KR20200009629A - Process for Compositions of Disinfectants for Viruses of Porcine Epidemic Diarrhea & Porcine Reproductive and Respiratory Syndrome Including Extract of Pleurotus nebrodensis - Google Patents

Abstract

The present invention relates to a disinfectant composition of porcine epidemic diarrhea virus (PEDV) and porcine reproductive and respiratory syndrome virus (PRRSV), composed by containing, in 95.5 parts by weight of purified water, 5.0 to 7.0 parts by weight of a Pleurotus nebrodensis extract, 3 parts by weight of a Perilla frutescens extract, 2 parts by weight of ionized mineral particles, 1 parts by weight of sodium cyanate (NaOCN), 0.5 parts by weight of glutaraldehyde (C_5H_8O_2), 0.5 parts by weight of guar hydroxypropyl-trimonium chloride, and 0.5 parts by weight of citric acid (C_6H_8O_7). The disinfectant composition of PEDV and PRRSV containing the Pleurotus nebrodensis extract, the Perilla frutescens extract, and minerals according to the present invention has an excellent disinfection effect for PEDV and PRRSV, and has little effect on environmental pollution.

Description

Process for Compositions of Disinfectants for Viruses of Porcine Epidemic Diarrhea & Porcine Reproductive and Respiratory Syndrome Including Extract of Pleurotus nebrodensis}

The present invention relates to a method for preparing a disinfectant composition of porcine diarrhea virus (PEDV) and respiratory virus (PRRSV) containing the king mushroom extract. More specifically, the preparation of antiseptic composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV), characterized in that the extract contains king mushroom extract, perilla extract and minerals ionized by electrolysis and nanonization by ultrasonic grinding A method and an antiseptic composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) prepared therefrom.

Swine diarrheal disease is one of the most important factors for productivity loss in the swine industry, such as not only high mortality rate but also low growth rate of piglets that have been infected and recovered, and increased susceptibility to other diseases. This has caused enormous damage. The major viral diarrheal diseases in pigs are known as transmissible gastroenteritis (TGE), porcine epidemic diarrhea (PED), and swine rotavirus infection.

Among these, swine pandemic diarrhea (PED) occurs regardless of age, and is characterized by vomiting and watery diarrhea, and is very similar to TGE, but mortality is lower than TGE in mammalian pigs less than 2 weeks old, and the incidence in finishing pigs and young pigs is lower. It is more common. It is an acute infectious disease that causes severe abdominal pain, loss of appetite, and depression in piglets by infection with the pandemic diarrhea virus belonging to the corona virus.

The biological and physicochemical properties of the swine epidemic diarrheal virus (PEDV) are similar to the swine infectious gastritis (TGE) virus, but belong to the coronavirus family, which has an unrelated crown. It is killed in ether and chloroform, and is contagious when heated for more than 30 minutes at a temperature of 60 ° C. or higher, but is generally stable at a temperature of 50 ° C. Although data on the propagation distance of PED viruses are insufficient, they are thought to have similar propagation power to TGE viruses belonging to the same virus family.

Porcine reproductive and respiratory syndrome (PRRS) was first reported in the United States in the late 1980s, and has been confirmed in many countries around the world, including Europe and Korea, and now in Asia. It occurs in almost every country. Pandemic PRRS is characterized by collective breeding disorders in gestational pigs, incidence of weak piglets and respiratory illness in all ages of pigs. After the epidemic, it is converted to endemic, causing reproductive dysfunction, increased respiratory disease and decreased growth of hog pigs in sows, and postweaning multisystemic wasting syndrome (PMWS). And one of the leading causes of porcine respiratory disease complex (PRDC). As such, PRRS is reported to cause about 600 million dollars in economic damage annually in the United States alone, causing various disease damages.

The PRRS virus (PRRSV), the cause of the genital respiratory syndrome (PRS) in pigs, has a variety of propagation paths, including contact infection with infected pigs, infection with a runny nose or urine, air infection, and transmission through infected semen. Because it is so fast, the virus spreads and spreads in a short period of time in a farm or area where an infection has occurred. In addition, because the virus is excreted from the infected pigs for a long time, it is difficult to break the circulatory loop of the virus, and it is difficult to prevent and eradicate the infection due to the increase in the subclinical type of infection that does not show a clear clinical symptom.

