is diluted with the addition of 10 times of water, and then mixed well in the same volume ratio, that is, in a volume ratio of 1: 1: 1: 1: 1 to prepare the first herbal component of the present invention.

Next, in the present invention, Baekchul (진), dermis (陳,), chajeon (저 前 子), ghost, Baekbokryeong, broiler (자), jasminoide, baekji (白芷), Daebokpi (大) The mixture of 반, 반 夏 (길 夏), Gil-kyeong (택) and taxa is used as the second herbal ingredient. The above 12 kinds of herbal medicines used as the second herbal medicine in the present invention may arbitrarily determine the usage amount of each component, but preferably the mixing ratio of each component is 1: 1. The second herbal ingredient of the present invention is prepared from fine powders, each of which is pulverized to a size that passes each of the above 12 kinds of herbal medicines through a grinder of 0.3 mm or less (1φ or less).

The first herbal ingredient prepared in the above-mentioned second herbal ingredient fine powder is mixed at a weight ratio of 1: 2 and sufficiently stirred so as not to form a lump in an automatic mixer.

Sodium bicarbonate (NaHCO _3, about 99% purity) is mixed with the mixture of the first herbal ingredient and the second herbal ingredient. At this time, the mixing ratio of each component is added in a weight ratio of first herbal component: second herbal component: sodium bicarbonate in a weight ratio of 1: 2: 0.2. The entire mixture to which sodium bicarbonate is added should preferably be stored in a cleanbench disinfection tank equipped with an ultraviolet fluorescent lamp for 24 hours sterilization to avoid moisture. When sodium bicarbonate is added in a weight ratio of 0.2 or more when mixed, it becomes difficult to absorb more moisture than necessary to maintain the proper viscosity of the entire mixture.

In the next step, Lactobacillus acidophilus , Aspergillus oryzae and Saccharin in a mixture of the first herbal ingredient and the second herbal ingredient comprising sodium bicarbonate sterilized as described above. Probiotics consisting of Saccharomyces cerevisiae are added at a weight ratio of 10: 1. The probiotic is preferably a microbial mixed agent containing at least 5 x 10 ⁵ CFU / kg of microbial strain. At this time, if the added amount of the probiotic is less than the above weight ratio, the effect by the microorganisms is weak, and when it is added more than the microbial population exceeded may cause problems in the breeding livestock.

The feed additive according to the present invention prepared as described above may be fed by adding 0.1 to 0.3% by weight relative to the daily feed amount according to the growth phase of the calf from 7 days of age to 7 months of age.

The characteristics and efficacy of the main herbal medicines used in the present invention are as follows.

Geumgeunhwa (金 銀花) is a dried stem or leaf of the stem of Lonicera japonica Thumb of the Caprifoliaceae , and its origin is Korea and China. The main ingredients are Tannin, Saponin, Lonicerin, Luteolin, Lnostiol, and the pharmacological effect is that the extract has a protective effect against peptic ulcer and especially Gram-positive (+) bacteria, Staphylococcus aureus, pneumonia, etc. It has a strong antimicrobial action against diplococci.

Pogongyoung is a plant of the genus Taraxacum, Compositae . It sprouts in early spring, the leaves are segmented, and flowers bloom at the end of one stem in the center. The flower is yellow and the stem is cut to produce white juice. It is usually harvested from April to May. It lives in Korea, China, and Japan, and its main ingredients are Taraxasterol, Choline, Inulin, Pectin, etc. The pharmacological effect is Staphylococcus, Hemolytic Streptococcus, Pneumococcus, Outeritis, Diphtheria, Pseudomonas aeruginosa, Modified bacillus, Tif It has bactericidal and antimicrobial effects in bacteria. It is also effective in diuretic, biliary, chronic gallbladder spasms and stones, indigestion, habitual constipation, anti-inflammatory, and health.

Local herb (초 之 草 or 玄 草), also called Nohhakcho, is a dried plant of Geranium Thunbergii Sieb of Geraniaceae . The leaves are long, lobes, viviparous, grayish green to green, and the side is long oval to iran, rusted and rusted. Its main ingredient contains tannins and its main ingredient is geranin, one of ellagitannin. In addition, it contains gallic acod and duccinic acid. Pharmacological action has bactericidal effect against E. coli, typhoid fever, Escherichia coli, Preslow gallbladder, and Geltnel gallbladder. In addition to the stomach, dressing is also favored.

Kwakhyang (藿香) is dried dried outpost or leaf of Pogostenon cablin Benth of Labiatae , also known as baecho incense, Kwangju Incense, or Bangalore ( Yangmyeongcho ) in Korea. Its mountainous area is Guangdong province of China, Philippines, Korea, etc. The stem is grey-green ark, gray-white hairs, leaves are leafy and viviparous, ovate or ovate oblong of 2.5-10 cm in length and 2.5-7 cm in width. Easy to break up There is a mount at the edge, the surface is dark brown, the back side is gray brown, and there is a triangular shape. It has an unusual scent, the taste is a bit bitter. Lots of fresh leaves, strong fragrance is good. The main ingredients are Pachoulic alcohol, Methylchvicol (esdragol), Cycoseychellene and Pogostol. Pharmacological action, methyl alcohol extract has a strong antimicrobial action against skin fungi.

