KR101948738B1 - Anti-freezing disinfectant composite and method for preventing disease and spread using the same - Google Patents

Abstract

The present invention relates to a disinfectant composition for a disinfectant, and a method of preventing and preventing disease using the disinfectant composition, and more particularly, to a disinfectant composition containing sodium dichloroisocyanurate as an active ingredient, The present invention relates to a method of disinfecting a virus. The disinfectant composition of the present invention is easy to transport and manage, is economical and environmentally friendly, and reduces the freezing point of the composition to -10 to -20 ° C, so that the disinfectant can be used even in the winter have.

Description

[0001] The present invention relates to a disinfectant composition and a method of preventing and spreading disease using the same,

The present invention relates to a flushing disinfectant composition and a method for preventing or preventing the spread of bacterial and viral diseases by using the flushing disinfectant composition. More particularly, the present invention relates to a flush disinfectant composition containing sodium dichloroisocyanurate And a method of using the same as an effective concentration.

Recent pathogens in the livestock industry are avian influenza virus, foot and mouth disease virus, swine cholera virus, salmonella, and streptococcus. In particular, avian influenza virus, which mainly affects poultry such as chickens and turkeys, is divided into three types: high pathogenic, weak pathogenic, and non-pathogenic according to the pathogenicity, The highest incidence of illness is classified as List A in the International Bureau of OIE (OIE) and as a livestock epidemic in Korea. Infection occurs mainly when direct contact with algae secretions occurs and is also spread by splashes and water, human feet, feed tea, utensils, equipment, and feces on the surface of eggs. When a bird flu outbreak occurs, most countries in the world will be slaughtered and no birds are exported. Thus, avian influenza virus is the leading cause of the development of the livestock industry. In addition, foot-and-mouth disease occurs when a domestic animal infects a foot-and-mouth disease virus. It has occurred in Korea and Japan in 2000, and has been occurring worldwide since 2001 in the UK. As a result of the above-mentioned foot-and-mouth disease, the foot-and-mouth disease is regarded as an international trade-regulated disease due to the economic damage caused by the decrease in animal productivity. Among the 15 diseases classified as Class A by the International Bureau of OIE (OIE) There is no special method at present except blocking and blocking. In order to develop the livestock industry, it is necessary to develop a safe wide disinfectant which can freely disinfect the body, not only the high sterilization power for destroying pathogens and viruses which cause various diseases, but also the body disinfection as well as the above listed pathogens.

Disinfectants are used to kill various products and air in homes and industrial sites, and to kill pathogens and viruses that cause various diseases. The components used in these disinfectants are mainly chlorine compounds, and chlorine compounds have excellent disinfecting power and are widely used for domestic and industrial use. Chlorine-based disinfectants are sodium hypochlorite (NaOCl), commonly known as lactose, chlorine dioxide (ClO ₂ ) It is used for sterilization in the form of chlorine gas (Cl ₂ ), hypochlorous acid (HOCl), hypochlorite ion (OCl ^- ).

On the other hand, when the outbreak of a common pathogenic infectious disease occurring in winter is carried out outdoors, the freezing of the disinfectant caused a great disruption in the prevention. In addition, from Jan. and Nov. 2010 to Jan. 2011, it is possible to prevent blockage in the case of foot-and-mouth disease that occurred nationwide including Gyeonggi Pocheon, Yeoncheon, Paju, Gyeongbuk Andong, Yeongju, Gangwon Pyeongchang and Chungcheong Cheonan It is difficult to spray the disinfectant by freezing the disinfectant. That is, when the conventional disinfectant is used, since the freezing point is high, the disinfectant can not be sterilized due to the freezing of the disinfectant at an average temperature of about -5 ° C. Therefore, it is inevitably used as a quicklime (granular type or powder type) There is a problem that the disinfecting effect is remarkably reduced when the disinfectant is sprayed.

Thus, sodium isocyanate chloride (NaDCC), sodium carbonate (Na ₂ CO ₃ ), chlorine dioxide (ClO ₂ ) Adipic acid etc. It is necessary to develop a disinfectant having a high disinfecting power and biodegradability and a combination of the components, establishing the optimal manufacturing method and processing, and facilitating long distance transportation and disinfecting at low temperature. In particular, other disinfectants may be unstable and danger of explosion, but sodium isocyanate chloride can safely be used in farms. Since various acute viral livestock infectious diseases are spread more actively at low outside temperature in winter, there is a great demand for the development of antiseptic sodium disinfecting isothiocyanate disinfectant in countries that have a long winter of low outside temperature. It is urgent to resolve disability problems caused by the freezing of disinfectants during preliminary disinfection and blocking prevention of foot-and-mouth disease and highly pathogenic infectious diseases that may occur in cold weather.

