WO2021162168A1 - Method for preparing disinfectant against african swine fever virus, and disinfectant against african swine fever virus, prepared thereby - Google Patents

Method for preparing disinfectant against african swine fever virus, and disinfectant against african swine fever virus, prepared thereby Download PDF

Info

Publication number
WO2021162168A1
WO2021162168A1 PCT/KR2020/003071 KR2020003071W WO2021162168A1 WO 2021162168 A1 WO2021162168 A1 WO 2021162168A1 KR 2020003071 W KR2020003071 W KR 2020003071W WO 2021162168 A1 WO2021162168 A1 WO 2021162168A1
Authority
WO
WIPO (PCT)
Prior art keywords
swine fever
african swine
fever virus
hypochlorous acid
acid water
Prior art date
Application number
PCT/KR2020/003071
Other languages
French (fr)
Korean (ko)
Inventor
조현승
Original Assignee
(주)엠씨씨바이오
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by (주)엠씨씨바이오 filed Critical (주)엠씨씨바이오
Publication of WO2021162168A1 publication Critical patent/WO2021162168A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N29/00—Biocides, pest repellants or attractants, or plant growth regulators containing halogenated hydrocarbons
    • A01N29/04—Halogen directly attached to a carbocyclic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents

Definitions

  • the present invention relates to a method for producing an African swine fever virus disinfectant and to an African swine fever virus disinfectant prepared thereby.
  • African swine fever virus is a first-class livestock contagious disease that occurs in all age groups of wild boars and domestic pigs.
  • pigs infected with African swine fever virus show 100% mortality within 2 to 10 days.
  • This African swine fever virus was first reported in Kenya in 1921, and started with the first infection in Europe in 1957, and then in China and Southeast Asia. Recently in Korea, efforts are being made to prevent the movement and quarantine of wild boars, which are vectors, in order to block the transmission.
  • African swine fever virus is a DNA virus, which is different from the RNA viruses of avian influenza and foot-and-mouth disease. Therefore, a vaccine has not been developed due to the lack of neutralizing antibodies and problems with genetic diversity.
  • the main cause of the spread of the African swine fever virus is through contact with infected individuals and the distribution and leftover feeding of infected individuals. It is reported that it can survive, has a long survival period at low temperatures, and is inactivated in strong acids with a pH of 3.9 or less and strong alkalis with a pH of 11.5 or more.
  • African swine fever virus disinfectants include citric acid, ortho-phenylphenol, iodine compound, potassium monopersulfate compound, sodium dichlori-s-trizinetrione ( NADCC), sodium hydroxide (NaOH), and aldehyde compounds.
  • 'Disinfectant having sterilization and disinfection synergistic effect and manufacturing method thereof (registration number: 10-1440375)', one or more organic acids of formic acid, fumaric acid, malic acid, citric acid, oxalic acid and ascorbic acid, phosphoric acid, boric acid and sulfa Disinfectants for sterilizing livestock pathogens by containing any one or more inorganic acids among the acids have been proposed.
  • the present invention was invented to solve the above problems, and a method for producing an African swine fever virus disinfectant comprising sodium-free hypochlorous acid water having a disinfecting effective concentration capable of killing African swine fever virus without cytotoxicity, and It is a technical solution to provide an African swine fever virus disinfectant manufactured by
  • the present invention comprises the steps of: preparing an electrolysis product containing chlorine gas by electrolyzing a hydrochloric acid solution into an electrolyzer; and preparing sodium-free hypochlorous acid water while mixing chlorine gas separated from the electrolysis product into distilled water; It provides a method for producing sodium-free hypochlorous acid water for African swine fever virus disinfectants, characterized in that by adjusting the sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm or 143 ppm.
  • the effective chlorine concentration of the hypochlorous acid water is 78 ppm, it is characterized in that the disinfection treatment by spraying directly on the pig.
  • the effective chlorine concentration of the hypochlorous acid water is 143 ppm, it is characterized in that the disinfection treatment by spraying the pig barn.
  • the present invention provides a disinfectant for African swine fever virus, characterized in that it comprises sodium-free hypochlorous acid water prepared by the above method.
  • the present invention kills the African swine fever virus by directly spraying the African swine fever virus disinfectant to pigs or swine barns, characterized in that the death rate is at least 99.97%, sodium
  • a method for killing African swine fever virus using non-containing hypochlorous acid water is provided.
  • the manufacturing method of the African swine fever virus disinfectant of the present invention by means of a solution to the above problem, effective chlorine in hypochlorous acid water without sodium through the control of the current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water Since the concentration can be kept constant at 78 ppm or 143 ppm, there is an effect of efficiently performing disinfection of African swine fever virus and preparing a disinfectant that can be used as a virucidal agent without cytotoxicity.
  • FIG. 1 is a flow chart according to the manufacturing method of the African swine fever virus disinfectant of the present invention.
  • FIG. 1 is a flowchart according to the manufacturing method of the African swine fever virus disinfectant of the present invention.
  • a hydrochloric acid solution is added to an electrolyzer and then electrolyzed to produce an electrolytic product containing chlorine gas ( It is prepared through S10) and the step (S20) of preparing sodium-free hypochlorous acid water while mixing chlorine gas separated from the electrolysis product into distilled water.
  • sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm or 143 ppm is produced by controlling the amount of current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water It is characterized in that, each of the above-described steps can be described as follows.
  • a hydrochloric acid solution is put into an electrolyzer and then electrolyzed to prepare an electrolysis product containing chlorine gas (S10).
  • electrolysis in general, a pair of electrodes forming a cathode and an anode are installed in an electrolyzer, and electricity is supplied to each electrode to electrolyze the material contained in the electrolyzer.
  • electrolysis may be divided into a diaphragm type having a diaphragm between the cathode and an anode, and a diaphragm-free type having no diaphragm depending on the presence or absence of the diaphragm.
  • the diaphragm-free type since there is an advantage in that the amount of generated electrolysis products can be increased by increasing the voltage and current and increasing the number of electrodes or the surface area of the electrodes, it is preferable to apply the diaphragm-free electrolysis in the present invention.
  • sodium chloride is mixed with a hydrochloric acid solution to further increase the electrolytic ability.
  • Sodium chloride has a large ionization, so the ratio of separation into cations and anions during electrolysis is close to 100%, so that the current flows well and the electrode
  • the toxicity is so strong that it is inconvenient to use it after diluting it in a large amount of water.
  • sodium chloride is mixed with the hydrochloric acid solution It is preferable not to
  • sodium-free hypochlorous acid water is prepared while mixing chlorine gas separated from the electrolysis product into distilled water (S20).
  • the distilled water introduced for the production of sodium-free hypochlorous acid water is supplied from an adjacent device, the amount of distilled water introduced can always change. Due to this, when the flow rate of distilled water is reduced or increased, the concentration of hypochlorous acid water generated can be increased or decreased.
  • the flow rate of the hydrochloric acid solution supplied to the electrolyzer is sensed and the current supplied to the electrolyzer is controlled by controlling the power supply during electrolysis based on the measured value, and generated in the electrolyzer by controlling the current
  • the effective chlorine concentration of hypochlorous acid water is maintained at 78 ppm or 143 ppm, so that it can be supplied constantly.
  • the current applied to the electrolyzer is controlled so that the effective chlorine concentration of the hypochlorous acid water is maintained at 78 ppm or 143 ppm, it has a certain effective chlorine concentration depending on the required place, so that the disinfecting power of the disinfectant can be constantly maintained.
  • the effective chlorine concentration of hypochlorous acid water is adjusted to be 78ppm.
  • distilled water is supplied at 1.0 ⁇ 1.5L/min.
  • the amount of distilled water is small and the effective chlorine concentration of the final product, hypochlorous acid water, exceeds 78ppm, which is not effective for convection prevention.
  • the amount of chlorous acid is too large, and the final product, hypochlorous acid water, is diluted, and the effective chlorine concentration is much lower than 78ppm, so the disinfection efficiency is not good.
  • the most preferable supply amount of distilled water is 1.25L/min so that the effective chlorine concentration of hypochlorous acid water becomes 78ppm.
  • the effective chlorine concentration of hypochlorous acid water generated by measuring the amount of current supplied to the electrolyzer through the power supply and controlling the power supplied from the power supply based on the measured amount of current is controlled.
  • the current is less than 18A, there is a disadvantage that the generation time of hypochlorite water needs to be further increased to meet the 78ppm concentration, and if it exceeds 22A, it is difficult to maintain a constant 78ppm concentration of hypochlorous acid.
  • the effective chlorine concentration of the hypochlorous acid water is adjusted to 143ppm.
  • hypochlorous acid water having an effective chlorine concentration of 143 ppm
  • distilled water is supplied at 0.8 to 0.9 L/min while flowing chlorine gas separated from the electrolysis product. If distilled water is supplied at less than 0.8L/min, the amount of distilled water is small and the effective chlorine concentration of the final product, hypochlorous acid water, exceeds 143ppm, which may increase toxicity.
  • the effective chlorine concentration of hypochlorous acid water, which is the final product is much lower than 143ppm, so even if it is sprayed where organic matter is present, the disinfection efficiency is not as good as expected.
  • the most preferable supply of distilled water is 0.88L/min so that the effective chlorine concentration of hypochlorous acid water becomes 143ppm.
  • the amount of current supplied to the electrolyzer through the power supply is measured, and the power supplied from the power supply is controlled based on the amount of current measured in this way. It controls the effective chlorine concentration of hypochlorous acid water produced by the If the current is less than 28A, there is a disadvantage that the amount of chlorine gas must be increased while increasing the electrolysis degree of the hydrochloric acid solution by increasing the current to meet the 143ppm concentration. There is this. In order to maintain the effective chlorine concentration of hypochlorous acid water at 143ppm, it is most desirable to flow a current of 30A.
  • hypochlorous acid water the effect on disinfection power is large depending on the pH.
  • it is suitable for application as a disinfectant in the range of pH 5.0 to 6.0, and the sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm of the present invention becomes pH 5.5, so it can be an appropriate disinfectant for convection prevention that can be directly sprayed on pigs .
  • the pH of hypochlorous acid water is lowered to 4.5 and the effective chlorine concentration is raised to 143 ppm, there is no cytotoxicity, so it is possible to kill the African swine fever virus by spraying it directly into the pig barn.
  • electrolysis was performed to prepare an electrolysis product containing chlorine gas.
  • 78 ppm of hypochlorous acid water was prepared by controlling a current of 20 A while mixing distilled water at 1.25 L/min under the supply of chlorine gas separated from the electrolysis product. At this time, the pH was 5.5.
  • electrolysis was performed to prepare an electrolysis product containing chlorine gas.
  • distilled water was mixed at 0.88 L/min and the current was adjusted to 30 A to prepare 143 ppm of hypochlorous acid water.
  • the pH of the hypochlorous acid water was 4.5.
  • the cytotoxicity and disinfection power of the African swine fever virus disinfectant was tested.
  • the inactivation of the virus was determined by finally reading the cytotoxicity of the cells inoculated with the African swine fever virus treated with the disinfectant.
  • Table 1 shows the virucidal test conditions for African swine fever.
  • Item condition active substance Hypochlorous acid (HOCl) Chlorine concentration of active substance 78ppm, 143ppm Virus processing time 30 minutes Virus test temperature 10°C Interfering substance (organic molar) concentration high concentration BSA 10g/L + yeast extract 10g/L low concentration BSA 3g/L additive-free MEM + 7% FCS cell culture temperature 37°C Virus species used African Swine Fever Virus (ASFV) strain BA71V
  • disinfectants having effective chlorine concentrations of 78 ppm and 143 ppm were prepared according to Examples 1 and 2 according to the African swine fever virus efficacy test method, respectively, and the African swine fever virus used in the experiment was African Swine Fever Virus (ASFV) strain BA71V was used.
  • Experimental treatment temperature was 10°C for 30 minutes.
  • the experimental group was divided into 3 groups, and for each of the experiments to verify the efficacy when organic substances were added, 3g/L bovine serum albumin was added to those containing low concentrations of organic substances, and high concentrations of organic substances were added.
  • 10g/L bovine serum albumin and 10g/L yeast extract were used together.
  • efficacy experiments were also conducted in the cell culture medium alone addition group.
  • the experimental method for verifying the virucidal virus for African swine fever was carried out in the same manner as in Table 2 below.
  • the experimental method as shown in Table 2, first, 1 ml of an organic material to be introduced as an interfering material was put into an experimental container, and 1 ml of a virus culture solution was added.
  • the experimental vessel is a thermostat maintained at 10 ⁇ 1°C, and the organic material and the virus culture solution were mixed for 2min ⁇ 10s. Then, 8ml of the disinfectant test solution was added and mixed, and treated at 10 ⁇ 1°C for 30min ⁇ 10s. Mix well just before the end of the treatment time.
  • test solution 0.5 ml was added to 4.5 ml of a mixture of MEM (minimum essential medium) + 2% FCS (fetal calf serum) maintained in a thermostatic water bath maintained at 4 ⁇ 1°C or crushed ice. added.
  • MEM minimum essential medium
  • FCS fetal calf serum
  • test substance is diluted to 10 -9 with a mixture of MEM (minimum essential medium) + 2% FCS (fetal calf serum) maintained in a constant temperature bath maintained at 4 ⁇ 1°C or crushed ice did.
  • MEM minimum essential medium
  • FCS fetal calf serum
  • the experimental dilutions after virus treatment were stored at 4°C or under crushed ice until inoculated into cell cultures.
  • the effective chlorine concentration of the disinfectant used in the experiment was tested at 78 ppm, and this will be denoted as MCC-A.
  • concentration of MCC-A actually used in the experiment was tested at the final 80% level according to the addition of virus solution, etc., and the treatment conditions were treated at 10° C. for 30 minutes.
  • Item density organic matter Cell cultured cells 50% infection concentration (lg TCID50) Difference between virus control and experimental group (log10) MCC-A 80% bovine albumin 10g/L+ yeast extract 10g/L 5.5 0.5 MCC-A 80% bovine albumin 3g/L 2.5 3.5 MCC-A 80% - ⁇ 1.5 ⁇ 4.5 Virus control N/a bovine albumin 10g/L+ yeast extract 10g/L 6 N/a Virus control N/a bovine albumin 3g/L 6 N/a Virus control N/a - 6 N/a N/a; not applicable
  • the high concentration organic matter addition group has a value of 0.5 log10, which means that when MCC-A is sprayed as a stock solution, there is a virucidal effect of 50%.
  • the low-concentration organic substance addition group it is 0.5441 when converted to a constant log value with a value of 3.5 log10.
  • the experimental group where no organic matter was added it is 0.6532 when converted to a constant log value with a value of ⁇ 4.5 log10.
  • the virucidal effect of 80% concentration of MCC-A under low concentration organic matter was 3.5 as a log value through comparison with the control group. Through this, it can be confirmed that it has a bactericidal effect with an approximately 5,400-fold dilution and has 99.98% virucidal power.
  • 80% MCC-A was checked for disinfection in the absence of organic matter, and the results are as follows. Table 7 shows.
  • Table 7 shows the results of the disinfection power test of 80% MCC-A in the absence of organic matter.
  • the virucidal effect is a log value through comparison with the control. ⁇ 4.5. Converting this to a constant log value, it is 0.6532, which means that there is a virucidal effect with a 6,500-fold dilution, so it has 99.99% virucidal power.
  • a secondary disinfection test was conducted to verify the virucidal virus of MCC-A against African swine fever.
  • the laboratory was conducted at the Polish National Academy of Veterinary Sciences in the same way as the first experiment.
  • the test procedure was performed according to "5.7 virucidal test" of PN-EN-14675_2015-06E, a European standard test regulation.
  • the effective chlorine concentration of MCC-A used in the experiment was tested at 143 ppm, and this will be referred to as MCC-A1.
  • the concentration of MCC-A1 actually used in the experiment was tested at the final 80% level according to the addition of virus solution. Treatment conditions were treated at 10° C. for 30 minutes.
  • 10g/L BSA + 10g/L yeast extract was added in the high concentration organic matter treatment condition, and 3g/L BSA was added in the low concentration organic matter treatment condition. used in the experiment.
  • the cytotoxicity of MCC-A1 on the morphology and growth of cells by comparison with the control group for each experimental group was also conducted.
  • the virus, culture medium, and toxicity according to the experimental method were performed, and it was confirmed that the test substance, MCC-A1, had no toxicity at all like MCC-A.
  • the results of the secondary virus efficacy test the results of the virucidal secondary efficacy test of the disinfectant having an effective chlorine concentration of 143 ppm were shown.
  • Item density organic matter Cell cultured cells 50% infection concentration (lg TCID50) Difference between virus control and experimental group (log10) MCC-A1 80% bovine albumin 10g/L+ yeast extract 10g/L 5.5 2.25 MCC-A1 80% bovine albumin 3g/L 2.5 4.25 Virus control N/a bovine albumin 10g/L+ yeast extract 10g/L 7.25 N/a Virus control N/a bovine albumin 3g/L 6.75 N/a
  • Table 9 shows the results of the secondary efficacy test for virucidal MCC-A1, and it can be seen that Table 9 shows the results of the efficacy test for the African swine fever virus of MCC-A1 at a concentration of 143ppm.
  • Table 9 shows the results of the efficacy test for the African swine fever virus of MCC-A1 at a concentration of 143ppm.
  • MCC-A1 at a concentration of 143ppm it is 0.3522 when converted to a constant log value as 2.25 log10 in the high concentration organic matter addition group, which has a virucidal effect with an approximately 3,500-fold dilution, and has a virucidal effect of 99.97%.
  • the virucidal effect of MCC-A1 under high-concentration organic matter was 2.25 as a log value through comparison with the control, and when converted to a constant log value, it was 0.3522, about 3,500 times dilution. It can be confirmed that it has a virucidal power of 99.97%. Then, 3 g/L of BSA was added to MCC-A1 of 80% concentration to create low-concentration organic matter treatment conditions, and then the disinfection power was tested, and the results are shown in Table 11 below. It was.
  • Table 11 is the experimental data of the disinfecting power effect of 80% MCC-A1 against 3g/L BSA. As shown in Table 11, the virucidal effect of MCC-A1 under low-concentration organic matter was compared with the control, and the log value was 4.25. appears, and when converted to a constant log value, it is 0.6284, which has a virucidal effect with an about 6,200-fold dilution, and it can be seen that it has 99.99% virucidal power.
  • Sodium-free hypochlorous acid water through the invention has not only no cytotoxicity, but also has more than 99.97% virucidal power against African swine fever, thereby confirming that it can kill the deadly viral hemorrhagic swine epidemic virus.
  • the present invention can produce sodium-free hypochlorous acid water that maintains an effective chlorine concentration of 78 ppm or 143 ppm constant by adjusting the amount of current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water. have.
  • the virucidal virus experiment on African swine fever attempted in the experimental example of the present invention has not been attempted before, and the effective chlorine concentration of hypochlorous acid water, which is the main effective substance, is adjusted to 78 ppm or 143 ppm to be used as a disinfectant for African swine fever virus. It is expected that the value of the present invention can be recognized in that it presents a new proposal for the application of

