KR20090059789A - Biocontrol agent containing live biological materials, its spraying apparatus and surface sterilization method with the same in preventing and treating pathogenic bacteria infection - Google Patents

Abstract

A sterilization composition containing an active biological material which enables eco-friendly sterilization using bateriophage is provided to prevent the harmfulness to human and animal, soil residue toxicity and accumulation in a body. A sterilization composition comprises an active biological material. The active biological material is bateriophage. The bacteriophage is selected from SAP-1(KCTC 11153BP) and SAP-2(KCTC 11154BP) which specifically cause the apoptosis of Staphylococcus aureus and SGP-1(KCTC 11174BP) and SEP-1(KCTC 11173BP) which specifically cause the apoptosis of salmonella.

Description

Biocontrol agent containing live biological materials, its spraying apparatus and surface sterilization method with the same in preventing and treating pathogenic bacteria infection}

The present invention is a bactericidal composition, bactericidal sprayer and the like comprising an active biological material bacteriophage having the ability to kill these pathogens for the purpose of reducing or eradicating pathogenic bacteria present in the surrounding environment. It relates to a pathogen sterilization method using the same.

Various fungicides and methods of sterilization exist. BACKGROUND OF THE INVENTION Sterilization methods using various chemical substances or various apparatuses such as electron beams and ultraviolet rays are known.

According to Korean Laid-Open Patent No. 2007-53742, sterilization treatment can be performed with a high sterilization capacity that can kill spores without causing thermal damage even to heat-treated objects such as plastic products. A sterilization method is provided, and in particular, a method of sterilizing a sterilization object by filling a process gas containing HCHO into a treatment chamber is proposed.

In addition, according to Korean Patent Publication No. 2007-103706, sterilizing water for spraying inside a livestock facility that is safe for livestock and workers and can achieve a sterilizing effect and the inside of the livestock facility using the sterilizing water. A method of sterilization is provided. For this purpose, water is mixed with an aqueous sodium hypochlorite solution and an acid to prepare sterilized water having a pH of 6.0 to 7.5 and an effective chlorine concentration of 25 ppm or more. A method has been proposed to sterilize infectious pathogens that adhere to the bottom surface of a livestock facility or to the livestock itself by spraying.

On the other hand, according to Korean Patent Publication No. 2000-49558, 'Environmental Improvement of Barn House Using Ozone Water', it is sprayed with fine water droplets in the contaminated barn room using ozone water having strong oxidizing and sterilizing power to remove odors, bacteria and Methods for virus sterilization and the like are known. Such ozone water can be understood as a kind of chemical that can be produced by chemical or physical methods.

However, the sterilization method using a conventional sterilizing agent made of a chemical substance has problems such as problems caused by soil residue, problems caused by accumulation in the living body, and danger to the living body living in the disinfectant spraying site.

The inventors noted an active biomolecule (active biological material) that can be contrasted with chemicals. Particular attention was paid to bacteriophages, a microorganism capable of killing pathogenic bacteria, a type of active biomaterial, and nature-friend antimicrobial agents or bioenvironmental agents that can reduce or eliminate certain pathogens. The present invention has been completed with the assurance that it can function as bio-control agents.

Bacteriophages are a type of virus that infects bacteria, often called abbreviations. Bacteriophages are simple organisms in which the protein envelope is wrapped around the core of a genetic material consisting of nucleic acids. Nucleic acids are composed of DNA or RNA, either single or double chains. Bacteriophages are essential for survival and all bacteria are known to contain specific bacteriophages. The bacteriophage penetrates the host, completes the replication process, and then expresses a group of enzymes necessary to degrade the cell wall of the host bacterium. These enzymes attack the peptidoglycan layer of the cell wall, which is responsible for the rigidity and mechanical strength of the cell wall, disrupting the cell wall.