KR10-2015-0030525 (2015.03.20.)

It is an object of the present invention to provide a disinfectant composition comprising a sodium dicarboxylate sodium salt as an active ingredient.

It is also an object of the present invention to provide a method of disinfecting bacteria or viruses using a disinfectant composition.

The present invention provides a disinfectant composition comprising a disodium hydrogenisocyanate as an active ingredient.

In addition, the present invention provides a method of disinfecting bacteria or virus using a flush disinfectant composition.

The antifreeze composition comprising sodium dichloroisocyanurate as an active ingredient according to the present invention is easy to transport and manage, is economical and environmentally friendly, and has a freezing point of the chlorine compound, By reducing the temperature to -20 ℃, the chlorine disinfectant can be used outdoors in winter without freezing under the freezing temperature.

The present invention relates to a flammable disinfectant composition comprising sodium dicarboxylate sodium as an active ingredient. Sodium hypochlorite (NaDCC) is about 100 times more effective than hypochlorous acid. It does not adversely affect the environment because it is 100% biodegradable and does not adversely affect the environment. As synergistic effect with sodium carbonate as a foaming agent, . ≪ / RTI >

The sodium sodium dicarboxylate may be contained in an amount of 0.1 to 37.5 parts by weight based on 100 parts by weight of the total composition. If the organic matter is less than 0.1, the disinfecting effect may not be obtained, and if it exceeds 37.5, the economical efficiency may be lowered.

The anti-corrosive composition may further comprise at least one selected from the group consisting of sodium ethoxide, adipic acid, sodium hydrogencarbonate, and sodium carbonate.

Sodium ethoxide is present as a solid salt, convenient to transport and manage, and can be used as an anti-frost agent for winter freeze prevention because of its excellent anti-dripping effect. In addition, unlike calcium chloride, it is environmentally friendly because it does not emit chlorine ions that damage the environment. Adipic acid, sodium hydrogencarbonate and sodium carbonate have high disinfecting power.

The anti-corrosive composition may have a freezing point of -10 to -20 [deg.] C. Preferably -13 to -17 < 0 > C. Accordingly, even when the outdoor air temperature reaches -15 DEG C in the cold weather, it can be disinfected by using the heat resistant disinfectant composition.

The anti-corrosive composition may be one used for bacterial or viral disinfection.

The bacterium may be selected from the group consisting of salmonella and apo-formed anaerobic bacteria, but is not limited thereto.

The virus may be selected from the group consisting of avian influenza, Newcastle virus, swine epidemic diarrhea virus, porcine reproductive / respiratory syndrome virus and foot-and-mouth disease virus, but is not limited thereto.

The present invention also relates to a method of disinfecting bacteria or viruses using a disinfectant composition.

The bacterium may be selected from the group consisting of salmonella and apo-formed anaerobic bacteria, but is not limited thereto.

The virus may be selected from the group consisting of avian influenza, Newcastle virus, swine epidemic diarrhea virus, porcine reproductive / respiratory syndrome virus and foot-and-mouth disease virus, but is not limited thereto.

The disinfection may be performed at -20 to 4 [deg.] C.

Example

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

material

My homosexuality  Disinfectant material

Sodium dichloroisocyanurate (Sigma, 96%), adipic acid (Sigma, 99%), sodium bicarbonate (99.9-100.5%), sodium carbonate (Wako, 99.8 %), Ethoxy antifreeze (sodium ethoxide vs. Refinement, 96%)

Bacteria and viruses

Salmonella typhimurium, Clostridium perfringens, Avian influenza virus H9N2, Newcastle diAaA virus LsSota, Porcine Epidemic Diarrhea virus PEDV 11.29, Porcine respiratory tract respiratory tract Syndrome virus (Porcine reproductive and respiratory syndrome LMY strain)

Test system

When the avian influenza virus was used as a pathogen, the developmental field of avian influenza virus was used as a test system. When using Newcastle virus as a pathogen, the Newcastle virus absence developmental cell line was used as a test system. Vero cells (Vero cell, KCLB 10081) were used as the test system when swine-based diarrhea virus was used as a pathogen. MARC-145 cells were used as a test system when pig genital respiratory syndrome virus was used as a pathogen.

Culture medium  And culture conditions

Salmonella was cultured in nutrient broth at 37 ° C.

Apo-formed anaerobic bacteria were cultured in anaerobic conditions at 37 ° C in NIH thioglycollate broth.

In the case of swine-diarrhea virus, cells were incubated at 37 ° C under 5% carbon dioxide, containing α-minimal essential medium (α-MEM), antibiotic-antimycotic and 5% heat inactivated FBS The virus inoculation was carried out in the presence of α-minimal essential medium (α-MEM), antibiotic-antifungal, 3% tryptose phosphate broth, 0.02% yeast extract and 2 μg trypsin / ml : 250).