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Pest Control & Pesticides (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Plant Pathology (AREA)
  • Toxicology (AREA)
  • Apparatus For Disinfection Or Sterilisation (AREA)

Abstract

The present invention relates to a method for preparing a disinfectant against African swine fever virus, and a disinfectant against African swine fever virus, prepared thereby. The technical subject matters of the present invention are a method for preparing sodium-free hypochlorous acid water for a disinfectant against African swine fever virus, and a disinfectant against African swine fever virus, prepared thereby, the method comprising the steps of: injecting a hydrochloric acid solution into an electrolytic bath, and then electrolyzing same, thereby preparing an electrolysis product containing chlorine gas; and preparing sodium-free hypochlorous acid water while mixing the chlorine gas, having been separated from the electrolysis product, into distilled water, wherein sodium-free hypochlorous acid water, of which the available chlorine concentration is 78 ppm or 143 ppm, is prepared by controlling the amount of current to be supplied to the electrolytic bath according to the amount of the chlorine gas and the amount of distilled water.

Description

아프리카 돼지열병 바이러스 소독제의 제조방법 및 이에 의해 제조되는 아프리카 돼지열병 바이러스 소독제Method for manufacturing African swine fever virus disinfectant and African swine fever virus disinfectant produced thereby
본 발명은 아프리카 돼지열병 바이러스 소독제의 제조방법 및 이에 의해 제조되는 아프리카 돼지열병 바이러스 소독제에 관한 것이다.The present invention relates to a method for producing an African swine fever virus disinfectant and to an African swine fever virus disinfectant prepared thereby.
아프리카 돼지열병 바이러스는 제1종 가축전염병으로 야생멧돼지와 사육돼지의 모든 연령층에서 발생하며, 특히 아프리카 돼지열병 바이러스에 감염된 돼지는 2~10일 사이에 폐사율 100%를 나타낸다. 이러한 아프리카 돼지열병 바이러스는 1921년 케냐에서 발병 사례가 처음 발표된 이후, 1957년 유럽에서 첫 감염을 시작으로 중국, 동남아 등에 발병하였다. 최근 우리나라에서도 발생하여 전파의 차단을 위하여 매개체인 야생멧돼지의 이동 방지 및 방역에 노력을 기울이고 있다.African swine fever virus is a first-class livestock contagious disease that occurs in all age groups of wild boars and domestic pigs. In particular, pigs infected with African swine fever virus show 100% mortality within 2 to 10 days. This African swine fever virus was first reported in Kenya in 1921, and started with the first infection in Europe in 1957, and then in China and Southeast Asia. Recently in Korea, efforts are being made to prevent the movement and quarantine of wild boars, which are vectors, in order to block the transmission.
아프리카 돼지열병 바이러스는 DNA 바이러스로써, RNA 바이러스인 조류독감 및 구제역 바이러스와는 차이점이 있기 때문에 현재 중화항체 결여 및 유전적 다양성의 문제 등으로 인하여 백신은 개발되어 있지 않은 상태이다.African swine fever virus is a DNA virus, which is different from the RNA viruses of avian influenza and foot-and-mouth disease. Therefore, a vaccine has not been developed due to the lack of neutralizing antibodies and problems with genetic diversity.
이러한 아프리카 돼지열병 바이러스의 전파 경로는 감염된 개체와의 접촉에 의한 것과 바이러스에 감염된 개체의 식자재로 유통 및 잔반급여에 따른 전파가 주요 원인인데, 아프리카 돼지열병 바이러스는 냉동에서 1,000일, 혈액에서 18개월간 생존이 가능하고, 저온에서 생존 기간이 길며, pH 3.9 이하의 강산과 pH 11.5 이상의 강알칼리에서 불활화되는 것으로 보고되고 있다.The main cause of the spread of the African swine fever virus is through contact with infected individuals and the distribution and leftover feeding of infected individuals. It is reported that it can survive, has a long survival period at low temperatures, and is inactivated in strong acids with a pH of 3.9 or less and strong alkalis with a pH of 11.5 or more.
현재 시판되고 있는 아프리카 돼지열병 바이러스 소독제로는 구연산(citric acid), 오르토-페닐페놀(ortho-phenylphenol), 요오드화합물(iodine compound), 3종염화합물(potassium monopersulfate compound), sodium dichlori-s-trizinetrione(NADCC), 수산화나트륨(Sodium hydroxide, NaOH) 및 알데하이드제재(aldehyde compound) 등이 있다.Currently marketed African swine fever virus disinfectants include citric acid, ortho-phenylphenol, iodine compound, potassium monopersulfate compound, sodium dichlori-s-trizinetrione ( NADCC), sodium hydroxide (NaOH), and aldehyde compounds.
예컨대 '살균 및 소독 상승효과를 갖는 소독제 및 이의 제조방법(등록번호: 10-1440375)'에서는 개미산, 푸마린산, 사과산, 구연산, 옥살산 및 아스코르브산 중 어느 하나 이상의 유기산과, 인산, 붕산 및 설파민산 중 어느 하나 이상의 무기산을 함유하여 가축 병원체를 살균하기 위한 소독제를 제시한바 있다.For example, in 'Disinfectant having sterilization and disinfection synergistic effect and manufacturing method thereof (registration number: 10-1440375)', one or more organic acids of formic acid, fumaric acid, malic acid, citric acid, oxalic acid and ascorbic acid, phosphoric acid, boric acid and sulfa Disinfectants for sterilizing livestock pathogens by containing any one or more inorganic acids among the acids have been proposed.
하지만 소독제의 pH가 강산성 또는 경우에 따라 수산화나트륨과 같은 알칼리물질이 사용되면 강알칼리성이 되므로 세포독성에 대한 명확한 기준이 설정되지 못할 뿐만 아니라 피부 접촉 및 흡입 노출에 대한 안정성의 우려가 초래되는 문제점이 있다.However, since the pH of the disinfectant is strongly acidic or, in some cases, when an alkaline substance such as sodium hydroxide is used, a clear standard for cytotoxicity cannot be set, and there is a problem that raises concerns about the safety of skin contact and inhalation exposure. have.
이를 해결해 보기 위해 pH 5.5~6.0의 미산성 상태에서 강력한 살균력을 갖는 차아염소산(HOCl)과 함께 염화나트륨(NaCl)을 혼합하여 소독제로 사용해보고자 한바 있다.In order to solve this problem, I tried to use it as a disinfectant by mixing sodium chloride (NaCl) with hypochlorous acid (HOCl), which has strong sterilizing power in a slightly acidic state of pH 5.5 to 6.0.
그러나 미산성 차아염소산에 나트륨이온(Na+)이 혼합되면 세포독성이 강하게 나타나 실제 돼지에 살포하기 어려운 한계점이 있을 뿐만 아니라, 독성이 강해 대용량 물에 희석해서 사용하여야 하기 때문에 아프리카 돼지열병 바이러스를 사멸시키기 위한 농도 조절이 어려운 한계점이 있다.However, when sodium ions (Na + ) are mixed with non-acidic hypochlorous acid, cytotoxicity is strong, and there is a limitation in that it is difficult to actually spray pigs, but also it is very toxic, so it must be diluted in a large amount of water, so it kills the African swine fever virus. There is a limitation in that it is difficult to control the concentration for this purpose.
따라서 아프리카 돼지열병 바이러스에 대한 소독 유효농도에 대한 보고가 아직 없는 차아염소산을 활용하여 치명적인 바이러스성 출혈성 돼지 전염병 바이러스를 소독할 수 있도록 하는 새로운 기술개발 연구가 절실히 요구되고 있는 시점이다.Therefore, research on the development of a new technology to disinfect the deadly viral hemorrhagic swine epidemic virus using hypochlorous acid, which has not yet been reported on the effective disinfection concentration for African swine fever virus, is urgently required.
본 발명은 상기한 문제점을 해소하기 위하여 발명된 것으로, 세포독성이 없으면서 아프리카 돼지열병 바이러스를 사멸시킬 수 있는 소독 유효농도를 갖는 나트륨 미함유 차아염소산수로 이루어진 아프리카 돼지열병 바이러스 소독제의 제조방법 및 이에 의해 제조되는 아프리카 돼지열병 바이러스 소독제를 제공하는 것을 기술적 해결과제로 한다.The present invention was invented to solve the above problems, and a method for producing an African swine fever virus disinfectant comprising sodium-free hypochlorous acid water having a disinfecting effective concentration capable of killing African swine fever virus without cytotoxicity, and It is a technical solution to provide an African swine fever virus disinfectant manufactured by
상기의 기술적 과제를 해결하기 위하여 본 발명은, 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조하는 단계; 및 상기 전해생성물로부터 분리된 염소가스를 증류수에 혼입하면서 나트륨 미함유 차아염소산수를 제조하는 단계;를 포함하여 이루어지고, 상기 염소가스의 양과 상기 증류수의 양에 따라 상기 전해조에 공급되는 전류의 양을 조절함으로써 유효 염소농도가 78ppm 또는 143ppm인 나트륨 미함유 차아염소산수를 제조하는 것을 특징으로 하는, 아프리카 돼지열병 바이러스 소독제용 나트륨 미함유 차아염소산수의 제조방법을 제공한다.In order to solve the above technical problem, the present invention comprises the steps of: preparing an electrolysis product containing chlorine gas by electrolyzing a hydrochloric acid solution into an electrolyzer; and preparing sodium-free hypochlorous acid water while mixing chlorine gas separated from the electrolysis product into distilled water; It provides a method for producing sodium-free hypochlorous acid water for African swine fever virus disinfectants, characterized in that by adjusting the sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm or 143 ppm.
본 발명에 있어서, 상기 차아염소산수의 유효 염소농도가 78 ppm이고, 돼지에 직접 분사하여 소독처리하는 것을 특징으로 한다.In the present invention, the effective chlorine concentration of the hypochlorous acid water is 78 ppm, it is characterized in that the disinfection treatment by spraying directly on the pig.
본 발명에 있어서, 상기 차아염소산수의 유효 염소농도가 143 ppm이고, 돼지 축사에 분사하여 소독처리하는 것을 특징으로 한다.In the present invention, the effective chlorine concentration of the hypochlorous acid water is 143 ppm, it is characterized in that the disinfection treatment by spraying the pig barn.
상기의 다른 기술적 과제를 해결하기 위하여 본 발명은, 상기 방법으로 제조되는 나트륨 미함유 차아염소산수를 포함하여 이루어지는 것을 특징으로 하는, 아프리카 돼지열병 바이러스 소독제를 제공한다.In order to solve the above other technical problems, the present invention provides a disinfectant for African swine fever virus, characterized in that it comprises sodium-free hypochlorous acid water prepared by the above method.
상기의 또다른 기술적 과제를 해결하기 위하여 본 발명은, 상기 아프리카 돼지열병 바이러스 소독제를 돼지 또는 돼지 축사에 직접 분사하여 아프리카 돼지열병 바이러스를 사멸하되, 사멸율이 적어도 99.97%인 것을 특징으로 하는, 나트륨 미함유 차아염소산수를 이용한 아프리카 돼지열병 바이러스의 사멸방법을 제공한다.In order to solve the above another technical problem, the present invention kills the African swine fever virus by directly spraying the African swine fever virus disinfectant to pigs or swine barns, characterized in that the death rate is at least 99.97%, sodium A method for killing African swine fever virus using non-containing hypochlorous acid water is provided.
상기 과제의 해결 수단에 의한 본 발명의 아프리카 돼지열병 바이러스 소독제의 제조방법에 따르면, 염소가스의 양과 증류수의 양에 따라 전해조에 공급되는 전류 조절을 통하여 나트륨을 함유하지 않으면서 차아염소산수의 유효 염소농도를 78ppm 또는 143ppm으로 일정하게 유지할 수 있으므로, 아프리카 돼지열병 바이러스에 대한 소독을 효율적으로 수행하면서 세포독성이 없는 살바이러스제로 활용 가능한 소독제를 제조할 수 있는 효과가 있다.According to the manufacturing method of the African swine fever virus disinfectant of the present invention by means of a solution to the above problem, effective chlorine in hypochlorous acid water without sodium through the control of the current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water Since the concentration can be kept constant at 78 ppm or 143 ppm, there is an effect of efficiently performing disinfection of African swine fever virus and preparing a disinfectant that can be used as a virucidal agent without cytotoxicity.
또한 상술한 아프리카 돼지열병 바이러스 소독제를 돼지에 직접 분사하거나, 돼지 축사에 분사하는 것만으로 아프리카 돼지열병 바이러스의 확산을 초기에 차단할 수 있는 방역 효과가 있다.In addition, there is a quarantine effect that can block the spread of the African swine fever virus in the early stage just by spraying the above-mentioned African swine fever virus disinfectant directly on pigs or on pig barns.