Licorice is a leguminosae mainly distributed in China, Russia, Spain, Iran, Afghanistan, and has a peculiar smell and sweet taste. The transverse section has a yellowish brown cork layer with 1 to 3 cell cork layers inside. In the flow cells of the cortex and the neck, there are monoclinic and starch granules. The quality is hard, powdery, and the cut surface is yellowish white. Glucuronic acid, a degradation product of Glycyrrhiza from Glycyrrhiza in licorice, binds harmful substances in the liver of the body and detoxifies it as glucuronide, and also has anti-allergic effect in dermatology. Another degradation product, glycyrrhetinic acid has anti-inflammatory and corticosteroid effects.

Sodium bicarbonate (NaHCO ₃ ) is also known as "heavy bath", "acidic sodium carbonate", "sodium bicarbonate", "sodium bicarbonate". Colorless crystalline powder, specific gravity 2.20. It is used in medicines such as antacids, alkalis, and side effects inhibitors when taking sulfa drugs. Applications such as gas generators are very diverse.

Feed additives having a diarrheal disease preventive activity of calf containing natural herbal medicines prepared as described above are fed by adding 0.1 to 0.3 parts by weight based on 100 parts by weight of the total amount of daily feed (rich) feed when used in farms or In addition, the feed is prepared by adding 0.1 to 0.3 parts by weight to 100 parts by weight of the total feed according to the growth stage to the total blending ratio of the calf feed on demand in the compound feed plant, and feed the calf.

The calf grown by feeding the feed additive according to the present invention shows a weight gain of 5.5 to 12.0% higher than the calf fed the normal calf feed in body weight to improve the productivity of the livestock industry and increase the immunoglobulin of livestock , The calf diarrhea rate was significantly reduced by 15 to 43%.

Therefore, the calf fed a feed additive containing a natural herbal medicine prepared by the method of the present invention has increased immunity and the treatment period is reduced by 50 to 60% even in the case of calf individuals suffering from diarrhea. As a result, treatment and production costs can be reduced.

Liver tissue damage, measured for toxicity and stability studies of feed, can be confirmed by measuring the outflow of enzymes present in cells into the blood or by monitoring pericentral necrosis. Thus, measuring the activity of enzymes released into the blood from the liver is one of the most useful methods in studying liver damage. GOT is an enzyme that transfers the amino group of glutamic acid to oxaloacetic acid. When the amino group is transferred, glutamic acid becomes α-ketoglutaric acid and oxaloacetic acid becomes aspartic acid. GPT is an enzyme that transfers the amino acid group of glutamic acid to alanine. When the amino group is transferred, glutamic acid becomes α-ketoglutaric acid and alanine becomes pyruvic acid. Therefore, when cells are destroyed, they are released into the blood and appear high. Elevation of GOT and GPT in serum may be an indicator of hepatic cell degeneration and necrosis because aminotransferase is released into the blood as the necrosis of liver cells and the destruction of liver tissues are caused by liver damage. .

In the toxicity test of the rats by adding the feed additive according to the present invention, the death of the rat was not observed even at the highest dose. The higher the amount of the feed additive according to the present invention, the higher the content of the immunoglobulin. It can be confirmed that the feed additive prepared by the method improves the immunity of the calf.

As described above, the present invention, by developing a feed additive based on natural herbal medicines, the calf fed the feed is grown healthy without diarrhea, and can increase feed intake to reduce treatment costs and ultimately improve productivity. have.

The present invention for achieving the above technical problem provides a feed additive for preventing the calf diarrhea produced by the method and the method of increasing the usability of the herbal medicine.

Feed additives for calf containing herbal medicines produced according to the method of the present invention is an eco-friendly product that does not accumulate antibiotic residues on the market. Can contribute to income

Hereinafter, the present invention will be described in more detail with reference to Examples. In the present invention, the examples are for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Test Example 1 Clinical efficacy test of feed additive containing natural herbal medicine

Example 1-1 Preparation of test animals and barn

The test publication began with 24 Korean calves of about 70 days of age. The stall type was carried out in an open-type barn of 4 × 6 m per compartment and Test Example 1 was placed in three treatments (8 heads x 3 treatments = 24 heads) by adding 8 cow calves. Winch curtains were installed on both sides to prevent wind in winter, and air was well ventilated in summer, and large fans were installed on the ceiling.

As the feed used for the test, the control group was used commercially available beef cattle calf feed containing the ingredients shown in Table 2, and the test feed was calf added with a feed additive containing a natural herbal medicine customized to the G-axis The feed was purchased and paid, but the feed rate was limited by growth stage, and the negative water freed the fresh water produced from the 250m underground rock well.

Example 1-2 Preparation of Feed Additives Comprising Natural Herbal Medicines

Gold, silver, pogongyoung, local herb, kwakhyang and licorice are shredded about 3 to 5 cm without crushing, 1 kg of each herb is placed in a large pot, and 10L of water is added and evaporated for 4 hours or more. When it was about L, the mixture was stopped and filtered with three layers of gauze to obtain the final hot water extract.