Porcine reproductive and respiratory syndrome viruses are grown in a medium containing DME medium, antibiotic-antifungal, 10% fetal bovine serum (FBS) and 5% nonessential amino acid solution minimal medium (MEM NEAA) at 37 ° C under 5% Lt; / RTI >

diluent

The following dilutions were prepared and used.

The hard water was prepared by adding 0.305 g of calcium chloride (CaCl ₂ ) and 0.139 g (w / v) of magnesium chloride (MgCl ₂ .6H ₂ O) to 1 liter of distilled water.

A 5% diluted solution was prepared by diluting with 20% (w / v) yeast extract in light water to make sterilized high-pressure water. The diluted solution was adjusted to pH 7.0 with 1N sodium hydroxide.

5% (w / v) fetal bovine serum (FBS) was used as an organic diluent for the virus.

Bacterial diluent

When using the standard test method, X1 Phosphate buffered saline was dissolved in distilled water and sterilized at 121 ° C for 15 minutes using a high-pressure sterilizer.

Neutralization badge

When using the standard test method, nutrient broth containing 5% non-assimilated serum was used for Salmonella. For the apo-formed anaerobic bacteria, nutrient medium (NIH thioglycollate broth) containing 5% non-assimilated serum was used. For avian influenza and Newcastle virus, phosphate buffered saline containing 10% fetal bovine serum was used. For porcine epidemic diarrhea virus, α-minimal essential medium (including α-MEM, antibiotic-antifungal agent) containing 10% unmixed fetal bovine serum and 10% unmixed fetal bovine serum for porcine respiratory syndrome virus Dulbecco's minimal medium (DMEM, antibiotic-antifungal agent) was used.

Production Example 1. Bacterial culture

Bacteria in the passage were cultured for 22 to 26 hours in a medium. The temperature was maintained at 37 ℃ until the end of use. The concentration of bacteria used was above 10 ⁸ CFU (colony forming unit) per ml.

Production Example 2. Dilution of disinfectant for hard water condition

Manufacturing example  2-1.

The disinfectant to be used in hard water condition was diluted 50 times with light water reactor and stored at 4 ℃.

Production Examples 2-2 to 2-9 were prepared by diluting with 100, 150, 200, 250, 500, 1000, 2000 and 3000 times, respectively, in Production Example 2-1.

Production Example 3. Dilution of disinfectant for organic condition

Manufacturing example  3-1.

The disinfectant to be used for the organic material condition was diluted 20 times with the organic material of the above material and stored at 4 캜.

Preparative Examples 3-2 to 3-10 were prepared by diluting with 30, 50, 60, 70, 80, 100, 150, 200, and 250 times in Preparation Example 3-1, respectively.

Manufacturing example  4. Pig epidemic diarrhea Virus solution  Produce

200 μl each of the cells was placed in a 96-well plate such that vero cells were 2 × 10 ⁵ cells / ml. The cells were cultured in an incubator at 37 ° C. under 5% carbon dioxide.

Bero cells were cultured in a T25 flask, and when a monolayer was formed, pig swine diarrhea virus (10 ⁵ -10 ⁷ TCID ₅₀ / ml) was inoculated and cultured in an incubator at 37 ° C under 5% carbon dioxide for 90 minutes. At this time, they were carefully shaken to distribute evenly every 15 minutes. After inoculation, the inoculum was removed, and the cells were cultured until a cytopathogenic effect (CPE) occurred by adding a virus inoculation medium. After cell denaturation, the cells were frozen and thawed three times and stored at 70 ° C. After that, they were kept in ice water for a short time until immediately before use.

Manufacturing example  5. Pigs Genital respiratory syndrome Virus solution  Produce

MARC-145 cells were seeded at a density of 2 × 10 ⁵ cells / ml in a 96-well plate and cultured in an incubator at 37 ° C. under 5% carbon dioxide.

MARC-145 cells were cultured in a T25 flask, and when a monolayer was formed, pig genital respiratory syndrome virus (10 ⁵ -10 ⁷ TCID ₅₀ / ml) was inoculated and cultured in an incubator for 90 minutes at 37 ° C under 5% . At this time, they were carefully shaken to distribute evenly every 15 minutes. After inoculation, the inoculum was removed, and the cells were cultured until a cytopathogenic effect (CPE) occurred by adding a virus inoculation medium. After cell denaturation, the cells were frozen and thawed three times and stored at 70 ° C. After that, they were kept in ice water for a short time until immediately before use.