Pig epidemic diarrheal disease (PED) and genital respiratory syndrome (PRRS) are classified as swine foot and mouth disease and classified as statutory infectious disease, and national efforts are being made to spread, prevent and spread the disease.

As a related art in this regard, Patent Document 0001 discloses a "functional composition containing baekhwangsacho and heatji mushroom extract." However, it is a substance derived from natural products and has no toxicity and side effects, and shows excellent bactericidal effect against PED virus, PPRS virus, etc., but it has only bactericidal power against various viruses, and in particular, it does not have excellent disinfection effect against PED virus and PPRS virus. There are disadvantages.

In addition, Patent Document 0002 discloses an "anti-viral composition for animals using octagonal fennel, persimmon, dried crushed powder and polar solvent extract", but this can contribute to the improvement of resistance to infection and health of PED virus and the like. As a result, it is only for the purpose of providing safe livestock that can reduce the mortality rate of livestock.

Therefore, the present inventors, unlike the prior art, while studying disinfectants for pig diarrhea virus (PEDV) and respiratory virus (PRRSV), the king mushroom extract and perilla leaf extract as an active ingredient in the ionized minerals by nanolysis and electrolysis by ultrasonic grinding Mixing was found to be very good disinfection effect for certain serotypes to complete the present invention.

Korean Unexamined Patent Publication No. 10-2006-0032733 (2006. 4. 18.) Korea Patent Publication No. 10-1197451 (2012. 10. 30.)

An object of the present invention is to provide a method for producing a disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) containing the king mushroom extract.

The present invention comprises the steps of: 1) cutting the king mushroom to an appropriate size, extracting using ethanol as a solvent, filtration and freeze drying to obtain the king mushroom extract; 2) a second step of cutting perilla leaves to an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract; 3) a third step of calcining the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral particles; 4) purified water, king mushroom extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ ); 5) provides a method for producing a disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) comprising a; a fifth step of nano-pulverizing the mixed solution made in the fourth step so as not to be precipitated by nanosonication. .

In addition, the present invention 95.5 parts by weight of purified water extract 5.0 ~ 7.0 parts by weight, perilla leaf extract 3.0 parts by weight, ionized mineral nanoparticles 2.0 parts by weight, sodium cyanate (NaOCN) 1.0 parts by weight, glutaraldehyde (C ₅ H ₈ O ₂ ) pig diarrhea virus (PEDV) and respiratory virus (PRRSV) containing 0.5 parts by weight, guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) 0.5 parts by weight, citric acid (C ₆ H ₈ O ₇ ) 0.5 parts by weight To provide an antiseptic composition.

Other specific examples for implementing the present invention are included in the following detailed description.

Method for preparing a disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) containing the king mushroom extract according to the present invention ionized various minerals obtained from king mushroom extract, perilla leaf extract and shellfish by electrolysis and nano process Method for preparing a disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV), which has excellent disinfection effect against swine diarrhea virus (PEDV) and respiratory virus (PRRSV) as well as environmental pollution The advantage is that it has little effect.

Hereinafter, a preferred embodiment and a physical property measuring method of the method for preparing a disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) of the present invention will be described in detail. The present invention can be better understood by the following examples, the following examples are intended to be described in detail so that those skilled in the art can easily carry out the invention, This does not mean that the technical spirit and scope of the present invention is limited.

In the present invention, "disinfection" means killing pathogens in order to prevent infection or transmission of a disease, and in the present invention, the "microorganism" means an active ingredient in the disinfectant composition of swine diarrhea and respiratory virus of the present invention. It is a concept that includes all bacteria, fungi, yeast, and algae that have a disinfecting effect.

Hereinafter, a method for preparing a disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) of the present invention will be described in more detail.

Method for producing a disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) according to an embodiment of the present invention 1) cut the king mushroom to the appropriate size, extracted using ethanol as a solvent, filtered and frozen First step of drying to obtain the king mushroom extract; 2) a second step of cutting perilla leaves to an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract; 3) a third step of calcining the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral particles; 4) purified water, king mushroom extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ ); 5) a fifth step of nanofiberization so as not to precipitate by ultrasonic grinding the mixed solution made in the fourth step; characterized in that it comprises a.