도 1은 본 발명의 아프리카 돼지열병 바이러스 소독제의 제조방법에 따른 순서도.1 is a flow chart according to the manufacturing method of the African swine fever virus disinfectant of the present invention.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
도 1은 본 발명의 아프리카 돼지열병 바이러스 소독제의 제조방법에 따른 순서도를 나타낸 것으로, 도 1을 참조하면, 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조하는 단계(S10)와, 전해생성물로부터 분리된 염소가스를 증류수에 혼입하면서 나트륨 미함유 차아염소산수를 제조하는 단계(S20)를 통하여 제조된다.1 is a flowchart according to the manufacturing method of the African swine fever virus disinfectant of the present invention. Referring to FIG. 1, a hydrochloric acid solution is added to an electrolyzer and then electrolyzed to produce an electrolytic product containing chlorine gas ( It is prepared through S10) and the step (S20) of preparing sodium-free hypochlorous acid water while mixing chlorine gas separated from the electrolysis product into distilled water.
특히 본 발명의 아프리카 돼지열병 바이러스 소독제의 제조방법에 따르면, 염소가스의 양과 증류수의 양에 따라 전해조에 공급되는 전류의 양을 조절함으로써 유효 염소농도가 78ppm 또는 143ppm인 나트륨 미함유 차아염소산수를 제조하는 것에 특징이 있으며, 상술한 각각의 단계에 대해서는 다음과 같이 설명될 수 있다.In particular, according to the production method of the African swine fever virus disinfectant of the present invention, sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm or 143 ppm is produced by controlling the amount of current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water It is characterized in that, each of the above-described steps can be described as follows.
이러한 제조방법에 의하면 먼저, 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조한다(S10).According to this manufacturing method, first, a hydrochloric acid solution is put into an electrolyzer and then electrolyzed to prepare an electrolysis product containing chlorine gas (S10).
통상 전기분해는 전해조 내에 음극과 양극을 형성하는 한 쌍의 전극이 설치되고, 각 전극에 전기를 공급하여 전해조 내에 들어있는 물질을 전기분해한다. 이러한 전기분해는 격막의 유무에 따라 음극과 양극 사이에 격막을 갖는 격막식과, 격막을 갖지 않는 무격막식으로 나뉠 수 있다. 무격막식의 경우, 전압과 전류를 높이고 전극의 개수 또는 전극의 표면적을 넓히게 되면 생성되는 전해생성물의 양을 늘릴 수 있는 장점이 있으므로, 본 발명에서는 무격막식 전기분해를 적용하는 것이 바람직하다.In general, in electrolysis, a pair of electrodes forming a cathode and an anode are installed in an electrolyzer, and electricity is supplied to each electrode to electrolyze the material contained in the electrolyzer. Such electrolysis may be divided into a diaphragm type having a diaphragm between the cathode and an anode, and a diaphragm-free type having no diaphragm depending on the presence or absence of the diaphragm. In the case of the diaphragm-free type, since there is an advantage in that the amount of generated electrolysis products can be increased by increasing the voltage and current and increasing the number of electrodes or the surface area of the electrodes, it is preferable to apply the diaphragm-free electrolysis in the present invention.
무격막식 전기분해에 의하면, 염산용액이 전기분해되면서 전해조의 양극에서 염소가스가 발생함과 동시에 음극에서 수소가스가 발생한다. 우선 염산용액이 전기분해되면 양극에서 다음과 같은 반응으로 염소가스가 생성된다.According to the diaphragm-free electrolysis, as the hydrochloric acid solution is electrolyzed, chlorine gas is generated at the anode of the electrolyzer and hydrogen gas is generated at the cathode. First, when the hydrochloric acid solution is electrolyzed, chlorine gas is generated at the anode through the following reaction.
HCl → H+ + Cl- HCl → H + + Cl -
2Cl- → Cl2 + 2e- 2Cl - → Cl 2 + 2e -
양극과 달리, 음극에서는 다음과 같은 반응으로 수소가스가 생성된다.Unlike the anode, at the cathode, hydrogen gas is generated by the following reaction.
HCl → H+ + Cl- HCl → H + + Cl -
2H+ + 2e- → H2 2H + + 2e - → H 2
상기와 같은 반응을 통하여 양극에서는 염소가스, 음극에서는 수소가스가 발생함을 확인할 수 있다. 이때 가볍고 많은 양으로 생성되는 수소가스의 경우 염소가스와 섞여 빠른 속도로 유출될 수 있기 때문에, 이를 해결하기 위해 전해조에 에어벤트를 설치하여 염소가스를 수소가스와 완전히 분리된 상태로 전해조의 외부로 방출하여 수소가스와 염소가스가 혼합되는 현상을 방지할 수 있다.Through the above reaction, it can be confirmed that chlorine gas is generated at the anode and hydrogen gas is generated at the cathode. At this time, in the case of light and large amount of hydrogen gas, it can be mixed with chlorine gas and flow out at a high speed. It is possible to prevent the mixture of hydrogen gas and chlorine gas.
특히 경우에 따라 염산용액과 함께 전기분해능을 더 높이기 위해 염화나트륨을 혼합하는 경우가 있는데, 염화나트륨은 이온화가 커서 전기분해되는 과정에서 양이온과 음이온으로 분리되는 비율이 100%에 가까워 전류를 잘 흐르게 하고 전극과 전극 사이의 저항을 작게 하는 장점이 있긴 하나, 염화나트륨의 나트륨이온이 차아염소산에 결합되면 독성이 너무 강하여 대용량의 물에 희석하여 사용해야만 하는 번거로움이 있기 때문에 본 발명에서는 염산용액에 염화나트륨을 혼합하지 않는 것이 바람직하다.In particular, in some cases, sodium chloride is mixed with a hydrochloric acid solution to further increase the electrolytic ability. Sodium chloride has a large ionization, so the ratio of separation into cations and anions during electrolysis is close to 100%, so that the current flows well and the electrode Although there is an advantage of reducing the resistance between the electrode and the sodium ion of sodium chloride, when the sodium ion of sodium chloride is combined with hypochlorous acid, the toxicity is so strong that it is inconvenient to use it after diluting it in a large amount of water. In the present invention, sodium chloride is mixed with the hydrochloric acid solution It is preferable not to
다음으로, 전해생성물로부터 분리된 염소가스를 증류수에 혼입하면서 나트륨 미함유 차아염소산수를 제조한다(S20).Next, sodium-free hypochlorous acid water is prepared while mixing chlorine gas separated from the electrolysis product into distilled water (S20).
나트륨이 함유되지 않은 차아염소산수 제조를 위해 유입되는 증류수는 인접된 위치의 장치에서 공급되기 때문에, 유입되는 증류수의 양은 항상 변할 수 있다. 이로 인해 증류수의 유량이 적어지거나 많아질 경우 생성되는 차아염소산수의 농도는 높아지거나 낮아질 수 있게 된다.Since the distilled water introduced for the production of sodium-free hypochlorous acid water is supplied from an adjacent device, the amount of distilled water introduced can always change. Due to this, when the flow rate of distilled water is reduced or increased, the concentration of hypochlorous acid water generated can be increased or decreased.
이의 해결수단으로, 본 발명에서는 전해조로 공급되는 염산용액의 유량을 감지하여 측정된 값을 바탕으로 전기분해 시 전력공급 조절을 통해 전해조에 공급되는 전류를 조절하고, 이러한 전류의 조절로 전해조에서 생성되는 전해생성물의 염소가스 양을 조절하여 차아염소산수의 유효 염소농도를 78ppm 또는 143ppm으로 유지하여 일정하게 공급할 수 있게 된다.As a solution to this, in the present invention, the flow rate of the hydrochloric acid solution supplied to the electrolyzer is sensed and the current supplied to the electrolyzer is controlled by controlling the power supply during electrolysis based on the measured value, and generated in the electrolyzer by controlling the current By controlling the amount of chlorine gas in the electrolysis product, the effective chlorine concentration of hypochlorous acid water is maintained at 78 ppm or 143 ppm, so that it can be supplied constantly.
즉 차아염소산수의 유효 염소농도가 78ppm 또는 143ppm으로 유지되도록 전해조 내에 인가되는 전류를 조절하기 때문에, 필요로 하는 장소에 따라 일정 유효 염소농도를 가져 소독제의 소독력을 항상 일정하게 유지할 수 있게 된다.That is, since the current applied to the electrolyzer is controlled so that the effective chlorine concentration of the hypochlorous acid water is maintained at 78 ppm or 143 ppm, it has a certain effective chlorine concentration depending on the required place, so that the disinfecting power of the disinfectant can be constantly maintained.
돼지에 직접 분사를 위한 대류 방역의 경우, 차아염소산수의 유효 염소농도가 78ppm이 되도록 조절한다.In the case of convection control for direct spraying on pigs, the effective chlorine concentration of hypochlorous acid water is adjusted to be 78ppm.
이때 전해생성물로부터 분리되는 염소가스를 흘려주면서 증류수는 1.0~1.5L/min로 공급한다. 증류수를 1.0L/min 미만으로 공급하면 증류수의 양이 적어 최종생성물인 차아염소산수의 유효 염소농도가 78ppm을 초과하게 되어 대류 방역을 하기에 효율적이지 못하고, 1.5L/min을 초과하여 공급하면 증류수의 양이 너무 많아져 최종생성물인 차아염소산수가 희석되어 유효 염소농도가 78ppm에 훨씬 못미치게 되어 소독효율이 좋지 못하다. 돼지에 직접 분사하여 소독처리함에 있어서 차아염소산수의 유효 염소농도가 78ppm이 되기 위해 가장 바람직한 증류수 공급량은 1.25L/min이라 할 수 있다.At this time, while flowing chlorine gas separated from the electrolysis product, distilled water is supplied at 1.0 ~ 1.5L/min. When distilled water is supplied at less than 1.0L/min, the amount of distilled water is small and the effective chlorine concentration of the final product, hypochlorous acid water, exceeds 78ppm, which is not effective for convection prevention. The amount of chlorous acid is too large, and the final product, hypochlorous acid water, is diluted, and the effective chlorine concentration is much lower than 78ppm, so the disinfection efficiency is not good. In the disinfection treatment by spraying directly on pigs, the most preferable supply amount of distilled water is 1.25L/min so that the effective chlorine concentration of hypochlorous acid water becomes 78ppm.
전류와 관련하여, 전원공급기를 통해 전해조로 공급되는 전원의 전류 량을 측정하여, 이렇게 측정된 전류의 량을 바탕으로 전원공급기에서 공급되는 전원을 제어해 생성되는 차아염소산수의 유효 염소농도를 제어하게 된다. 전류가 18A 미만이면 78ppm 농도를 맞추기 위해 차아염소수 생성시간을 더욱 늘여야 하는 단점이 있고, 22A를 초과하면 차아염소산수를 78ppm 농도를 일정하게 유지해 주기에 어려운 단점이 있다. 차아염소산수의 유효 염소농도를 78ppm으로 유지시켜주기 위해서는 20A의 전류를 흘려주는 것이 가장 바람직하다.In relation to the current, the effective chlorine concentration of hypochlorous acid water generated by measuring the amount of current supplied to the electrolyzer through the power supply and controlling the power supplied from the power supply based on the measured amount of current is controlled. will do If the current is less than 18A, there is a disadvantage that the generation time of hypochlorite water needs to be further increased to meet the 78ppm concentration, and if it exceeds 22A, it is difficult to maintain a constant 78ppm concentration of hypochlorous acid. In order to maintain the effective chlorine concentration of hypochlorous acid water at 78ppm, it is most desirable to flow a current of 20A.
한편, 축사 바닥이나 기구 등 유기물이 함유된 돼지 축사에 직접 분사하기 위해서는 차아염소산수의 유효 염소농도가 143ppm이 되도록 조절한다.On the other hand, in order to directly spray the pig barn containing organic matter such as the barn floor or equipment, the effective chlorine concentration of the hypochlorous acid water is adjusted to 143ppm.
143ppm의 유효 염소농도를 갖는 차아염소산수를 제조하기 위해서는 전해생성물로부터 분리되는 염소가스를 흘려주면서 증류수는 0.8~0.9L/min로 공급한다. 증류수를 0.8L/min 미만으로 공급하면 증류수의 양이 적어 최종생성물인 차아염소산수의 유효 염소농도가 143ppm을 초과하게 되어 독성이 강해질 수 있으며, 0.9L/min을 초과하여 공급하면 증류수의 양이 너무 많아져 최종생성물인 차아염소산수의 유효 염소농도가 143ppm에 훨씬 못미쳐 유기물이 존재하는 곳에 살포하게 되더라도 소독효율이 기대에 못미치게 된다. 유기물이 존재하는 장소에 살포하여 소독함에 있어 차아염소산수의 유효 염소농도가 143ppm이 되기 위해 가장 바람직한 증류수 공급량은 0.88L/min이라 할 수 있다.In order to produce hypochlorous acid water having an effective chlorine concentration of 143 ppm, distilled water is supplied at 0.8 to 0.9 L/min while flowing chlorine gas separated from the electrolysis product. If distilled water is supplied at less than 0.8L/min, the amount of distilled water is small and the effective chlorine concentration of the final product, hypochlorous acid water, exceeds 143ppm, which may increase toxicity. The effective chlorine concentration of hypochlorous acid water, which is the final product, is much lower than 143ppm, so even if it is sprayed where organic matter is present, the disinfection efficiency is not as good as expected. In order to sterilize by spraying the place where organic matter exists, the most preferable supply of distilled water is 0.88L/min so that the effective chlorine concentration of hypochlorous acid water becomes 143ppm.