Twelve herbal medicines such as Baekchul, Dermis, Chajeon, Garyeong, Baekbokryeong, Broiler, Japonica, Baekji, Daebokpi, Banja, Gilgyeong, and Taeksa were first crushed with an electric grinder and the size of the grinder was 30 (3.5Φ = 3mm). After the first crushing, and then again passed through the 10 neck (1.0Φ = 0.3 mm) crusher and pulverized into fine powder, it was put into a clean bench disinfection tank equipped with ultraviolet fluorescent lamp and sterilized for 24 hours.

The hot water extract was diluted 10 times with water, and then mixed well in a volume ratio of 1: 1: 1: 1: 1. 3L of the mixed extract and 0.5 kg of each of the twelve herbal medicine powders were mixed and mixed for at least 1 hour so as not to form a lump in the automatic mixer. 0.6 kg of sodium bicarbonate (NaHCO ₃ , Prenatal Pure Chemical Industries, Ltd.) was added to the mixed herbal medicine mixture, followed by mixing in an automatic mixer.

Then, in the mixture, strains of Lactobacillus acidophilus , Aspergillus oryzae, and Saccharomyces cerevisiae were more than 5 × 10 ⁵ CFU / kg. The included probiotic was added at a weight ratio of 10: 1 and mixed to prepare a feed additive.

Example 1-3 Formulation of Test Feed and Calf Breeding Method

Feeding the diet was fed from 70 days of age to 220 days of age as shown in Table 1, and the mixing ratios and general ingredients of the control and feed fed diets are shown in Tables 2 and 3.

The feed of the control group was fed the young calf feed produced in the G-axis, the test group feed the feed additives prepared by the method of Example 1-2 containing natural herbal medicines in the young calf feed produced in the G-axis Order feed prepared by adding 0.1 kg and 0.3 kg to 100 kg of feed was fed. At this time, the feed composition is shown in Table 2, and the analysis of the feed composition was analyzed according to the AOAC (1998) method. In addition, the control was carried out according to the Farm Practices Act, and the survey fee was imported from T.Mich (from the start of the test up to 3 months of age) and rice straw (from the test of 4 months of age), and the water was free from drinking water. In order to freeze the rinkal block was suspended separately.

TABLE 1

division 70-160 days 161-220 days Remarks Control General feed General feed 8 batches T1
(Test 1 ball) Custom Test Feed
(0.1 parts by weight of feed additive) Custom Test Feed
(Added 1 part by weight of feed additive) 8 batches T1
(Test 1 ball) Custom Test Feed
(0.1 parts by weight of feed additive) Custom Test Feed
(Added 2 parts by weight of feed additive) 8 batches Investigation
Free vegetarian Free vegetarian Timothy Hay Crested straw

※ Each treatment group has a ratio of female (50%) and male (50%).

TABLE 2

[Table 3]

Example 1-4 Measurement of Feed Intake and Weight Gain

Feed intake and weight gain

Body weight was measured monthly, and feed was divided into two daily meals at 8:00 am and 17:00 pm, and the remaining amount was weighed before weighing the morning of the next day, and the remaining daily amount was subtracted from the total daily feed. The intake was calculated and the feed demand rate was calculated by dividing the total weight gain by the feed intake.

2. Blood collection and ingredient analysis

At the beginning of the trial, in the growing and finishing periods, 20 ml of blood was collected through the calf jugular vein using heparin-treated vacumtainer ^® (Becton Dicknson, Franklin Lakes, NJ, USA) at 3 months of age. 10ml each was divided into the untreated tube. After standing for 2 hours at room temperature to coagulate the blood, centrifuged for 15 minutes at 4 ℃, 3,000rpm to separate pure serum only and stored in an ultra-low temperature freezer at -70 ℃ until analysis, the automatic blood analyzer (Hema Vet 950) The analysis was carried out.

3. Sample Collection and Temperature Measurement

The test axis was collected by rectal extraction method, in which the subjects were calibrated on the test day every month, and their hands were pulled out into the anus, and then buried in a sterile swab previously put in a vacuum tube and immediately transferred to an analysis room to isolate and identify bacteria. At the same time, the thermometer was pushed into the anus and the temperature was measured.

Example 1-5 Component Analysis of Feces

1.Isolation and analysis of bacteria (E. coli)

5-10 g of calf feces were collected in a fecal collection container (Greiner, Germany), and enriched in 45 ml of MEC broth (Merck, Germany) at 37 ° C. for 18 to 24 hours, and the isolated culture was blood medium (Asan Pharmaceutical). And EMB (Merck, Germany), which were plated directly on MacConkey agar (Merck, Germany) and subjected to selective culture at 37 ° C. for 18 to 24 hours, followed by 3 to 5 bacteria causing β-hemolysis and suspected red colonies. ), Incubated at 37 ° C for 24 hours, and then subjected to gram staining, biochemical characterization by selecting metallic gloss colony [4-methylumbellifervl-β-d-glucuronide (MUG) test, indole test, methyl red (MR) test , Voges-Proskaur (VP) test, citrate test, gas production test from lactose], API 20E kit (BioMerieux, France) or microbial isolation identification (Vitek system, BioMerieux, France) test. Separated.