First, the first step is a preparation step in the preparation method of the disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) of the present invention, cut the king mushroom to the appropriate size and extract using ethanol as a solvent After filtering, freeze-drying is a step of obtaining the extract.

The second step is to cut perilla leaves to an appropriate size, extract using ethanol as a solvent, filtered and freeze-dried to obtain perilla leaves extract.

Pleurotus nebrodensis (Pleurotus nebrodensis) used in the present invention is a type of Awi mushroom called Awi nectar of the genus Pleurotus eryngii, and is called by the name of the king mushroom utilizing the genetic characteristics different from the existing Awi mushroom. Among the edible mushrooms, the stem is 2 ~ 3㎝ and the diameter of the lampshade is 10 ~ 15㎝, which is known to be the largest. It is 10 ~ 40 times more nutritious than similar mushrooms. In particular, large mushrooms contain a large amount of amino acids, vitamins, minerals and the like and are medicinal natural foods, and are known to remove harmful oxygen causing degenerative diseases such as dementia, stroke, rheumatism and diabetes.

The term "perilla perilla" refers to a leaf of a year-old herb belonging to the Lamiaceae, and the perilla leaves may be any perilla leaves, preferably domestic perilla leaves, which are commercially available in Korea.

King mushroom extract or perilla leaf extract of the present invention can be extracted by room temperature or warmed under the condition that the active ingredient is not destroyed or minimized. The king mushroom extract or perilla leaf extract may be obtained by extracting the giant mushroom or perilla leaf with reflux for 70 minutes at 80 ° C. for 50 minutes and 30 minutes, respectively.

The king mushroom extract, perilla leaf extract is preferably extracted by adding an extract solvent of 2-20 times, preferably 5-15 times the dry weight of king mushroom, sesame leaf, and most preferably 10 times the weight. . Although water is generally used as the extraction solvent, other organic solvents may be used in addition to water, if necessary. Organic solvents that may be used include, but are not limited to, C ₁ to C ₄ lower alcohols. Preferably ethanol and its aqueous solution are mentioned, Most preferably, 70 vol% ethanol. In the present invention, the "great mushroom extract", "perilla perilla extract" is preferably a great mushroom extract, perilla leaf extract prepared using the extracting solvent, respectively, the addition amount thereof in the disinfectant composition of the present invention is 5.0 ~ It is preferable to set it as 7.0 weight part and 3.0 weight part. At this time, if the addition of the king mushroom extract less than 5.0 parts by weight, the bird flu virus inhibitory effect is somewhat inadequate, when added to 7.0 parts by weight or more excellent virus inhibitory effect, but the composition manufacturing cost increases.

In the present invention, the "wang mushroom extract" or "perilla perilla leaf extract" is preferably a king mushroom extract or perilla leaf extract prepared using the extraction solvent. The giant mushroom extract or perilla leaf extract extracted using an ethanol solvent may be filtered to remove debris and the like to prevent deterioration and freeze drying for preservation.

The third step is the step of sintering the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanonized, followed by electrolysis to ionize the mineral particles.

First, the shellfish is not limited because any kind of shellfish that can be obtained from the beach can be used, but it is preferable to use a scallop shell or cockle shells. The calcined mineral obtained by calcining the shellfish at 1,200 ° C. for 2 hours may be pulverized with a pulverizer, and may contain Mg, Na, K, P, S, Fe, and Zn as constituents thereof. It is preferable that the particle size is a powder of 30 ~ 300 mesh, and if the particle size is less than 30 mesh, the price is expensive and there is an economic problem, and if the particle size exceeds 300 mesh, a problem occurs that does not melt.

In addition, the ultrasonic grinding process of applying ultrasonic wave 25W three times for 10 minutes to the meshed mineral particles is first performed. 100 nm-sized nanoparticles were obtained through the ultrasonic grinding process, and the size of the particles did not change even when kept at room temperature for 7 days (see Table 1). After the nano-ization process through the ultrasonic pulverization process, while stirring the mineral particles at 100rpm in the state dissolved in water to increase the temperature slightly to 45 ~ 50 ℃ condition, and then to the (+) and (-) Electrolyte leads to electrolysis of 0.1 ~ 5V electricity for 3 ~ 5 times in a few minutes. Electrolysis of water in the state of dissolving mineral particles promotes ionization of minerals, thereby inhibiting precipitation of each mineral particle and maintaining an ionized state. At this time, the following reaction occurs in the cathode (Cathode) and the anode (Anode), the ionization of the minerals in this process proceeds.