전류와 관련해서는, 유효 염소농도를 78ppm으로 조절하는 경우와 마찬가지로, 전원공급기를 통해 전해조로 공급되는 전원의 전류 량을 측정하여, 이렇게 측정된 전류의 량을 바탕으로 전원공급기에서 공급되는 전원을 제어해 생성되는 차아염소산수의 유효 염소농도를 제어하게 된다. 전류가 28A 미만이면 143ppm 농도를 맞추기 위해 전류를 더 높여 염산용액의 전기분해도를 높이면서 염소가스 발생량을 늘여야 하는 단점이 있고, 32A를 초과하면 차아염소산수를 143ppm 농도를 일정하게 유지해 주기에 어려운 단점이 있다. 차아염소산수의 유효 염소농도를 143ppm으로 유지시켜주기 위해서는 30A의 전류를 흘려주는 것이 가장 바람직하다.Regarding the current, as in the case of adjusting the effective chlorine concentration to 78ppm, the amount of current supplied to the electrolyzer through the power supply is measured, and the power supplied from the power supply is controlled based on the amount of current measured in this way. It controls the effective chlorine concentration of hypochlorous acid water produced by the If the current is less than 28A, there is a disadvantage that the amount of chlorine gas must be increased while increasing the electrolysis degree of the hydrochloric acid solution by increasing the current to meet the 143ppm concentration. There is this. In order to maintain the effective chlorine concentration of hypochlorous acid water at 143ppm, it is most desirable to flow a current of 30A.
차아염소산수의 경우 pH에 따라 소독력에 미치는 영향이 크다. 보통 pH 5.0~6.0 범위에서 소독제로 적용하기에 적합한데, 본 발명의 유효 염소농도가 78ppm인 나트륨 미함유 차아염소산수는 pH 5.5가 되어 돼지에 직접 분사가 가능한 대류 방역에 적절한 소독제가 될 수 있다. 특히 차아염소산수의 pH를 4.5로 낮춰 유효 염소농도를 143ppm으로 높이게 되더라도 세포독성이 없어 돼지 축사에 직접 분사를 통해 아프리카 돼지열병 바이러스를 사멸시킬 수 있게 된다.In the case of hypochlorous acid water, the effect on disinfection power is large depending on the pH. Usually, it is suitable for application as a disinfectant in the range of pH 5.0 to 6.0, and the sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm of the present invention becomes pH 5.5, so it can be an appropriate disinfectant for convection prevention that can be directly sprayed on pigs . In particular, even if the pH of hypochlorous acid water is lowered to 4.5 and the effective chlorine concentration is raised to 143 ppm, there is no cytotoxicity, so it is possible to kill the African swine fever virus by spraying it directly into the pig barn.
이하, 본 발명의 실시예를 더욱 상세하게 설명하면 다음과 같다. 단, 이하의 실시예는 본 발명의 이해를 돕기 위하여 예시하는 것일 뿐, 이에 의하여 본 발명의 범위가 한정되는 것은 아니다.Hereinafter, an embodiment of the present invention will be described in more detail as follows. However, the following examples are merely illustrative to aid the understanding of the present invention, and the scope of the present invention is not limited thereby.
<실시예 1><Example 1>
0.1% 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조하였다. 전해생성물로부터 분리된 염소가스 공급 하에 증류수를 1.25L/min으로 혼입하면서 20A의 전류로 조절하여 78ppm의 차아염소산수를 제조하였다. 이때 pH는 5.5였다.After adding 0.1% hydrochloric acid solution to the electrolyzer, electrolysis was performed to prepare an electrolysis product containing chlorine gas. 78 ppm of hypochlorous acid water was prepared by controlling a current of 20 A while mixing distilled water at 1.25 L/min under the supply of chlorine gas separated from the electrolysis product. At this time, the pH was 5.5.
<실시예 2><Example 2>
0.1% 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조하였다. 전해생성물로부터 분리된 염소가스 공급 하에 증류수를 0.88L/min으로 혼입하면서 30A의 전류로 조절하여 143ppm의 차아염소산수를 제조하였다. 이때 차아염소산수의 pH는 4.5였다.After adding 0.1% hydrochloric acid solution to the electrolyzer, electrolysis was performed to prepare an electrolysis product containing chlorine gas. Under the supply of chlorine gas separated from the electrolysis product, distilled water was mixed at 0.88 L/min and the current was adjusted to 30 A to prepare 143 ppm of hypochlorous acid water. At this time, the pH of the hypochlorous acid water was 4.5.
<실험예 1><Experimental Example 1>
본 실험예에서는 아프리카 돼지열병 바이러스 소독제의 세포독성 및 소독력을 실험하였다. 세포독성은 소독제로 처리된 아프리카 돼지열병 바이러스를 접종한 세포에 나타나는 세포독성을 최종 판독하여 바이러스의 불활화 유무를 판정하였으며, 소독력은 아래의 실험조건에 따라 실험을 통해 분석을 진행하였다.In this experimental example, the cytotoxicity and disinfection power of the African swine fever virus disinfectant was tested. For the cytotoxicity, the inactivation of the virus was determined by finally reading the cytotoxicity of the cells inoculated with the African swine fever virus treated with the disinfectant.
실험조건Experimental conditions
아프리카 돼지열병에 대한 살바이러스 실험조건은 표 1과 같다.Table 1 shows the virucidal test conditions for African swine fever.
항목Item 조건condition
유효물질active substance 차아염소산(HOCl)Hypochlorous acid (HOCl)
유효물질의 염소농도Chlorine concentration of active substance 78ppm, 143ppm78ppm, 143ppm
바이러스 처리시간Virus processing time 30분30 minutes
바이러스 실험온도Virus test temperature 10℃10℃
방해물질(유기몰) 농도Interfering substance (organic molar) concentration 고농도high concentration BSA 10g/L + yeast extract 10g/LBSA 10g/L + yeast extract 10g/L
저농도low concentration BSA 3g/LBSA 3g/L
무첨가군additive-free MEM + 7% FCSMEM + 7% FCS
세포배양 온도cell culture temperature 37℃37℃
사용된 바이러스 종Virus species used African Swine Fever Virus(ASFV) strain BA71VAfrican Swine Fever Virus (ASFV) strain BA71V
표 1에 나타낸 것처럼, 아프리카 돼지열병 바이러스 효력실험법에 준하여 실시예 1 및 실시예 2에 따라 유효 염소농도가 78ppm 및 143ppm인 소독제를 각각 준비하고, 실험에 사용되는 아프리카 돼지열병 바이러스는 African Swine Fever Virus(ASFV) strain BA71V을 사용하였다.실험 처리온도는 10℃에서 30분간 처리하였다. 실험군은 3가지로 구분하였으며, 각각은 유기물이 첨가되었을 때의 효력 검증 실험을 위하여 저농도의 유기물이 함유된 것에 해당하는 것은 3g/L 소혈청 알부민(bovine serum albumin)을 첨가하고, 고농도의 유기물이 함유된 것에 해당하는 것은 10g/L 소혈청 알부민과 10g/L 효모추출물(yeast extract)을 함께 사용하였다. 또한 세포배양배지 단독 첨가군에서도 효력실험을 실시하였다.As shown in Table 1, disinfectants having effective chlorine concentrations of 78 ppm and 143 ppm were prepared according to Examples 1 and 2 according to the African swine fever virus efficacy test method, respectively, and the African swine fever virus used in the experiment was African Swine Fever Virus (ASFV) strain BA71V was used. Experimental treatment temperature was 10°C for 30 minutes. The experimental group was divided into 3 groups, and for each of the experiments to verify the efficacy when organic substances were added, 3g/L bovine serum albumin was added to those containing low concentrations of organic substances, and high concentrations of organic substances were added. Corresponding to what was contained, 10g/L bovine serum albumin and 10g/L yeast extract were used together. In addition, efficacy experiments were also conducted in the cell culture medium alone addition group.
아프리카 돼지열병 바이러스에 효력실험을 실시하기 전에 실험에 사용되는 모든 물질(유기물, 세포배양액 및 소독제 등)은 10±1℃ 유지되는 항온조에서 취급하였다.All materials (organic material, cell culture solution, disinfectant, etc.) used in the experiment were handled in a thermostat maintained at 10±1℃ before performing the efficacy test on African swine fever virus.
실험방법Experimental method
아프리카 돼지열병에 대한 살바이러스 검증을 위한 실험방법은 아래의 표 2에서와 같은 과정으로 진행하였다.The experimental method for verifying the virucidal virus for African swine fever was carried out in the same manner as in Table 2 below.
순서order 내용(유럽 표준형: PN-EN-14675_2015-06E "5.7 virucidal test")Content (European standard: PN-EN-14675_2015-06E "5.7 virucidal test")
1One 유기물 1ml + 바이러스 배양액 1ml1ml organic material + 1ml virus culture medium
22 10±1℃ 유지되는 항온조에서 2min ± 10s 간 혼합Mix for 2min ± 10s in a thermostat maintained at 10±1℃
33 실험용액 8ml을 첨가 혼합하고,10±1℃에서 30 min ± 10 s 간 처리8ml of the test solution is added and mixed, and treated at 10±1℃ for 30 min±10 s
44 실험액 0.5ml을 4±1℃유지한 MEM(minimum essential medium) +2 % FCS(fetal calf serum; 소태아 혈청) 혼합액 4.5ml에 첨가0.5ml of the test solution was added to 4.5ml of MEM (minimum essential medium) +2% FCS (fetal calf serum) mixture maintained at 4±1℃
55 4±1℃ 유지한 MEM + 2% FCS 혼합액으로실험물질이 10-9까지 희석된 실험 표본을 준비Prepare a test sample in which the test substance is diluted to 10 -9 with a mixture of MEM + 2% FCS maintained at 4±1℃
66 실험 희석액은 세포배양에 접종할 때까지 4℃ 유지The experimental dilution is maintained at 4°C until inoculated into cell culture.
77 실험 희석액을 배양세포가 함유된 배양 용기 단위로 접종Inoculate the experimental dilution solution into a culture vessel containing cultured cells.
88 각 희석액은 8개 단위별로 접종Each dilution was inoculated in units of 8
99 세포배양 후 희석별로 바이러스 감염도 측정Measurement of virus infectivity by dilution after cell culture
1010 정량적 실험의 적정 결과는 희석 배율에 따른 CPE(cytopathic effect; 세포병변효과)의 존재 유무에 따라 10~100% 비율로 측정[Spearman and Karber 의 방법에 따라 측정]The titration result of the quantitative experiment is measured at a ratio of 10 to 100% depending on the presence or absence of CPE (cytopathic effect) according to the dilution factor [measured according to the method of Spearman and Karber]
1111 Plaque assay(바이러스의 존재 유무 확인법)도 병행하여 수행Plaque assay (a method for confirming the presence or absence of a virus) is also performed in parallel.
1212 각 실험구 및 대조구의 세포변성 및 세포성장 측정 등을 통하여 세포독성 판별Determination of cytotoxicity by measuring cell degeneration and cell growth of each experimental group and control group
실험방법은 표 2에서와 같이, 우선 방해물질로써 투입되는 유기물 1ml을 실험용기에 넣고, 바이러스 배양액 1ml을 투입하였다. 실험용기는 10±1℃ 유지되는 항온조이고, 유기물과 바이러스 배양액을 2min ± 10s 간 혼합하였다.그후, 소독제 실험용액 8ml을 첨가 혼합하고, 10±1℃에서 30min ± 10s 간 처리하였다. 처리시간이 끝나기 바로 직전 잘 섞어주었다.As for the experimental method, as shown in Table 2, first, 1 ml of an organic material to be introduced as an interfering material was put into an experimental container, and 1 ml of a virus culture solution was added. The experimental vessel is a thermostat maintained at 10±1°C, and the organic material and the virus culture solution were mixed for 2min±10s. Then, 8ml of the disinfectant test solution was added and mixed, and treated at 10±1℃ for 30min±10s. Mix well just before the end of the treatment time.
처리시간이 종료된 후, 실험액 0.5ml을 4±1℃ 유지되는 항온수조 또는 잘게 부순 얼음에서 유지한 MEM(minimum essential medium) + 2% FCS(fetal calf serum; 소태아 혈청) 혼합액 4.5ml에 첨가하였다.After the treatment time was over, 0.5 ml of the test solution was added to 4.5 ml of a mixture of MEM (minimum essential medium) + 2% FCS (fetal calf serum) maintained in a thermostatic water bath maintained at 4±1°C or crushed ice. added.
4±1℃로 유지되는 항온수조 또는 잘게 부순 얼음에서 유지한 MEM(minimum essential medium) + 2% FCS(fetal calf serum; 소태아 혈청) 혼합액으로 실험물질이 10-9까지 희석된 실험 표본을 준비하였다.Prepare an experimental sample in which the test substance is diluted to 10 -9 with a mixture of MEM (minimum essential medium) + 2% FCS (fetal calf serum) maintained in a constant temperature bath maintained at 4±1°C or crushed ice did.