Manufacturing example  6. Foot and mouth disease Virus solution  Manufacturing and disinfectant dilution

Both kidney cells were divided into 6 well plates so as to be 3 × 10 ⁵ cells / ml in Eagle minimum essential medium (EMEM) supplemented with L-glutamine, sodium bicarbonate, non essential amino acid, sodium pyruvate, fetal bovine serum and antibiotics And cultured in an incubator at 37 ° C under 5% carbon dioxide.

The foot-and-mouth disease virus (O ₁ BFS) with a viral titer of 10 ^{7 0} TCID ₅₀ / ml or more was stored at 4 ° C for a short time until immediately before use.

Dilution of disinfectant for organic conditions was performed by adding 0.304 g of calcium chloride, 0.139 g (w / v) of magnesium chloride and 1% of fetal calf serum to 1 L of distilled water.

Example 1. Antimicrobial disinfectant composition

375 grams of sodium dichloroisocyanurate, 168.75 grams of adipic acid, 62.5 grams of sodium bicarbonate, 18.75 grams of sodium carbonate and 375 grams of sodium ethoxide To prepare a solid anti-corrosive composition. Then, they were mixed and diluted by the method of Production Example 2 or 3 and used.

Experimental Example 1. Standard Test Method (Liquid Phase Test Method)

Experimental Example 1-1. Disinfectant efficacy test for Salmonella (at 4 ℃)

4 ml of salmonella prepared according to Preparation Example 1 was mixed with 96 ml of the prepared hard water at a temperature of 4 캜 and 96 ml of the organic substance diluent for bacteria. 2.5 ml of the mixed bacterial dilutions were mixed with 2.5 ml of the disinfectant diluent prepared according to Preparation Examples 2 and 3 at a temperature of 4 占 폚. The reaction was carried out at 4 ° C for 5 minutes or 30 minutes. When the reaction was carried out for 30 minutes, stirring was carried out every 10 minutes. Immediately after the reaction of the mixture was completed, 1 ml was taken out and mixed in 9 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was inoculated into 5 nutrient media test tubes in 0.1 ml increments for each disinfectant dilution step. And cultured in a 37 ° C incubator for 48 hours. It was visually confirmed to determine whether the bacteria were proliferating. The experiment was repeated three times, and in comparison with Comparative Example 1 below, a final disinfection dilution step in which no more than 4 growths were prolonged in the nutrient medium of five identical disinfectant dilutions was determined as an effective dilution factor.

Experimental Example 1-2. Disinfectant efficacy test for Salmonella (-20 ℃)

The same method as in Experimental Example 1-1 was used to mix the same amount of each of organic bacteria diluted with bacteria, hard water and bacteria at 4 ° C. Thereafter, the mixture was dispensed into 2.5 ml tubes and frozen at -20 캜. 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 was added to the diluted solutions of the frozen bacteria and mixed. The reaction was carried out at -20 DEG C for 5 minutes or 30 minutes. After the reaction of the mixture was completed, 45 ml of the neutralization medium at 37 ° C was added to a disinfectant reaction test tube (5 ml) and dissolved. The neutralized solution was inoculated into 5 nutrient media test tubes in 0.1 ml increments for each disinfectant dilution step. And cultured in a 37 ° C incubator for 48 hours. It was visually confirmed to determine whether the bacteria were proliferating. The experiment was repeated three times, and in comparison with the comparative example 1, the final disinfection dilution step in which no more than 4 pieces of proliferation could be performed in the nutrient medium of five identical disinfectant dilutions was determined as the effective dilution factor.

Experimental Examples 1-3. Test for disinfectant effect on apo-formed anaerobic bacteria (at 4 ℃)

A mixture of bacteria and disinfectant diluent was prepared in the same manner as in Experimental Example 1-1 except that the apo-formed anaerobic bacteria prepared in Preparation Example 1 were used. Immediately after the reaction of the mixture was completed, 1 ml was taken out and mixed in 9 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was treated and cultured in the same manner as in Experimental Example 1-1, and visually confirmed. 3 times repeatedly, and in comparison with Comparative Example 2, the final disinfection dilution step in which four or more non-multiplying dilutions were performed in the nutrient medium of five identical disinfectant dilutions was determined as an effective dilution factor.

Experimental Examples 1-4. Test for disinfectant effect on apo-formed anaerobic bacteria (-20 ℃)

A mixture of bacteria and disinfectant diluent was prepared in the same manner as in Experimental Example 1-2, except that the apo-formed anaerobic bacteria prepared in Preparation Example 1 were used. After the reaction of the mixture was completed, 45 ml of the neutralization medium at 37 ° C was added to a disinfectant reaction test tube (5 ml) and dissolved. The neutralized solution was treated and cultured in the same manner as in Experimental Example 1-2, and visually confirmed. In contrast to the comparative example 2-2, the final disinfection dilution step in which no more than 4 growths were made in the nutrient medium of five identical disinfectant dilutions was judged as an effective dilution factor.