(1) cathode _{(Cathode): 2H 2 O +} 2e - -> 2OH - + H 2

(2) The positive electrode ^{(Anode): 2OH - ->} H 2 O + 2e - + 1 / 2O 2

On the other hand, the nano- and electrolysis carried out in the above raises the solubility of water and minerals, causing the effect of maintaining solubility even after a considerable time (about 4 weeks or more) time after dissolution. As a result of experimenting with the aqueous solution of mineralized particles after the nano-ization and electrolysis separately, the nanoparticle size change was observed within the error range of less than 10% by maintaining 105 to 110 nm even after 270 days at room temperature. It can be seen that the relatively stable state (see Table 1). In addition, the addition amount in the disinfectant composition of the present invention is preferably 2.0 parts by weight.

Particle Size by Preservation Period of Mineral Particles Storage temperature Retention period (days) Particle size (nm) 25 ± 2 ℃ 0 100 7 101-103 Room temperature 30 100 60 101 90 102 180 104 240 108 270 110

The fourth step is purified water, king mushroom extract, wild perilla leaf extract obtained in the first and second steps, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ) to the ionized mineral nanoparticles of the third step , Guar Hydroxypropyltrimonium Chloride (Guar Hydroxypropyltrimonium Chloride), citric acid (C ₆ H ₈ O ₇ ) is a step of making a mixed solution. At this time, 95.5 parts by weight of purified water extract 5.0 ~ 7.0 parts by weight, perilla leaf extract 3.0 parts by weight, mineral particles 2.0 parts by weight, sodium cyanate (NaOCN) 1.0 parts by weight, glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 weight In addition, 0.5 parts by weight of guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) may be contained to form a mixed solution.

The sodium cyanate (NaOCN) is a white crystalline solid, which is used as an oxidizing agent in the disinfectant composition of the present invention to destroy proteins of viruses by oxidation. In addition, the addition amount in the disinfectant composition of the present invention is preferably 1.0 parts by weight.

Since glutaraldehyde (C ₅ H ₈ O ₂ ) is toxic, it should not be in direct contact with humans and livestock, and the amount is preferably 0.5 parts by weight. In addition, this is a good disinfecting effect even if some organic matters, but the expensive side has a disadvantage that takes a lot of disinfection costs to use in large quantities. This also means that the disinfection effect is maintained even without addition.

The citric acid (citric acid) is contained in the seed or juice of many plants as a free acid, and is widely used in various applications, mainly fruit flavor supplements, preservatives, pH regulators. It is safe for people and clothes, so the effect is good, but the penetration is weak, so the effect is very low in the presence of organic matter. The addition amount is preferably 0.5 parts by weight, which means that the disinfection effect is maintained even without addition.

In the fifth step, the mixed solution made in the fourth step is ultrasonically pulverized so as not to precipitate. It is preferable to help the solution of the mixture by nanosonication by performing ultrasonic sonication process of applying ultrasonic wave of 5W three times for 10 minutes again to the mixed solution prepared in the fourth step for 10 minutes, and preventing the precipitate from occurring.

By performing the first step to the fifth step sequentially 95.0 parts by weight purified water extract 5.0 ~ 7.0 parts by weight, wild perilla leaf extract 3.0 parts by weight, ionized mineral nanoparticles 2.0 parts by weight, sodium cyanate (NaOCN) 1.0 parts by weight , Glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 parts by weight, guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) 0.5 parts by weight, citric acid (C ₆ H ₈ O ₇ ) 0.5 parts by weight to prevent precipitation Disinfectant compositions of swine diarrhea virus and respiratory virus can be prepared.

On the other hand, the disinfectant composition of porcine diarrhea virus and respiratory virus containing the extract of King Mushroom of the present invention, additives such as dispersants, carriers and other antimicrobial agents in addition to the components, so long as the disinfectant effect of the active ingredient is not inhibited It may be prepared further comprising a.