바이러스 처리가 끝난 실험 희석액은 세포배양에 접종할 때까지 4℃ 또는 잘게 부순 얼음 하에서 보관하였다.The experimental dilutions after virus treatment were stored at 4°C or under crushed ice until inoculated into cell cultures.
실험 희석액은 배양세포가 함유된 배양 용기 단위로 접종하였으며, 각 희석액은 8개 단위별로 접종하였다.Experimental dilutions were inoculated in units of culture vessels containing cultured cells, and each dilution was inoculated in units of 8 units.
세포배양 후 희석별로 감염도를 측정하였다. 정량적 실험의 적정 결과는 희석 배율에 따른 CPE(cytopathic effect; 세포병변효과)의 존재 유무에 따라 0~100% 비율로 측정하였다. 이 값은 Spearman and Karber의 방법에 따라 측정한 것이다. 이 실험에서는 또한 plaque assay(바이러스의 존재 유무 확인법)도 병행하여 수행하였다.After cell culture, infectivity was measured for each dilution. The titration result of the quantitative experiment was measured at a ratio of 0-100% depending on the presence or absence of CPE (cytopathic effect; cytopathic effect) according to the dilution factor. This value was measured according to the method of Spearman and Karber. In this experiment, plaque assay (a method to confirm the presence or absence of virus) was also performed in parallel.
아프리카 돼지열병 바이러스 1차 소독력 실험African swine fever virus primary disinfection power test
소독제의 아프리카 돼지열병에 대한 살바이러스 효과 검증을 위하여 정량적 실험을 실시하였다. 실험기관은 폴란드 국립수의과학원에서 수행하였으며, 실험절차는 유럽 표준 실험 규정인 PN-EN-14675_2015-06E의 "5.7 virucidal test"에 따라 수행하였다.A quantitative experiment was conducted to verify the virucidal effect of the disinfectant on African swine fever. The laboratory was conducted at the National Academy of Veterinary Medicine in Poland, and the experimental procedure was performed according to "5.7 virucidal test" of PN-EN-14675_2015-06E, a European standard laboratory regulation.
실험에 사용된 소독제의 유효 염소농도는 78ppm으로 실험하였으며, 이를 MCC-A로 표기하기로 한다. 실험에 실제 사용된 MCC-A의 농도는 바이러스액 첨가 등에 따라 최종 80% 수준에서 실험하였다, 처리조건은 10℃에서 30분간 처리하였다.The effective chlorine concentration of the disinfectant used in the experiment was tested at 78 ppm, and this will be denoted as MCC-A. The concentration of MCC-A actually used in the experiment was tested at the final 80% level according to the addition of virus solution, etc., and the treatment conditions were treated at 10° C. for 30 minutes.
유기물의 함량에 따른 MCC-A의 살바이러스 효과 검증을 위하여 고농도 유기물 처리조건에서는 10g/L BSA + 10g/L yeast extract를 첨가하여 실험에 실시하였으며, 저농도 유기물 처리조건은 3g/L BSA를 첨가하여 실험에 사용하였다. 그리고 유기물을 첨가하지 않은 실험구로 구분하여 실험을 실시하였다.In order to verify the virucidal effect of MCC-A according to the content of organic matter, 10g/L BSA + 10g/L yeast extract was added in the high concentration organic matter treatment condition, and 3g/L BSA was added in the low concentration organic matter treatment condition. used in the experiment. And the experiment was carried out by dividing into the experimental group to which no organic matter was added.
우선, 각 실험구별로 대조구와의 비교를 통하여 세포의 형태 및 성장에 MCC-A가 영향을 미치는지에 대한 세포독성에 관한 조사를 진행하여 아래의 표 3에 나타내었다.First, by comparing with the control group for each experimental group, the cytotoxicity of whether MCC-A affects the cell morphology and growth was investigated, and it is shown in Table 3 below.
조항article 수행 여부Whether to perform
실험 바이러스 현탁액의 적정(실험물질의 처리 전후 로그수치 4 감소의 결정에 따른다.)Titration of the test virus suspension (subject to the determination of the logarithmic 4 decrease before and after the treatment of the test substance). 수행Perform
실험물질의 세포독성이 없음의 판별 여부(세포의 변성, 증식 또는 바이러스 감수성)Whether the test substance is non-cytotoxic (cell denaturation, proliferation, or virus susceptibility) 수행Perform
배양세포에 접종된 바이러스 비교 검토(실험 혼합액 또는 배양배지의 희석에 따른 비교)Comparative review of virus inoculated into cultured cells (comparison according to dilution of experimental mixture or culture medium) 수행Perform
표 3을 참조하면, 실험방법에 따른 바이러스, 배양배지, 독성여부를 실시하였으며, 실험물질인 MCC-A는 독성이 전혀 없는 것을 확인할 수 있었다.이어서 표 4는 MCC-A의 살바이러스 1차 효력실험결과를 나타낸 것으로, 78ppm의 유효 염소농도를 갖는 소독제의 살바이러스 1차 효력실험결과를 알 수 있다.Referring to Table 3, the virus, culture medium, and toxicity were tested according to the experimental method, and it was confirmed that the test material, MCC-A, had no toxicity. As a result of the experiment, it is possible to know the results of the first virucidal efficacy test of a disinfectant having an effective chlorine concentration of 78ppm.
항목Item 농도density 유기물organic matter 세포배양세포50% 감염농도(lg TCID50)Cell cultured cells 50% infection concentration (lg TCID50) 바이러스 대조구와실험구의 차이값(log10)Difference between virus control and experimental group (log10)
MCC-AMCC-A 80%80% bovine albumin 10g/L+ yeast extract 10g/Lbovine albumin 10g/L+ yeast extract 10g/L 5.55.5 0.50.5
MCC-AMCC-A 80%80% bovine albumin 3g/Lbovine albumin 3g/L 2.52.5 3.53.5
MCC-AMCC-A 80%80% -- ≤1.5≤1.5 ≥4.5≥4.5
Virus controlVirus control N/aN/a bovine albumin 10g/L+ yeast extract 10g/Lbovine albumin 10g/L+ yeast extract 10g/L 66 N/aN/a
Virus controlVirus control N/aN/a bovine albumin 3g/Lbovine albumin 3g/L 66 N/aN/a
Virus controlVirus control N/aN/a -- 66 N/aN/a
N/a; not applicableN/a; not applicable
표 4에 나타나 있듯이, 78ppm의 MCC-A에서는 고농도 유기물 첨가군에서는 0.5 log10 수치로써 MCC-A를 원액으로 살포할 시 50%의 살바이러스 효과가 있음을 의미한다. 저농도 유기물 첨가군에서는 3.5 log10 수치로 상수 로그값으로 환산하면 0.5441인데, 약 5천4백배 희석액으로의 살바이러스 효과가 있다는 것으로 99.98% 살바이러스력을 가지는 것을 의미한다. 유기물이 첨가되지 않은 실험구에서는 ≥4.5 log10 수치로 상수 로그값으로 환산하면 0.6532인데, 6천5백배 희석액으로 살바이러스 효과가 있다는 것으로 99.99% 살바이러스력을 가지는 것을 의미한다.표 4를 기반으로, 80% 농도의 MCC-A에 bovine albumin 10g/L + yeast extract 10g/L의 고농도 유기물이 첨가된 경우와, 대조군의 살바이러스 효능을 실험해 보았으며, 그 결과를 아래의 표 5에 정리하였다.As shown in Table 4, in the MCC-A of 78ppm, the high concentration organic matter addition group has a value of 0.5 log10, which means that when MCC-A is sprayed as a stock solution, there is a virucidal effect of 50%. In the low-concentration organic substance addition group, it is 0.5441 when converted to a constant log value with a value of 3.5 log10. In the experimental group where no organic matter was added, it is 0.6532 when converted to a constant log value with a value of ≥4.5 log10. , when a high concentration organic material of 10 g/L of bovine albumin + 10 g/L of yeast extract was added to MCC-A at a concentration of 80%, and the virucidal efficacy of the control group, the results are summarized in Table 5 below. .
항목Item 농도density 유기물organic matter 처리시간(분)Processing time (minutes) 희석농도 (lg)Dilution Concentration (lg)
22 33 44 55 66 77 88 99
MCC-AMCC-A 80%80% bovine serumalbumin 10g/L+yeast extract10g/Lbovine serumalbumin 10g/L+yeast extract10g/L 3030 11111111 11111111 11111111 11111111 00000000 00000000 00000000 00000000
ViruscontrolVirus control N/aN/a bovine serumalbumin 10g/L+yeast extract10g/Lbovine serumalbumin 10g/L+yeast extract10g/L 3030 11111111 11111111 11111111 11111111 10101010 00000000 00000000 00000000
1; virus present - 4개의 세포배양내에서의 세포변성효과(CPE)0; no virus presentN/a; not applicableOne; virus present - cytopathic effect (CPE)0 in 4 cell cultures; no virus presentN/a; not applicable
이러한 표 5를 참조하면, 80% 농도의 MCC-A가 10g/L BSA + 10g/L yeast extract의 고농도 유기물 하에서 소득력이 있는지의 여부를 나타낸 것으로, 표 5에서 보여지듯이, 고농도 유기물 하에서 MCC-A의 살바이러스 효과는 대조구와의 비교를 통하여 로그값으로 0.5를 나타냄에 따라, 대조구에 비해 강한 소독력을 가짐을 알 수 있다.이어서 80% 농도의 MCC-A에 BSA 3g/L을 첨가하여 저농도 유기물 처리조건을 만들어 준 후 소독력을 실험하여, 그 결과를 아래의 표 6에 나타내었다.Referring to Table 5, it indicates whether or not the 80% concentration of MCC-A has income under the high concentration organic matter of 10g/L BSA + 10g/L yeast extract, and as shown in Table 5, MCC- As the virucidal effect of A was compared with the control, it was found to have a stronger disinfecting power compared to the control, as it showed a log value of 0.5. Then, 3g/L of BSA was added to 80% concentration of MCC-A After making the organic matter treatment conditions, the disinfection power was tested, and the results are shown in Table 6 below.
항목Item 농도density 유기물organic matter 처리시간(분)Processing time (minutes) 희석농도 (lg)Dilution Concentration (lg)
22 33 44 55 66 77 88 99
MCC-AMCC-A 80%80% bovine serum albumin3g/Lbovine serum albumin3g/L 3030 10111011 00100010 00000000 00000000 00000000 00000000 00000000 00000000
ViruscontrolVirus control N/aN/a bovine serum albumin3g/Lbovine serum albumin3g/L 3030 11111111 11111111 11111111 11111111 00110011 00000000 00000000 00000000
1; virus present - 4개의 세포배양내에서의 세포변성효과(CPE),0; no virus presentN/a; not applicableOne; virus present - cytopathic effect (CPE) in 4 cell cultures, 0; no virus presentN/a; not applicable
즉 표 6을 참조하면, 80% 농도의 MCC-A가 저농도 유기물 하에서 살바이러스 효과는, 대조구와의 비교를 통하여 로그값으로 3.5를 나타내었다. 이를 통해 약 5천4백배 희석액으로의 살바이러스 효과가 있어 99.98% 살바이러스력을 가짐을 확인할 수 있다.또한 80% MCC-A를 유기물이 없는 상황에서 소독력을 확인해 보았으며, 그 결과는 아래의 표 7과 같다.That is, referring to Table 6, the virucidal effect of 80% concentration of MCC-A under low concentration organic matter was 3.5 as a log value through comparison with the control group. Through this, it can be confirmed that it has a bactericidal effect with an approximately 5,400-fold dilution and has 99.98% virucidal power. In addition, 80% MCC-A was checked for disinfection in the absence of organic matter, and the results are as follows. Table 7 shows.
항목Item 농도density 유기물organic matter 처리시간(분)Processing time (minutes) 희석농도 (lg)Dilution Concentration (lg)
22 33 44 55 66 77 88 99
MCC-AMCC-A 80%80% NoneNone 3030 00000000 00000000 00000000 00000000 00000000 00000000 00000000 00000000
ViruscontrolVirus control N/aN/a NoneNone 3030 11111111 11111111 11111111 11111111 10011001 00000000 00000000 00000000
1; virus present - 4개의 세포배양내에서의 세포변성효과(CPE)0; no virus presentN/a; not applicableOne; virus present - cytopathic effect (CPE)0 in 4 cell cultures; no virus presentN/a; not applicable
즉 표 7은 유기물이 없는 조건에서 80% MCC-A의 소독력 실험결과를 나타낸 것으로, 표 7을 참조하면, MCC-A의 유기물이 없는 상황에서는 살바이러스 효과가 대조구와의 비교를 통하여 로그값으로 ≥4.5를 나타내었다. 이를 상수 로그값으로 환산하면 0.6532인데, 6천5백배 희석액으로의 살바이러스 효과가 있음을 의미하므로, 99.99% 살바이러스력을 가지게 된다.That is, Table 7 shows the results of the disinfection power test of 80% MCC-A in the absence of organic matter. Referring to Table 7, in the absence of organic matter of MCC-A, the virucidal effect is a log value through comparison with the control. ≥4.5. Converting this to a constant log value, it is 0.6532, which means that there is a virucidal effect with a 6,500-fold dilution, so it has 99.99% virucidal power.
아프리카 돼지열병 바이러스 2차 소독력 실험African Swine Fever Virus Secondary Disinfecting Power Test