의 세균학자 d'Herelle이 이질환자 변의 여과액 중에 적리균(Shigella disentriae)

을 녹이는 작용을 가진 것이 있다는 것을 발견하고 이에 대한 연구를 통해 독립적으로 박테리오파지를 발견하였으며, 세균을 잡아먹는다는 뜻에서 박테리오파지라고 명명하였다. 이후 이질균, 장티푸스균, 콜레라균 등 여러 병원균에 대한 박테리오파지가 계속적으로 발견되었다. 그러나 1950년 Flemming에 의해 페니실린이 발견된 이후, 항생제 사용의 보급화로 인해 일부 동유럽 국가에 한정되어서만 박테리오파지에 대한 연구가 계속되었으며 그 밖의 지역에서는 다소 시들하였다. 그러나 2000년 이후에 항생제의 오남용으로 인해 다재 내성(Multidrug-resistant)을 지닌 병원성 세균의 출현빈도가 높아지고 기존 항생제의 많은 문제점들이 부각되면서 기존 항생제의 대체 물질로의 개발 가능성 때문에 박테리오파지에 대한 연구가 선진국들을 중심으로 많은 관심을 받으며 다시 활발하게 진행되고 있다.The bacteriophage was discovered in 1915 by a British bacteriologist Twort, in a study of the transparent melting of micrococcus colonies by something. In 1917, France

Of bacteriologist d'Herelle in Shigella disentriae

They found that they had a function of dissolving the bacteria, and through this research, they independently discovered bacteriophages and named them as bacteriophages in order to eat bacteria. Since then, bacteriophages have been found for several pathogens such as dysentery, typhoid, and cholera. However, after penicillin was discovered by Flemming in 1950, research on bacteriophage continued to be limited to some Eastern European countries due to the widespread use of antibiotics, and withered elsewhere. However, since 2000, due to the misuse of antibiotics, the development of multidrug-resistant pathogenic bacteria has increased, and many problems of conventional antibiotics have emerged. The center has received a lot of attention and is actively progressing again.

Bacteriophages have been used to alleviate or treat certain infectious diseases until now mainly by oral administration or injection. However, the present inventors have applied to the disinfectant application made of conventional chemicals if they are applied to the surrounding environment to reduce or eliminate pathogens or prevent contamination by these pathogens in places including barns, homes, hospital rooms, cooking places, slaughterhouses and slaughterhouses. It has been confirmed that many problems that have emerged can be solved.

Therefore, the inventors of the present invention, if prepared and sprayed active biomaterials, especially bacteriophage as an active ingredient, and sprayed, it is natural, harmless to animals, including humans, there is no fear of causing any problems due to soil residue, It was confirmed that sterilization treatment without problems such as internal accumulation was possible.

Accordingly, an object of the present invention is to propose a bactericidal composition containing bacteriophage as an active ingredient for the purpose of reducing and eradicating pathogens present in the surrounding environment. Another object of the present invention is to provide a nebulizer including the composition, and to propose a surface sterilization method for applying the composition to a surface.

The present invention is used as a fungicide of a composition comprising bacteriophage as an active ingredient for the purpose of reducing and eradicating pathogens present in the surrounding environment, preventing the growth of pathogens present in the surrounding environment, and preventing the contamination of the surrounding environment by pathogens, Provided is a sprayer comprising the sterilization composition, and a surface sterilization method for automatically spraying the sterilant and regularly applying it to the surface.

It is intended to define the terminology used herein. The sterilizing composition according to the present invention is applied to the 'surface' and exerts its sterilizing ability, and the term 'surface' means the surface of an organism such as skin of an animal including a human; And inanimate surfaces, including hard surfaces such as walls, tiles, glass, and floors, and soft surfaces such as textiles, clothing, and carpets in places including barns, homes, hospital rooms, cooking places, slaughterhouses, and slaughterhouses. 'Applied' on the 'surface' means that the fungicide is deposited on the surface in the form of liquid droplets (mist) and sprayed to reduce or kill pathogens by exerting bactericidal activity. The term “spray dispenser” means a spray dispenser, and should be understood to mean a manual or automatic spray container, particularly suitable for vertical surfaces, including trigger sprayers or pump dispenses. 'Automatic atomizer' is understood as a nebulizer capable of automatically and periodically spraying a composition filled in a nebulizer pressure vessel by a mechanical actuation mechanism.