2. Parasite (Coccidium) Test (Saturated Saline Floating Method)

Take about 20 ml of saturated saline solution in a small beaker, add 2 g of test material, and stir well.Place a metal funnel or a layer of gauze on a small funnel, filter the fecal fluid into a test tube, place it in a test tube, and apply the surface tension to the spout. Saturated saline was slowly added until a swelling surface was formed. After that, it is left for 30 to 40 minutes and then the cover glass is held horizontally and carefully contacted with the swelling surface to attach the suspended matter. This was placed on a slide glass, and the coccidium protozoa was examined with an optical microscope (100 times).

3. Viral (BVDV PCR) method (RNA extraction and RT-PCR method)

RNA was extracted from feces using the RNeasy mini kit (Qiagen, Germany) according to the manufacturer's instructions. PCR was performed using a VeTek ^™ BVDV Detection Kit (iNtRON, D60060). In the PCR reaction, 2ul of extracted RNA and 18ul of water without DNase / RNase were added to the kit so that the total amount was 20ul. After reacting for 30 minutes at 45 ℃ synthesized cDNA and then reacted for 5 minutes at 94 ℃, 30 times at 94 ℃, 30 seconds at 58 ℃, 40 times each 40 seconds at 72 ℃ and then 5 at 72 ℃ The reaction was carried out for a minute. After completion of the PCR reaction, 10 ul of each reaction solution was taken and subjected to electrophoresis on 1.5% agarose gel containing ethidium bromide, and then the presence or absence of 297bp band under ultraviolet light was observed.

Example 1-6 Statistical Analysis

Statistical analysis was performed by T-test according to GLM (General Linear Model) analysis method of SAS program (2002), and the comparison between means of treatment was Duncan (1955) 's multiple test method at 5% level. It was.

Results of Test Example 1

1. Changes in Body Weight, Daily Weight Gain and Feed Demand Rate of Test Calves

Feed intake and weight gain measured from the Example was confirmed to be excellent in weight gain and feed intake for the entire test period (150 days), as shown in Tables 4 and 5 below.

Table 4 lists changes in body weight, weight gain, and daily weight gain during the test period, and Table 5 describes changes in feed intake during the test period.

[Table 4]

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) adding 0.1 parts by weight of the feed additive of Example 1-2; 3) standard error of the mean; 4) Statistical T-test test of mean values according to treatment Pr> │t │

The results of feeding the feed additives prepared by the method of Example 1-2 to the Korean calf during the whole test period (150 days). The initial body weight of the control, test 1 and test 2 were 46.10 kg, 43.70 kg and 44.60 kg, respectively, and the same age and weight were 2 to 3 kg less than the control, but the overall weight was similar. The test was undertaken. The control, test 1, and test 2 groups at the age of 5 months were 183.9 kg, 186.5 kg, and 176.8 kg, respectively, and the highest growth rate was found in the test 1 group (P <0.0465), followed by the control, test 2 The results were in order of order, and these results were similar in the total weight gain and daily weight gain.

As such, the test 1 was significantly higher than the other treatments at the same age compared to the control or the test 2, and it was determined that the addition of the feed additive according to the present invention had not only a therapeutic effect on diarrhea but also an immune and digestive effect.

TABLE 5

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) adding 0.3 parts by weight of the feed additive of Example 1-2; 3) Standard Error for Mean

a-b: Statistical T-test test of mean value by treatment

The results of the change in feed intake fed the feed additive prepared according to the method of Example 1-2 to the Korean calf during the whole test period (150 days). At the start of the test, the feed intake per head was 32.7kg, 39.8kg, and 32.0kg in control, test 1, and test 2, respectively. At this time, feed intake per head was 1.09kg, 1.33kg, and 1.07kg, respectively. There was a tendency to be somewhat higher than control and test 2. However, the statistically significant difference was found in the control diet, control group, test 1 group, and test 2 group at 5 months of age (176.3kg, 164.6kg, 162.6kg, respectively) (P <0.0436). 5.88kg, 5.49kg and 5.42kg showed that the first test was slightly higher than the control and the second test.

2. Blood Characteristics and Characteristics of Test Calf

Total Cholesterol content in the blood tended to decrease as the test progressed, but overall, the lower total cholesterol content in the 3 month-old test than the 1-month old test was due to the lower milk absorption.

On the other hand, test 1 showed a higher tendency than other treatments due to higher feed intake and weight gain than other treatments.

Triglyceride, meanwhile, was in the treatment range (4.29 to 20.14 mg / dl), and GOT, which represents the criteria for liver function, was in the range of 69.14 to 85.86 mg / dl, and gradually increased as the test progressed. It appeared to decrease. In the case of GPT, the range of 17.71 to 22.71 mg / dl, as in the GOT, tended to decrease as the test proceeds. In particular, GOT and GPT of the test group showed a stable response with little difference compared to the control group. Glucose and blood urea nitrogen (BUN) showed little difference between treatments.

Physicochemical properties in the blood taken from the jugular vein of Hanwoo calves were found in the range of 7.08 to 12.62 k / ul for WBCs (Leukocytes) and 8.86 to 19.45 M / for RBCs (Erythrocytes). was in the range of ul. Hemoglobin (Hb) levels ranged from 9.76 to 12.07 g / dl with little difference between treatments.