For example, in the case of dispersants, water, alcohols (e.g. methyl alcohol, ethyl alcohol, ethylene glycol, propylene glycol, diethylene glycol, glycerin, etc.), ketones (e.g. acetone, methyl ethyl ketone, etc.), ethers (e.g. Dioxane, tetrahydrofuran, cellosolve, diethylene glycol dimethyl ether, etc., aliphatic hydrocarbons (e.g. hexane, kerosene, etc.), aromatic hydrocarbons (e.g. benzene, toluene, xylene, naphthalene, methyl naphthalene, etc.), halogenated hydrocarbons (E.g. chloroform, carbon tetrachloride, etc.), acid amides (e.g. dimethyl formamide, etc.), esters (e.g. methyl acetate ester, ethyl acetate ester, butyl acetate ester, fatty acid glycerine ester, etc.), nitrole (e.g. Acetonitrile, etc.), surfactants (higher alcohol sulfate esters, alkyl sulfonic acids, alkyl allyl sulfonic acids, quaternary ammonium salts, oxy alkylamines, fatty acids) Thermal switch, a polyalkylene oxide compound, anhydro-sorbitol compounds), but can be one or two or more selected from, but is not limited thereto.

Also in the case of carriers, clays (e.g. kaolin, bentonite, acid clays, etc.), talc (e.g. talc powders, feldspar powders, etc.), silicas (e.g. diatomaceous earth, silicic anhydrides, mica powders, etc.), alumina, sulfur powders and activated carbon One or two or more of them may be selected, but is not limited thereto.

The disinfectant composition of swine diarrhea virus and respiratory virus containing the extract of King Mushroom of the present invention may be prepared in liquid or solid phase, and the disinfectant composition of the present invention may be administered in any manner such as oral or parenteral. Oral dosage forms include tablets, pills, powders, solutions, and food forms. Parenteral dosage forms include injections, topical dosages (creams, ointments, etc.), suppositories, and sprays (if applied to plants). Etc. can be mentioned.

The disinfectant composition of the present invention is preferably administered in a topical manner, and is applied to swine diarrhea virus (PEDV) and respiratory virus (PRRSV), it can be administered / distributed in and around the swine colony and breeding area.

In addition, a tissue made of a mixed material of pulp and nonwoven fabric having a size of 20 cm × 10 cm in width and width is immersed in the disinfectant composition according to the present invention, and the disinfectant composition is allowed to soak into the tissue for 1 to 2 hours. An antiviral cleaning tissue impregnated with the disinfectant composition according to the invention can be prepared. In the case of cleaning the face and hands using the cleaning tissue prepared in this way, the antiviral effect was significantly improved compared to the case of washing the exposed skin using the conventional hand cleaner. Iii) it was confirmed by skin irritation test that no skin damage shape appeared.

Hereinafter, the present invention can be better understood by the following preparation examples and examples, the following preparation examples and examples are for illustrative purposes of the present invention and are not intended to limit the protection scope of the present invention.

<Production example>

  1. Experimental Materials

    In order to verify the antiviral effects of the test substance on swine diarrhea virus (PEDV) and respiratory virus (PRRSV), the experimental animals were selected and used piglets at 7 weeks of age.

  2. King Mushroom Extract Manufacturing

     Giant king mushroom was purchased from a pharmaceutical store. After cutting 1000 g of king mushrooms into suitable sizes, 10 L of 70 Vol% ethanol was used as a solvent, and extracted at 80 ° C. for 50 minutes with a reflux extractor. The extract was filtered through Whatman filter paper and 31.3 g of each dry weight extract was obtained using a lyophilizer.

  3. Perilla Leaf Extract Manufacturing

     Perilla leaves were purchased at hypermarkets. 1000g of perilla leaves were cut to a suitable size, and then extracted with a reflux extractor at 80 ° C. for 30 minutes using 10L of 70Vol% ethanol as a solvent. The extract was filtered through Whatman filter paper and 32.2 g of dry weight extract was obtained using a lyophilizer.

  4. Preparation of Ionized Mineral Nanoparticles

     1) The scallops or cockle shells were calcined at 1,200 ° C. for 2 hours, and the calcined minerals were pulverized with a grinder to obtain a powder having a particle size of 30 to 300 mesh.