1) Bacteriophage Sterilizer

Any kind of bacteriophage to which the present invention can be applied does not matter. In the present invention, for the convenience of description, the present invention shows the result of using four types of bacteriophages that have been separated and deposited by the present inventors, but the present invention is merely presented by way of example and the present invention is not limited to the type of bacteriophage. Not restricted by In the present invention, bacteriophage SAP-1 (KCTC 11153BP; Deposited July 18, 2007) and SAP-2 (Korea Life) can specifically kill Staphylococcus aureus . Depository KCTC 11154BP; Deposited July 18, 2007), Bacteriophage SGP-1 (KCTC 11174BP, Korea Biotechnology Center), which can specifically kill Salmonella (2007); Deposits of August 21) and SEP-1 (KCTC 11173BP; Deposit of August 21, 2007) were selected as an example.

Of course, the bacteriophage that can be utilized in the present invention is not limited thereto, but is presented as follows. Bacteriophage belonging to the Myoviridae family include bacteriophage A, EW, K, Ph5, Ph9, Ph10, Ph13, P1, P2, P3, P4, P8, P9, P10, RG, SB-1, S3K, Thiwort , φSK311, φ812, 06, 40, 58, 119, 130, 131, 200, 1623, and the like, and bacteriophages belonging to the Siphoviridae family include bacteriophage AC1, AC2, AC3, A6 "C". , A7, A8, A9 "C", A10, b581, b595n, B3, B33, B39, BI-1, C22, CA-1, CA-2, CA-3, CA-4, CA-5, D, D3, D11, D37, D40, D62, D3112, F7, F10, g, gd, ge, gf, HK2, Hw12, Jb19, KF1, L39X35, L54a, M42, N1, N2, N3, N4, N5, N7, N8, N9, N10, N11, N12, N13, N14, N15, N16, OXN-32P, O6N-52P, P52, P87, Ph6, Ph12, Ph14, PCH-1, PC13-1, PC35-1, PH2, PH51, PH93, PH132, PMW, PM13, PM57, PM61, PM62, PM63, PM69, PM105, PM113, PM681, PM682, PO4, PP1, PP4, PP5, PP64, PP65, PP66, PP71, PP86, PP88, PP92, PP401, PP711, PP891, Pssy41, Pssy42, Pssy403, Pssy404, Pssy420, Pssy923, PS4, PS-10, Pz, S1, S6, SD1, SL1, SL3, SL5, SM, UC-18, U4, U15, S1, S2, S3, S4, S5, Z1, Z4, X2, φC5, φC11, φC11-1, φC13, φC15, φMO, φX, φ04, φ11, φ240, φB5-2, φD, φRE, ω, 2, 2F, 3A, 3B, 3C, 5, 6, 7, 7m, 11, 13, 13/441, 14, 15, 16, 20, 21, 24, 28, 28A, 29, 31, 31B, 37, 40, 42B, 42C, 42D, 42E, 44, 44A, 45, 47, 47A, 47C, 48, 49, 51, 52, 52A, 52B, 53, 54, 54X1, 55, 61, 69, 70, 71, 71A, 72, 73, 75, 76, 77, 78, 79, 80, 80α, 81, 82, 82A, 83A, 84, 85, 86, 88, 88A, 89, 90, 92, 93, 94, 95, 96, 101, 102, 105, 107, 108, 110, 111, 115, 129/16, 129-26, 130, 130A, 148, 155, 157, 157A, 160, 165, 174, 187, 198, 218, 222, 236, 242, 246, 249, 258, 269, 275, 275A, 275B, 295, 297, 309, 318, 342, 350, 351, 356, 357-1, 400-1, 456, 459, 471, 471A, 489, 581, 594n, 676, 898, 1139, 1154A, 1259, 1314, 1363/14, 1380, 1405, 1563, 2148, 2460, 2638A, 2638B, 2638C, 2731, 2792A, 2792B, 2818, 2835, 2848A, 3619, 5841, 12100, etc., and the bacteriophage belonging to the Podoviridae family, such as bacteriophage A856, B26. , CI-1, CI-2, C5, D, gh-1, F116, HF, H90, K5, K6, K104, K109, K166, K267, N4, N5, O6N-25P, PE69, Pf, PPN25, PPN35 , PPN89, PPN91, PP2, PP3, PP4, PP6, PP7, PP8, PP56, PP87, PP114, PP206, PP207, PP306, PP651, Psp231a, Pssy401, Pssy9220, ps1, PTB2, PTB20, PTB42, PX1, PX3, PX10 , PX12, PX14, PYO70, PYO71, R, SH6, SH133, tf, Ya5, Ya7, φBS, φKf77, φ-MC, φmnF82, φPLS27, φPLS743, φS-1, 1, 2, 3, 4, 5, 6 , 8, 9, 10, 11, 12, 12B, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 31, 53, 73, 119X, 145, 147 , 170, 267, 284, 308, 525 and the like can be presented. Available bacteriophages can also be purchased from the American Type Culture Collection under the following access number. ATCC 12055-B1, ATCC 12055-B2, ATCC 12055-B3, ATCC 14205-B1, ATCC 14206-B1, ATCC 14207-B1, ATCC 14209-B1, ATCC 14210-B1, ATCC 14211-B1, ATCC 14212-B1, ATCC 14213-B1, ATCC 14214-B1, ATCC 15692-B2, ATCC 15692-B3, ATCC 25102-B1, ATCC BAA-26-B1, ATCC BAA-27-B1, ATCC BAA-28-B1, ATCC BAA-28 -B2, ATCC BAA-29-B1, ATCC BAA-30-B1, ATCC BAA-31-B1, ATCC BAA-47- B1, ATCC BAA-79-B1, ATCC BAA-81-B1, ATCC BAA-81- For example, B2.