In particular, in the case of IgG, an immunoglobulin, the average control was 574.38 mg / dL, but the first test was 605.19 mg / dL, and the second test was 601.81 mg / dL. There were no significant differences between 673 and 688 mg / dL at the start of the trial, but at 3 months of age, the control was 559 mg / dL, but the first trial was 598 mg / dL, and the second trial was 602 mg / dL. The test group was about 6-7% higher than the control group. In addition, the control group was 476 mg / dL at 5 months of age, but the test group was 544 mg / dL, and the test group was 525 mg / dL, which was about 8 to 14% higher than the control group.

These results indicate that the immunity of the test group in which the feed additive including the natural herbal medicine is added to the feed is improved, and as a result, it is expected to significantly affect calf diarrhea prevention and shorten the treatment period.

Table 6 lists the results of physicochemical properties in serum, and Table 7 lists the results of physicochemical properties in blood.

TABLE 6

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) adding 0.3 parts by weight of the feed additive of Example 1-2; 3) Standard Error for Mean

TABLE 7

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) adding 0.3 parts by weight of the feed additive of Example 1-2; 3) Standard Error for Mean

3. Characteristics of Calf Fecal and Calves Diarrhea

Fecal samples were collected from 1 month of age to 5 months of age (150 days), and the causative organisms were separated and divided into parasites (coccidium), bacteria (hemolytic Escherichia coli) and viruses (BVD). And the result of the treatment days. Overall, the causative agent was high at 1 month of age, and as the test progressed, the causative agent was lower.

In particular, hemolytic Escherichia coli contained more than control group (average 1.8) than test group (average 0.8 to 1.0), and virus (BVD) control group (average 3.2) and test group (average 0.8 to 1.2). Parasite (coccidium) was also higher in the control group (mean 0.6) than in the test group (mean 0.2-0.4).

In addition, the treatment period of the affected individual is shorter than the control group (0.8-0.9 days) than the control group (1.5 days), thereby reducing the treatment cost and the production cost. The rectal temperature of the diarrheal calves ranged from 38.10 to 38.40 ° C on average.

Also, test 1 group had the shortest incidence of diarrhea and duration of treatment compared to other treatment groups.

Therefore, the test and the results of the test calf feces and calf diarrhea incidence and treatment status results are shown in Table 8.To summarize the feed additives containing the natural herbal medicine according to the present invention for the prevention of diarrhea of the calf It was found to be more useful as a feed additive.

[Table 8]

※ The treatment period is calculated to be 1.0 days combined with morning (0.5 days) and afternoon (0.5 days) treatment.

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) adding 0.3 parts by weight of the feed additive of Example 1-2; 3) Standard Error for Mean

a-b: The statistically significant difference (p <0.05) of the mean value according to treatment.

The incidence of diarrheal lesions and respiratory disease was 7 in the control, 3 in the test 1, 6 in the test 2, and the incidence of the control was higher than the test. As a result, the treatment costs were 43,940 won for the control group and 27,780 won for the test 1 group and 30,280 won for the test 2 group, which was much more expensive than the control group. In addition, even during the treatment period, the test group showed a faster recovery by shortening the treatment period by about 40 to 50% than the control group. Calf disease treatment history for the entire trial (150 days) is shown in Table 9 below.

These results suggest that the disease resistance, or immunity, has increased.

TABLE 9

※ The number of heads: counts as one head even if the same individual repeatedly occurs at intervals of 10 days

※ Cost of Treatment

① Ephedrine: 50㏄-1,000 won. Bite reel 50 ㏄-20,000 won. Anapa 100 won-25,000 won.

   Copamycin costs 50-4,000 won. Jesrong 10-3,000 won. Combined buscopan 100 ㏄-22,000 won.

   Comityl 50-2,000 won. Antihistamine 100㏄-1,500 won. Vitamin 50-6,000 won.

   King Bellin 50㏄-1,200. Apramycin 50 ㏄-10,000 won. Jesrong 20 ㏄-2,000 won.

② Labor cost is not included in the total treatment cost calculation.

1) Add 0.1 part by weight of the feed additive of Example 1-2; 2) Add 0.3 parts by weight of the feed additive of Example 1-2

a-b: The statistically significant difference (p <0.05) of the mean value according to treatment.

To summarize the above test 1, the feed additive of Example 1-2 was added to the calf calf from 2 months of age to 7 months of age, and 0.1 parts by weight of the feed additive of Example 1-2 was added. One test group gained 142.8 kg, while the control group gained 137.8 kg, resulting in a 3.6% gain. In addition, the test group which added 0.1 parts by weight of immunoglobulin (IgG) for diarrheal disease was 605.19 mg / dL, but the general control group was 574.38 mg / dL, which improved the immunity of about 5%. Therefore, the number of diarrheal heads and the number of treatment days during the entire test period were treated with three heads for six days, while the control group treated with seven heads for 12 days, and the two treatments were treated with six heads for 7 days.

As a result, the cost of treatment was higher in the control group (43,940 won), and the test group was 27,780 won and the test group 2 was 30.280 won. .