Experimental Examples 1-5. Disinfectant efficacy test for avian influenza virus (at 4 ℃)

The frozen avian influenza virus (10 ⁵ to 10 ⁷ EID ₅₀ / ml) was dissolved and stored in ice water for a short time until just before use. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. 2.5 ml of the mixed virus solutions were mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at a temperature of 4 캜. The reaction was carried out at 4 ° C for 5 minutes or 30 minutes. When the reaction was carried out for 30 minutes, stirring was carried out every 10 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was diluted with phosphate buffered saline, 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^-5 times in phosphate buffered saline, . After inoculation, the cells were cultured at 37 ° C for 5 days, and the cells were harvested every day, and dead cells within 24 hours of inoculation were evaluated as accidents. All the inoculated dead cells within 24 hours of inoculation within 5 days were stored at 4 ℃. After 5 days of inoculation, the urogenital fluid was collected from all the surviving bovine oocytes and dead oocytes stored at 4 ℃, and the presence or absence of virus and viral titers were determined by hemagglutination using 1% chicken erythrocytes. In comparison with Comparative Example 3, when a decrease of 10 ⁴ or more was confirmed in the pathogen (bacterial spot formation unit, pfu), it was judged to be effective, and this was regarded as an effective dilution factor.

Experimental Example  1-6. Disinfectant efficacy test for avian influenza virus (-20 ℃)

The frozen avian influenza virus (10 ⁵ to 10 ⁷ EID ₅₀ / ml) was dissolved and stored in ice water for a short time until just before use. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. Mixed viral solutions were dispensed and frozen at -20 ° C. 2.5 ml of the diluted solution of the disinfectant prepared in Preparation Examples 2 and 3, at a temperature of 4 캜, were mixed. The reaction was carried out at 4 DEG C for 5 minutes or 30 minutes. After the reaction of the mixture was completed, 45 ml of the neutralization medium at 37 ° C was added to a disinfectant reaction test tube (5 ml) and dissolved. The neutralized liquid was tested, evaluated and evaluated in the same manner as in Experimental Example 1-5.

Experimental Example 1-7. Disinfectant efficacy test for Newcastle virus (at 4 ° C)

Frozen Newcastle virus (10 ⁵ to 10 ⁷ EID ₅₀ / ml) was dissolved and stored in ice water for a short time until just before use. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. 2.5 ml of the mixed virus solutions were mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at a temperature of 4 캜. The reaction was carried out at 4 ° C for 5 minutes or 30 minutes. When the reaction was carried out for 30 minutes, stirring was carried out every 10 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was tested and evaluated in the same manner as in Experimental Example 1-5, and it was judged to be effective when a decrease of 10 ⁴ or more pathogen was confirmed as compared with Comparative Example 4, and this was used as an effective dilution factor.

Experimental Examples 1-8. Disinfectant efficacy test for Newcastle virus (at -20 ° C)

Frozen Newcastle virus (10 ⁵ to 10 ⁷ EID ₅₀ / ml) was dissolved and stored in ice water for a short time until just before use. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. The combined viral solutions were frozen at -20 < 0 > C. 2.5 ml of the diluted solution of the disinfectant prepared in Preparation Examples 2 and 3, at a temperature of 4 캜, were mixed. The reaction was carried out at 4 DEG C for 5 minutes or 30 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized liquid was tested, evaluated and evaluated in the same manner as in Experimental Example 1-7.

Experimental Example 1-9. Disinfectant efficacy test for swine diarrhea virus (at 4 ℃)

Porcine epidemic diarrhea virus was prepared by the method of Production Example 4 above. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. 2.5 ml of the mixed virus solutions were mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at a temperature of 4 캜. The reaction was carried out at 4 ° C for 5 minutes or 30 minutes. When the reaction was carried out for 30 minutes, stirring was carried out every 10 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was diluted with a virus-inoculating medium to a stock solution of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^-5 times, and 8 layers of monolayers (80 to 90 %) Were formed. After inoculation, the cells were incubated at 37 ° C under 5% carbon dioxide for 5 days in an incubator to observe daily cytopathic effect (CPE). In comparison with Comparative Example 2-5, when a decrease of 10 ⁴ or more was confirmed in pathogen (bacterial spot formation unit, pfu), it was judged to be effective, and this was regarded as an effective dilution factor.