Pathogens that can be applied by the composition containing the bacteriophage as an active ingredient in the present invention are all bacteria having a cell wall. In the present invention, as a representative example, Staphylococcus aureus and Salmonella was selected as a pathogen, but this is only representative because it can not be seen for all pathogens, Enterobacteriacae, Staphylococcus genus, Enterococcus, Streptococcus, Staphylococcus epidermidis , coagulase-negative staphylococci, Pseudomonas aeruginosa , Klebsiella pneumoniae , Escherichia coli , Enterococcus faecalis , Enterococcus faecium , Providencia stuartii , Proteus mirabilis , Morganella morganii , Acinetobacter calcoaceticus , Enterobacter aerogenes, Streptococcus agalactiae, Streptococcus avium, Streptococcus bovis, Streptococcus durans, Streptococcus faecalis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus suis, Streptococcus viridans, Streptococcus salivarius, Burkholderia cepacia , Stenotrophomonas maltophilia , Acaligenes xylosoxidans , non-tuberculous mycobacteria, Mycobacterium bovis , Mycobacterium smegmatis , Mycobacterium tuberculosis , Burkholderia multivorans , Burkholderia stablis , Burkholderia vietnamesis and the like may be the target pathogen of the present invention, and the target pathogen of the present invention is not limited thereto.

The composition containing the bacteriophage as an active ingredient may be mainly composed of a water-soluble form, but an addition form of an organic solvent or a suspension form may be possible. The bactericidal composition of the present invention comprises 1 × 10 3 -1 × 10 15 pfu / ml of bacteriophages, preferably 1 × 10 6 -1 × 10 10 pfu / ml of bacteriophages. Of course, the concentration of this bacteriophage can also be arbitrarily adjusted to suit the purpose, which can be generally carried out by those skilled in the art.