Test Example 2 Comparison of effects between feed additives containing natural herbal medicines and commercial feeds

Example 2-1 Preparation of test animals and barn

The test began with 28 Korean cow calves from birth (1 day) to 3 months (100 days of age). The stall type was carried out by placing 4 cows (7 cows × 4 cows = 28 cows) by adding 7 cow calfs in the open barn of 4 × 6 m per square.

Winch curtains were installed on both sides to prevent wind in winter, and air was well ventilated in summer, and large fans were installed on the ceiling.

The feed used for the test was a commercial Hanwoo calf feed (Nonghyup feed) for the control, and the feed for the test was added to the feed additive containing a natural herbal medicine prepared in Example 2-2, which was custom-made in G-axes. Purchasing exclusive feed (Table 2) was provided, but the feed was limited by growth stage, and water was freed.

Example 2-2 Preparation of Test Feed

Gold, silver, pogongyoung, local herb, kwakhyang and licorice are shredded about 3 to 5 cm without crushing, 1 kg of each herb is placed in a large pot, and 10L of water is added and evaporated for 4 hours or more. When it was about L, the mixture was stopped and filtered with three layers of gauze to obtain the final hot water extract.

Twelve herbal medicines such as Baekchul, Dermis, Chajeon, Garyeong, Baekbokryeong, Broiler, Japonica, Baekji, Daebokpi, Banja, Gilgyeong, and Taeksa were first crushed with an electric grinding machine and the size of grinder was 30 (3.5Φ = 3mm). After this first crushing, and then again passed through the 10 wood (1.0 Φ = 0.3 mm) crushing sieve and pulverized into fine powder, it was sterilized for 24 hours in a clean bench disinfection tank with an ultraviolet fluorescent lamp.

The hot water extract was diluted 10 times with water, and then mixed well in a volume ratio of 1: 1: 1: 1: 1. 3L of the mixed extract and 0.5 kg of each of the twelve herbal medicine powders were mixed and mixed for at least 1 hour so as not to form a lump in the automatic mixer. 0.6 kg of sodium bicarbonate (NaHCO ₃ , Prenatal Pure Chemical Industries, Ltd.) was added to the mixed herbal medicine mixture, followed by mixing in an automatic mixer.

Then, in the mixture, strains of Lactobacillus acidophilus , Aspergillus oryzae, and Saccharomyces cerevisiae were more than 5 × 10 ⁵ CFU / kg. The included probiotic was added at a weight ratio of 10: 1 and mixed to prepare a feed additive.

The feed additive thus prepared was added to 0.1 kg and 0.3 kg for the total 100 kg of customized calf feed at the feed compound to prepare the feed and fed the calf.

Example 2-3 Breeding Method

For the rich feed of the control group, the feed for the young calf produced in G-axes was supplied, and for the C-1, the commercial probiotics (Product Name-ISPI, CTC Bio) + Commercial IgY (Product Name-Azide Drink, Ed Biotech Co., Ltd.) ) C-2, commercially available IgY (product name-Azidelink, EDBIOTECH Co., Ltd.), C-3, commercially available natural herbal herbal medicine 0.1% + commercially available IgY (product name-Azidrink, EDBIOTECH Co., Ltd.), The test zone was fed a test diet feed 0.1 parts by weight of a feed additive containing a natural herbal medicine prepared by the method of Example 2-2.

At this time, the feed composition is shown in Table 2, and the feed ingredient analysis was analyzed according to the AOAC (1998) method. In addition, the control was carried out according to the Farm Practices Act, and the control fee was imported from Klein (from the start of the test until May). It was forever.

※ Each treatment group has a ratio of female (50%) and male (50%).

※ Example 2-2 in the above table means a feed additive prepared by the method of Example 2-2.

Example 2-4 Measurement of Feed Intake and Weight Gain

Body weight was measured monthly, and feed was divided into two daily meals at 8:00 am and 17:00 pm, and the remaining amount was weighed before weighing the morning of the next day, and the remaining daily amount was subtracted from the total daily feed. The intake was calculated and the feed demand rate was calculated by dividing the total weight gain by the feed intake.

Example 2-5 Statistical Analysis

Statistical analysis was performed by T-test according to GLM (General Linear Model) analysis of SAS program (2002), and the comparison between means of the treatment was performed by Duncan (1955) at 5% level. .

Result of Test Example 2

The following table 10 shows the change in weight of the calf and the weight gain per day.

TABLE 10

1) Standard Error for Mean

a-b: The statistically significant difference of the mean values according to treatments (p <0.05)

2) 'Example 2-2' in the table means a feed additive prepared by the method of Example 2-2-.

Hanwoo calves were reared from birth until weaning, 100 days (about 3 months). The initial body weight was the highest at 28.57 ㎏ C-1, 30.3 ㎏ C-2, 28.3 ㎏ C-3 and 30.4 ㎏ test, but the weight of 100 days of age, weaning period, was 118.6 ㎏. The rate was good, followed by C-3, C-2, and C-1.

Therefore, the total weight gain and daily weight gain over 100 days were 73.57kg and 0.74kg in C-1, 76.75kg and 0.77kg in C-2, 82.58kg and 0.83kg in C-3, and 88.20kg and 0.88kg in Test. Compared to farm practices, the addition of the feed additive containing the natural herbal medicine according to the present invention showed a significantly higher growth rate.