     2) The calcined minerals contained Mg, Na, K, P, S, Fe, Zn as constituents. Ultrasound of 25 W was repeatedly applied to the mesh-sized mineral particles three times for 10 minutes, and the nanoparticles obtained by grinding were electrolyzed to obtain ionized mineral nanoparticles.

<Example>

In order to prepare a disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) according to the present invention, the composition is uniformly blended in the composition ratio shown in the following [Table 2] to make a mixed solution and then ultrasonic wave of 25W 3 for 10 minutes The mixture was repeatedly added and pulverized, and the mixed solution was electrolyzed again to obtain an ionized nanomixed solution to obtain a disinfectant composition.

Ingredients and Ingredient Ratios Ingredient Component ratio (part by weight) Purified water 95.5 Giant Mushroom Extract 6.0 Perilla Leaf Extract 3.0 Ionized mineral nanoparticles 2.0 Sodium cyanate 1.0 Glutaraldehyde 0.5 Guar hydroxypropyltrimonium chloride 0.5 Citric acid 0.5

Experimental Example Evaluation of Experimental Methods and Inhibitors

  1. Experiment Method

    In order to test the disinfecting effect of the disinfectant composition of the <Example> was tested according to the National Veterinary Research and Quarantine Service Notification No. 2016-91 [Disinfectant Effectiveness Test Guide] and the results are shown in the following [Table].

  2. Evaluation of Inhibitors

   1) Each virus stock was diluted 1: 10 times, 1.5 ml ep tube was used, and 1 µg / ml TPCK in DMEM was used as the dilution medium.

   2) The test substance and the virus reaction were kept at 4 ℃ by storing all the test solution and reagents, ep tube was used.

     2-1) 450 μl of test substance and 100 μl of virus were mixed.

     2-2) The material control group was mixed with 450 µl of PBS and 100 µl of virus.

     2-3) After mixing, the mixture was reacted at 4 ° C. for 30 minutes, and mixed well every 10 minutes.

   3) 450 μl 1M Tris-HCl (pH 7.2) was added, and 1M Tris-HCl (pH 7.2) for neutralizing the test substance pH was used as a diluent.

   4) Viral titer evaluation was performed on the mixed solution, and a deep well plate was used.

      4-1) 10-fold step dilution was performed at a ratio of 100 µl of the mixed solution and 900 µl of the medium.

4-2) The dilution factor was 10 ⁻¹ , 10 ⁻² , 10 ⁻³ , 10 ⁻⁴ , 10 ⁻⁵ , 10 ⁻⁶ , and 100 μl of the cells was inoculated, and 8 replicates were performed for each dilution factor.

       Dilution medium: 1 μg / ml TPCK in DMEM

     * In the case of fertilized eggs 0.2ml was inoculated into the allantoic cavity, incubated for 5 days in a 37 ℃ incubator and inspected every day, and the presence of virus growth was examined.

   5) The reaction was carried out for 1 hour at 37 ℃.

   6) After the reaction, the inoculum was removed.

   7) 200 ml of culture medium was added.

     Medium: 1 µg / ml TPCK in DMEM

8) Cultured in a 37 ℃ CO ₂ incubator, Carbapenemase production Enterobacteriaceae (CPE) was observed, the results are shown in the table below.

<Experimental Results> Evaluation of Antiviral Inhibitory Activity against Swine Diarrheal Disease and Respiratory Virus

<Comprehensive Analysis of Experiment Results>

In summary, the disinfectant composition of the pig diarrhea (PED) virus and respiratory (PRRS) virus of the present invention is excellent in inhibiting ability to the pig diarrhea (PED) virus and respiratory (PRRS) virus as well as by the virus It has been evaluated to protect birds from disease and to use them safely without concern about side effects to drugs and safety of livestock products by residual drugs.

As mentioned above, although this invention was demonstrated by the limited Example, this invention is not limited by this and it should be described below by the person of ordinary skill in the art, and the following. Of course, various modifications and variations are possible within the scope of the claims.