In addition, the sterilization composition of the present invention may further include other components in addition to the bacteriophage. Acceptable carriers included in the compositions of the present invention are commonly used in the preparation of water-soluble preparations, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. It is not. The composition of the present invention may further include flavoring agents, suspending agents, preservatives and the like in addition to the above components.

2) Input Type of Sterilization Composition

The sterilizing composition according to the present invention can be put and packaged in various suitable packages known to those skilled in the art. The sterilization composition implemented in water-soluble form, organic solvent addition form or suspension form, etc. may be preferably added to a nebulizer usually made of plastic, and may be preferably provided in a nebulized form into an automatic nebulizer. In the nebulizer, the water-soluble disinfectant is divided into fine liquid droplets and sprayed towards the surface to be treated, and the disinfectant contained in the nebulizer is actually delivered to the pumping mechanism when the pumping mechanism is activated by the user if it is manual or by the drive if it is automatic. The droplets are sprayed through the nebulizer head by the energized energy.

The automatic atomizer is a device designed to inject a specific component by using a high pressure, and is used to fill the pressure vessel together with the gas and the spray composition required for spraying, and the filled pressure vessel (also called aerosol can) inside the automatic injection device The nozzle is projected to the head of the pressure vessel by the operation pressurizing means and then pressurized by a predetermined time interval so that the sterilization composition is simultaneously sprayed with the gas by using the high pressure of the gas in the can vessel. In general, such an automatic spraying machine is composed of a battery, a pressure vessel filled with a spraying component, and an operation unit for pressurizing and controlling the head nozzle of the pressure vessel in the inner space formed by the front and rear cases. A timer is usually installed to control the spray cycle, but recently, various control means such as an infrared detector may be applied instead of the timer. In the embodiment of the present invention, the liquid propane gas (liquified propane gas; LPG) is used as the gas filled together in the atomizer pressure vessel, but is not limited thereto, and this may be replaced with liquid carbon dioxide or liquid nitrogen. Yes is very apparent to those skilled in the art. In addition, although the embodiment was described as a ratio of 9: 1 to the sterilization composition of the filling liquefied gas, which is related to the degree of aerosol to be injected, it can be applied to various modifications such as 7: 3, 6: 4.

3) Sterilization Method

The present invention includes a sterilization method for spray application on the surface of the sterilization composition comprising bacteriophage as an active ingredient. The present invention may be applied to the surface of the user manually, but is preferably periodically applied to the surface by an automatic spray method can effectively sterilize various pathogens on the surface.

The sterilization method of the present invention may vary depending on the size of the place requiring the sterilization treatment, the strength of the wind, the nature of the place, and there may be an increase or decrease in the effect according to the installation of the automatic atomizer for the sterilization. You may install it. In general, it is suitable to install one automatic injection device per 1 m 2-10,000 m 2, preferably one per 4 m 2-900 m 2, most preferably one per 9 m 2-100 m 2. Is suitable. In addition, this may vary depending on the concentration of the bacteriophage included in the sterilizing composition, the amount of single injection, and the injection pressure. In other words, when spraying a composition containing a high concentration of bacteriophage, or spraying a large amount of injection and powerful spraying, a small number of automatic atomizers may be installed in a larger area, and a spraying or single injection amount of a composition containing a low concentration of bacteriophage If this is small or the spray strength is weak, it is desirable to increase the number of installations even if the same area.

The single injection amount and the injection cycle of the sterilization composition including bacteriophage as an active ingredient in the automatic spraying machine can be arbitrarily adjusted according to the degree of contamination of pathogenic bacteria in the surrounding environment and the degree of contamination of the pathogen desired by the user. In addition, the single injection amount and the injection cycle may vary depending on the concentration of the bacteriophage included in the composition. Usually, 0.01 ml-10 ml is suitable as a single injection amount, Preferably 0.05 ml-1 ml is suitable, Most preferably, 0.1 ml-0.5 ml is suitable. The injection cycle is usually suitable for a period of 15 minutes to 1 day, preferably 15 to 12 hours, most preferably 15 to 6 hours.