These results indicate that the feed additives containing natural herbal medicines had a significant effect on the immunity and rumen stabilization of Hanwoo calves.

Table 11 below describes the onset of diarrhea of the calf fed the feed containing the feed additive of Example 2-2 during the whole test period (100 days).

 1) Standard Error for Mean

 a-b: The statistically significant difference (p <0.05) of the mean value according to treatment.

 2) 'Example 2-2' in the table means a feed additive prepared by the method of Example 2-2.

This is the result of analyzing the current condition and causative organisms of diarrhea in Korean cattle calf during the whole test period. In farming practices, probiotics and IgY were fed, and 4 cases of C-1, 2 cases of C-2, and 1 case of C-3 occurred in the number of calf diarrhea, but were prepared by the method of Example 2-2. No test group was added with 0.1 kg of feed additive.

The above results can be seen that the calf diarrhea prevention effect of the feed additive containing a natural herbal medicine.

Test Example 3 Toxicity and Stability Test

Example 3-1 Experimental Animals and Test Preparation

Subacute toxicity test was conducted to confirm the safety of feed additives containing natural herbal medicines prepared by the method of the present invention.

70 SD rats with a weight of 80 to 90 g were purchased for 7 days, and then visually observed for general symptoms, and 64 healthy rats and males were used (♀ 32, ♂ 32) for the experiment. Breeding conditions are the temperature 22 ± 2 ℃, relative humidity 55 ± 5%, contrast time 12 hours (light / dark cycle), the feed is prepared in Example 1-2 by powdering a rat feed for rats dedicated experimental animals 1 g, 3 g, and 5 g of mixed feed additives were applied to 100 g of feed, and water was freed.

Example 3-2 Symptom Observation, Body Weight, Feed Intake Measurement

Observation of the general symptoms according to the administration of the feed additives of Example 1-2 was made freely for the experimental animals, paying attention to the symptoms that may appear at a certain time every day change of general symptoms, poisoning symptoms and the presence of dead animals Observed. In addition, the weight change was measured before and after the daily diet, and the weight before the start of the experiment was subtracted from the final weight and expressed as the weight gain during the experiment. Feed intake was measured every day at a fixed time and then subtracted from the remaining amount.

Example 3-3 GOT, GPT and Immunogen (IgG) Irradiation

Observation of the general symptoms according to the feed additive administration of Example 1-2 was made freely for the experimental animals, and carefully observed daily for symptoms that may occur at a certain time, blood GOT (glutamic oxalacetic transaminase) and GPT ( Activity of glutamic pyruvic transaminase was measured by assay kit (Asan Pharmaceuticals) by Reitman-Frankel method (1), and the activity unit was expressed as IU / L serum.

On the other hand, immunoglobulin measurement was commissioned by neodine Co., Ltd. to measure IgG, IgA, IgE.

Example 3-4 Statistical Analysis

All experimental results were expressed as mean ± standard deviation using SPSS statistical program, and Duncan's multiple test at p <0.05 level tested the significance of treatment interval.

Result of Test Example 3

Table 9 shows the daily weight gain and feed intake of rats during the experiment.

In the increase of body weight, the control group (3.23g) was the lowest in the female and male groups, and the experimental group (4.06-4.59g) added 1 to 5 parts by weight of the feed additives of Example 1-2 showed similar body weight. . In addition, the control group (12.34g) was the smallest in the feed intake of both males and females, and the experimental group (13.5-14.59g) to which the feed additive of Example 1-2 was added 1-5 parts by weight showed similar intake.

As can be seen in Table 12, in the experimental group fed the feed containing the feed additive including the natural herbal medicine according to the present invention, it can be seen that the weight gain is higher and the feed intake is also increased.

TABLE 12

1) a-b : 처리구별에 따른 평균치의 통계적인 유의차(p<0.05).

1) ab: The statistically significant difference of the mean values according to treatments (p <0.05).

2) 'Example 1-2' in the table means a feed additive prepared by the method of Example 1-2.

TABLE 13

※ 'Example 1-2' in the above table means a feed additive prepared by the method of Example 1-2.

As shown in Table 13 above, the rats were mixed with the feed additive prepared in Example 1-2 in powdered feed with 1 part by weight, 3 parts by weight, and 5 parts by weight, based on 100 parts by weight of the whole feed, for 28 days. As a result of observing the general clinical symptoms, no special findings were found to be due to natural herbal feed additives such as changes in exercise status and appearance.

 ※ Deaths / total experimental animals

 ※ 'Example 1-2' in the above table means a feed additive prepared by the method of Example 1-2.

Table 14 shows the mortality rates of test mice. As shown in Table 14 above, the feed additive prepared in Example 1-2 was added to the powdered rats by adding 1 part by weight, 3 parts by weight, and 5 parts by weight to 100 parts by weight of the whole feed, and then fed the SD system. As a result of investigating mortality due to toxicity for 28 days in rats, both males and females did not die during the experimental period. No mortality was observed even at the highest dose of 5%. LD ₅₀ values could not be calculated. LD ₅₀ values of male and female were estimated to be 5% or more.

TABLE 15

※ a-b: The statistically significant difference of the mean value according to treatment group (p <0.05).