MCC-A의 아프리카 돼지열병에 대한 살바이러스 검증을 위한 2차 소독력 실험을 실시하였다. 실험기관은 1차 실험과 동일하게 폴란드 국립수의과학원에서 수행하였다. 실험절차는 유럽 표준 실험 규정인 PN-EN-14675_2015-06E의 "5.7 virucidal test"에 따라 수행하였다.A secondary disinfection test was conducted to verify the virucidal virus of MCC-A against African swine fever. The laboratory was conducted at the Polish National Academy of Veterinary Sciences in the same way as the first experiment. The test procedure was performed according to "5.7 virucidal test" of PN-EN-14675_2015-06E, a European standard test regulation.
실험에 사용된 MCC-A의 유효 염소농도는 143ppm으로 실험하였으며, 이를 MCC-A1으로 표기하기로 한다. 실험에 실제 사용된 MCC-A1의 농도는 바이러스액 첨가 등에 따라 최종 80% 수준에서 실험하였다. 처리조건은 10℃에서 30분간 처리하였다. 유기물의 함량에 따른 MCC-A1의 살바이러스 효과 검증을 위하여 고농도 유기물 처리조건에서는 10g/L BSA + 10g/L yeast extract를 첨가하여 실험에 실시하였으며, 저농도 유기물 처리조건은 3g/L BSA를 첨가하여 실험에 사용하였다. 이와 함께 1차 실험과 마찬가지로, 각 실험구별로 대조구와의 비교를 통하여 세포의 형태 및 성장에 MCC-A1이 영향을 미치는지에 대한 세포독성에 관한 조사도 진행하였다.The effective chlorine concentration of MCC-A used in the experiment was tested at 143 ppm, and this will be referred to as MCC-A1. The concentration of MCC-A1 actually used in the experiment was tested at the final 80% level according to the addition of virus solution. Treatment conditions were treated at 10° C. for 30 minutes. In order to verify the virucidal effect of MCC-A1 according to the content of organic matter, 10g/L BSA + 10g/L yeast extract was added in the high concentration organic matter treatment condition, and 3g/L BSA was added in the low concentration organic matter treatment condition. used in the experiment. Along with this, as in the first experiment, the cytotoxicity of MCC-A1 on the morphology and growth of cells by comparison with the control group for each experimental group was also conducted.
조항article 수행 여부Whether to perform
실험 바이러스 현탁액의 적정(실험물질의 처리 전후 로그수치 4 감소의 결정에 따른다.)Titration of the test virus suspension (subject to the determination of the logarithmic 4 decrease before and after the treatment of the test substance). 수행Perform
실험물질의 세포독성이 없음의 판별 여부(세포의 변성, 증식 또는 바이러스 감수성)Whether the test substance is non-cytotoxic (cell denaturation, proliferation, or virus susceptibility) 수행Perform
배양세포에 접종된 바이러스 비교 검토(실험 혼합액 또는 배양배지의 희석에 따른 비교)Comparative review of virus inoculated into cultured cells (comparison according to dilution of experimental mixture or culture medium) 수행Perform
표 8에 나타난 것처럼, 실험방법에 따른 바이러스, 배양배지 및 독성여부를 실시하였으며, 실험물질인 MCC-A1은 MCC-A와 마찬가지로 독성이 전혀 없는 것으로 확인되었다.이어서 표 9에서는 MCC-A1의 살바이러스 2차 효력실험결과를 나타낸 것으로, 143ppm의 유효 염소농도를 갖는 소독제의 살바이러스 2차 효력실험결과를 나타내었다.As shown in Table 8, the virus, culture medium, and toxicity according to the experimental method were performed, and it was confirmed that the test substance, MCC-A1, had no toxicity at all like MCC-A. As the results of the secondary virus efficacy test, the results of the virucidal secondary efficacy test of the disinfectant having an effective chlorine concentration of 143 ppm were shown.
항목Item 농도density 유기물organic matter 세포배양세포 50% 감염농도(lg TCID50)Cell cultured cells 50% infection concentration (lg TCID50) 바이러스 대조구와 실험구의 차이값(log10)Difference between virus control and experimental group (log10)
MCC-A1MCC-A1 80%80% bovine albumin 10g/L+ yeast extract 10g/Lbovine albumin 10g/L+ yeast extract 10g/L 5.55.5 2.252.25
MCC-A1MCC-A1 80%80% bovine albumin 3g/Lbovine albumin 3g/L 2.52.5 4.254.25
Virus controlVirus control N/aN/a bovine albumin 10g/L+ yeast extract 10g/Lbovine albumin 10g/L+ yeast extract 10g/L 7.257.25 N/aN/a
Virus controlVirus control N/aN/a bovine albumin 3g/Lbovine albumin 3g/L 6.756.75 N/aN/a
즉 표 9는 MCC-A1의 살바이러스 2차 효력실험결과로써, 표 9에 143ppm 농도의 MCC-A1가 아프리카 돼지열병 바이러스에 대한 효력실험의 결과를 나타낸 것임을 알 수 있다.표 9에 나타났듯이, 143ppm 농도의 MCC-A1에서는 고농도 유기물 첨가군에서 2.25 log10 수치로 상수 로그값으로 환산하면 0.3522인데, 이는 약 3천5백배 희석액으로의 살바이러스 효과가 있어 99.97% 살바이러스 효과를 있다.That is, Table 9 shows the results of the secondary efficacy test for virucidal MCC-A1, and it can be seen that Table 9 shows the results of the efficacy test for the African swine fever virus of MCC-A1 at a concentration of 143ppm. As shown in Table 9 , in MCC-A1 at a concentration of 143ppm, it is 0.3522 when converted to a constant log value as 2.25 log10 in the high concentration organic matter addition group, which has a virucidal effect with an approximately 3,500-fold dilution, and has a virucidal effect of 99.97%.
저농도 유기물 첨가군에서는 4.25 log10 수치로 상수 로그값으로 환산하면 0.6284인데, 이는 약 6천2백배 희석액으로의 살바이러스 효과가 있어 99.99% 살바이러스력을 가진다.In the low-concentration organic substance addition group, when converted to a constant log value as 4.25 log10, it is 0.6284, which has a virucidal effect of about 6,200-fold dilution and has 99.99% virucidal power.
표 9를 기반으로, 80% 농도의 MCC-A1에 bovine albumin 10g/L + yeast extract 10g/L의 고농도 유기물이 첨가된 경우와, 대조군의 살바이러스 효능을 실험해 보았으며, 그 결과를 아래의 표 10에 정리하였다.Based on Table 9, when high concentration organic matter of 10g/L + yeast extract 10g/L was added to MCC-A1 at 80% concentration, and the control group's virucidal efficacy was tested, and the results are shown below. It is summarized in Table 10.
항목Item 농도density 유기물organic matter 처리시간(분)Processing time (minutes) 희석농도 (lg)Dilution Concentration (lg)
22 33 44 55 66 77 88 99
MCC-A1MCC-A1 80%80% bovine serum albumin10g/L+yeast extract10g/Lbovine serum albumin10g/L+yeast extract10g/L 3030 11111111 11111111 11111111 11111111 00000000 00000000 00000000 00000000
ViruscontrolVirus control N/aN/a bovine serum albumin10g/L+yeast extract10g/Lbovine serum albumin10g/L+yeast extract10g/L 3030 11111111 11111111 11111111 11111111 11111111 10101010 00100010 00000000
1; virus present - 4개의 세포배양내에서의 세포변성효과(CPE)0; no virus presentN/a; not applicableOne; virus present - cytopathic effect (CPE)0 in 4 cell cultures; no virus presentN/a; not applicable
표 10에서 보듯이, MCC-A1의 고농도 유기물 하에서 살바이러스 효과는 대조구와의 비교를 통하여 로그값으로 2.25를 나타났으며, 이를 상수 로그값으로 환산하면 0.3522로, 약 3천5백배 희석액으로의 살바이러스력 99.97%를 가지게 됨을 확인할 수 있다.이어서 80% 농도의 MCC-A1에 BSA 3g/L을 첨가하여 저농도 유기물 처리조건을 만들어 준 후 소독력을 실험하여, 그 결과를 아래의 표 11에 나타내었다.As shown in Table 10, the virucidal effect of MCC-A1 under high-concentration organic matter was 2.25 as a log value through comparison with the control, and when converted to a constant log value, it was 0.3522, about 3,500 times dilution. It can be confirmed that it has a virucidal power of 99.97%. Then, 3 g/L of BSA was added to MCC-A1 of 80% concentration to create low-concentration organic matter treatment conditions, and then the disinfection power was tested, and the results are shown in Table 11 below. It was.
항목Item 농도density 유기물organic matter 처리시간(분)Processing time (minutes) 희석농도 (lg)Dilution Concentration (lg)
22 33 44 55 66 77 88 99
MCC-A1MCC-A1 80%80% bovine serum albumin 3g/Lbovine serum albumin 3g/L 3030 11111111 00000000 00000000 00000000 00000000 00000000 00000000 00000000
ViruscontrolVirus control N/aN/a bovine serum albumin 3g/Lbovine serum albumin 3g/L 3030 11111111 11111111 11111111 11111111 11111111 01000100 00000000 00000000
1; virus present - 4개의 세포배양내에서의 세포변성효과(CPE)0; no virus presentN/a; not applicableOne; virus present - cytopathic effect (CPE)0 in 4 cell cultures; no virus presentN/a; not applicable
즉 표 11은 80% MCC-A1가 3g/L BSA에 대해 갖는 소독력 효력실험자료로써, 표 11에서 보여지듯이 MCC-A1의 저농도 유기물 하에서 살바이러스 효과는 대조구와의 비교를 통하여 로그값으로 4.25로 나타나며, 이를 상수 로그값으로 환산하면 0.6284인데, 이는 약 6천2백배 희석액으로의 살바이러스 효과가 있으며, 99.99% 살바이러스력을 가짐을 알 수 있다.상술한 실시예 및 실험예의 결과로부터, 본 발명을 통한 나트륨 미함유 차아염소산수는 세포독성이 없을 뿐만 아니라 아프리카 돼지열병에 대하여 99.97% 이상의 살바이러스력을 가짐으로써, 치명적인 바이러스성 출혈성 돼지 전염병 바이러스를 사멸할 수 있음이 확인된다.That is, Table 11 is the experimental data of the disinfecting power effect of 80% MCC-A1 against 3g/L BSA. As shown in Table 11, the virucidal effect of MCC-A1 under low-concentration organic matter was compared with the control, and the log value was 4.25. appears, and when converted to a constant log value, it is 0.6284, which has a virucidal effect with an about 6,200-fold dilution, and it can be seen that it has 99.99% virucidal power. From the results of the above-described Examples and Experimental Examples, Sodium-free hypochlorous acid water through the invention has not only no cytotoxicity, but also has more than 99.97% virucidal power against African swine fever, thereby confirming that it can kill the deadly viral hemorrhagic swine epidemic virus.
이와 같이 본 발명은 염소가스의 양과 증류수의 양에 따라 전해조에 공급되는 전류의 양을 조절함으로써, 유효 염소농도를 78ppm 또는 143ppm으로 일정하게 유지하는 나트륨 미함유 차아염소산수를 제조할 수 있는데 특징이 있다.As such, the present invention can produce sodium-free hypochlorous acid water that maintains an effective chlorine concentration of 78 ppm or 143 ppm constant by adjusting the amount of current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water. have.
특히 본 발명의 실험예에서 시도한 아프리카 돼지열병에 관한 살바이러스 실험은 기존에 시도한 적이 없는 것으로, 주요 효능물질인 차아염소산수의 유효 염소농도를 78ppm 또는 143ppm으로 조절하여 아프리카 돼지열병 바이러스에 대한 소독제로의 적용에 대한 새로운 제안을 제시한다는 점에서, 본 발명의 가치가 인정될 수 있을 것으로 기대된다.In particular, the virucidal virus experiment on African swine fever attempted in the experimental example of the present invention has not been attempted before, and the effective chlorine concentration of hypochlorous acid water, which is the main effective substance, is adjusted to 78 ppm or 143 ppm to be used as a disinfectant for African swine fever virus. It is expected that the value of the present invention can be recognized in that it presents a new proposal for the application of
이상의 설명은 본 발명의 기술 사상을 예시적으로 설명한 것에 불과한 것으로, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자라면 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 다양한 수정 및 변형이 가능할 것이다.The above description is merely illustrative of the technical idea of the present invention, and various modifications and variations will be possible without departing from the essential characteristics of the present invention by those skilled in the art to which the present invention pertains.
따라서 본 발명에 개시된 실시예는 본 발명의 기술 사상을 한정하기 위한 것이 아니라, 설명하기 위한 것이고, 이러한 실시예에 의하여 본 발명의 기술 사상의 범위가 한정되는 것도 아니다.Therefore, the embodiments disclosed in the present invention are not intended to limit the technical spirit of the present invention, but to illustrate, and the scope of the technical spirit of the present invention is not limited by these embodiments.
본 발명의 보호 범위는 특허청구범위에 의하여 해석되어야 하며, 그와 동등한 범위 내에 있는 모든 기술사상은 본 발명의 권리범위에 포함되는 것으로 해석되어야 할 것이다.The protection scope of the present invention should be construed by the claims, and all technical ideas within the equivalent range should be construed as being included in the scope of the present invention.