The sterilizing composition of the present invention does not cause problems due to soil accumulation or bioaccumulation and does not harm the living body, so there is no need to worry about the injection amount. In addition, there is no need to worry more because the growth of the bacteriophage is adjusted according to the degree of contaminated pathogens due to its self-proliferative capacity.

The present invention provides a sterilization composition containing a bacteriophage capable of killing pathogens as an active ingredient, a sprayer to which the composition is applied, and a sterilization method applied to a peripheral surface for the purpose of reducing or eradicating pathogens present in the surrounding environment. do. The sterilization method of the present invention is particularly effective when the composition containing the bacteriophage as an active ingredient is used as a natural friendly disinfectant or a bioenvironmental purifier. In particular, their useful value may be high in barns, homes, hospital rooms, cooking sites, slaughterhouses and slaughterhouses. The present invention can reduce the burden on the care required for spraying management, and does not require labor to be applied for spraying operations, especially with automatic spraying. In addition, the present invention does not cause a problem due to the soil residue of the disinfecting component due to the continuous spraying with a conventional disinfectant, problems caused by the accumulation of disinfecting components in vivo, and problems related to the risk to the living body inhabiting the disinfecting component spraying site. Do not. In addition, the present invention utilizes the mechanism of inhibiting and killing pathogens by viable living organisms having self-proliferation ability, thereby providing the advantage that the growth of bacteriophages is automatically adjusted according to the degree of contaminated pathogens. In addition, when spraying live bacteriophage, bacteriophage can be delivered through the pathogens and animals as well as the spraying site, and thus the spraying effect can be expected in a wider area compared to the case of spraying with the main ingredient. This is based on the characteristics of bacteriophages that can reach effective concentrations that, even when very small amounts are delivered, will be effective in killing pathogens if they are contaminated.

Hereinafter, although an Example demonstrates this invention more concretely, these Examples are only illustrations of this invention, The scope of the present invention is not limited to these Examples.

Example  1: Preparation of a composition comprising a bacteriophage as an active ingredient

Bacteriophage SAP-1 and SAP-2 separated by the present inventors were used to prepare a first bactericidal composition to be applied to Staphylococcus aureus, and also bacteriophage SGP-1 and SEP- separated by the present inventors. Using 1 to prepare a second sterilized composition to be applied to Salmonella.

The preparation of the SAP-1 bacteriophage solution will be described in detail. First, Tryptic Soy Broth (TSB) medium (casein digest, 17 g / l; soybean digest, 3 g / l; dextrose, 2.5 g / l; NaCl, 5 10 ml of dipotassium phosphate (2.5 g / l) was inoculated with Staphylococcus aureus and shaken overnight at 37 ° C. The next day, 500 ml of Staphylococcus aureus culture and 1 ml of SAP-1 bacteriophage solution were inoculated in 4 ml of fresh TSB medium and incubated at 37 ° C. for at least 5 hours. After incubation, the culture was centrifuged at 5,000 rpm for 10 minutes to recover the supernatant. The recovered supernatant was filtered using a 0.45 μm filter, and then 1.5 ml of Staphylococcus aureus cultured overnight was added to the recovered filtrate. This was shaken overnight at 37 ° C. The next day, the supernatant was recovered by centrifuging the culture at 5,000 rpm for 10 minutes, and the recovered supernatant was filtered using a 0.45 μm filter, and then 2 ml of Staphylococcus aureus cultured overnight was added to the recovered filtrate. . This was incubated at 37 ° C. for 4 hours. After incubation, the culture solution was centrifuged at 5,000 rpm for 10 minutes to recover the supernatant, and the recovered supernatant was filtered using a 0.45 μm filter to obtain a primary bacteriophage solution. The bacteriophage solution is a state in which the bacteriophage is suspended in TSA medium, and the following steps were performed to exchange it with a more physiologically suitable medium. 6-fold volume of polyethylene glycol (PEG) precipitation buffer [20% (w / v) PEG 8,000; 2.5 M NaCl] was added. It was mixed well and left at 4 ° C. overnight. The next day, it was centrifuged at 8,000 rpm for 15 minutes to obtain bacteriophage precipitate. The resulting bacteriophage precipitate was dissolved using phosphate buffer saline (PBS). Diluted solution (10-14-10-18) was prepared from the bacteriophage solution thus prepared using PBS at an appropriate ratio, and then subjected to a conventional plaque assay to determine the bacteriophage concentration in the bacteriophage solution prepared above. . Based on the bacteriophage concentration in the bacteriophage solution thus determined, the bacteriophage solution was finally prepared such that the bacteriophage concentration was 1010 pfu / ml using PBS. The thus prepared bacteriophage solution is the final SAP-1 bacteriophage solution.