※ 'Example 1-2' in the above table means a feed additive prepared by the method of Example 1-2.

Table 15 lists the immunoglobulin measurement results. In females, IgG was 13.67, 17.00, and 30.33 mg / dL in the experimental group in which the control group was 12.67, 1 part by weight, 3 parts by weight, and 5 parts by weight of the feed additive of Example 1-2. The addition of 1 part by weight of feed additive did not show a significant increase, and the highest increase was found in the addition of 5 parts by weight. In IgA, the control group was 2.00 mg / dL, and in the experimental group, 1 part, 3 parts, and 5 parts by weight, 0.67, 0.33, and 0 mg / dL, respectively, showed no obvious enhancement. In IgE, the control group was 0.23 mg / dL, and in the experimental group, 1 part, 3 parts, and 5 parts by weight were 0.6, 0.33, and 0.57 mg / dL. In males, IgG was 7.33 mg / dL in the control group, 1 part by weight, 3 parts by weight, and 5 parts by weight of 7.67, 13.67, 9.00 mg / dL in the control group. In the group of 1 part by weight of the feed additive according to Example 1-2, there was no obvious increase and there was the highest increase in 3 parts by weight. In IgA, the control group was 0.33 mg / dL, and the experimental group 1 part, 3 parts, and 5 parts by weight were 0.33, 1.00, and 2.00 mg / dL, respectively, in proportion to the concentration of feed additives. In IgE, the control group was 0.23 mg / dL, and the experimental group was 1 part by weight, 3 parts by weight, and 5 parts by weight of 0.33, 0.47, 0.24 mg / dL.

In the test, the normal group was used as a control group, and the feed additives of Example 1-2 were mixed with 1 part by weight, 3 parts by weight, and 5 parts by weight of the feed, respectively, and fed to rats, and then the degree of liver damage in the experimental group was examined. In order to see the changes in the enzyme activity of GOT, GPT in the blood was measured and the results are shown in Table 16 below.

TABLE 16

  ※ a-b: The statistically significant difference of the mean value according to treatment group (p <0.05).

  ※ 'Example 1-2' in the above table means a feed additive prepared by the method of Example 1-2.

As a result of toxicity test by adding 1 to 5 parts by weight of a feed additive comprising a natural herbal medicine prepared according to the method of the present invention to rats in Test Example 3 above, the toxicity was obtained even at the highest dose of 5 parts by weight. Death did not occur, and no general symptoms were found.

In addition, the immunoglobulin showed a tendency to increase the IgG content as the amount of the feed additive according to Example 1-2 increases, so that it is possible to estimate the improvement of immunity, and the symptoms of liver damage did not occur in GOT and GPT.

The present invention is not limited to the technical spirit of the specific examples, test examples, drawings or tables described above, and the general knowledge in the technical field to which the invention belongs without departing from the spirit of the invention claimed in the claims. Of course, any person having a variety of modifications can be made, and such changes are within the scope of the claims.

Figure 1 shows the manufacturing process of the feed additive having a diarrheal disease prevention activity of calf containing a natural herbal medicine according to the present invention.

Figure 2 is a graph comparing the change in weight gain of the Hanwoo cow calf fed the feed comprising a feed additive prepared by the method according to the present invention and the control group.

Figure 3 is a graph comparing the diarrheal incidence of the Hanwoo cow calf fed the feed comprising a feed additive prepared by the method according to the present invention and the control group.

Figure 4 is a graph comparing the number of days of treatment for diarrheal disease of the Korean cattle calf fed the feed containing a feed additive prepared by the method according to the invention and the control group.

Figure 5 is a photograph showing an example of the production of a feed additive having a calf diarrheal disease preventive activity produced by the method according to the present invention.

Claims (3)

1) finely cut gold silver, pogongyoung, local vinegar, kwakhyang and licorice, and boiled by adding water in a weight ratio of 1: 10 to evaporate water to obtain respective hot water extracts; 2) diluting each of the hot water extracts of step 1) with 10 times of water and then mixing them in the same volume ratio; 3) Grind the white powder, dermis, chajeon, seonbi, baekbokyeong, broiler, pomegranate, white paper, daebokpi, halves, gilkyung, and tack with fine powder of 0.3 mm or less and take each fine powder in the same weight ratio and mix powder Obtaining step; 4) the hot water extract mixture of step 2): the mixed powder of step 3): sodium bicarbonate prepared in a weight ratio of 1: 2: 0.2 by first adding the mixed mixture of the hot water extract solution and step 3) to the automatic mixer After stirring, mixing sodium bicarbonate there; And 5) The probiotic consisting of Lactobacillus acidophilus , Aspergillus oryzae and Saccharomyces cerevisiae was added to the mixture obtained in step 4). A method for preparing a feed additive having a diarrheal disease prevention activity of a calf containing a natural herbal medicine, characterized in that the step of mixing in a weight ratio. Feed additive having a diarrheal disease preventive activity of the calf containing a natural herbal medicine prepared by the method according to claim 1. Feed prepared by adding a feed additive having a diarrheal disease preventive activity of the calf containing the natural herbal medicine according to claim 2 with respect to the total feed amount.

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