Claims (5)

  1. 염산용액을 전해조에 투입한 후 전기분해하여 염소가스를 포함하는 전해생성물을 제조하는 단계; 및preparing an electrolysis product containing chlorine gas by electrolyzing the hydrochloric acid solution into an electrolyzer; and
    상기 전해생성물로부터 분리된 염소가스를 증류수에 혼입하면서 나트륨 미함유 차아염소산수를 제조하는 단계;를 포함하여 이루어지고,Preparing sodium-free hypochlorous acid water while mixing chlorine gas separated from the electrolysis product into distilled water;
    상기 염소가스의 양과 상기 증류수의 양에 따라 상기 전해조에 공급되는 전류의 양을 조절함으로써 유효 염소농도가 78ppm 또는 143ppm인 나트륨 미함유 차아염소산수를 제조하는 것을 특징으로 하는,Characterized in that by adjusting the amount of current supplied to the electrolyzer according to the amount of chlorine gas and the amount of distilled water to prepare sodium-free hypochlorous acid water having an effective chlorine concentration of 78 ppm or 143 ppm,
    아프리카 돼지열병 바이러스 소독제용 나트륨 미함유 차아염소산수의 제조방법.Method for producing sodium-free hypochlorous acid water for African swine fever virus disinfectant.
  2. 제1항에 있어서,According to claim 1,
    상기 차아염소산수의 유효 염소농도가 78 ppm이고, 돼지에 직접 분사하여 소독처리하는 것을 특징으로 하는,The effective chlorine concentration of the hypochlorous acid water is 78 ppm, characterized in that the disinfection treatment by spraying directly on the pig,
    아프리카 돼지열병 바이러스 소독제용 나트륨 미함유 차아염소산수의 제조방법.Method for producing sodium-free hypochlorous acid water for African swine fever virus disinfectant.
  3. 제1항에 있어서,According to claim 1,
    상기 차아염소산수의 유효 염소농도가 143 ppm이고, 돼지 축사에 분사하여 소독처리하는 것을 특징으로 하는,The effective chlorine concentration of the hypochlorous acid water is 143 ppm, characterized in that the disinfection treatment by spraying the pig barn,
    아프리카 돼지열병 바이러스 소독제용 나트륨 미함유 차아염소산수의 제조방법.Method for producing sodium-free hypochlorous acid water for African swine fever virus disinfectant.
  4. 제1항 내지 제3항 중 어느 한 항의 방법으로 제조되는 나트륨 미함유 차아염소산수를 포함하여 이루어지는 것을 특징으로 하는, 아프리카 돼지열병 바이러스 소독제.A disinfectant for African swine fever virus, characterized in that it comprises sodium-free hypochlorous acid water prepared by the method of any one of claims 1 to 3.
  5. 제4항에 따른 아프리카 돼지열병 바이러스 소독제를 돼지 또는 돼지 축사에 직접 분사하여 아프리카 돼지열병 바이러스를 사멸하되, 사멸율이 적어도 99.97%인 것을 특징으로 하는, 나트륨 미함유 차아염소산수를 이용한 아프리카 돼지열병 바이러스의 사멸방법.The African swine fever virus disinfectant according to claim 4 is directly sprayed on pigs or pig pens to kill the African swine fever virus, but the killing rate is at least 99.97% African swine fever using sodium-free hypochlorous acid water How to kill the virus.
PCT/KR2020/003071 2020-02-14 2020-03-04 Method for preparing disinfectant against african swine fever virus, and disinfectant against african swine fever virus, prepared thereby WO2021162168A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2020-0018025 2020-02-14
KR20200018025 2020-02-14
KR10-2020-0025078 2020-02-28
KR1020200025078A KR102144428B1 (en) 2020-02-14 2020-02-28 Manufacturing method of African swine fever virus disinfectant and African swine fever virus disinfectant