The SAP-2 bacteriophage solution was also prepared in the same manner as the preparation of the SAP-1 bacteriophage solution. That is, the step where the SAP-1 bacteriophage was applied in the method was replaced with the SAP-2 bacteriophage.

Also, bacteriophage SGP-1 and SEP-1 for Salmonella were prepared in the same manner as in the preparation of SAP-1 bacteriophage solution, but Salmonella was used instead of Staphylococcus aureus. That is, in the method for preparing the SAP-1 bacteriophage solution, the step of applying the SAP-1 bacteriophage was replaced with SGP-1 bacteriophage or SEP-1 bacteriophage, and the step of applying Staphylococcus aureus was replaced with Salmonella.

The bacteriophage solution thus prepared was used to prepare a final sterilization composition. The first sterilization composition to be applied to Staphylococcus aureus was prepared by mixing the SAP-1 bacteriophage solution prepared above and SAP-2 bacteriophage solution in a 1: 1 ratio, and the second sterilization composition to be applied to Salmonella was SGP-1 prepared above. The bacteriophage solution and SEP-1 bacteriophage solution were prepared by mixing 1: 1.

Example  2: injection into automatic atomizer

The compositions were filled in an automatic atomizer pressure vessel (52φ) and LPG was used as the filling liquefied gas. The used amount of the first sterilization composition or the second sterilization composition was 10% of the total volume.

Example  3: Application to Staphylococcus aureus

The automatic spraying machine in which the first sterilization composition was added was installed at a cattle breeding barn at 5 m intervals. And automatic spraying machine was set so that the sterilization composition is sprayed every 30 minutes. On the other hand, a container containing 100 ml of TSB medium in the center of the barn was left without a lid. As a control experiment, only pure PBS containing no bacteriophage was prepared in the same manner, put into an automatic spraying machine, and installed in another barn. The distance between the two houses was approximately 1 km and the other experimental conditions were the same. The experiment was conducted for 3 days and the number of Staphylococcus aureus in each TSB medium was examined after 3 days. The investigation of the number of Staphylococcus aureus was carried out as follows. 0.5 ml of TSB medium left for 3 days was spread on Baird-Parker agar solid medium, which is a selective medium of Staphylococcus aureus, and then cultured overnight in a 37 ° C. incubator and the colonies formed on the solid medium the next day ( colony) was counted.

As a result, compared to the control experiments, it was confirmed that the staphylococcus aureus significantly reduced in the experiments were sprayed with the composition containing the bacteriophage as an active ingredient. The results are shown in FIG.

Example  4: Application to Salmonella

An automatic spraying machine capable of periodically spraying the second sterilizing composition was installed at a poultry farm at 5 m intervals and set to spray at intervals of 30 minutes. Meanwhile, a vessel containing 100 ml of TSB medium in the center of the poultry farm was left without a lid. As a control experiment, only pure PBS containing no bacteriophage was prepared in the same manner, put into an automatic spraying machine, and installed in another poultry farm. The distance between the two poultry farms was approximately 1 km and the other experimental conditions were the same. The experiment was conducted for 3 days and the number of Salmonella present in each TSB medium was examined after 3 days. Salmonella count was investigated as follows. 0.5 ml of TSB medium left for 3 days was plated in Salmonella-Shigella (SS) agar solid medium, which is a selective medium of Salmonella, and then cultured overnight in a 37 ° C. incubator, and the number of colonies formed on the solid medium was examined the next day.