Experimental Example  1-10. Disinfectant efficacy test for swine diarrhea virus (-20 ℃)

Porcine epidemic diarrhea virus was prepared by the method of Production Example 4 above. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. The mixed virus solution was frozen at -20 占 폚. 2.5 ml of the mixed virus solutions were mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at a temperature of 4 캜. The reaction was carried out at 4 DEG C for 5 minutes or 30 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized liquid was inoculated by the method of Experimental Example 1-9, and observed and evaluated.

Experimental Example  1-11. pig Genital respiratory syndrome  Disinfectant efficacy test for virus (at 4 ℃)

Porcine reproductive tract respiratory syndrome virus was prepared by the method of Production Example 5 above. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. 2.5 ml of the mixed virus solutions were mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at a temperature of 4 캜. The reaction was carried out at 4 ° C for 5 minutes or 30 minutes. When the reaction was carried out for 30 minutes, stirring was carried out every 10 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized solution was diluted with a virus-inoculating medium to a stock solution of 10 ^-1 , 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^-5 , and a monolayer of 80 wells (80 wells per well) %), Respectively. After inoculation, the cells were incubated at 37 ° C under 5% carbon dioxide for 5 days in an incubator to observe daily cytopathic effect (CPE). Compared with Comparative Example 6, when a decrease of 10 ⁴ or more was confirmed in the pathogen (bacterial spot formation unit, pfu), it was judged to be effective, and this was regarded as an effective dilution factor.

Experimental Example  1-12. pig Genital respiratory syndrome  Disinfectant efficacy test for virus (-20 ℃ condition)

Porcine reproductive tract respiratory syndrome virus was prepared by the method of Production Example 5 above. 1.0 ml of the virus solution at 4 占 폚 was mixed with 19 ml each of diluted solution for hard water and virus. The combined viral solutions were frozen at -20 < 0 > C. 2.5 ml of the mixed virus solution was mixed with 2.5 ml of the disinfectant diluent prepared in Preparation Examples 2 and 3 at 4 캜. The reaction was carried out at 4 DEG C for 5 minutes or 30 minutes. Immediately after the reaction of the mixture was completed, 1 ml of the mixture was taken out and added to 1 ml of the neutralization medium at 37 ° C and mixed. The neutralized liquid was inoculated by the method of Experimental Example 1-11, and observed and evaluated.

Experimental Example 1-13. Disinfectant efficacy test for foot-and-mouth disease virus (at 4 ° C)

Foot-and-mouth disease virus was prepared by the method of Preparation Example 6 above. 1.8 ml of the virus solution at 4 占 폚 was added to 0.2 ml of the diluted organic solution prepared in Preparation Example 6 and mixed. The mixed virus solution was reacted at 4 ° C for 5 minutes or 30 minutes, and the mixture was mixed well before the reaction was completed. Upon completion of the reaction, 0.05 ml of the mixture was added to 5 ml of neutralization medium (Eagle minimum essential medium containing 2% of antibiotics and 5% of fetal bovine serum) at 4 ° C and mixed. The neutralized solution was diluted to 10 ^-2 , 10 ^-3 , 10 ^-4 and 10 ^-5 times and 0.2 ml of each final dilution of the diluted solution containing the negative control (inoculated with distilled water) and the positive control (1% sodium hydroxide) Lt; RTI ID = 0.0 > 6 < / RTI > The cells were incubated at 37 ° C under 5% carbon dioxide for 1 hour and shaken well every 8 to 10 minutes. After 1 hour of inoculation, 2.5 ml of a medium containing 1% methylcellulose was added and further incubated for 2 days at 37 ° C under 5% carbon dioxide wet condition. The cells were then washed with 0.01-0. 5% citric acid and stained with amido black to yield plague forming unit (PFU) values. Compared with Comparative Example 7, when a decrease of 10 ⁴ or more was confirmed in the pathogen (bacterial spot formation unit, pfu), it was judged to be effective, and this was regarded as an effective dilution factor.

COMPARATIVE EXAMPLE 1 Effectiveness of Salmonella Antiseptics Test Treatment Control group

As a control group for the above Experimental Examples 1-1 and 1-2, the pathogen control group was tested in the light water condition without the disinfectant, and the neutralization reaction and the propagation step were carried out with a titer of 2 × 10 ⁵ CFU or more per ml.

Comparative Example 2. Effectiveness of disinfectant of apo-formed anaerobic bacteria Tested control group

As a control group for Experimental Examples 1-3 and 1-4, the pathogen control group was experimented without the disinfectant under the hard water condition, and the neutralization reaction and the proliferation step were performed with the activity value of 2 × 10 ⁵ CFU or more per ml.

Comparative Example 3. Effect of disinfectant on avian influenza test Control group

As a control group for Experimental Examples 1-5 and 1-6, the pathogen control group was experimented without water disinfectant under light water conditions, and had a potency of 2 × 10 ⁵ EID ₅₀ / ml or more per ml in the neutralization reaction and proliferation step.