Publications (1)

Publication Number Publication Date
WO2021162168A1 true WO2021162168A1 (en) 2021-08-19

Family

ID=72039162

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2020/003071 WO2021162168A1 (en) 2020-02-14 2020-03-04 Method for preparing disinfectant against african swine fever virus, and disinfectant against african swine fever virus, prepared thereby

Country Status (2)

Country Link
KR (1) KR102144428B1 (en)
WO (1) WO2021162168A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070103706A (en) * 2006-04-19 2007-10-24 가부시키가이샤 하셋퍼 엔지니어링 Disinfecting liquids to spray into livestock facilities and the methods of using the liquids to disinfect the insides of livestock facilities
KR20160001059A (en) * 2014-06-26 2016-01-06 대한민국 (식품의약품안전처장) Disinfection Method of Dressed Carcass
KR101938121B1 (en) * 2018-10-17 2019-01-15 주식회사 에코바이론 Disinfection and disinfection system inside and outside of the pig house using sodium hypochlorite
KR20190023209A (en) * 2017-08-28 2019-03-08 케이테크전해 주식회사 Electrolysis apparatus for producing between germicide and detergent

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101440375B1 (en) 2012-12-05 2014-09-17 주식회사 우성양행 Disinfectant with synergistic effect and method for manufacturing thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070103706A (en) * 2006-04-19 2007-10-24 가부시키가이샤 하셋퍼 엔지니어링 Disinfecting liquids to spray into livestock facilities and the methods of using the liquids to disinfect the insides of livestock facilities
KR20160001059A (en) * 2014-06-26 2016-01-06 대한민국 (식품의약품안전처장) Disinfection Method of Dressed Carcass
KR20190023209A (en) * 2017-08-28 2019-03-08 케이테크전해 주식회사 Electrolysis apparatus for producing between germicide and detergent
KR101938121B1 (en) * 2018-10-17 2019-01-15 주식회사 에코바이론 Disinfection and disinfection system inside and outside of the pig house using sodium hypochlorite

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KIM, EUN SANG: "Cover Story / Excellent hypochlorous acid water catches bacteria and odors", 31 December 2020 (2020-12-31), pages 1 - 4, XP055835831, Retrieved from the Internet <URL:http://www.hyunchuk.co.kr/news/8849> *

Also Published As

Publication number Publication date
KR102144428B1 (en) 2020-08-12

Similar Documents

Publication Publication Date Title
Blaser et al. Inactivation of Campylobacter jejuni by chlorine and monochloramine
US7011739B2 (en) Method for sanitizing shells of eggs using electrolyzed oxidizing water
EP1214081B1 (en) Superoxidized water based on hypochlorous acid for the treatment of wounds
Matsunaga et al. Electrochemical disinfection of bacteria in drinking water using activated carbon fibers
DE202009019172U1 (en) Stabilized composition of an antimicrobial solution of hypochlorous acid
US7001493B1 (en) Apparatus for preparing sterilizing water and process for sterilizing water
US7651704B2 (en) Use of an aqueous solution in the treatment of live animals
KR20120092056A (en) The electrolytic apparatus for clo2 gas and sterilization system of room
CN101053330A (en) Disinfectant for improving hydrogen peroxide bactericidal activity
CN105836860B (en) A kind of stable type dioxygen aqueous disinfectant and its application in drinking water disinfection
KR101974874B1 (en) Vehicle disinfection system and vehicle disinfection method using the same
CN101746857A (en) Method and equipment of electrochemical disinfection for water
CN108633888B (en) A kind of sodium dichloroisocyanuratepowder powder and preparation method thereof
WO2021162168A1 (en) Method for preparing disinfectant against african swine fever virus, and disinfectant against african swine fever virus, prepared thereby
CN1978612A (en) Disinfecting detergent powder, and its preparing method
KR101857736B1 (en) Composition of chlorine disinfections free of chlorine smell
CN113018498A (en) Photoelectrocatalysis radio frequency heating sterilizer that disinfects
WO2010137852A2 (en) Real-time sterilizing and disinfecting apparatus, and real-time sterilizing and disinfecting method using same
CN112640912A (en) Active oxygen composite bactericide and preparation method thereof
WO2022045535A1 (en) Natural disinfectant based on hypochlorous acid solution, and manufacturing method therefor
WO2023277439A1 (en) Environment-friendly disinfectant composition, for preventing infectious disease, having excellent antimicrobial and deodorizing performance by containing liquid sodium silicate, and environment-friendly disinfectant, for preventing infectious disease, comprising same
CN111493086A (en) Acidic oxidation potential aqueous solution capable of killing coronavirus, production device, production process and application thereof
Haas et al. Repeated exposure of Escherichia coli to free chlorine: Production of strains possessing altered sensitivity
CN113082261A (en) Method for disinfecting livestock and poultry farm by utilizing strongly alkaline electrolyzed water
CN217149328U (en) Ozone water preparation device and sterilizer with same

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20918780

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20918780

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 20918780

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 22/06/2023)

122 Ep: pct application non-entry in european phase

Ref document number: 20918780

Country of ref document: EP

Kind code of ref document: A1