Claims (15)

1) the first step of cutting the king mushroom to the appropriate size, extract using ethanol as a solvent, filtered and lyophilized to obtain the king mushroom extract;
2) a second step of cutting perilla leaves to an appropriate size, extracting using ethanol as a solvent, filtering and freeze drying to obtain perilla leaves extract;
3) a third step of calcining the shells, pulverizing and meshing, and then ultrasonically pulverizing and nanoning, followed by electrolysis to ionize the mineral particles;
4) purified water, king mushroom extract obtained in the first step, perilla leaf extract obtained in the second step, sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ), A fourth step of preparing a mixed solution by mixing guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) and citric acid (C ₆ H ₈ O ₇ );
5) a fifth step of nano-sizing to prevent precipitation of the mixed solution made in the fourth step so as not to precipitate; the method of producing a disinfectant composition of swine diarrhea virus and respiratory virus comprising a.
The method according to claim 1,
In the first step, ethanol was added 70 vol% to the king mushroom 10 times by weight, 50 minutes at 80 ℃, ethanol was added 70 vol% by weight 10 times to perilla leaves in the second step, at 80 ℃ 30 minutes each Method of producing a disinfectant composition of swine diarrhea virus and respiratory virus to extract reflux.
The method according to claim 2,
In the third step, the shellfish is a method of producing a disinfectant composition of swine diarrhea virus and respiratory virus that is a scallop shell or cockle shell.
The method according to claim 3,
In the third step, the firing is 2 hours at 1,200 ℃, pulverization is a mesh of 30 ~ 300 mesh to prepare a disinfectant composition of swine diarrhea virus and respiratory virus.
The method according to claim 4,
In the third step, the nanonization is a method of producing a disinfectant composition of swine diarrhea virus and respiratory virus that the particle size is 100nm by repeating the ultrasonic wave of 25W for 10 minutes, three times.
The method according to claim 5,
In the fourth step, the mixed solution is 95.5 parts by weight of purified water, 5.0-7.0 parts by weight of king mushroom extract, 3.0 parts by weight of perilla leaf extract, 2.0 parts by weight of ionized mineral nanoparticles, 1.0 part by weight of sodium cyanate (NaOCN), glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 parts by weight of guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride) 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ) is a mixture of porcine diarrhea virus and respiratory virus Method for producing the disinfectant composition of the.
The method according to claim 6,
In the fifth step, the mixed solution is nanowires repeated 25 times for 10 minutes, 25W ultrasonic wave diarrhea virus and respiratory virus disinfectant composition manufacturing method.
95.5 parts by weight of purified water extract 5.0 ~ 7.0 parts by weight, perilla leaf extract 3.0 parts by weight, ionized mineral nanoparticles 2.0 parts by weight, sodium cyanate (NaOCN) 1.0 parts by weight, glutaraldehyde (C ₅ H ₈ O ₂ ) 0.5 A disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV), which comprises by weight, guar hydroxypropyltrimonium chloride (Guar Hydroxypropyltrimonium Chloride), 0.5 parts by weight of citric acid (C ₆ H ₈ O ₇ ).
The method according to claim 8,
The king mushroom extract is a disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) that is obtained by refluxing the king mushroom with 70Vol% ethanol at 80 ℃ for 50 minutes.
The method according to claim 9,
The perilla leaf extract is a disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) that is obtained by reflux extract perilla leaves at 80 ℃ for 30 minutes with 70Vol% ethanol.
The method according to claim 10,
The mineral particles are obtained by firing shellfish at 1,200 ℃ for 2 hours disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV).
The method according to claim 11,
The shellfish is a disinfectant composition of scallop shell or pigtail diarrhea virus (PEDV) and respiratory virus (PRRSV).
The method according to claim 12,
Disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV) is the ionized mineral nanoparticles containing Mg, Na, K, P, S, Fe, Zn as a component.
The method according to any one of claims 8 to 13,
The disinfectant composition of swine diarrhea virus (PEDV) and respiratory virus (PRRSV) further comprises one or more additives selected from a dispersion, a carrier disinfectant composition of pig diarrhea virus (PEDV) and respiratory virus (PRRSV).
The method according to any one of claims 8 to 13,
Antiviral cleaning tissue, characterized in that the impregnated disinfectant composition of the pig diarrhea (PED) virus and respiratory (PRRS) virus.