Comparative Example 4. Effectiveness of Newcastle virus disinfectant test Control group

As a control for Experimental Examples 1-7 and 1-8, the pathogen control was tested in the water-free condition without the disinfectant, and in the neutralization reaction and the proliferation stage, the titer was 2 × 10 ⁵ EID ₅₀ / ml or more per ml.

Comparative Example 5. Effectiveness of disinfectant test for swine epidemic diarrhea virus Control group

As a control group for Experimental Examples 1-9 and 2-10, the pathogen control group was experimented without the disinfectant under the hard water condition, and the titer was 2 × 10 ⁵ TCID ₅₀ / ml or more per ml in the neutralization reaction and the proliferation step.

Comparative Example  6. Pigs Genital respiratory syndrome  Antiseptic effect test of virus Treatment  Control group

As a control group for the Experimental Examples 2-11 and 2-12, the pathogen control group was tested in the light water condition without the disinfectant, and the neutralization reaction and the proliferation step were carried out with a titer of 2 × 10 ⁵ TCID ₅₀ / ml or more per ml.

Comparative Example 7. Disinfectant efficacy test of foot-and-mouth disease virus Control group

As a control group for Experimental Example 2-13, the pathogen control group was experimented without disinfectant under the organic condition, and the neutralization reaction and the proliferation step were performed at a concentration of 2 × 10 ⁵ TCID ₅₀ / ml or more per ml.

Analysis of disinfectant efficacy test results

Experimental Examples 1-1, 1-3, 1-5, 1-7, 1-9. Table 1 shows the effective dilution ratios of the antidote disinfectant pathogens when the reaction time is 5 minutes in 1-11 and 1-13. Disinfectant diluent is mixed with the final cells in a ratio of 1: 1, the actual effective dilution factor described in the following table is twice the dilution factor described in Production Examples 2 and 3. Multiples in parentheses represent 80% of the test dilution, which is the dilution factor specified in the quarantine notice.

When the number of effective dilutions is less than "effective", it was shown that there was no disinfection effect when the effective dilution ratio was more than the effective dilution ratio. When the effective dilution factor is described as "exceeding", if it exceeds the dilution factor, it may have the effect of disinfection but it means that the effect of anti-inflammation is lowered.

Pathogen type Dilution condition Antibacterial disinfectant Effective dilution drainage Salmonella
(Salmonella typhimurium) Organic matter Less than 100 times (40) Water More than 400 times (160) Apo-forming anaerobic bacteria
(Clostridium perfringens) Organic matter Less than 140 times (56) Water More than 200 times (80) Avian influenza
(Avian influenza virus H9N2) Organic matter More than 400 times (160) Water More than 400 times (160) Newcastle virus
(Newcastle diAaA virus LsSota) Organic matter More than 400 times (160) Water More than 400 times (160) Swine flu epidemic virus
(Porcine Epidemic Diarrhea virus PEVDV 11.29) Organic matter More than 400 times (160) Water More than 500 times (200) Pig Genital Respiratory Syndrome Virus
(Porcine reproductive and respiratory syndrome LMY strain) Organic matter More than 400 times (160) Water More than 500 times (200) Foot and mouth disease Organic matter More than 100 times (40)

Experimental Examples 1-1, 1-3, 1-5, 1-7, 1-9. 1-11, and 1-13, the effective dilution factor for the antidote agent pathogen in the case of a reaction time of 30 minutes is shown in Table 2 below.

Pathogen type Dilution condition Antibacterial disinfectant Effective dilution drainage Salmonella
(Salmonella typhimurium) Organic matter Less than 40 times (16) Water Less than 6000 times (2400) Apo-forming anaerobic bacteria
(Clostridium perfringens) Organic matter Less than 120 times (48) Water More than 4000 times (1600) Avian influenza
(Avian influenza virus H9N2) Organic matter More than 400 times (160) Water More than 400 times (160) Newcastle virus
(Newcastle diAaA virus LsSota) Organic matter More than 400 times (160) Water More than 400 times (160) Swine flu epidemic virus
(Porcine Epidemic Diarrhea virus PEVDV 11.29) Organic matter More than 500 times (200) Water More than 500 times (200) Pig Genital Respiratory Syndrome Virus
(Porcine reproductive and respiratory syndrome LMY strain) Organic matter More than 500 times (200) Water More than 500 times (200) Foot and mouth disease Organic matter More than 200 times (80)

Experimental Examples 1-2, 1-4, 1-6, 1-8, 1-10. Table 1 shows the effective dilution ratios of the antidote disinfectant pathogens when the reaction time is 5 minutes in 1-12 days.