As a result, compared to the control experiment, Salmonella was found to be significantly reduced in the experiments that sprayed the composition containing the bacteriophage as an active ingredient. The results are shown in FIG.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

1 is a result showing the effect of spraying the first bactericidal composition containing a bacteriophage having a specific killing ability of Staphylococcus aureus according to the present invention as an active ingredient, using an automatic nebulizer, FIG. 2. Is a result showing the effect of spraying a second bactericidal composition including a bacteriophage having Salmonella specific killing ability as an active ingredient in Salmonella according to the present invention. In Figures 1 and 2 <-bacteriophage> is the case of spraying the composition consisting of PBS does not contain bacteriophage, <+ bacteriophage> is the case of spraying the composition containing the bacteriophage according to the present invention as an active ingredient. The number at the top of the bar graph is the proportional% value when the number of bacteria in the case of <-bacteriophage> is 100%.

Claims (12)

Sterilization composition containing active biological material as an active ingredient. According to claim 1, wherein the active biomaterial is a bacteriophage, characterized in that the sterilized composition. The method according to claim 2, wherein the bacteriophage specific for SAP-1 (Accession No. KCTC 11153BP), SAP-2 (Accession No. KCTC 11154BP), Salmonella (Salmonella) that can specifically kill Staphylococcus aureus Sterilizing composition, characterized in that one or more selected from the group consisting of bacteriophage SGP-1 (Accession No. KCTC 11174BP), and SEP-1 (Accession No. KCTC 11173BP) that can be killed. A sterilizing sprayer containing a sterilizing composition containing an active biological material as an active ingredient. The sterilizing sprayer of claim 4, wherein the active biomaterial is bacteriophage. According to claim 5, The bacteriophage SAP-1 (Accession No. KCTC 11153BP), SAP-2 (Accession No. KCTC 11154BP), Salmonella (Salmonella) that can specifically kill Staphylococcus aureus A sterilizing sprayer, characterized in that one or more selected from the group consisting of bacteriophage SGP-1 (Accession No. KCTC 11174BP), and SEP-1 (Accession No. KCTC 11173BP) that can be specifically killed. Staphylococcus aureus SAP-1 capable of killing (Staphylococcus aureus) specifically (Accession No. KCTC 11153BP), SAP-2 (Accession No. KCTC 11154BP), bacteriophages capable of killing Salmonella (Salmonella) specifically SGP- A sterilizing composition containing one or more bacteriophages selected from the group consisting of 1 (Accession No. KCTC 11174BP), and SEP-1 (Accession No. KCTC 11173BP) as an active ingredient. A surface sterilization method comprising applying a sterilization composition containing an active biological material as an active ingredient to a surface. The method of claim 8, wherein the active biomaterial is bacteriophage. 10. The method of claim 9, wherein the bacteriophage specific for SAP-1 (Accession No. KCTC 11153BP), SAP-2 (Accession No. KCTC 11154BP), Salmonella (Salmonella) that can specifically kill Staphylococcus aureus Surface killing method characterized in that one or more selected from the group consisting of bacteriophage SGP-1 (Accession No. KCTC 11174BP), and SEP-1 (Accession No. KCTC 11173BP) that can be killed. Staphylococcus aureus SAP-1 capable of killing (Staphylococcus aureus) specifically (Accession No. KCTC 11153BP), SAP-2 (Accession No. KCTC 11154BP), bacteriophages capable of killing Salmonella (Salmonella) specifically SGP- Surface sterilization method which periodically sprays a sterilization composition including one or more bacteriophages selected from the group consisting of 1 (Accession No. KCTC 11174BP) and SEP-1 (Accession No. KCTC 11173BP) as an active ingredient to the surface periodically. . 12. The method of claim 11, wherein the automatic spraying is implemented by an automatic nebulizer.

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