Pathogen type Dilution condition Antibacterial disinfectant Effective dilution drainage Salmonella
(Salmonella typhimurium) Organic matter More than 60 times (24) Water More than 200 times (80) Apo-forming anaerobic bacteria
(Clostridium perfringens) Organic matter More than 100 times (40) Water More than 200 times (80) Avian influenza
(Avian influenza virus H9N2) Organic matter More than 400 times (160) Water More than 400 times (160) Newcastle virus
(Newcastle diAaA virus LsSota) Organic matter More than 400 times (160) Water More than 400 times (160) Swine flu epidemic virus
(Porcine Epidemic Diarrhea virus PEVDV 11.29) Organic matter More than 400 times (160) Water More than 500 times (200) Pig Genital Respiratory Syndrome Virus
(Porcine reproductive and respiratory syndrome LMY strain) Organic matter More than 400 times (160) Water More than 500 times (200)

Experimental Examples 1-2, 1-4, 1-6, 1-8, 1-10. Table 1 shows the effective dilution ratios of the antidote disinfectant pathogens when the reaction time is 30 minutes in 1-12 days.

Pathogen type Dilution condition Antibacterial disinfectant Effective dilution drainage Salmonella
(Salmonella typhimurium) Organic matter More than 60 times (24) Water More than 200 times (80) Apo-forming anaerobic bacteria
(Clostridium perfringens) Organic matter More than 100 times (40) Water More than 200 times (80) Avian influenza
(Avian influenza virus H9N2) Organic matter More than 400 times (160) Water More than 400 times (160) Newcastle virus
(Newcastle diAaA virus LsSota) Organic matter More than 400 times (160) Water More than 400 times (160) Swine flu epidemic virus
(Porcine Epidemic Diarrhea virus PEVDV 11.29) Organic matter More than 400 times (160) Water More than 500 times (200) Pig Genital Respiratory Syndrome Virus
(Porcine reproductive and respiratory syndrome LMY strain) Organic matter More than 400 times (160) Water More than 500 times (200)

Floating effect of antiseptic agent

The anti-corrosive composition of Example 1 was diluted 100-fold according to Production Example 2-2 to confirm the immobility by temperature. The results are shown in Table 5 below, as compared with the case where sodium ethoxide (SE) was diluted to various concentrations.

-10 ° C -15 ℃ -20 ° C SE 1% 2 hours full floating 1.5 hour partial floating
Freeze for 2 hours 0.5 hour partial floating
Freeze for 1 hour SE 0.5% 2 hours full floating 1 hour partial floating
Freeze 1.5 hours 0.5 hour partial floating
Freeze for 1 hour SE 0.4% 2 hours full floating 1 hour partial floating
Freeze 1.5 hours 0.5 hour partial floating
Freeze for 1 hour SE 0.3% 2 hours full floating 1 hour partial floating
Freeze 1.5 hours Freeze for 0.5 hours SE 0.2% 2 hours full floating Freeze for 0.5 hours Freeze for 0.5 hours SE 0.1% 2 hours full floating Freeze for 0.5 hours Freeze for 0.5 hours Example 1
(SE 0.375%) 3 hours full floating Freeze 1.5 hours 1.0 hour freeze

The present invention has been described by way of examples. It will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention is defined by the appended claims rather than the detailed description and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

Claims (11)

Sodium dicarboxylate, sodium ethoxide, adipic acid, sodium hydrogencarbonate, and sodium carbonate. The method according to claim 1,
Wherein the sodium disodium isocyanurate is contained in an amount of 0.1 to 37.5 parts by weight based on 100 parts by weight of the composition. delete The method according to claim 1,
Wherein the freezing point is -10 to -20 占 폚. The method according to claim 1,
Wherein the composition is used for bacterial or viral disinfection. 6. The method of claim 5,
Wherein the bacterium is selected from the group consisting of salmonella and apo-formed anaerobic bacteria. 6. The method of claim 5,
Wherein the virus is selected from the group consisting of avian influenza, Newcastle virus, swine epidemic diarrhea virus, porcine genital respiratory syndrome virus and foot-and-mouth disease virus. A method for disinfection of bacteria or viruses, comprising disinfecting animals other than humans using the anti-corrosive composition according to claim 1. 9. The method of claim 8,
Wherein the bacterium is selected from the group consisting of salmonella and apo-formed anaerobic bacteria. 9. The method of claim 8,
Wherein the virus is selected from the group consisting of avian influenza, Newcastle virus, swine epidemic diarrhea virus, porcine reproductive-respiratory syndrome virus, and foot-and-mouth disease virus. 9. The method of claim 8,
Wherein the disinfection is carried out at -20 to < RTI ID = 0.0 > 4 C. < / RTI >